OBJECTIVE:To develop an HPLC method for the determination of L-Carnosine in Polaprezine granules.METHODS:The Chromatographic column was Agilent ODS with detection wavelength of 215 nm and flow rate of 1 mL?min-1.The mobile phase consisted of sodium subsulfite buffer solution(pH=3)-acetonetrile(77:23).The column temperature was set at 30 ℃ and injection size was 10 ?L.RESULTS:The good linearity was obtained in the range of 0.1559～3.1584 ?g for L-Carnosine(r=1.0000).The average recovery was 99.65%(RSD=0.078%).CONCLUSION:The method is simple,accurate and sensitive,and it is suitable for content determination of L-Carnosine in Polaprezine granules.

Objective Toinvestigate the relationship between red blood cell volume distribution width (RDW) and acute coronary syndrome (ACS),and the correlation between RDW and degree of coronary artery stenosis. Methods A total of 221patients accepting coronary angiography were involved as ACS group and another 50 patients also accepting coronary angiography as normal control group. ACS group wasfurther divided into unstable angina (UA) group of 77 cases and acute myocardial infarction (AMI) group of 144 cases ,then their level of RDW was detected. According to Gensini score ,we divided the 221 positive cases of coronary angiography into ≤ 30 point group ,31-60 point group and > 60 point group and their level of RDW was detected. Results The level of RDW in ACS group(13.02 ± 0.68)was significantly higher than that innormal control group(12.62 ± 0.38),and the difference was statisticallysignificant(P < 0.05). And the difference between UA group(12.84 ± 0.68)and AMI group(13.12 ± 0.66)also showed statistical significance(P < 0.05). With the increase of Gensini score ,the level of RDW was also increased ,and the difference between each two groups was statistically significant (P < 0.05) except that between control group and ≤ 30 point group. Conclusions The level of RDW in ACS group and AMI group is significantly higher ,suggesting that RDW may be involved in the pathogenesis of acute coronary syndrome ,and the RDW level could be a potential indicator of ACS. RDW level is closely related to the severity of coronary stenosis and could be one of its predictors.

Objective To investigate the clinical characteristics,peripheral insulin sensitivity,and β-cell function in patients with ketosis-prone diabetes(KPD).Methods Thirty-one patients with newly diagnosed ketosisprone diabetes were admitted to West China Hospital from January 2004 to December 2009.They were divided into 2 groups according to their body mass index (BMI):OB-KPD (BMI ≥ 25 kg/m2,n =22) and Lean-KPD (BMI ＜ 23 kg/m2,n =9).10 patients with newly-onset type 2 diabetes free from ketosis (OB-DM:BMI ≥ 25 kg/m2,n =10) were enlisted as control.Detailed assessments of medical history and symptoms of hyperglycemia were performed.The islet cell antibody (ICA),insulin autoantibody (IAA),anti-glutamic acid decarboxylase antibody (GAD-Ab),fasting plasma glucose,serum insulin,C-peptide and free fat acids concentrations were measured.All of the subjects underwent oral and intravenous glucose tolerance tests,euglycemic-hyperinsulinemia and hyperglycemia clamp test,to evaluate the insulin secretion and insulin sensitivity respectively.Insulin sensitivity was determined by glucose disposal rate (GDR) of steady state during euglycemic clamp and acute insulin secretion was calculated by insulin area under curve(AUCins 0-10 min) during IVGTT.Maximal insulin secretion was determined by glucose infusion rate (GIR) and serum insulin concentration of steady state during hyperglycemic clamp test.Results Age,sex,duration of diabetes were matched among groups.A family history of diabetes was strongly associated with those patients with obesity,compared with lean ketosis prone diabetes(16/22 vs 1/9).GDR was (4.91 ± 1.82) mg · kg 1 · min-1 in subjects with OB-KPD,being lower than that in Lean-KPD patients[(6.26 ± 1.89) mg · kg 1 · min-1] and OB-DM group[(6.78-± 1.69) mg · kg 1 · min-1,P＜0.01].Serum insulin and C-peptide in OB-KPD patients were higher than Lean-KPD patients.Area under the insulin curve [AUCins0-10min (183.86 ± 31.1) mIU/L] and GIR[(2.65 ±1.53) mg · kg-1 · min-1] in OB-KPD patients were lower than those in OB-DM group[(697.06-± 231.9) mIU/L,(6.53 ± 2.21)mg · kg 1 · min-1,P＜0.0 1],but slightly higher than the Lean-KPD group [AUCins0 10min (92.1 ±29.8) mUU/L,GIR (2.55 ± 1.49) mg · kg 1 · min-1,P＜0.05].Glucose disposal rate (GDR) was strongly associated with casual plasma glucose (r =-0.502,P＜0.01),HbA1C(r =-0.553,P＜0.0 1) and FFA eoneentrations (r=-0.504,P＜0.01) on admission.Conclusions Insulin resistance and β-cell dysfunction coexist in all KPD patients.OB-KPD patients exhibit more severe insulin resistance,while Lean-KPD patients have lower insulin secretion.KPD patients had severe hyperglycemia,hypertriglyceridemia,and high plasma FFA levels on admission,suggesting that hyperglycemia and elevated FFA levels could result in serious insulin resistance,β-cell dysfunction,and diabetic ketosis in patients with KPD.

Objective To investigate the response of cryopreserved ovarian tissue after autologous implantation in mouse stimulated with gonadotrophin.Methods Thirty six female mice were randomly divided into three groups,with 12 mice in each group.In group of fresh ovarian tissue,fresh ovarian tissue was implanted into kidney capsule of mice:in group of cryopreserved ovarian tissue,ovarian tissue was implanted into kidney capsule of mice after cryopreserved by vitrification for two weeks.We investigated the response of ovarian tissue two weeks later after autologous implantation stimulated with gonadotrophin.Immunohistochemistry staining method was used to observe the expression of follicle stimulating hormone receptor.Results Before and after stimularian with gonadotrophin,the mature follicle rate of group of fresh ovarian tissue was 2.3%and 4.2%.that of group of cryopreserved ovarian tissue was 2.3%and 4.0%,and that of group of control was 2.6%and 5.8%.Regarding the percentages of mature follicle.there were significant differences after stimulation with gonadotrophin(P＜0.05).After stimulating with gonadotrophin the percentages of mature follicle were the same in the fresh tissue group,cryopreserved tissue group and control group(P＞0.05).The integrated optical density of follicle stimulating hormone receptor of fresh ovarian tissue in antrofollicle and pre-antrofollicle were 9408±2777 and 4531±1903.that of cryopreserved ovarian tissue were 9175±3093 and 4808±1386.and that of the control ovarian tissue were 8838±2064and 5516±1136 respectively.There was no significant difference between any two groups(P＞0.05).Conclusion The follicle stimulating hormone receptor is preserved by cryopreservation and transplantation,small pieces of ovarian tissue response to gonadotropin stimulation is normal.

Objective To investigates factors affecting the positive rate of blocking antibody treated by paternal lymphocyte immunotherapy in patients with recurrent spontaneous abortion (RSA).Methods From January 2008 to August 2012,326 RSA cases undergoing treatment in Infertility Center of Qilu Hospital were studied retrospectively.Those patients were divided into 2 groups randomly:260 cases in intradermal injection group were administered via bilateral forearm intradermal injections for immunotherapy once 21 days,then the blocking antibody was determined after 2 (23 cases),3 (73 cases),4 (74 cases),5(90 cases) times respectively,while in subcutaneous injection group,the 66 cases were administered via subcutaneous injection once 21 days,the blocking antibody measured after 3 times; In both cases,the blocking antibody was all determined 2 weeks later.The positive rate of blocking antibodies and the rate of successful pregnancy was recorded,and then followed up after the blocking antibody turning positive.Results (1)Positive rate of blocking antibodies:the positive rate of blocking antibodies were 17％ (4/23),58％ (42/73),72％ (53/74) and 84％ (76/90) in the 2,3,4,and 5 times of intradermal injection group,respectively (P ＜ 0.05).In subcutaneous injection group,the positive rate of blocking antibodies was 38 ％ (25/66),which was significantly lower than that in group intradermal injection receiving 3 times immunotherapy (P ＜0.05).(2) The rate of pregnancy:the 176 patients out of 200 patients were pregnant when antibody was positive after immunotherapy,with 71.6％ (126/176)of patients gained successful pregnancy(the length of pregnancy more than 5 months).Conclusions The route and frequency of administration of immunotherapy could influence the positive rate of blocking antibody.The rate of successful pregnancy will be increased after blocking antibody turning positive.

Objective To assess the effect of recombinant human leukemia inhibitory factor (rhLIF) on mouse embryo development in vitro Methods Mice were randomly divided to three groups, one in vivo control (group Ⅰ) and two in vitro (group Ⅱ and Ⅲ) Mice were sacrificed at 116 120 hours (group Ⅰ) and 44-48 hours (group Ⅱ and Ⅲ) subsequent human chorionic gonadotropin (hCG) injection Two cell embryos (group Ⅱ and Ⅲ) and blastocysts (group Ⅰ) were obtained Embryos in group Ⅱwere cocultured with human tubal fluid (HTF) + 10% human serum and in group Ⅲ with HTF + 10% human serum+rhLIF (1 000 U/ml) The number of embryo in different stage was recorded and compared Results Embryo in four, eight cell and morula was noted in group Ⅱ and Ⅲ, 87 7% versus 91 2% and 75 0% versus 85 4% respectively There was no significant difference. However, further embryo development to the blastocyst, expanded blastocyst, and hatching blastocyst in group Ⅱ (48 1%, 32 1% and 18 4%) was lower than that in group Ⅲ (82 3%, 59 7% and 36 3%) There was no difference between blastocyst in group Ⅰ and group Ⅲ (86 0% vs 82 3%). Conclusion RhLIF does not provide obvious stimulation in early mouse embryo, however, rhLIF can promote the growth, differentiation, and hatching of preimplantion blastocyst

Objective To investigate the efficacy of the indocyanine green (ICG) clearance test (ICGR15) combined with the model for end-stage liver disease (MELD) for assessing the short-term prognosis of patients with liver failure.Methods Eighty patients with liver failure were analyzed retrospectively.ICGR15 and relevant clinical data within 24 hours of diagnosis were analyzed.Meanwhile,the MELD score and King's College Hospital (KCH) were evaluated.All findings were tested for correlation with 3-month mortality.Quantitative data were analyzed with analysis of variance and Student's t-test.Count data were analyzed with chi-square test.Correlation analysis was performed with Pearson's coefficient test.Results Among 80 patients with liver failure,39 patients survived and 41 died.The mortality rate of all patients was 51.2％.The serum total bilirubin,creainine concentrations,ICGR15,MELD scores and patient number in accordance with KCH criteria of surviving patients were (288.0±109.1) μmol/L,(63.3±24.4) μmol/L,(48.1±10.2)％,20.6±4.4,and 6 cases,respectively,which were lower than those in dead patients [(340.7 ± 108.2) μmol/L,(98.8 ± 59.1) μmol/L,(60.2 ± 10.6) ％,26.9 ± 7.1 and 19 cases,respectively] (P =0.033,P＝ 0.001,P＝ 0.000,P＝ 0.000 and P =0.003,respectively).There was no significant difference of ICGR15 among four types of liver failure.A positive correlation was observed between ICGR15 and MELD score (r=0.289,P=0.009).The ICGR15-MELD model was created by subjecting ICGR15 and MELD scores to Logistic regression analysis.The following ICGR15-MELD model,Logit (P) =0.105 × ICGR15 + 0.178 × MELD score-9.734,was constructed by Logistic regression analysis.The area under the receiver operating characteristic (ROC) curve was 0.860 and the cut offpoint of 0.3 had sensitivity of 85.40％ and specificity of 74.40％.The area under the curve of the ICGR15-MELD model was significantly higher than those of ICGR15 (0.791),MELD score (0.770) and KCH criteria (0.655).Conclusions ICGR15 and MELD scores perform better than the KCH criteria in predicting the prognosis of liver failure.The ICGR15-MELD model is superior to ICGR15,MELD score,and KCH in predicting the short term prognosis of patients with liver failure.

Objective: To evaluate the efifcacy and safety on clopidogrel combining proton pump inhibitor (PPI) for treating the patient after percutaneous coronary intervention (PCI) by Meta analysis.
Methods: We searched MEDLINE, EMBASE, Cochrane Library and conference databanks, the retrieval time ended at 2014-03 and 14 references were selected for Meta analysis by RevMan 5.2 software. A total of 52274 patients were enrolled and divided into 2 groups, Control group, the patients received clopidogrel, n=43809 and Combination group, the patients received clopidogrel and PPI n=8465. The efifcacy and safety were compared between 2 groups.
Results: Compared with Control group, the patients in Combination group showed increased all cause death rate (OR=1.20, 95% CI 1.05-1.37), re-myocardial infarction (MI) (OR=1.19, 95% CI 1.07-1.33) and in-steut re-vascularization (OR=1.22, 95% CI 1.08-1.39), all P<0.05; while the MACE (OR=1.29, 95% CI 0.98-1.69), in-stent thrombosis (OR=1.22, 95% CI 0.97-1.54) and gastro- intestinal bleeding (OR=0.95, 95% CI 0.55-1.67) were similar between 2 groups, all P>0.05. Further analysis found that PPI (such as omeprazole and esomeprazole) could compete the CYP2C19 enzyme location of clopidogrel, increase the risk of cardiovascular events and decrease the gastrointestinal protection.
Conclusion: Clopidogrel combining PPI may increase the risk of all cause death, MI, in-stent revascularization and decrease the gastrointestinal protection in patients after PCI, especially for omeprazole and esomeprazole which may compete the CYP2C19 enzyme location of clopidogrel.

A high performance liquid chromatographic method with fluorimetric detector was developed for the simultaneous determination of three sulfonamides in vegetable samples. Vegetable samples were extracted with methanol for three times, and then the combined extracts were evaporated to dryness under reduced pressure at 45 ℃. The residue was dissolved in 0.1 mol/L HCl and the analytes were derivatized with fluorescamine. The chromatographic separation was performed on an ODS column with a gradient elution program using mobile phases based on mixtures of acetonitrile and 0.5% acetic acid aqueous solution. The derivatized compounds were detected with fluorimetric detector. The limit of detection was 1.02-1.29 μg/L and the limit of quantification was 3.4-4.3 μg/kg(fresh weight, F.W.) for three sulfonamides in vegetable. The average recoveries were higher than 87%, inter and intra RSDs were lower than 10% for all samples spiked with 0.2-1.0 μg/g of sulfonamides. The proposed method has been applied to the analysis of vegetables sold in Hefei markets. The result indicated that 3 SAs were found at different degree in the practical vegetable samples with the total concentrations between 0.0726-0.3709 μg/g(F.W.).

Objective To establish a rapid, accurate and sensitive chemiluminescence method for determining sulfydryl-containing drugs. Methods In sulfuric acid solution, glyoxal could be oxidized by potassium permanganate, and weak chemiluminescence could be observed. Chemiluminescence signal could be enhanced in the presence of sulfydryl-containing drugs. Thus, on this basis we established a new method of determining the concentration of sulfydryl-containing drugs with flow injection chemiluminescence analysis. Results Under the optimized conditions, the linear range of methimazole, captopril and acetylcysteine was 1.0×10~(-8)- 5.0×10~(-6), 7.0×10~(-8)-1.0×10~(-6) and 3.0×10~(-8)-1.0×10~(-6)g/mL, respectively. The limit of detection of methimazole, captopril and acetylcysteine was 1.0, 3.9 and 3.7ng/mL, respectively. Conclusion The method was successfully applied to determine the three drugs that contain sulfydryl. Compared with the results of pharmacopeia methods, the results we obtained were satisfactory.