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 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 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 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 value of brachial-ankle pulse wave velocity (baPWV) in the prediction of hypertension in middle-aged men with prehypertension.Methods A total of 2580 middle-aged (35 to 55 years old ) prehypertensive individuals who underwent health check-up during September 2006 and December 2007 in our hospital were recruited for this prospective cohort study.After a 4-year follow-up,2451subjects entered final analysis.Logistic regression analysis was used to assess the value of baPWV in the prediction of hypertension.Results(1) Two hundred and eight subjects (8.5％ )developed hypertension after 4-year follow-up study.( 2 ) At baseline,no significant differences of family history of hypertension,heart rate and total cholesterol were found between normotensive and hypertensive subjects (P ＞0.05 ).(3) Logistic regression analysis showed that in age-adjusted model (Model1),the odds ratio (OR) and 95％ confidence interval (CI) of baPWV (140 cm/s) was 2.20 (1.78 to 2.62 )( P ＜0.01) ; while in multi-factor adjusted model ( Model 3 ),OR and 95c％c CI of baPWV was I.49 (1.15 to 1.73) (P ＜ 0.01).(4) Subjects were stratified by quartiles of baPWV at baseline.OR and 95％ CI of hypertension in those of highest quartile was higher than those of lowest quartile ( Model1:OR =10.9,95％CI 5.1-22.7,P＜0.01; Model 3:0R=2.6,95％CI1.2 -6.1,P＜0.05).Conclusion baPWV could be an independent predictor of hypertension among prehypertensive populations.

Objective To study the effect of high intensity focused ultrasound(HIFU)ablation on microvessel density(MVD)and vascular endothelial growth factor(VEGF)in rabbit hepatcocellular carcinoma (HCC)models.Methods The VX2 t umor cells were planted in the hepatic left-central lobe on 30 rabbits.All animals were divided into two groups randomly:HIFU group(n=25)and control group(n=5).The MVD and the expression of VEGF in the HCC was detected by immunohistochemical SABC methods in control group and 0,1,3.7,16 d after the operation in HIFU group.Results The MVD and the expression of VEGF in the HCC tissue after HIFU treatment in the HIFU group were decreased significantly as compared with those in the control group (P＜0.01).However,there was no significant difference in MVD and the expression of VEGF in the HCC tissue after HIFU treatment in the HIFU group among the different time points(P＞0.05).Conclusions HIFU treatment can effectively destroy the micovessel,repress the neovascularization and reduce the blood supply of the HCC.

Components of free fatty acids (FFA) were examined by reverse high performance liquid chromatography in 25 patients with type 2 diabetes and 25 control subjects.The changes in components of FFA were observed before treatment, at 3 months and 1 year after treatment with pioglitazone in type 2 diabetic patients.The serum lauric acid, myristic acid and stearic acid levels were much higher in the diabetic patients than those in control subjects.Pioglitazone could decrease the levels of these fatty acids in the patients with type 2 diabetes mellitus.

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.).