Objective To investigate the effects of metformin on serum testosterone (T)levels and ovulation in hyperinsulinemic women Methods Twenty nine patients with hyperinsulinemic anovulation were treated with metformin for 12 weeks Among them 19 had history of clomiphene(CC) treatment with 11 cases failure CC was given again after one month of metformin treatment Serum T, luteinizing hormone,follicle stimulating hormone, insulin(Ins) levels were measured after 4 weeks of treatment Basic body temprature was also observed Results Serum T and Ins levels declined significantly after 4 weeks of metformin treatment [(2 7?1 6) nmol/L Vs (1 9?1 0) nmol/L, (49 2?34 1) mU/L Vs (25 3?23 9) mU/L, P

Objective To evaluate the effect of hepatic resection for liver malignancies. Method The clinical data including the patterns and effect of operation in 108 patients with malignant liver disease underwent hepatic resection in recent 5 years in our hospital were analyzed retrospectively.Result Tweent-eight cases occurred complications and four died after operation . The 1-,3-year survival rate were 87.5% and 43.8% respectively. Conclusions Surgical resection is still an effective procedure for the liver malignancies .

Objective To compare the efficacy and safety of long-term treatment (48 weeks) with tiotropium bromide (5 μg) via Respimat(R) with placebo in patients with chronic obstructive pulmonary disease (COPD).Methods A total of 338 patients were randomized in this double-blind,placebo controlled,parallel study.All patients received either tiotropium bromide or placebo.Tiotropium bromide solution 5 μg (2 × 2.5 μg/puff) or matching placebo was delivered via Respimat(R) at a dosage of once daily for 48 weeks.Co-primary endpoints were trough forced expiratory volume in one second (FEV1) and the time to first exacerbation.Results Statistically significant improvements of both trough FEV1 and trough forced vital capacity (FVC) in the tiotropium group were achieved at weeks 4,24,and 48 compared with those in the placebo group(P ＜ 0.000 1).Tiotropium treatment delayed the time to first exacerbation.The time was 157 days in the tiotropium group and 85 days in the placebo group.A statistically significant difference (P =0.002 7) in favor of tiotropium was also observed.The total numbers of exacerbation during treatment were 90 and 128 in the tiotropium and placebo groups,respectively.The Poisson regression analysis gave a mean exacerbation rate per patient year exposure of 0.67 in the tiotropium group compared to 0.98 in the placebo group with a rate ratio of 0.69 (95％ CI O.50-0.93,P =0.016 4).A much larger improvement from baseline in St.George's respiratory questionnaire (SGRQ) total score was observed for the tiotropium group than in the placebo group(P =0.036 7),SGRQ symptom and activity scores of patients in the tiotropium group were also superior to those of patients receiving placebo.The drugs-related adverse events in the tiotropium and placebo groups were 12 cases and 11 cases,respectively.Conclusions Tiotropium significantly improved lung function and quality of life,delayed the time to first exacerbation,reduced the number of exacerbation.Overall,tiotropium was well tolerated.

Objective To compare blood flow perfusion conditions of renal allograft of different renal function with conventional ultrasound and contrast enhanced ultrasonography (CEUS). Methods Sixty patients underwent kidney transplatation were divided into normal group (SCr≤110 μmol/L) and abnormal group (SCr＞110 μmol/L). Renal structure and inner diameter of renal artery were messured with two-dimensional ultrasound. The hemodynamic parameters (PSV, EDV and RI) were messured with CDFI. Data of renal blood flow perfusion (AT, TTP, △I and Increase), data of renal blood flow perfusion (AT, TTP, △I and Increase) were quantitatively analyzed with CEUS combined with time intensity curve. All the data were compared between two groups.Results There was no significantly difference of above indexes for B-mode ultrasound and CDFI between two groups, except EDV of arcuate arteries. CEUS parameters of patients with normal renal function were better than those with abnormal function.Conclusion Microcirculation perfusion changes of transplanted kidney with abnormal function could be detected with CEUS. The quantitative indexes of transplanted kidney with normal function were better than those with abnormal function.

Objective To explore the effect of depakine in the treatment of traumatic epilepsy relapse,and compared carbamazepine and lamotrigine combined with depakine in the treatment of traumatic epilepsy.Methods Open trial in 131 patients with traumatic epilepsy relapse treated with depakine,and all patients were aderministered with carbamazepine or lamotrigine combined with depakine.3 months before treatment,the epilepsy seizures frequency was compared,and 6 months after the treatment,the efficacy,adverse reactions and safety was compared between two mothods.Results Application of carbamazepine or lamotrigine combined with depakine treatment for 6 months,the seizure frequency significantly decreased compared with before treatment;The seizure frequency reduction( ≥50％ ) were 72.6％ and 91.3％,the difference was statistically significant between before and after treatment( P ＜ 0.01 ) ;The adverse reactions of carbamazepine combined with depakine was 37.1％,and it of lamotrigine combined with depakine was 8.7％.Conclusion Lamotrigine combined with depakine in the treatment of traumatic epilepsy recurrence was effective,and its side effects was light.

Objective To establish a flow cytometry-based drug susceptibility test for the rapid de-tection of antifungal susceptibility or resistance of Candida isolates.Methods The gate selection and opti-mal experimental conditions of flow cytometry-based drug susceptibility test were determined by using Candi-da albicans strain ATCC90029 as the test strain and propidium iodide ( PI) as the fluorescent dye .The es-tablished flow cytometry-based drug susceptibility test was used to detect the susceptibility or resistance to fluconazole or voriconazole of 110 isolates belonging to Candida species, and the obtained results were com-pared with those by using typical M 27-A3 constant dilution method .Results The killed and viable Candida albicans ATCC90029 strains were clearly divided into two groups on the figure of SS /log (FL3) by regulating voltages.There was a high correlation between the results of susceptibility test and the proportions of killed and viable fungi in mixture (r=0.999).The flow cytometry-based drug susceptibility test could provide the results within 30 min and its optimal concentration of fungal suspension , time of drug-fungus incubation , dyeing method and time were 1.0×106/ml, 3 h incubation and sodium deoxycholate-pretreated plus PI dye-ing for 5 min, respectively .The total coincident rates between the established test and the constant dilution method were 98.2%and 87.3%in the detection of drug susceptibility of 110 fungal isolates to fluconazole and voriconazole .Conclusion The flow cytometry-based drug susceptibility test shows advantages of rapidi-ty, accuracy and high sensitivity compared with the constant dilution method .It has a great potential for clin-ical application .

Aim To observe the effect of BAPTA-AM on extracellular acid-induced autophagy in rat articular chondrocytes and its possible mechanisms.Methods Rat articular chondrocytes were isolated from Sprague-Dawley rats and incubated with different pH medium. The states of autophagy were examined by acridine or-ange (AO ) staining .Moreover,the expressions of LC3 ,Beclin-1 ,ULK1 ,CaMKKβ,AMPK and mTOR were detected using Western blot or quantitative real-time PCR (qRT-PCR ). Intracellular calcium ([Ca2+]i )was analyzed by a Ca2+-imaging method. Results Compared with pH 6.0 group,BAPTA-AM could significantly decrease the activation of autophagyinduced by acid exposure,and the expressions of autophagy markers including LC3 Ⅱ,Beclin1 and ULK1were also decreased,accompanied with reduced acidinduced [Ca2 +]i influx,decreased proteins expressionof CaMKKβand phosphorylatedAMPK,and increasedphosphorylation of mTOR.Conclusion BAPTAAMcan significantly restrain acidinduced autophagy in ratarticular chondrocytes,the mechanism of which may beassociated with decreased Ca2 + influx.

Objective To investigate the effect of occult HBV infection (OBI) on carcinogenesis of cryptogenic hepatocellular carcinoma.Methods Samples of hepatocellular carcinoma (HCC) and pericarcinomatous tissues obtained after hepatectomy from January 2011 to November 2013 at the Third Affiliated Hospital of Sun Yat-Sen University were collected.They were divided into two groups:the cryptogenic HCC group (the CH group,n =26) and the HBV related HCC group (the HH group,n =40).These samples were compared with the normal liver tissues obtained in 30 patients.HBV DNA was identified by the nested polymerase chain reaction and the immunohistochemical method was taken to examine the hepatitis B virus X protein (HBx) and Yes-associated protein (YAP) expressions.Results OBI was identified in 20 (77.8％) cryptogenic HCC patients and 8 (26.7％) in the control group.There was a significant difference between the two groups (x2 =14.072,P ＜ 0.05).HBV DNA was detected in all the HBV-related HCC patients.The HBx protein expression was mainly located in the cytoplasm of liver cells and liver cancer cells,but YAP was expressed in the nucleus.Both of them showed diffuse brown or tan particles.In the HH group and the CH group,the positive expression rates of HBx protein in the tumorous tissues were 80.0％ and 90.0％,respectively,and 85.0％ and 82.5％ in the nontumorous tissues,but only in 40.0％ in the control group.The positive expression rates of YAP in the tumorous tissues were 65.0％ and 67.5％,respectively,15.0％ and 20.0％ in the nontumorous tissues,respectively,but only in 12.5％ in the control group.The HBx expression in the cancerous tissues and para-cancerous tissues of the HH group and the CH group showed no significant difference (P ＞ 0.05),but the YAP expression in the tumor tissues was significantly higher than that in the nontumorous tissues (P ＜ 0.05).The HBx and YAP expressions in the HH group were comparable to the CH group (P ＞ 0.05).However,their expressions in the cancerous tissue of the HH group and the CH group were significantly higher than in the normal liver tissues (P ＜ 0.05).Conclusion A high prevalence of HBV infection was observed in HBsAg-negative HCC and the high expressions of HBx and YAP might be involved in the process of cryptogenic hepatocellular carcinoma.

Aim To study the effects of extracellular acidosis on articular chondrocytes pyroptosis and its possible mechanisms. Methods Primary articular chondrocytes were incubated in different pH and NAC. The expression of proinflammatory cytokines IL-1β, IL-18, ASC, NLRP3, caspase-1 were detected by Western blot and real-time PCR. The state of pyropto-sis was identified by AO/EB staining and LDH con-tents. The expression of ROS was observed by DCFH-DA, and ELISA was used to detect the IL-1β,IL-18 in cultured supernatants. Results Compared with the normal cell, extracellular acidosis could increase the expression of IL-1β, IL-18, ASC, NLRP3 and caspase-1 , upregulate the fluorescence intensity of in-tercellular ROS, accompanied with the promoted release of LDH. Moreover, it is observed that extra-cellular acidosis could also induce chondrocytes death by AO/EB staining. NAC,the scavenger of ROS could inhibit these effects of extracellular acidosis on chon-drocytes. Conclusion Extracellular acidosis may in-duce chondrocyte pyroptosis via upregulating the intra-cellular ROS content.

Objective To study the effect of Tspan 8 on metastasis and invasion of human hepatocellular carcinomas(HCC).Methods RT-PCR and western blot were used to detect the expressions of Tspan 8 in HCC cell lines,HCC and matched nontumorous tissues.The expression of Tspan 8 was then down-regulated by LV/GFP/Tspan 8 in HCC cells.The expressions of Tspan 8 mRNA and protein were determined by RT-PCR and Western blot assay,respectively.The proliferation was examined by MTT,the expression of AMDM12 was assessed by Western blot,and the invasion ability of HCC cells was evaluated by transwells.Results A high level of Tspan 8 was found in high metastatic potential HCC cells,and the expression of Tspan 8 in HCC tissues was much higher than that in the matched nontumorous tissues. Down-regulation of Tspan 8 had no influence on the proliferation of HCC cells (P＞0.05),while it inhibited the expression of ADAM12 and the invasive ability of HCC cells (P＜0.01,P＜0.01 respectively).Conclusion Tspan 8 played an important role in invasion and metastasis of human hepatocellular carcinomas and down-regulation by LV/GFP/Tspan 8 inhibited the invasiveness of human hepatocellular carcinoma cells.