Potential chronic liver toxicity in rats orally administered an ethanol extract of Huangqin(Radix Scutellariae Baicalensis)

INTRODUCTION

No obvious abnormal findings such as swelling,effusion,capillary hemorrhage,or conglutination were seen upon macroscopic examination of heart,liver,spleen,lung,adrenal gland,pancreas,stomach,duodenum,small intestine(jejunum,ileum),large intestine(cecum,colon,rectum),testicle,epididymis,prostate,seminal vesicle,uterus,ovary,thoracic cavity,abdominal cavity and pelvic cavity in any SBE treated group.

In ICR mice,acute(14-d),subchronic(28-d),and chronic(90-d)safety trials of refined extracts of Huangqin(Radix Scutellariae Baicalensis)have been performed and there were no significant differences between the extract treated and placebo groups.18Studies confirmed that in rats,dogs,and even human receiving long-term(rats and dogs for up to 26 weeks,human for up to ten days)oral high-doses of the major bioactive Huangqin(Radix Scutellariae Baicalensis)-derived flavonoids(including baicalin,baicalein and wogonoside),no drug-related adverse reactions were observed.19-24Therefore,the main bioactive constituents of Huangqin(Radix Scutellariae Baicalensis)were considered safe,well tolerated and without serious accumulation effect.In contrast,hepatotoxicity of the extract of this herb have been reported in the clinic:four patients from the United States who had used Huangqin(Radix Scutellariae Baicalensis)in a dietary supplement displayed hepatotoxicity,25-28while nineteen Japanese patients developed liver injury after the use of the herb.29Thus,whether Huangqin(Radix Scutellariae Baicalensis)is hepatotoxic is controversial,and the systematic studies of its toxicity are also lacking.Studies in HepG2 cells have suggested that baicalin is more cytotoxic than baicalein,so baicalin may have the potential to induce hepatotoxicity,but whether it is the cause of Huangqin(Radix Scutellariae Baicalensis)-induced hepatotoxicity or whether there are other liver toxic components involved is not yet clear.30

Thus far,previous studies have primarily focused on the safety of the aqueous extract of Huangqin(Radix Scutellariae Baicalensis),with little investigation regarding the safety of oral administration of the ethanol extract.Considering the reported clinical adverse reactions,especially the hepatotoxicity of the extract when taken orally,the present study investigated a 26-week repeated oral administration of the ethanol extract of Huangqin(Radix Scutellariae Baicalensis)(SBE)in Wistar rats to evaluate its potential chronic toxicity,especially liver toxicity.

MATERIALS AND METHODS

Reagents and instruments

The root of Huangqin(Radix Scutellariae Baicalensis)as identified by the authors was collected in Hebei province,China,and the voucher specimen was deposited at the Institute of Chinese Materia Medica China Academy of Chinese Medical Sciences(ICMM).Baicalin,baicalein,wogonoside and wogonin(＞98.0%)were all purchased from the National Institute for the Control of Pharmaceutical and Biological Products(Beijing,China).Hematological parameters were analyzed using an automatic blood cell analyzer(MEK 6318K;Nihon Kohden Co.,Ltd.,Tokyo,Japan),and the dilution and hemolytic reagents were supplied by Yantai B&E Scientific Instrument Co.,Ltd.,(Shandong Province,China).All serum biochemical reagents andQC serum were purchased from BioSino Bio-Technology and Science Co.,Ltd.,(Beijing,China)and biochemical analysis was performed by an automatic biochemistry analyzer(Dimension AR;Siemens,Munich,Germany).Easy Lyte plus Na/K/CL analyzer kit and quality controls were purchased from Medica Co.,(Bedford,MA,USA)and tested by electrolytes analyzer(Easy Lyte PLUS Na/K/Cl analyzer,Medica Co.,).Prothrombin time(PT)and activated partial thromboplastin time(APTT)kits were purchased from Shanghai Sun Bio Tech Co.,Ltd.,(Shanghai,China).PT and APTT were determined using a blood coagulation meter(LG-PABER,Beijing Shidi Science&Technology Instrument Co.,Ltd.,Beijing,China).Urine analysispapers(URS-10T)werepurchased from Changchun Wancheng Bio-Electron Co.,Ltd.(Changchun,China),and a Clinitek 50 urine chemistry analyzer was used(Bayer,Leverkusen,Germany).The histomorphological examination was conducted using a automation-tissue-dehydrating machine(Shandon Excelsior ES,Shandon,Runcorn,UK),paraffin embedding station(MICROM EC350,Thermo Scientific,Waltham,MA,USA),slicing machine(Finesse 325,Shandon,China),automatic dyeing machine(MICROM HMS740,Thermo Fisher Scientific,Massachusetts,USA)and microscope and image analysis system(LEICA DM500,Leica Biosystems GmbH,Nussloch,Germany).

Preparation and analysis of SBE

The dried,powdered root of Huangqin(Radix Scutellariae Baicalensis)(80 kg)was extracted twice under reflux with 70%EtOH(480 L for 2 h,400 L for 2 h).The EtOH solutions were combined and concentrated to yield a dried extract(20 kg),with a yield of 25%.The extract batch used was analyzed by ICMM.The chemical structure of the flavonoids is shown in Figure 1.Analysis was performed on a Waters Alliance system(Waters,Milford,MA,USA),equipped with a binary pump,an auto sampler,a column oven and a Waters 2996 DAD.An Agilent Zorbax C18 column(5 μm,4.6 mm×250 mm;Agilent Technologies,Santa Clara,CA,USA)was used and the column temperature was maintained at 30℃.For the analysis of SBE,a solution of CH3OH(A)and H2O-THF-H3PO4(80∶10∶0.2)(B)(A∶B,55∶45)was used as the mobile phase.The flow rate was 1.0 mL/min.The standard solution was prepared by dissolving each standard in methanol to the concentrations of 77.20 μg/mL (baicalin),11.50μg/mL(baicalein),52.70μg/mL(wogonoside)and 13.50 μg/mL(wogonin).The EtOH extract(3.98 mg)was extracted with 10 mL methanol in an ultrasonic bath for 15 min.After cooling down,the extraction solution was adjusted to the original weight,and then filtered through a 0.22 μm filter before injection.

Figure 1 Structure of the tested flavonoids

Animal husbandry

The experiments were conducted in line with the Guide for Care and Use of Laboratory Animals issued by the National Institutes of Health(NIH Publications No.8023,revised 1978),and the procedures were approved by the Research Ethics Committee of the Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing,China.All experiments were carried out in accordance with the ethical guidelines and regulations for the care and use of laboratory animals.Male and female specific pathogen free Wistar rats at an initial age of 5 weeks,were purchased from the Institute of Laboratory Animal Science,Chinese Academy of Medical Sciences&Peking Union Medical College(CAMS&PUMC)(Certificate of quality No.SCXK[jing]2012-0001).The animals were kept in the Medical Experimental Animal Center of the China Academy of Chinese Medical Sciences.Standardized rat husbandry procedures were in keeping with the State Standard for Animal Administration of the People's Republic of China,with 12 h of electrical lighting and 12 h of dark daily,room temperature 20-22℃,relative humidity of 40%-70%,air change approximately 15 times per hour.

2017年11月7日，中国人大网公布了由第十二届全国人大常委会第二十八次会议审议通过的《中华人民共和国监察法(草案)》，向社会公开征求意见。当即就有学者提出，《监察法(草案)》缺乏宪法依据，同时该草案中的部分内容不当限制了公民基本权利，调整了人大制度下的权力关系，呼吁全国人大主导对草案进行合宪性审查[16]。另有学者在《监察法(草案)》公开征求意见不久，就以公民身份对该草案提出合宪性审查请求，引起宪法学界广泛关注。

Treatment groups

The results of blood electrolyte analysis showed that in male rats after 13 weeks'administration of 1250 mg/kg SBE,the concentration of K ion was markedly lower than that in the control group(P＜0.05),and at theend of 26 weeks treatment,the value of Na ion in male rats treated with 300 mg/kg SBE also decreased significantly(P＜0.05).

Observation and clinical examination

Physical condition of rats:the physical condition of rats including behavior,gait and general activity,appearance,hair luster,stool,and urine was observed daily.Record the onset,duration,recovery,etc.of any abnormalities caused by the drug(including death or death).

Body weight and food consumption:the body weight of all rats was measured once a week during the study and the dosing volume for each rat was adjusted according to the most recent body weights.Before euthanasia,all rats in each group were fasted for 17 h,so the body weights for the calculation of relative organ weight are fasted weights.The remaining granules in each cage were weighed before feed was quantitatively added per week.The amount of remaining feed subtracted from the total amount of feed added was equal to the food consumption.

Urinalysis:before euthanasia,each rat was placed individually into a metabolic cage and subjected to fasting overnight,but with free to access water.The 24 h-urine output of each rat was measured,and a quantitative analysis of urine pH,protein,glucose,occult blood,urine specific gravity,urobilinogen,nitrite,ketone body,bilirubin,and leukocytes were conducted with the Urine Analyzer.

Hematological and serum biochemical analysis:the rats were anesthetized by intraperitoneal injection(i.p)of 30 mg/kg phenobarbital sodium,and blood was taken from the abdominal aorta to test the hematological,coagulation and biochemical parameters.Hematological and coagulation tests were performed with plasma isolated from the blood.Hematological testing included the following parameters:red blood cell count(RBC),white blood cell count(WBC),hemoglobin(HGB),hematocrit(HCT),blood platelet number(PLT),mean corpuscular volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration(MCHC),and leukocyte differential count.PT and APTT were determined to assess coagulation function.In addition,the blood was centrifuged to isolate the serum,and levels of serum total protein(TP),albumin(ALB),aspartate transaminase(AST),alanine transaminase(ALT),alkaline phosphatase(ALP),γ-glutamyl-transferase(γ-GT),creatine kinase(CK),blood urea nitrogen(BUN),creatinine(CRE),glucose(GLU),total bilirubin(TBIL),total cholesterol(CHO),triglyceride(TG),and the concentration of Na+,K+,Cl－were determined by the electrolyte analyzer.

Autopsy and Histomorphology Observation:autopsy was performed after rats were exsanguinated,and the following organs were removed for macroscopic observation:brain,pituitary,thyroid,salivary gland(submaxillary and sublingual salivary gland),thymus,lung with bronchi,heart,liver,spleen,kidney,adrenal gland,testicle,epididymis,prostate,seminal vesicle,ovary,uterus,stomach,duodenum,small intestine(je-junum,ileum),large intestine(cecum,colon,rectum),urinary bladder,sternum,femur,eyeballs,optic nerve,pancreas and mesenteric lymph nodes.The organ weights of brain,pituitary,salivary gland,thymus,lung with bronchi,heart,liver,spleen,kidney,stomach,adrenal gland,testicle,prostate,seminal vesicle,ovary,uterus and the fasted-body weight were measured,and then the relative organ weight was calculated as follows:relative organ weight(ROW)(%)=[organ weight(g)/body weight(g)]×100.Finally,all removed organs were fixed in 10%formalin solution,embedded in paraffin,sectioned and hematoxylin and eosin(HE)stained,and then histomorphology observation was performed under light microscopes.

Statistical analysis

10 Wang D,Wang L,Gu J,Yang H,Liu N,Lin Y.Scutellarin inhibits high glucose-induced and hypoxia-mimetic agent-induced angiogenic effects in human retinal endothelial cells through reactive oxygen species/hypoxia-inducible factor-1α/vascular endothelial growth factor pathway.J Cardiovasc Pharmacol 2014;64(3):218-227.

RESULTS

Baicalin is the major flavonoid present in the SBE

The typical HPLC profiles of the sample solutions from SBE are shown in Figure 2.The content of flavonoids in SBE was as follow:baicalin 18.9%,wogonoside 5.56%,baicalein 1.45%and wognin 0.57%.

SBE does not affect rat behavior,body weight or food consumption

No animal deaths occurred during the experiment.Behavior,gait,general activity,fecal and urine output of rats in all groups showed no obvious abnormalities.There was no significant difference in body weight and food consumption between all SBE treatment and control groups(Figures 3,4).

SBE does not affect urine production

There was no significant difference in 24 h urine output between the three SBE treatment groups and the control group.After SBE treatment,urine was browner in a dose-depended manner,which may be related to the color of SBE.Compared with rats in the control group,no significant differences were observed in the qualitative analysis of urine parameters in SBE treatment groups after 13 weeks and 26 weeks'oral administration(data not shown).

SBE has no consistent effect on hematological parameters

At the end of 13 weeks,although the number of WBC in the 300 and 1250 mg/kg SBE treatment groups'male rats were significantly lower than those in the control group(P＜0.01),they were still in the normal range.There were no differences in other hematological parameters between the three SBE groups and the control group(Table 1).After 26 weeks'treatment,compared with the control group,MCH and MCHC were decreased in female rats of all SBE groups(P＜0.01),however no other treatment-related changes were observed in hematological parameters(Table 2).The hematological parameters in the three SBE groups had no significant differences compared with the control group after 4 weeks'recovery time(Table 3).

SBE may be prothrombogenic

Compared with the control group,after 13 and 26 weeks'treatment,the PT of female rats in SBE treatment groups was significantly shorter(P＜0.01),in a dose-dependent fashion.Similarly,the APTT of male rats in SBE groups was markedly shorter than that in the control group after 26 weeks'treatment(P＜0.001).However,the PT and APTT values of all SBE groups returned to normal by the end of the recovery time,with no significant difference from the control group(Tables 1-3).

SBE may influence renal function and lipid metabolism

Figure 2 Chromatogram of the sample solution ethanol extracted from the root of Huangqin(Radix Scutellariae Baicalensis)a:baicalin;b:wogonoside;c:wogonin;d:baicalein.

Figure 3 Mean body weights of male and female rats during 26 weeks'oral administration of SBE followed by a 4-week recovery period

SBE:ethanol extract of Huangqin(Radix Scutellariae Baicalensis).

Figure 4 Mean food consumption of male and female rats during 26 weeks'oral administration of SBE followed by a 4-week recovery period

As shown in Tables 4-6,the concentration of BUN of male rats in the 300 and 1250 mg/kg SBE treatment groups was lower(P＜0.05),while that in 2500 mg/kg treatment group was higher than the control group(P＜0.05).Except for the above,there was no significant difference in other biochemical parameters between all SBE treated groups and the control group after 13 weeks'treatment.After 26 weeks'treatment,compared with the control group,in male rats treated with SBE,the levels of CK,CRE and BUN were significantly lower(P＜0.05),and TG markedly higher(P＜0.05);and in female rats,the level of CHO and TG were also increased(P＜0.05).At the end of recovery time,in male and female rats of the SBE treated groups,the levels of CK,BUN,ALT and CHO were lower(P＜0.05),and the level of GLU in female rats was higher than that in the control group(P＜0.05).The decrease in CK,CRE,ALT and CHO were not effective for clinical diagnosis.Except the above,no significant differences in the other biochemistry parameters between SBE treatment groups and the control group were observed.

After acclimating for 7 d,the rats were about 6weeks old and then were randomly divided into 4 groups by random number table method:control group and 3 dose-treated groups.Each group contained 20 males and 20 female rats.The control group rats were treated with distilled water by oral gavage while the three treatment groups were given SBE at a dose of 300,1250 or 2500 mg·kg－1·d－1respectively.Rats were orally administered SBE once a day and were observed before and after administration.The rats were euthanized after 13 and 26 weeks of daily oral gavage dosing(5 male and 5 female rats per group).The remaining rats had a recovery period of 4 weeks without SBE administration and were euthanized at the end of recovery time.

最令人担心的是，根据爱因斯坦的理论，这颗以每秒5000英里的速度沿着鸡蛋形轨道疾驰的恒星，应该经历了宇宙中的所有奇异之处。这颗恒星表面遭受的强烈引力会减缓光波的振动，将其拉长。于是，从地球上看来，它会变得比正常状态更红一些。

Table 1 Hematological data for male and female rats orally administered SBE for 13 weeks(ˉ±s)

Notes:WBC:white blood cells;RBC:red blood cells;HGB:hemoglobin;HCT:hematocrit;MCV:mean corpuscular volume;MCH:mean corpuscular hemoglobin;MCHC:mean corpuscular hemoglobin concentration;PLT:blood platelet number;GR:neutrophilic granulocyte ratio,LY:lymphocyte ratio;MO:monocytes ratio;PT:prothrombin time;APTT:activated partial thromboplastin time.Significantly different from the control group by Dunnett's test,aP＜0.01,bP＜0.001,cP＜0.05.

Item Dose(mg·kg－1·d－1)2500 15.92±2.57 7.84±0.23 14.05±0.73 37.46±1.43 48.15±1.04 18.06±0.63 37.46±0.94 250.37±19.36 17.06±7.58 77.64±7.96 5.34±1.15 13.16±1.34 15.34±2.72 9.13±3.28 6.87±0.56 13.31±0.54 34.16±2.22 50.24±1.38 19.35±1.10 38.34±1.56 253.40±31.03 9.38±7.06 86.43±7.74 4.42±1.17 13.35±0.60a18.76±2.12aMales Females WBC(109/L)RBC(1012/L)HGB(g/dL)HCT(%)MCV(fL)MCH(pg)MCHC(g/dL)PLT(109/L)GR(%)LY(%)MO(%)PT(S)APTT(S)WBC(109/L)RBC(1012/L)HGB(g/dL)HCT(%)MCV(fL)MCH(pg)MCHC(g/dL)PLT(109/L)GR(%)LY(%)MO(%)PT(S)APTT(S)0 15.71±1.22 8.02±0.58 14.53±0.87 38.94±1.65 48.56±1.95 17.95±0.85 36.84±1.62 297.80±42.14 15.85±5.74 78.45±6.02 5.84±1.23 12.34±2.91 16.34±2.96 8.80±2.32 7.04±0.64 13.73±1.07 35.34±2.48 50.47±1.72 19.12±0.98 37.82±1.54 248.75±41.15 9.32±7.67 85.57±7.75 5.25±1.82 14.53±0.34 21.75±1.34 300 12.10±1.24a7.70±0.56 14.28±1.24 37.10±2.38 48.24±2.66 18.28±1.04 37.82±1.07 295.31±50.02 15.13±6.66 78.83±6.51 6.12±1.54 11.75±2.47 18.28±3.49 7.29±1.81 7.32±0.43 14.04±0.65 36.72±1.46 50.26±1.94 18.96±0.95 37.71±0.88 262.20±29.27 14.54±3.21 79.00±2.93 6.64±0.75 14.24±0.92 22.17±1.33 1250 11.36±2.13b7.16±1.44 13.34±2.22 34.42±5.84 48.73±1.75 18.64±0.87 38.08±0.62 250.28±39.23 19.02±8.32 73.92±8.87 7.23±1.42 12.16±1.06 13.07±5.43 8.15±2.14 7.00±0.52 12.85±1.06 34.10±1.94 48.93±1.36 18.49±0.70 37.50±1.40 263.44±32.18 9.16±5.38 86.12±5.98 4.91±1.28 13.73±0.64c21.88±1.34

Macroscopic observations

Huangqin(Radix Scutellariae Baicalensis)is a widely used herb in both Eastern and Western traditional medicine.In China,it has a long medicinal history of 2000 years,and is usually used as the main raw material to prepare traditional herbal formulations and modern Chinese medicines.1-3Huangqin(Radix Scutellariae Baicalensis)has various pharmacological effects and is mainly used in the clinical treatment of respiratory infections,pulmonary heat,enteritis,dysentery,jaundice,purulent infection and gastrointestinal infections.4,5Additionally,it was used as a nerve tonic,sedative and anticonvulsant by Native Americans and Europeans.6,7Recent research has revealed that Huangqin(Radix Scutellariae Baicalensis)also possesses potent anticancer,antidiabetic and fat decreasing activities.8-12Based on these beneficial properties,Huangqin(Radix Scutellariae Ba-icalensis)has been used as an ingredient in botanical formulations of health foods which promote health functions such as relieving the inflammatory immune response,combating pathogen and so on.13Its effective components are mostly flavonoids,which decrease inflammation,stop tumor growth,and have antibacterial,antiviral,antiplatelet aggregation,anti spasmogenic and antioxidant properties.14-17Dozens of flavonoids compounds have been extracted and identified from Huangqin(Radix Scutellariae Baicalensis),including baicalein,wogonin,oroxylin A,baicalin,wogonoside,and oroxylin A-7-glucuronide.1,2

这部小说把那一段历史作为表现内容，把路线的形成过程作为描写主线，就深入到抗日战争的骨子里去了。小说的题目是《共赴国难》，关键在“共赴”二字上。写了策略上的交流、商讨、演变，就使原来各自为政、互不联系的分散状态扭成一股绳，真正变成了一种合力，使“共赴”二字得到了有力的表现。

ROW

After administration of SBE for 13 weeks,the ROW of the adrenal gland in female rats was higher than that in the control group(P＜0.05),while after 26 weeks,in male rats treated with SBE,the ROW of the adrenal gland was lower than that of the control rats(P＜0.001).13 weeks after administration of SBE in female rats,and 26 weeks after administration of SBE in male rats,their thymus ROWs were significantly lower than that of the control rats(P＜0.05).The liver index increased significantly in female SBE treated rats after 26 weeks'treatment(P＜0.001).After the recovery period,in the SBE treated groups,the ROW of kidneys in male rats and ROW of hearts in female rats increasedsignificantly(P＜0.05),however there was no significant change in the ROW of other organs compared with the control group(Tables 7-9).

实习报告评分标准：“思路清晰，条理清楚，能运用专业理论知识分析具体问题及按要求完成报告”为优秀；“能结合实习情况描述实习内容及解决的具体问题，按要求完成报告”为良好；“罗列材料，缺少分析，基本完成实习报告”为合格；“实习报告与实习内容关系不大或未能按要求完成报告”为不合格。

小学生的数学学习能力普遍不足。这一方面是因为应试教育的影响，一方面是教师在平时的教学中忽视学生的主体学习地位，学生感受不到足够的学习乐趣，进而也就无法在数学学习中形成探索精神和思考能力。学生跟着教师的节奏走，跟着课堂走，跟着作业走，跟着考试走的数学学习习惯依然是主要表现形式。再有就是学生上课提问少，互动交流少，做题创新少，考试反思少，也严重限制了学生数学学习能力的提升。

Table 2 Hematological data for male and female rats orally administered SBE for 26 weeks(ˉ±s)

Notes:WBC:white blood cells;RBC:red blood cells;HGB:hemoglobin;HCT:hematocrit;MCV:mean corpuscular volume;MCH:mean corpuscular hemoglobin;MCHC:mean corpuscular hemoglobin concentration;PLT:blood platelet number;GR:neutrophilic granulocyte ratio;LY:lymphocyte ratio;MO:monocytes ratio;PT:prothrombin time;APTT:activated partial thromboplastin time.Significantly different from the control group by Dunnett's test,aP＜0.01,bP＜0.001.

Item Dose(mg·kg－1·d－1)Males Females WBC(109/L)RBC(1012/L)HGB(g/L)HCT(%)MCV(fL)MCH(pg)MCHC(g/L)PLT(109/L)GR(%)LY(%)MO(%)PT(S)APTT(S)WBC(109/L)RBC(1012/L)HGB(g/L)HCT(%)MCV(fL)MCH(pg)MCHC(g/L)PLT(109/L)GR(%)LY(%)MO(%)PT(S)APTT(S)0 9.5±1.8 7.6±0.8 13.2±1.1 35.3±2.8 47.1±1.6 17.3±0.7 36.8±0.8 271.4±85.0 17.5±5.0 75.0±4.9 7.6±0.9 14.9±1.1 28.4±2.1 6.7±1.7 6.8±0.6 12.6±0.9 33.4±2.4 49.8±1.1 18.5±1.2 37.4±2.3 267.1±41.1 13.2±7.4 79.9±7.6 7.1±1.3 16.4±1.5 15.5±2.8 300 9.2±1.3 7.4±0.6 12.8±1.1 34.6±2.4 46.8±1.3 17.2±0.9 36.8±2.1 278.8±45.1 19.1±11.3 71.3±10.9 9.7±2.5 15.6±2.2 26.9±5.4 7.0±0.7 7.2±0.6 12.8±1.2 35.0±3.2 49.7±1.3 18.1±0.8 36.5±0.8 247.4±84.0 14.2±11.3 78.2±11.3 7.6±1.2 14.8±1.0a15.5±3.8 1250 11.1±1.8 7.6±0.5 13.3±0.6 35.6±1.6 47.0±2.2 17.2±1.0 36.3±1.2 267.6±55.2 16.0±3.1 76.1±4.1 8.0±2.4 16.2±0.9 21.1±2.8b8.3±2.6 7.2±0.4 12.6±0.7 35.0±2.2 49.4±1.3 17.5±0.7a34.9±1.0a261.2±37.2 15.3±13.0 76.8±12.8 8.0±1.8 13.8±1.3b18.3±1.5 2500 10.6±2.4 8.0±0.6 13.1±1.0 35.6±2.7 47.1±1.2 17.0±0.6 36.3±0.9 276.1±23.2 22.5±10.8 69.1±10.0 8.5±1.8 15.8±0.5 22.2±2.0b8.2±2.3 6.7±0.8 12.2±1.0 33.7±3.0 50.7±2.1 18.2±1.1 35.7±1.6a271.9±66.4 13.2±7.8 78.2±7.6 8.7±1.8 14.0±1.0b16.3±3.3

Histomorphological observations

Under a light microscope,we did not find treatment-related toxicological changes in the heart,liver,spleen,kidney,adrenal,bronchial,thymus,thyroid,brain,pituitary,stomach,duodenum,small intestine,large intestine,pancreas,bladder,testis,epididymis,uterus,ovary and other organs of rats.The pathological changes in the liver of male and female rats at 26 weeks are shown in Figure 5.We can see that high-dose SBE treated-group rats at 26 weeks with a large number of leukocytes infiltrating in the liver and no other pathological changes.However,at week 13 high-dose SBE treatment,at 4 weeks of recovery from high-dose SBE treatment and at weeks 26 of low-dose and medium-dose SBE treatment,we found no significant pathological changes in rat livers(pathological images not shown).The histological structure of the other major organs in control and high-dose SBE group rats at 26 weeks are shown in Figure 6.

DISCUSSION

The aim of this study was to confirm the safety of 26 weeks'oral intake of SBE and to further explore its effect on the liver in rats.The results demonstrated that during the SBE administration period,even at the highest dose of 2500 mg·kg－1·d－1,no deaths and no overall adverse effects were observed.In the SBE treatment groups,some hematological and serum biochemical parameters underwent changes:after 13 weeks'treatment,the WBC of male rats in the 300 and 1250 mg/kg SBE groups were slightly decreased but still within the normal range,while at the end of 26 weeks,the WBC in the SBE groups showed no deviation from the control rats.Thus,it was considered that the changes in WBC were not treatment-related toxicity or abnormality.MCH and MCHC are parameters used to detect anemia,and they were decreased in female rats in the 1250 and 2500 mg/kg SBE groups after treatment for 26 weeks,although at the same time other parameters indicative of anemia such as RBC,Ht,and Hb showed no change.Thus,the changes of MCH and MCHC in this study were not sufficient to support the hypothesis that SBE induces anemia.The prolongation or shortening of APTT and PT are associated with increased risk of coagulation disorders or thrombosis respectively.It was previously found that SBE has certain hemostatic effects in pharmacodynamics studies,therefore we surmised that the shortening of PT and APTT in the rats of the SBE treatment groups may be related to these pharmacological effects.17At the end of the recovery time,there was no significant difference in the hemato-logical parameters between the three SBE groups and the control group,all changes returned to normal,which further confirmed that SBE did not have long term toxic effects on the blood of rats.

Table 3 Hematological data for male and female rats after 4 weeks'recovery(ˉ±s)

Notes:WBC:white blood cells;RBC:red blood cells;HGB:hemoglobin;HCT:hematocrit;MCV:mean corpuscular volume;MCH:mean corpuscular hemoglobin;MCHC:mean corpuscular hemoglobin concentration;PLT:blood platelet number;GR:neutrophilic granulocyte ratio;LY:lymphocyte ratio;MO:monocytes ratio;PT:prothrombin time;APTT:activated partial thromboplastin time.Significantly different from the control group by Dunnett's test,aP＜0.05.

Item Dose(mg·kg－1·d－1)Males Females 2500 10.60±3.15 7.95±1.05 13.81±2.20 38.12±5.15 48.06±0.93 17.35±0.93 35.84±2.05 225.18±56.26 10.64±4.46 79.94±5.76 9.50±2.45 15.56±0.88 17.69±3.85 5.18±1.08 7.46±1.45 12.87±2.38 36.74±6.20 50.15±1.65 17.54±0.46 34.88±0.75 242.33±55.12 11.84±9.07 79.61±8.88 8.77±2.02 16.76±0.86 18.76±1.73 WBC(109/L)RBC(1012/L)HGB(g/L)HCT(%)MCV(fL)MCH(pg)MCHC(g/L)PLT(109/L)GR(%)LY(%)MO(%)PT(S)APTT(S)WBC(109/L)RBC(1012/L)HGB(g/L)HCT(%)MCV(fL)MCH(pg)MCHC(g/L)PLT(109/L)GR(%)LY(%)MO(%)PT(S)APTT(S)0 11.40±1.56 9.02±0.54 15.49±1.55 42.70±3.12 47.63±1.04 16.91±0.75 35.84±0.86 289.90±28.02 13.01±5.29 78.17±4.73 8.92±2.08 16.19±1.63 19.67±2.27 6.50±0.92 8.23±0.22 13.81±0.49 40.05±0.91 48.96±1.34 16.97±0.62 34.82±0.84 271.07±37.00 7.74±3.75 81.85±4.04 10.63±0.93 17.65±1.21 18.90±1.24 300 12.07±1.58 8.54±0.31 15.23±1.16 41.53±1.26 49.24±0.82a17.82±0.52 36.28±1.17 289.43±25.12 16.42±6.95 75.30±8.74 8.38±2.67 15.82±0.64 21.13±2.74 5.73±1.91 7.91±0.43 13.83±0.87 39.10±2.02 49.64±0.37 17.52±0.94 35.45±1.88 235.18±34.33 11.50±7.01 78.44±7.92 10.14±1.72 16.52±0.35 21.71±2.15 1250 10.64±1.94 9.01±0.58 16.02±0.72 42.94±1.64 47.53±1.16 17.67±1.04 37.20±1.68 304.22±64.22 14.26±4.83 76.73±5.95 9.15±1.63 14.91±0.45 21.34±2.53 4.76±0.70 7.82±0.44 13.86±0.84 39.23±2.20 50.12±0.58 17.37±0.29 34.63±0.92 248.29±33.01 19.12±12.03 71.80±11.15 9.12±1.74 16.53±1.47 21.93±3.60

Table 4 Serum biochemistry data for male and female rats orally administered SBE for 13 weeks(ˉ±s)

Notes:ALB:total albumin;ALP:alkaline phosphatase;ALT:alanine aminotransferase;AST:aspartate aminotransferase;CHO:cholesterol;CK:creatine kinase;CRE:creatinine;GGT:γ-glutamyl transpeptidase;GLU:glucose;TBIL:total bilirubin;TG:triglycerides;TP:total protein;BUN:blood urea nitrogen;Na+:sodium;K+:potassium;Cl-:chloride;SBE:ethanol extract of Huangqin(Radix Scutellariae Baicalensis).Significantly different from the control group by Dunnett's test:aP＜0.05.

2500 33.91±1.65 122.53±24.24 31.23±5.13 83.04±13.53 1.41±0.31 587.36±137.36 62.03±5.58 0.94±4.97 9.23±1.28 6.72±1.40 0.91±0.37 57.73±2.98 9.87±1.24a141.36±2.17 5.07±0.32 115.06±3.37 35.90±1.73 64.05±13.36 23.24±2.88 75.82±10.89 1.64±0.45 548.27±159.36 56.71±4.32 3.84±0.46 8.98±2.03 7.42±1.31 0.62±0.14 60.34±2.67 7.32±1.13 142.25±2.01 3.92±0.14 115.37±2.34 Item Dose(mg·kg－1·d－1)Males Females ALB(g/L)ALP(IU)ALT(U/L)AST(U/L)CHO(mmol/L)CK(U/L)CRE(mmol/L)γ-GT(U/L)GLU(mmol/L)TBIL(μmol/L)TG(mmol/L)TP(g/L)BUN(mmol/L)Na(mmol/L)K(mmol/L)Cl(mmol/L)ALB(g/L)ALP(IU)ALT(U/L)AST(U/L)CHO(mmol/L)CK(U/L)CRE(μmol/L)γ-GT(U/L)GLU(mmol/L)TBIL(μmol/L)TG(mmol/L)TP(g/L)BUN(mmol/L)Na(mmol/L)K(mmol/L)Cl(mmol/L)0 33.90±1.92 93.05±11.04 31.34±9.08 93.77±20.06 1.82±0.54 628.07±129.23 62.31±3.92 1.64±4.71 9.28±1.37 4.62±3.67 0.83±0.42 58.23±4.30 8.32±0.86 141.25±3.07 5.40±0.63 114.38±1.09 37.31±2.08 60.08±15.02 23.80±1.93 71.53±13.20 1.50±0.21 506.08±183.23 58.12±2.84 4.57±0.92 7.72±0.58 6.74±1.15 0.51±0.20 62.91±2.73 8.02±1.01 142.37±2.10 4.00±0.32 115.08±1.12 300 34.23±2.33 97.28±19.47 30.22±4.61 86.53±10.92 1.63±0.42 654.35±113.14 58.06±2.03 2.43±2.30a9.23±1.10 6.41±1.98 1.27±0.65 56.82±6.01 6.81±0.50a138.34±7.18 5.11±0.72 113.65±7.14 36.73±1.76 54.73±11.14 21.74±2.36 87.06±32.51 1.42±0.20 736.14±466.12 53.16±5.05 3.14±1.93 7.31±0.56 6.50±0.82 0.42±0.13 61.32±2.80 7.63±1.20 144.28±2.05 4.14±0.52 116.27±3.02 1250 34.04±1.30 98.42±17.22 27.51±3.64 80.50±15.24 1.65±0.38 669.27±254.47 57.37±3.46 1.92±3.55 10.56±1.97 6.24±1.06 1.13±0.30 56.08±3.04 6.92±1.26a139.06±3.21 4.66±0.31a113.29±3.23 35.92±2.08 68.02±10.24 24.06±3.21 82.04±10.07 1.43±0.22 505.38±160.06 57.62±5.74 4.72±0.28 7.52±0.74 6.84±0.63 0.54±0.11 59.91±2.92 7.34±0.71 143.37±3.24 4.04±0.31 114.36±2.24

As for the biochemistry indices investigated,after SBE treatment for 13 weeks,the concentration of BUN in the male rats of the 2500 mg/kg group were obviously higher than that in control rats.Although the increase of BUN and CRE indicates impaired renal function,after 26 weeks treatment and the recovery time,the value of BUN and CRE in the male rats of the 2500 mg/kg SBE treatment group decreased.The relative kidneyweight is another indicator of kidney abnormalities,although it has some limitations that do not fully reflect pathological changes in the kidney.In the SBE treatment period,the relative kidney weight of the rats treated with SBE was the same as control rats,while the relative kidney weight markedly increased in male rats of the 2500 mg/kg SBE group at the end of the recovery time after treatment had ceased.The increase in relative kidney weight may indicate an acute kidney lesion,however,other kidney lesion indices such as urine parameters,serum level of BUN and CRE,and the gold standard of histopathological examination showed noabnormalities.Considering all the above outcomes,long-term use of SBE does not induce renal toxicity.

联盟共举办两届“基于HIM的广义BIM矩阵(GIM)论坛”、两届“中国BIM经理高峰论坛”，第一届全国建筑业“互联网+”技术展示会，2016建筑施工BIM应用项目观摩会，参展“第十三届国际绿色建筑与建筑节能大会暨新技术与产品博览会”，协办“buildingSMART 2017年香港国际BIM大赛暨高峰论坛”，协办第四届BIM国际技术交流会等，通过论坛展览展示活动不断扩大联盟影响力。

Table 5 Serum biochemistry data for male and female rats orally administered SBE for 26 weeks(ˉ±s)

Notes:ALB:total albumin;ALP:alkaline phosphatase;ALT:alanine aminotransferase;AST:aspartate aminotransferase;CHO:cholesterol;CK:creatine kinase;CRE:creatinine;GGT:γ-glutamyl transpeptidase;GLU:glucose;TBIL:total bilirubin;TG:triglycerides;TP:total protein;BUN:blood urea nitrogen;Na+:sodium;K+:potassium;Cl-:chloride.Significantly different from the control group by Dunnett's test,aP＜0.05.

Item Dose(mg·kg－1·d－1)Males Females ALB(g/L)ALP(IU)ALT(U/L)AST(U/L)CHO(mmol/L)CK(U/L)CRE(μmol/L)γ-GT(U/L)GLU(mmol/L)TBIL(μmol/L)TG(mmol/L)TP(g/L)BUN(mmol/L)Na(mmol/L)K(mmol/L)Cl(mmol/L)ALB(g/L)ALP(IU)ALT(U/L)AST(U/L)CHO(mmol/L)CK(U/L)CRE(μmol/L)γ-GT(U/L)GLU(mmol/L)TBIL(μmol/L)TG(mmol/L)TP(g/L)BUN(mmol/L)Na(mmol/L)K(mmol/L)Cl(mmol/L)0 36.84±1.62 114.16±14.20 87.23±21.42 100.42±56.33 1.94±0.46 696.64±155.12 64.85±7.04 1.94±3.62 6.08±1.54 8.60±1.03 0.87±0.31 64.54±4.62 8.47±1.42 139.36±2.12 5.73±0.55 110.07±2.11 39.21±1.53 69.18±19.10 35.91±17.96 96.5±24.4 1.84±0.32 649.38±284.02 62.41±3.72 2.82±2.03 4.91±0.40 8.11±2.53 0.67±0.38 69.33±2.91 8.47±1.18 140.37±3.28 4.31±0.32 111.08±2.04 300 35.63±1.31 101.24±13.12 33.31±4.45 82.56±11.33 1.73±0.32 738.28±268.20 58.52±4.63a1.04±6.05 5.34±0.66 9.12±1.14 1.13±0.34a62.43±3.55 6.64±0.81a136.47±3.21a5.64±0.52 107.28±2.13 42.92±3.84b79.37±23.22 27.80±4.02 72.6±10.5 2.57±0.76 510.37±143.12 63.85±8.63 4.45±1.21 5.15±0.83 9.54±1.42 0.74±0.12 78.48±11.51 8.62±1.24 141.46±7.31 4.24±0.45 109.19±6.15 1250 36.50±0.96 111.34±22.32 39.04±11.53 92.21±13.24 1.95±0.46 683.31±152.33 55.81±3.94b2.22±1.03 5.32±0.45 9.95±1.23 1.35±0.46c64.42±3.61 6.73±0.49a139.05±2.24 5.50±1.81 109.49±3.07 41.56±3.08 61.46±11.25 28.08±4.33 73.2±12.2a2.13±0.35 484.08±136.10 62.74±6.15 4.36±0.74 5.14±0.47 9.42±1.14 0.9±0.30a74.07±6.35 9.09±1.35 143.29±10.10 4.47±0.48 111.34±6.08 2500 36.02±0.93 120.08±28.46 38.91±7.60 77.50±9.76 1.74±0.21 445.28±129.07b58.32±8.07a2.91±0.90 5.48±0.56 9.16±0.84 1.12±0.43a63.33±3.56 6.54±0.72b139.61±2.08 5.54±0.50 108.16±3.24 40.27±2.54 70.08±10.09 30.71±9.85 69.5±23.5 2.20±0.46a626.47±930.35 61.92±6.73 3.73±1.42 5.45±0.42 9.95±2.63 1.04±0.45 72.94±5.96 9.04±1.26 141.46±7.07 4.36±0.49 110.56±5.24

Table 6 Serum biochemistry data for male and female rats after 4 weeks'recovery(ˉ±s)

Notes:results are shown as the mean±standard deviation for 10 rats per sex in each group.Significantly different from the control group by Dunnett's test:aP＜0.001,bP＜0.05,cP＜0.01.ALB:total albumin;ALP:alkaline phosphatase;ALT:alanine aminotransferase;AST:aspartate aminotransferase;CHO:cholesterol;CK:creatine kinase;CRE:creatinine;GGT:γ-glutamyl transpeptidase;GLU:glucose;TBIL:total bilirubin;TG:triglycerides;TP:total protein;BUN:blood urea nitrogen.

Dose(mg·kg－1·d－1)Item Males Females ALB(g/L)ALP(IU)ALT(U/L)AST(U/L)CHO(mmol/L)CK(U/L)CRE(μmol/L)γ-GT(U/L)GLU(mmol/L)TBIL(μmol/L)TG(mmol/L)TP(g/L)BUN(mmol/L)ALB(g/L)ALP(IU)ALT(U/L)AST(U/L)CHO(mmol/L)CK(U/L)CRE(μmol/L)γ-GT(U/L)GLU(mmol/L)TBIL(μmol/L)TG(mmol/L)TP(g/L)BUN(mmol/L)0 35.41±0.92 107.25±14.13 36.81±5.15 83.66±7.23 2.14±0.51 551.27±74.19 68.11±3.47 3.08±0.81 5.85±0.53 9.82±1.01 1.34±0.61 62.07±2.34 6.24±0.60 39.12±1.23 77.38±20.19 34.21±6.10 90.20±21.88 1.61±0.30 583.37±261.46 72.20±10.41 4.01±0.40 4.52±0.51 9.80±0.92 0.82±0.25 68.10±1.42 7.60±1.64 300 20.65±13.26 113.09±33.45 34.42±2.64 77.23±6.94 0.45±0.33a191.43±194.47b71.65±3.60 3.10±0.95 5.83±0.61 8.63±1.28 1.15±0.48 63.21±2.15 4.77±0.64c29.52±15.55 58.51±27.03b29.83±6.27 73.69±4.45a1.15±0.79 292.07±148.31b74.24±11.85 4.92±0.74 4.80±0.54 9.23±0.90 1.08±0.35 65.93±3.40 5.75±3.61 1250 35.73±1.34 107.28±21.07 31.66±1.73b82.05±12.17 2.15±0.52 459.02±310.10 72.94±3.93 2.65±1.52 5.84±1.32 9.94±0.82 1.45±0.52 65.66±3.63 4.46±2.73 41.30±3.61 73.46±8.08 31.20±4.85 74.47±20.43 1.82±0.34 352.49±165.28 69.78±4.17 3.81±0.32 5.23±0.61b9.62±1.61 0.66±0.27 70.64±7.32 6.14±2.10 2500 31.22±3.80 72.46±23.58 27.47±2.91a82.02±18.95 1.34±0.75a202.26±151.41b67.95±6.28 3.91±0.70 5.76±0.50 9.73±0.85 1.21±0.20 62.42±2.84 2.80±1.84b32.17±5.08 62.29±21.31 28.03±2.96b79.21±15.30 1.16±0.77 132.36±108.47c71.09±5.06 5.03±1.70 5.60±0.64c9.61±1.10 0.85±0.18 68.16±3.30 1.16±1.10a

After SBE administration for 26 weeks,the relative liver weight of female rats was significantly higher than that of control rats and tended to remain increased even after the 4 weeks'withdrawal.Notably,exposure to drugs or toxins can cause hepatomegaly.To determine whether long-term administration of SBE had caused the rats,especially female rats,liver injury,we referred to the histopathological examination and observed that after 2500 mg/kg SBE treated rats for 26 weeks,the liver appeared inflammatory changes,which is mainly manifested as inflammatory cell infiltration,Thus,in this study,there was no significant change in the rats'serum enzymes associated with liver injury such as AST,ALT,ALP and γ-GT after long-term treatment with high-dose SBE,but pathological findings showed mild inflammation change in the liver,so that long-term and high-dose SBE may cause liver damage.However,the structural damage of the liver will be self-healing after the ethanol extract stopping,that is,the damage is reversible.

It is notable that the TG levels of male and female animals in the SBE treated groups were significantly higher,indicating that long-term SBE treatment may affect lipid metabolism,but the level of TG returned to normal after 4 weeks'recovery,which indicates lipid metabolism can be restored after SBE withdrawal.At the end of recovery period,the level of GLU in female rats of 1250 and 2500 mg/kg SBE groups werehigher than that of the control rats,but still within the normal range and without pathological significance.Therefore,it is notable that after prolonged use of SBE,blood GLU levels may increase even after ceasing to take the drug.In addition,the K and Na ion value of male rats in SBE groups decreased slightly,which suggests that during the long-term usage of SBE the electrolyte levels may need to be monitored periodically.

Table 7 Relative organ weights of male and female rats orally administered SBE for 13 weeks(%,ˉ±s)

Notes:results are shown as the mean±standard deviation for 5 rats per sex in each group.Significantly different from the control group by Dunnett's test:aP＜0.05.

Item Males Dose(mg·kg－1·d－1)Females Heart Liver Spleen Lung Kidney Brain Stomach Adrenal gland Thymus Testis Epididymis Prostate gland Heart Liver Spleen Lung Kidney Brain Stomach Adrenal gland Thymus Ovary Uterus 0 0.286±0.022 2.738±0.283 0.196±0.024 0.403±0.037 0.600±0.037 0.396±0.041 0.526±0.059 0.017±0.001 0.122±0.027 0.784±0.057 0.342±0.036 0.204±0.038 0.335±0.024 2.853±0.337 0.261±0.031 0.501±0.031 0.668±0.038 0.654±0.055 0.613±0.036 0.031±0.005 0.134±0.021 0.212±0.045 0.058±0.014 300 0.304±0.031 2.545±0.279 0.183±0.025 0.359±0.029 0.585±0.054 0.403±0.026 0.483±0.039 0.018±0.002 0.117±0.026 0.763±0.050 0.325±0.041 0.191±0.062 0.321±0.022 2.694±0.194 0.264±0.018 0.465±0.012 0.686±0.035 0.629±0.031 0.587±0.059 0.029±0.006 0.104±0.009a0.202±0.046 0.052±0.011 1250 0.287±0.033 2.734±0.142 0.204±0.020 0.370±0.034 0.613±0.057 0.401±0.026 0.466±0.051 0.017±0.002 0.109±0.027 0.760±0.076 0.329±0.023 0.215±0.032 0.335±0.019 2.700±0.181 0.245±0.027 0.503±0.025 0.689±0.037 0.663±0.040 0.610±0.044 0.039±0.004a0.147±0.034 0.225±0.048 0.068±0.005 2500 0.294±0.025 2.697±0.208 0.194±0.013 0.341±0.019 0.673±0.062 0.363±0.028 0.473±0.038 0.015±0.002 0.101±0.031 0.710±0.053 0.317±0.064 0.177±0.051 0.341±0.031 2.811±0.270 0.250±0.027 0.483±0.061 0.700±0.052 0.629±0.030 0.664±0.052 0.034±0.005 0.134±0.020 0.232±0.033 0.056±0.004

Moreover,compared with the control group,the ROWs of the adrenal gland and thymus in the SBE treatment groups were decreased and/or increased during the administration period and the relative heart weight increased after stopping drug administration,but none of these organs showed his to pathological changes.Therefore,it is considered that SBE did not cause significant toxicity in the above organs,although there is a need for adequate monitoring of these organs,especially heart function,when an SBE is used long term.

In conclusion,when SBE at a dose of up to 2500 mg·kg－1·d－1was fed to male and female rats for 26 weeks,it did not produce general systemic or major organ toxicity except for liver tissue showing some reversible inflammatory change(recovery after withdrawal).In addition,high-dose SBE treatment of 26 weeks in rats,GLU,electrolyte and lipid levels also have some changes.We think that SBE will be well-tolerated for long-term use as a drug or health food.However,consider that long-term high-dose SBE may cause liver damage,as well as disorders of blood glucose,blood lipids and electrolytes,therefore,in order to ensure drug safety,liver and heart function,and serum glucose,electrolyte and lipid levels should be monitored when using SBE long term.

①汛前来水多、水位高。受开江水位高和冬春降水多影响，嫩江、松花江、黑龙江干流5月份来水量比常年同期偏多4～6成，6月上旬河道水位比常年同期偏高1～3m。

REFERENCES

9 Song KH,Lee SH,Kim BY,Park AY,Kim JY.Extracts of Scutellaria baicalensis reduced body weight and blood triglyceride in db/db mice.Phytother Res 2013;27(2):244-250.

我喜欢会安人的生活方式和乐观的精神。会安是一个小镇，周围是稻田和海滩，一整年都阳光普照，最低温度是20℃，这在我的老家诺曼底就是夏天的气温。而且越南的经济形势稳定，在这里开画廊比较容易。

Table 8 Relative organ weights of male and female rats orally administered SBE for 26 weeks(%,±s)

Notes:results are shown as the mean±standard deviation for 10 rats per sex in each group.Significantly different from control by Dunnett's test:aP＜0.05,bP＜0.001,cP＜0.01.

Item Dose(mg·kg－1·d－1)Males Females Heart Liver Spleen Lung Kidney Brain Stomach Adrenal gland Thymus Testis Epididymis Prostate gland Heart Liver Spleen Lung Kidney Brain Stomach Adrenal gland Thymus Ovary Uterus 0 0.261±0.024 2.547±0.317 0.167±0.015 0.357±0.037 0.571±0.081 0.405±0.033 0.457±0.065 0.012±0.002 0.087±0.010 0.639±0.096 0.304±0.040 0.242±0.047 0.323±0.012 2.375±0.158 0.225±0.024 0.488±0.028 0.616±0.034 0.683±0.036 0.571±0.093 0.023±0.005 0.088±0.019 0.235±0.042 0.064±0.010 300 0.254±0.020 2.526±0.503 0.188±0.022 0.393±0.032 0.552±0.066 0.392±0.044 0.436±0.043 0.012±0.001 0.072±0.018 0.650±0.060 0.291±0.035 0.188±0.035 0.319±0.019 2.712±0.338a0.233±0.031 0.514±0.053 0.655±0.048 0.674±0.052 0.647±0.086 0.022±0.004 0.081±0.018 0.240±0.041 0.062±0.009 1250 0.259±0.021 2.459±0.166 0.175±0.016 0.366±0.033 0.599±0.072 0.384±0.019 0.431±0.054 0.009±0.002a0.065±0.017a0.660±0.076 0.296±0.025 0.186±0.045 0.329±0.022 2.921±0.398b0.241±0.026 0.498±0.037 0.671±0.064 0.693±0.041 0.644±0.064 0.021±0.004 0.083±0.012 0.281±0.075 0.063±0.013 2500 0.272±0.012 2.454±0.094 0.171±0.018 0.397±0.023 0.627±0.046 0.409±0.030 0.456±0.028 0.010±0.002a0.057±0.015c0.708±0.117 0.318±0.035 0.194±0.040 0.312±0.028 2.836±0.237c0.248±0.032 0.517±0.063 0.656±0.048 0.676±0.049 0.648±0.077 0.024±0.007 0.087±0.018 0.257±0.079 0.055±0.013

2 Li C,Lin G,Zuo Z.Pharmacological effects and pharmacokinetics properties of Radix Scutellariae and its bioactive flavones.Biopharm Drug Dispos 2011;32(8):427-445.

3 Gaire BP,Moon SK,Kim H.Scutellaria baicalensis in stroke management:nature's blessing in Traditional Eastern Medicine.Chin J Integr Med 2014;20(9):712-720.

4 4 Muluye RA,Bian Y,Alemu PN.Anti-inflammatory and antimicrobial effects of heat-clearing chinese herbs:a current review.J Tradit Complement Med 2014;4(2):93-98.5 Ji S,Li R,Wang Q,et al.Anti-H1N1 virus,cytotoxic and Nrf2 activation activities of chemical constituents from Scutellaria baicalensis.J Ethnopharmacol 2015;176:475-484.

6 Zhang Z,Lian XY,Li S,Stringer JL.Characterization of chemical ingredients and anticonvulsant activity of American skullcap(Scutellaria lateriflora).Phytomedicine 2009;16(5):485-493.

7 Kuroda M,Iwabuchi K,Mimaki Y.Chemical constituents of the aerial parts of Scutellaria lateriflora and their alpha-glucosidase inhibitory activities.Nat Prod Commun 2012;7(4):471-474.

8 Li-Weber M.New therapeutic aspects of flavones:the anticancer properties of Scutellaria and its main active constituents Wogonin,Baicalein and Baicalin.Cancer Treat Rev 2009;35(1):57-68.

1 Cole IB,Cao J,Alan AR,Saxena PK,Murch SJ.Comparisons of scutellaria baicalensis,scutellaria lateriflora and scutellaria racemosa:genome size,antioxidant potential and phytochemistry.Plant Med 2008;74(4):474-481.

The parameters of body weight,food consumption,organ weight,urinalysis parameters,hematological parameters and serum biochemistry parameters are expressed as the mean±standard error and analyzed using SPSS 16.0 software(IBM SPSS,Chicago,IL,USA).Dunnett's test was used for comparison between the control group and each test group.A difference was considered statistically significant atP＜0.05.

11 Kim DS,Kim SH,Cha J.Antiobesity effects of the combined plant extracts varying the combination ratio of phyllostachys pubescens leaf extract and scutellaria baicalensis root extract.Evid Based Complement Alternat Med 2016;2016(5):9735276.

12 Wu X,Zhang H,Salmani JM,Fu R,Chen B.Advances of wogonin,an extract from Scutellaria baicalensis,for thetreatment of multiple tumors.Onco Targets Ther 2016;9(1):2935-2943.

Table 9 Relative organ weights of male and female rats after 4 weeks'recovery(%,ˉ±s)

Notes:results are shown as the mean±standard deviation for 5 rats per sex in each group.Significantly different from the control group by Dunnett's test:aP＜0.05,bP＜0.001.

Items Dose(mg·kg－1·d－1)Males Females Heart Liver Spleen Lung Kidney Brain Stomach Adrenal gland Thymus Testis Epididymis Prostate gland Heart Liver Spleen Lung Kidney Brain Stomach Adrenal gland Thymus Ovary Uterus 0 0.259±0.009 2.440±0.170 0.161±0.017 0.347±0.034 0.519±0.034 0.387±0.044 0.407±0.042 0.010±0.002 0.058±0.020 0.628±0.051 0.280±0.040 0.193±0.036 0.316±0.012 2.596±0.186 0.234±0.021 0.483±0.021 0.662±0.040 0.684±0.043 0.558±0.067 0.022±0.003 0.074±0.014 0.210±0.048 0.065±0.016 300 0.269±0.010 2.494±0.211 0.172±0.018 0.353±0.015 0.547±0.036 0.398±0.023 0.442±0.035 0.010±0.001 0.080±0.006 0.668±0.044 0.301±0.045 0.221±0.034 0.320±0.019 2.943±0.439 0.265±0.032 0.481±0.049 0.682±0.052 0.724±0.054 0.669±0.094 0.026±0.005 0.081±0.012 0.258±0.060 0.066±0.003 1250 0.274±0.027 2.582±0.307 0.155±0.024 0.340±0.039 0.564±0.054 0.377±0.043 0.419±0.018 0.016±0.007 0.048±0.011 0.659±0.090 0.283±0.040 0.201±0.030 0.349±0.021a2.821±0.272 0.213±0.033 0.510±0.043 0.680±0.048 0.669±0.050 0.618±0.086 0.025±0.003 0.068±0.012 0.230±0.024 0.056±0.011 2500 0.282±0.023 2.628±0.118 0.189±0.012 0.355±0.041 0.638±0.030b0.433±0.028 0.454±0.055 0.014±0.002 0.060±0.016 0.687±0.043 0.307±0.041 0.194±0.025 0.355±0.028a2.811±0.149 0.229±0.033 0.465±0.049 0.746±0.184 0.688±0.074 0.622±0.063 0.021±0.004 0.069±0.005 0.210±0.037 0.068±0.012

Figure 5 Representative HE-stained liver sections of rats after 26 weeks'treatment with SBE

Chronic oral toxicity test showed no gross pathological changes(HE stain,×400).A:liver section of female control rat after 26 weeks distilled water treatment;B:liver section of male control rat after 26 weeks distilled water treatment;C:liver section of female rat after 26 weeks'treatment with 2500 mg·kg－1·d－1SBE;D:liver section of male rat after 26 weeks'treatment with 2500 mg·kg－1·d－1SBE.The black arrow indicates leukocytes infiltrating in the liver.HE:hematoxylin and eosin;SBE:ethanol extract of Huangqin(Radix Scutellariae Baicalensis).

13 Yimam M,Brownell L,Pantier M,Jia Q.UP446,analgesic and anti-inflammatory botanical composition.Pharmacognosy Res 2013;5(3):139-145.

23 Pang H,Xue W,Shi A,et al.Multiple-ascending-dose pharmacokinetics and safety evaluation of baicalein chewable tablets in healthy chinese volunteers.Clin Drug Investig 2016;36(9):713-724.

15 Chen H,Gao Y,Wu J,et al.Exploring therapeutic potentials of baicalin and its aglycone baicalein for hematological malignancies.Cancer Lett 2014;354(1):5-11.

16 Ku SK,Bae JS.Antithrombotic activities of wogonin and wogonoside via inhibiting platelet aggregation.Fitoterapia 2014;98:27-35.

17 Li Y,Tu M,Cheng C,et al.Wogonoside induces apoptosis in Bel-7402,a hepatocellular carcinoma cell line,by regulating Bax/Bcl-2.Oncol Lett 2015;10(3):1831-1835.

Figure 6 Representative HE-stained sections of other vital organs of rats treated with 2500 mg·kg－1·d－1SBE for 26 weeks(HE stain,×200)

A,B:representative sections of heart;C,D:representative sections of spleen;E,F:representative sections of lung;G,H:representative sections of kidney;I,J:representative sections of brain;K,L:representative sections of stomach;M,N:representative sections of intestinal;O,P:representative sections of testis;Q,R:representative sections of uterus;S,T:representative sections of ovary.A,C,E,G,I,K,M,Q,S:female control group;B,D,F,H,J,L,N,R,T:female high-dose SBE group;O:male control group;P:male high-dose SBE group.HE:hematoxylin and eosin;SBE:ethanol extract of Huangqin(Radix Scut ellariae Baicalensis).

18 Burnett BP,Silva S,Mesches MH,Wilson S,Jia Q.Safety evaluation of a combination,defined extract of Scutellaria baicalensis and Acacia catechu.J Food Biochem 2007;31(6):797-825.

19 Qi Q,Peng J,Liu W,et al.Toxicological studies of wogonin in experimental animals.Phytother Res 2009;23(3):417-422.

20 Yimam M,Zhao Y,Ma W,Jia Q,Do SG,Shin JH.90-day oral toxicity study of UP446,a combination of defined extracts of Scutellaria baicalensis and Acacia catechu,in rats.Food Chem Toxicol 2010;48(5):1202-1209.

21 Lee YC,Hyun E,Yimam M,Brownell L,Jia Q.Acute and 26-week repeated oral dose toxicity study of up446,a combination of scutellaria extract and acacia extract in rats.Food Nutr Sci 2013;4(7):14-27.

与他们交往的过程中，我们不光提升了自己的英语能力，也了解了不少国家的人文风情，更收获了一份友谊。这就是我游学最重要的目的：在这里，我有了很多和外国人交流的机会，也让自己的英语得到了很好的锻炼。

22 Li M,Shi A,Pang H,et al.Safety,tolerability,and pharmacokinetics of a single ascending dose of baicalein chewable tablets in healthy subjects.J Ethnopharmacol 2014;156(28):210-215.

14 Gasiorowski K,Lamer-Zarawska E,Leszek J,et al.Flavones from root of Scutellaria baicalensis Georgi:drugs of the future in neurodegeneration?CNS Neurol Disord Drug Targets 2011;10(2):184-191.

24 Yimam M,Lee YC,Qi J.26-week repeated oral dose toxicity study of UP446,a combination of defined extracts of Scutellaria baicalensisand Acacia catechu,in beagle dogs.Regul Toxicol Pharmacol 2016;78:66-77.

25 Linnebur SA,Rapacchietta OC,Vejar M.Hepatotoxicity associated with chinese skullcap contained in Move Free Advanced dietary supplement:two case reports and review of the literature.Pharmacotherapy 2010;30(7):258-262.

26 Chalasani N,Vuppalanchi R,Navarro V,et al.Acute liver injury due to flavocoxid(Limbrel®),a medical food for osteoarthritis:A case series.Ann Intern Med 2012;156(12):297-300.

27 Yang L,Aronsohn A,Hart J,Jensen D.Herbal hepatoxicity from Chinese skullcap:A case report.World J Hepatol 2012;4(7):231-233.

28 Dhanasekaran R,Owens V,Sanchez W.Chinese skullcap in Move Free arthritis supplement causes drug induced liver injury and pulmonary infiltrates.Case Reports Hepatol Article 2013;(2013-4-14),2013:965092.

29 Morgan S L,Baggott J E,Moreland L,Desmond R,Kendrach A C.The safety of flavocoxid,a medical food,in the dietary management of knee osteoarthritis.J Med Food 2009;12(5):1143-1148.

高管内部薪酬差距、高管与员工薪酬差距与公司绩效的关系 ………………………… 刘春旭，丁 鹏（5．22）

30 Khanal T,Kim H G,Choi J H,et al.Protective role of intestinal bacterial metabolism against baicalin-induced toxicity in HepG2 cell cultures.J Toxicol Sci 2012;37(2):363-371.

战乱时期，父亲把工厂迁到武昌区法租界里。1941年太平洋战争爆发，日军立即没收了英法美等租界的全部资产，我父亲的企业落在汪伪政权手中，家中生活来源断绝。父亲经常与老乡躲在一边听收音机短波，盼望把日本鬼子赶出中国，但他没能看到这一天。生活的苦难摧残父亲的健康，他的肺病日渐严重。