AIM: To establish an effective and convenient method for applying HPLC fingerprints to quality control in the production of Xiasangju Granules(Spica prunellae,Folium mori,Flos chrysanthemi). METHODS: Komasil Sunfrie C_(18)(150 mm?4.6 mm,5 ?m) analytical column was used and eluted with a gradient program consisted of phase A(1% acetic acid) and phase B(methanol) and detected at 290 nm.The fingerprints of aqueous extract,alcohol-precipitated extract,concentrate and finished product were compared with. RESULTS: The fingerprint method for Xiasangju Granules was established.The similarity among 10 batches of Xiasangju Granules was no less than 0.970.The difference between extracts and finished product of Xiasangju Granules was obvious. CONCLUSION: This validated method is available for quality evaluation and quality control in Xiasangju Granule's production.

AIM:To establish the fingerprint chromatogram of amino acids compounds of Xiasangju Granules(Spica Prunellae,Flos Chrysanthemi indici,Folium mori.).METHODS:To apply 6-aminoqtiinoly-N-hydroxysuccinimdyl carbamate(AQC) pre-column derivatization HPLC method.Separation was performed on SYMMETRY C_(18)(150 mm?(4.6) mm,5 ?m) analytical column with mobile phase consisting of acetate(pH=(5.05)) and 60% acetonitrile with gradient elution with the flow rate(1.0) mL/min and the column temperature at 37 ℃.The Fluorescence wavelength used for detection was set at 250 nm(Excitation wavelength) and 395 nm(Emission wavelength) and the analysis time was 50 min.RESULTS:12 co-peaks on the HPLC fingerprints of Xiasangju Granules were indicated.The similarities were determined by the coefficients of cosine and correlation.The results of similarity analysis were(0.95)-(1.00).CONCLUSION:Perfect fingerprints were obtained which can be used for the quality control of Xiasangju Granules.

Carcinoma ; genetics ; metabolism ; pathology ; surgery ; China ; Humans ; Male ; Oncogene Proteins, Fusion ; genetics ; Prostatic Hyperplasia ; genetics ; metabolism ; pathology ; surgery ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-ets ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serine Endopeptidases ; genetics ; metabolism

Objective To investigate the clinical effect of renal transplantation in elderly patients.Methods Retrospective analysis was held on 49 elderly patients aged over 65 years who underwent renal transplantation,and compared with 87 patients under the age of 50 years who underwent the renal transplantation during the same time.Results The survival rate at one year was 85.7% in elderly patients,and 93.1% in the non-elderly patients(P＜0.05).The graft survival rate at one year was 89.8% in elderly patients,and 90.8% in the non-elderly patients,but no significant difference was found between the two groups.The rate of acute rejection response was 10.2% in elderly patients,significantly lower than that in non-elderly patients(28.7%).The rate of complication of cardio-cerebrovascular diseases in elderly patients(51.1%)were higher than those in non-elderly patients(19.5%,P＜0.05).The dosages of immunosuppressive agent cyclosporine in 3 months,3-6 months and 6-12 months were(3.6?1.1),(3.1?0.9)mg?kg~(-1)?d~(-1) and (2.9?0.7) mg?kg~(-1)?d~(-1)in elderly group,significantly lower than those[(5.4?1.4),(4.6?1.3)mg?kg~(-1)?d~(-1)and(4.1?1.0)mg?kg~(-1)?d~(-1)]in non-elderly group(P＜0.05).Conclusions Renal transplantation in elderly patients is effective and reliable.

Background T helper cell 17 (Th17),a newly discovered subset of CD4+ T cells,have been found to play an important role in dry eye disease in animal model.Further investigation should be done on the immunopathogenesis of Th17 cells in dry eye patients.Objective This study was to analyze the expression status of interleukin-17 (IL-17) and IL-22 in tear and supernatant of peripheral blood mononuclear cells (PBMCs) of dry eye patients and their correlation with clinical symptom and sign.Methods Twenty Sj(o)gren syndrome (SS)patients,twenty non-Sj(o)gren syndrome (NSS) patients were included in Wuxi Second Hospital from 2010 to 2011,and twenty healthy volunteers were recruited at the same period.All of subjects understood the purpose and procedure of research and written informed consent was obtained form each subject initial of this study.Dry eye symptom questionnairs were self-answered and multiple dry eye disease-related clinical tests,including the breakup time of tear film (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescein staining were performed.The periphery blood of 3 ml and tear were collected in all the subjects,and IL-17 and IL-22 levels in supernatant of PBMCs and tear were detected by enzyme-linked immunosorbent assay (ELISA).The correlations between levels of IL-17 and IL-22 with BUT,S Ⅰ t,corneal fluorescein staining and dry eye scores were analyzed.Results The dry eye scores reduced,BUT prolonged,S Ⅰ t increased and corneal fluorescein dye decreased from SS group,NSS group to normal control group,with significant differences among the three groups (dry eye scores:H =40.81,P＜0.01 ; BUT:H =40.15,P＜0.01 ;S Ⅰ t..H=50.07,P＜0.01 ;corneal dye scores:H=40.52,P＜0.01).The concentration of IL-17 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (964.92±124.83)ng/L,(718.85± 115.89)ng/L and (341.95±85.08) ng/L,showing a statistically significant difference among them (F=162.95,P＜0.01).The levels of IL-17 in the tear were (440.69±126.09) ng/L,(364.33±126.85) ng/L and (61.16±11.60) ng/L in the SS group,NSS group and normal control group respectively,exhibiting an elevated level in the SS group and NSS group compared with the control group (F=75.27,P＜0.01).In addition,the levels of IL-22 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (98.77± 11.27) ng/L,(79.65 ± 11.01) ng/L and (32.78±9.34) ng/L,and those in the tear were (22.22 ± 8.96) ng/L,(14.92 ±4.35) ng/L and (10.47 ± 2.67) ng/L,with significant differences among the three groups (F =206.27,P＜0.01 ;F =19.87,P＜0.01).The significant correlations were found between the IL-17 and IL-22 concentration in the supernatant of PBMCs and tear with corneal fluorescein staining scores and S Ⅰ t.Conclusions The contents of IL-17 and IL-22 in PBMCs and tear upregulate in the SS and NSS patients,indicating that Th17 plays a key role in the immunity mechanism of dry eye.

Objective To investigate the effect of S-Nitrosoglutathione (GSNO)on acute ischemia reperfusion (I/R)induced in-testinal barrier function lesion in a mouse model.Methods Twenty-four 6-8-year-old C57BL/6 mice were divided into 3 groups,8 for each:(1)the sham group;(2)the I/R group;(3)the I/R+GSNO group.The mouse intestine I/R model was established by the occlusion of the superior mesenteric artery temporarily followed by reperfusion for 6 h.Histological changes in the small intestine were observed after HE staining;the expression of claudin-1 protein in the intestine epithelium was assessed by immunohistochem-istry staining as well as western blot analysis.Results Both HE staining and immunohistochemistry results showed the integrate intestinal villi with the continuous Claudin-1 expression alone the villi in the sham group;the intestinal villi of the I/R group partial-ly detached,thickened,crooked and fractured,with the obvious disconnection of Claudin-1 staining alone the top of the villi;while the intestinal villi of the I/R+GSNO group were neatly arranged and damage to intestinal mucosa was much alleviated,accompanied with the marked restoration of the continuity of claudin-1 staining.Compared to the sham group,claudin-1 protein level of for the I/R group and the I/R+GSNO group decreased by 32.5% and 13.8% respectively (P <0.05);and compared to the I/R group,clau-din-1 protein level of the I/R+GSNO group increased by 27.8% (P <0.05).Conclusion Protein level of claudin-1 would decrea-ses after I/R,and pretreatment with GSNO can effectively relieve the damage of intestinal mucosal structure as well as intestinal tight junction barrier through upregulating the expression of claudin-1 protein.

Objective To observe the effect of recombinant human mannose-binding lectin ( rhMBL) on apoptosis of intestinal epithelial cells. Methods Fourteen C57 mice were randomly divided into two groups. The LPS mice received intra-peritoneal injection of LPS for indu-cing acute intestinal barrier injury. The expression of MBL protein and mRNA were observed by immuno-histochemisty and RT-PCR respec-tively. The apoptosis of intestinal epithelial cells was detected by tunel staining. In vitro, the Caco-2 cells were treated with 0、 5、10、20 μg/mLrhMBL, flow cytometry was used to detect the apoptosis rate of Caco-2 cells. The Bax and Bcl-2 protein and mRNA were detected by Western blotting and Real-Time PCR methods between various groups. MAPK signal proteins were detected by Western blotting method. Results The MBL and TUNEL staining were apparently increased under LPS condition. The apoptotic rates of Caco-2 cells which were trea-ted by different rhMBL concentrations after 48 h were increased in 20μg/mL rhMBL when compared with other groups. The apoptotic rates of Caco-2 cells which were treated by 20 μg/mL rhMBL after different time were increased in time-dependent manner. The results of Western blotting and Real-Time PCR assay showed that levels of Bax/Bcl-2 was increased in Caco-2 cells which were treated by 20μg/mL rhMBL af-ter 48h. Meanwhile, the phosphorylation of P38 and ERK were obviously increased in Caco-2 cells. The inhibitory effect of ERK and P38 in-hibitors were studied by Western blotting. Our current study showed that a block in P38 MAPK dependent cell death might contribute to the decrease in 20μg/mL MBL-mediated Caco-2 apoptosis. Conclusion High concentration of rhMBL may play an important role in the intesti-nal epithelial cell apoptosis, and P38 pathway may be involved in this regulation.

OBJECTIVETo explore the access to tobacco and exam the predictors of successful tobacco purchase attempts among Chinese minors.

METHODSA simulative trial of purchasing cigarettes was participated by 201 sixth grade students to assess the prevalence of illegal cigarette sales to minors in Guangzhou. Methods of Chi-square and unconditional logistic regression were used to identify the significant predictors,with the result of tobacco purchase as the dependent variable and the characteristics of stores, retailers and minors as the independent variables.

RESULTSA total of 165 students succeeded in purchasing cigarettes but 36 failed, and the percentage of successful purchase attempts was 82. 1% . Data from univariate analysis indicated that 9 factors were significantly associated with students' success in purchasing cigarettes. They were age and height of the purchasers, types of stores, seller's gender and age, posting cigarette advertisements,showing warning signs of 'no cigarette selling to minors' ,asking buyer's age,and asking whom you buy the cigarettes for. The results of multivariable analysis showed that only three variables entering the final logistic regression: the age of students, the type of stores, and showing warning signs of 'no cigarette selling to minors'.

CONCLUSIONChinese minors have easy access to purchasing cigarettes, especially in groceries and small markets. Selling cigarettes by sellers to minors should be monitored and managed in the future.

Adolescent ; Child ; China ; Commerce ; legislation & jurisprudence ; statistics & numerical data ; Female ; Humans ; Male ; Minors ; statistics & numerical data ; Smoking ; epidemiology ; legislation & jurisprudence ; Students ; statistics & numerical data

Objective To investigate the effects of tumor necrosis factor-alpha (TNF-α) on insulin sensitivity and glucose-lipid metabolism in TNF-α-induced IR mice. Methods Male C57BL/6J mice were given an intraperitoneal injection of TNF-α (H group,6μg/kg; M group,3μg/kg; L group,1μg/kg;twice daily) and saline (NC group) for 7 days. The plasma glucose and insulin were assayed during intravenous glucose tolerance test (IVGTT) and hyperinsulinemic-euglycemic clamp combined with 3-[3H] glucose as a tracer was carried out. Results After TNF-α treatment,fasting blood glucose (FBG),plasma insulin and free fatty acids (FFA) were significantly elevated in H group compared with NC,L and M groups (P<0.01 and P<0.05,respectively). There was a lower glucose tolerance in H group versus other three groups during IVGTT. The insulin release by glucose stimulation was higher in H group versus NC and L groups (P<0.01 and P<0.05). Basal glucose disappearance rate (GDR) and hepatic glucose production (HGP) were significantly increased in H group compared with NC group (P<0.01). During the steady-state of clamp,plasma insulin levels were significantly increased in H group versus NC group (341.7±17.7 vs 84.7±5.5mU/L,P<0.01). The suppressive effect of insulin on FFA was significantly blunted in H group compared with NC group (0.82±0.03 vs 0.43±0.07mmol/L,P<0.01). Steady-state glucose infusion rate (GIR) was significantly decreased in H group compared with NC group (39.1±2.3 vs 54.2±2.2 mg·kg-1·min-1,P<0.01). Although GDR was increased in both group,but it was still lower in H group than in control group(47.9±0.8 vs 53.9±2.0 mg.kg-1.min-1,P<0.01).As compared with baseline,HGP in the controls was almost completely suppressed during steady state of clamp,but in H group suppressed by approximately 41%. Conclusions High-dose TNF-α treatment induces the abnormality of glucose-lipid metabolism and the insulin resistance of hepatic and peripheral tissue in mice

Objective To investigate the effects of extracellular ATP on the expression of(Mi- crotubule associated proteins MAP-2)and the recovery of motor function after spinal cord injury in rats. Methods Six rats were selected randomly from 66 adult healthy Wistar rats as the normal control group,the rest animals were divided into two groups after contusion injury was performed by drop weight method with Allen impactor:Group A(ATP group)and Group B(control group),each group contained thirty rats.At days 1,3,7,14,and 28 after injury,the rats were killed,the expression of MAP-2 was detected with immunohistochemistry.The expression of MAP-2 in the adjacent area was quantitatively an- alyzed with a computer image analysis system.The recovery of motor function after spinal cord injury was assessed with improved Tarlov scores.Results The expression of MAP-2 was higher in Group A than in Group B after spinal cord injury in rats.Significant difference was revealed by the expression of MAP- 2 between the two groups at days 14 and 28 after injury(P