OBJECTIVE: To evaluate the quality of Gentiana scabra in Qingre jiedu oral solution. METHODS: HPLC method was established to determine the content of gentiamarin in the oral solution. 299 batches of Qingre jiedu oral solution in circulation were measured and evaluated. RESULTS: Developed HPLC method is simple and reliable. The quality of Qingre jiedu oral solution in circulation is not satisfied. CONCLUSION: Great importance should be attached to the manufacture procedures monitoring of Qingre jiedu oral solution and quality control of G. scabra.

[Objective]To compare the effects of continuous femoral nerve block(CFNB) and epidural analgesia(CEA) on rehabilitation after total knee arthroplasty(TKA) surgery. [Methods]Fifty patients undergoing unilateral TKA surgery were randomly divided into group CFNB(n=25) and group CEA(n=25).All patients were received unilateral spinal anesthesia with 0.5% dicaine and given analgesia with 0.2 % ropivacaine with 1ug/ml sufentanil.VAS pain scores during rest and continuous passive movement(CPM) at each time point was recorded.Other parameter such as the angle of initiative flections,blood loss at 6 h post operation,the concentration of serum hemoglobin at 24 h,48 h and side effect were recorded.[Results]The VAS pain scores during test had no significant difference between two groups.The VAS scores of CPM at 24 h and 48 h in group CFNB were obvious lower(3.68?0.93 and 3.93?0.78) than those in group CEA.The initiative flections were similar at 12 and 24 hours after operation.The concentration of hemoglobin at 6 h after operation was lower than that before operation.After reinfusion it was increased but did not reach the level before operation.The incidence of side effects was not noted in both groups.[Conclusion]After TKA surgery,the continuous femoral nerve block can provide better pain relief.No adverse impact on rehabilitation movement of operated legs or blood loss and has less side effect.Therefore,it should be considered an alternative analgesia method after TKA.

Objective To observe the effect of fluvastatin on the activity of extracellular signal-regulated kinase(ERK)1/2,expression of connective tissue growth factor(CTGF) in glomerular mesangial cells stimulated by high glucose,and explore the pathogenic mechanism of diabetic nephropathy(DN) and its early prevention and treatment.Methods The study consisted of two parts,the first part was to repeat the existing experienment of stimulatory effect of high glucose on mesangial cells,and validated that high glucose could up-regulated the expression of CTGF through stimulating ERK,and the change was unrelated with hypertonic conditions.There were five groups:①normal control group(NG);②high glucose group(HG);③mannitol group(MN);④NG+ERK inhibitor PD98059 group(NG+PD);⑤HG+ ERK inhibitor PD98059 group(HG+PD).The second part was to observe the changes of the expressions of CTGF protein and mRNA in mesangial cells at high glucose concentration before and after fluvastatin intervention.There were also five groups:①normal control group(NG);②high glucose group(HG);③normal glucose and fluvastatin group(NG+F);④high glucose + fluvastatin(HG+F);⑤high glucose + fluvastatin and mevalonic acid group(HG+F+M).The activity of ERK1/2 and the expressions of CTGF protein and mRNA in cultured glomerular mesangial cells stimulated by high glucose were detected with Western blotting and RT-PCR.The amount of fibronectin(FN) in the supernatant of cells was analyzed by ELISA.Intracellular total protein level was measured by Coomassie brilliant blue method.Results The first part showed that the activity of ERK1/2 and the expression of CTGF protein in HG were more higher than those in NG(P

AIM:To investigate the influence of high-mobility group box 1 (HMGB1) on the proliferation of neural stem cells in peri-infarction cortex of focal cerebral ischemia/reperfusion model rats .METHODS: Male SD rats (n=48) were randomly divided into sham group , ischemia/reperfusion (I/R) group, RNA interference group and nega-tive interference group .The rat middle cerebral artery was blocked to establish focal cerebral I /R model ( ischemia for 1 h and reperfusion for 7 d).Lentivirus vector of HMGB1 shRNA was used to suppress the HMGB1 protein expression in the rat brain.The effect of RNA interference was evaluated by the methods of double-immunofluorescence labeling of HMGB 1/GFAP and Western blotting .The proliferation of neural stem cells in the peri-infarction cortex was assessed by double labe-ling of BrdU/nestin.RESULTS: The protein expression of HMGB1 in I/R group was much higher than those in sham group (P<0.05).RNA interference effectively inhibited the HMGB1 expression (P<0.05).Double labeled BrdU/nestin positive cells in I/R group were more than that in sham group (P<0.05).The double labeled BrdU/nestin positive cells were significantly decreased in RNA interference group (P<0.05).CONCLUSION:Focal cerebral ischemia/reperfusion injury promotes the proliferation of neural stem cells in peri-infarction cortex by increasing HMGB 1 protein level .

Objective: To detect circulating tumor cells (CTCs) in patients with gastric cancer and evaluate the relationship among CTCs, clinico-pathological characteristics, and prognosis of gastric cancer. Methods: Peripheral blood samples (10 mL in EDTA) were obtained from 45 patients with gastric cancer. CTCs were detected using density-gradient centrifugation and immunofluo-rescence staining. The clinical significance of the two methods were also compared and investigated. Results:CTC-positive case was defined by the presence of at least one CK19 (+)-CTC per 10 mL of the sample. CTCs were found in 27 of the 45 patients with gastric cancer. The presence of CTCs was significantly correlated with lymph node metastasis, distant metastasis, and recurrence (P=0.007, 0.035, 0.035, respectively). However, CTCs were not significantly correlated with sex, age, tumor location, TNM staging, and tumor differentiation (P>0.05). Conclusion:CTCs were associated with poor prognosis of gastric cancer.

Objective To observe the effect of autophagy inhibitor on the activation of alcohol induced hepatic stel-late cells, and the mechanisms thereof. Methods HSC-T6 cells were cultured in vitro and divided into four groups, includ-ing blank control group, alcohol group, 5 mmol/L 3-MA+alcohol group (low alcohol group) and 10 mmol/L 3-MA+alcohol group (high alcohol group). RT-PCR was used to detect the expression levels ofα-smooth muscle actin (α-SMA) and typeⅠcollagen. The levels of LC3Ⅱ,α-SMA and typeⅠcollagen were detected by Western blot assay. The cell viability of HSC-T6 was detected by MTT assay. Results The mRNA expressions ofα-SMA, typeⅠcollagen and the protein of expressionsα-SMA, typeⅠcollagen and LC3Ⅱwere significantly up-regulated in alcohol group compared with those of control group (P<0.05), while the expressions of those parameters were significantly down-regulated in 10 mmol/L 3-MA+alcohol group (P<0.01). The mRNA and protein levels ofα-SMA and typeⅠcollagen were significantly decreased in two 3-MA-treated groups compared with those in alcohol group (P<0.05). Meanwhile, compared with the 5 mmol/L 3-MA+alcohol group,the protein expressions ofα-SMA, typeⅠcollagen and LC3Ⅱwere significantly decreased in10 mmol/L 3-MA+alcohol group (P < 0.05 ). Compared with the alcohol group,there was significantly lower proliferation activity in all two 3-MA-treated groups (P<0.05). Conclusion 3-MA can inhibit the protein expression of LC3Ⅱ,α-SMA and typeⅠcollagen induced by alcohol in HSC-T6 cells, and inhibit the proliferation of HSC cells.

Objective To explore the clinical effects of different regimens of letrozole in the treatment of PCOS in patients with infertility.Methods A total of 87 cases of PCOS infertility were selected as the research subjects.The patients were divided into the group A(44 cases of menstrual cycle 3rd~7th day oral letrozole 5mg/d), the group B 43 cases(3D of the menstrual cycle to take a one-time oral letrozole 20mg),ultrasound monitoring was used in the treatment process,when the maximum follicle diameter greater than or equal to 18mm,HCG intramuscular injection of human chorionic 10 000IU(HCG).The changes of endocrine hormones in the two groups were compared and the effect of ovulation induction was compared.Results Menstruation the fifth day serum T,FSH,LH,between the two groups had no statistically significant differences (P>0.05 ).The serum E2 in the group B [(106.1 ± 43.2)nmol/L]was lower than that in the group A[(183.2 ±69.4)nmol/L](t=6.204,P<0.05).Injection of HCG on the same day,the serum T,FSH and E2 between the group A and group B showed no significant differences(P>0.05),serum LH level in group B[(31.1 ±9.2)IU/L]was lower than that in the group A[(34.6 ±8.9)IU/L](t=2.010,P<0.05 ).In group B,the endometrial thickness,the number of mature follicles,the ovulation rate,the pregnancy rate within 12 months were (0.94 ±0.11)mm,(1.46 ±0.41),95.35%,35.26% respectively,which were significantly higher than those of the group A[(0.83 ±0.13)mm,(1.14 ±0.25),70.45%,13.64%](t=4.256,t=4.407,χ2 =9.445,4.398,all P<0.05).Conclusion The 3rd day at menstrual cycle,taking a single oral dose of letrozole 20mg program is favor of an early lowering E2 levels,and can promote follicular maturation and development,improve the pregnancy rate in patients with PCOS.

Objective To investigate the expression pattern of growth arrest-specific transcript 5 ( GAS5 ) in hepatocellular carcinoma and assess its pre-operation and post-operation levels.Methods Totally 243 patients were collected in Zhongnan Hospital in 2015, and were divided into 4 groups: pre-operation (117 cases), 1 weeks after operation (39 cases), patients with hepatitis B (55 cases) and cirrhosis ( 71 cases ) .Meanwhile, 129 controls were collected.The expression of GAS5 in plasma was detected by real-time quantitative PCR. The levels of GAS5 in subgroups were compared by Oneway ANOVA.The relationship between GAS5 and the clinical pathologic features, and its pre-operation and post-operation expression were analyzed by Student′s t test.Results GAS5 was downregulated in HCC plasma and the levels of GAS5 were associated with tumor differentiation ( R2 =0.219,P=0.011) and TNM stage (R2 =0.036,P=0.044).Compared with the levels in pre-operation group (3.958 ±0.282), GAS5 were upregulated after surgery (3.843 ±0.223),t=2.283, P=0.028.In addition, GAS5 expression in well and moderately differentiated HCC was higher than poorly differentiated ones before operation[(3.873 ±0.191) and(4.151 ±0.365),t=2.271,P=0.035], but there was no significant difference after operation[(3.880 ±0.154) and (3.879 ±0.246),t=0.032, P=0.975].The ROC curves indicated that GAS5 had a good sensitivity to differentiate HCC from the healthy ( 88%, 88/100 ) and the cirrhosis ( 90%, 90/100 ) . Conclusion GAS5 might be used to assess the treatment of HCC.

Objective To investigate the role of OMA1 in acute kidney injury (AKI) induced by lipopolysaccharide (LPS).Methods OMA1 wild-type and knocked out mice (8 week old) were injected with 10 mg/kg body weight of LPS.The model was confirmed by testing mouse serum creatinine and blood urea nitrogen.The apoptosis in mouse kidney cortex was examined by TUNEL staining and cleaved caspase 3.In vitro,in humam kidney proximal tubular cells (HK2) were knocked down OMA1 by transfecting OMA1 shRNA,with the scramble shRNA being used as negative control of transfection.HK2 cells were cultured with 5 μg/ml of LPS for 24 hours to induce apoptosis.DAPI staining of cells and caspase-3 activity were applied to test apoptosis.The images of mitochondria in cells were obtained by transfection of mito-green plasmid and OMA1 shRNA.Western blotting was used to exam the OMA1 and Cytochrome C expressions.Resudts Compared with OMA1 KO mice,LPS induced more severe AKI of WT mice with higher Scr [(97.2±26.5) μmol/L vs (53.0±17.7) μmol/L,P ＜ 0.05] and BUN [(43.3± 13.7) mmol/L vs (29.7±7.7) mmol/L,P ＜ 0.05].Moreover,there were more apoptosis cells in kidney cortex in WT mice than in OMA1 KO mice [(75.4± 26.1)/ram2 vs (38.3± 14.4)/mm2,P＜ 0.05].About 46％ of OMA1 expressions in HK2 cells were inhibited by OMA1 shRNA transfection (P ＜ 0.05).Further,OMA1 shRNA cells with LPS stimulation had decreased mitochondria fragmentation [(29.8±10.9)％ vs (43.2±6.8)％,P ＜ 0.05],Cytochrome C release [(37.0±12.3)％ vs (76.0±26.2)％,P ＜ 0.05],and cell apoptosis [(13.2±3.9)％ vs (25.0±7.1)％,P ＜ 0.05] as compared with control cells.Conclusion Knockdown of OMA1 alleviated septic AKI through inhibition of cell apoptosis,mitochondria fragmentation,and Cytochrome C release.

Objective To investigate clinical significance of Homer expression in peripheral blood leukocytes of patients with ischemic stroke (IS).Methods It was a retrospecive study.The gene expression levels of Homer were measured by RT-qPCR.266 patientscollected in Zhongnan Hospital from September 2015 to June 2016were divided into 5 groups:large-artery atherosclerosis (LAA,100 cases),cardioembolism (CE,42 cases),small vessel occlusion (SVO,68 cases),stroke of other demonstrated etiology (SOE,23 cases) and stroke of undemonstrated etiology (SUE,33 cases).Meanwhile,age and sex matched 126 healthy controls were also collected.IS diagnostic criteria for cerebral infarctionwas in accordance with the guideline for acute ischemic stroke in China in 2010.The levels of Homers in subgroups were compared by Oneway ANOVA.The area under curve (AUC) and 95％ confidence interval (CI) were calculated using ROC analyses.The odds ratio (OR) and 95％ CI were calculated using the multivariate logistic regression analyses.Results The levels of Homer1 [2.01 ± 0.15] and Homer2 [1.81 ± 0.31] in LAA patients were significantly higher than othergroups [Homer1 CE:2.40 ± 0.34;SVO:2.38 ± 0.35;SOE:2.36 + 0.33;SUE:2.40 ± 0.30;control group:2.35 ± 0.28;Homer2 CE:2.09 ± 0.38;SVO:2.08 ± 0.30;SOE:2.09 ± 0.41;SUE:2.10 ± 0.34;control group:2.12 ± 0.31] (Homer1 CE:t =9.353,P＜0.001;SVO:t =9.258,P＜0.001;SOE:t =5.396,P＜0.001;SUE:t=9.644,P＜0.001;control group:t =11.882,P＜0.001;Homer2 CE:t =4.725,P＜0.001;SVO:t =5.545,P＜0.001;SOE:t=3.640,P ＜ 0.001;SUE:t =4.669,P ＜ 0.001);There was no significant difference in the expression of Homer1 (F =0.940,P =0.441) and Homer2 (F =0.336,P =0.854) between non-LAA groupsand healthy controls.There was no significant difference in the expression of Homer3among the groups (F =0.641,P =0.669).Multinomial logistic regression analyses revealed that,higher Homerl (adjusted OR =8.62,95％ CI:4.13-18.00,P＜0.001) and Homer2 (adjusted OR=2.42,95％ CI:1.75-3.36,P ＜ 0.001) levels showed significant associations with increased odds of having LAA stroke,compared with the controls.ROC curves showed that the AUC of the combination of Homer1 and Homer2 for differentiating LAA and controls was 0.896 (95％ CI:0.862-0.929,P ＜0.001) and the AUCfor differentiating LAAand non-LAA was 0.847 (95％ CI:0.800-0.894,P ＜ 0.001).Conclusion The expression of Homer1 and Homer2 in peripheral blood leukocytes could be used as novel biomarkers for LAA stroke.