Objective To discuss whether it is necessary to integrate metabolic indices into diagnosis of polycystic ovary syndrome(PCOS).Methods Taking ESHRE /ASRM diagnosis standard as gold standard,the case group composed of 51 women with PCOS and the control group composed of 47 women without PCOS were selected.By using classification tree C5.0,significant variables chosen by single factor analysis were used to establish a new diagnostic model which combined reproductive indices and metabolic indices.The validity and reliability of the new diagnostic model by using ROC curve analysis were evaluated.Finally,the consistence and difference between the new diagnostic model and the gold standard were analyzed.Results Single factor analysis chose 13 significant variables. ROC analysis showed an area under the curve of 0.930(P =0.000)and the optimal cut -off point was 0.190 with a sensitivity of 94.10%,a specificity of 91.50% and a consistency of 92.86%,which told us the new diagnostic model had superior validity and reliability.The two diagnostic methods had excellent consistence (Kappa =0.857,P =0.000)and there was no statistically significant difference (P =1.000).Conclusion Considering that whether metabolic indices are integrated into diagnosis of PCOS doesn′t change the diagnosis result,and it is unnecessary to integrate metabolic indices into diagnosis of PCOS.

Objective To investigate the state of perceived discrimination,mental health knowledge and self-discrimination of the public,and analyze the relationship among these variables,and explore the mediating effect of mental health knowledge between the perceived discrimination and self-discrimination.Methods A total of 1 088 residents in Guangzhou City were assessed with the Perceived Devaluation-Discrimination Scale (PDDS),Attitudes about Mental Illness Associated Stigma Scale-Chinese Version (AMIASS-C) and Mental Heahh Knowledge Questionnaire (MHKQ).Results The average score of PDDS was 2.63±0.35.The rate of mental health knowledge was 72.4％(788/1 088) and the score of AMIASS-C was 2.58±0.45.The perceived discrimination,mental health knowledge and self-discrimination were significantly correlated with each other,r=0.320,-0.240,P＜0.01.The mental health knowledge played a negative mediating role between perceived discrimination and self-discrimination.Conclusions The mental health knowledge was a mediator between perceived discrimination and self-discrimination.

Objective To investigate the relationship between vaspin,Apelin,leptin and polycystic ovary syndrome.Methods We determined the serum vaspin,Apelin and leptin levels of 40 cases of PCOS patients and 30 cases of control group,and divided the PCOS group into three subgroups (normal weight,overweight and obesity) according to WHO criteria for obesity.Then we compared the serum vaspin,Apelin and leptin levels in PCOS and control group,and the differences in body weight index (body mass index,BMI) in women with polycystic ovary syndrome,and analyzed roles of adipokine vaspin,Apelin and leptin in the pathogenesis of PCOS.Results In PCOS patients adipokine vaspin,Apelin,leptin,insulin resistance index (Homeostasis model assessment for insulin index in resistence,HOMA-IR),fasting insulin (INS) and blood glucose,blood lipid were higher than those of the control group and increased with the increase of body mass index (BMI),and three kinds of adipokines and PCOS patients with HOMA-IR were positively correlated,there were statistically significant differences (P ＜ 0.05).Conclusion The results of vaspin,Apelin and leptin may be involved in the occurrence of PCOS.

Objective To investigate the effect of small interfering RNA (siRNA) interference in the expression of epidermal growth factor receptor (EGFR) on the radiosensitivity of esophageal squamous carcinoma (Eca-109) and esophageal adenocarcinoma (OE-19) cell lines.Methods Human Eca-109 and OE-19 cell lines were selected as study subjects.Various EGFR-siRNA and negative siRNA were synthesized chemically through lipofection.Reverse transcription-polymerase chain reaction and Western blot were applied to measure the expression of EGFR before and after transfection,and the CCK8 assay was applied to analyze the influence of transfection on cell proliferation.Blank control groups of Eca-109 and OE-19 cells (O1 and O2 groups),simple irradiation groups (R1 and R2 groups),and EGFR-siRNA irradiation groups (E-R1 and E-R2 groups) were set,and the doses for single irradiation were 0,2,4,6,and 8 Gy.The colony-forming assay was applied to calculate survival fraction (SF) and sensitization enhancement ratio (SERD0 ratio),and flow cytometry was applied to evaluate the influence of EGFR-siRNA combined with radiotherapy on cell cycle distribution and apoptosis rate,and the dose for single irradiation was 6 Gy.Results The expression of EGFR in both cell lines was significantly down-regulated by EGFR-siRNA,and the inhibition rate of cell proliferation by transfection was＜5％ (4.9％ and 4.5％,respectively).The results of colony-forming assay showed that the cells in the E-R1 and E-R2 groups had a lower SF than those in the O1 and O2 groups,with an SERD0ratio of 1.40 and 1.01,respectively.The results from flow cytometry showed that compared with the E-R2 group,the E-R1 group had a higher proportion of cells in G2/M phase and a lower proportion of cells in S phase after irradiation (P=0.016 and 0.028),as well as a higher apoptosis rate (P=0.007).Conclusions Compared with the cell line OE-19,the cell line Eca-109 has a significantly increased radiosensitivity when treated with siRNA interference in EGFR expression.

Objctive To study the effects of traditional Chinese medicine S Kangdu mixture on functions of immune system and bone marrow in mice. Methods Animal model, which marrow and immune system were inhibited, was established by 60 Co or cyclophosphamide injection. The mixture was administrated orally. The white blood cell counts, spleen colony formed unites (CFU S), and the index of carbon clean up were measured. Results The mixture obviously improved leukopenia in peripheral induced by cyclophosphamide administration, and facilitated marrow karyocyte proliferation and formation of CFU S in 60 Co treated mice and reinforced phagocytosis of mononuclear phagocytic system in cyclophosphamide treated mice. Conclusion Traditional Chinese medicine S Kangdu mixture can significantly ameliorate immunological function and stimulate hemopoietic function in mice.

AIMTo clone the gene of staphylococcal enterotoxin C2 and express it in the form of a soluble fusion protein in E. coli. Then the activation of SEC2 on mice lymphocyte and its lethal effects on tumor cells were studied.

METHODSStaphylococcus aureus SEC2 gene was cloned into GST gene fusion vector pGEX-4T-1. The resultant plasmid pGEX-4T-SEC2 was used to transform E. coli BL21, where the GST-SEC2 fusion protein was expressed efficiently. The rSEC2 protein was purified with Glutathione Sepharose 4B affinity column and digested with thrombin. The in vitro culture system was utilized to observe the activation of the SEC2 on mice lymphocyte and the lethal effects on tumor cells of the activated mice lymphocyte.

RESULTSThe proper gene of SEC2 was cloned and purified rSEC2 was obtained. The MTT results indicated that rSEC2 have strong ability to stimulate mice lymphocyte to proliferate with a dose-dependent manner. With the proliferation of mice splenic lymphocyte, rSEC2 has a strong lethal effect on tumor cells B16, K562 and K562-AD.

CONCLUSIONIn this study, the gene of SEC2 was cloned and the rSEC2 protein was obtained, which had strong lethal effect on tumor cells B16, K562 and K562-AD.

Animals ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cloning, Molecular ; Enterotoxins ; genetics ; metabolism ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Female ; Genetic Vectors ; Glutathione Transferase ; genetics ; Lymphocyte Activation ; drug effects ; Lymphocytes ; cytology ; immunology ; Male ; Mice ; Mice, Inbred ICR ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Spleen ; cytology ; Transfection

To establish a UPLC-MS/MS method for the simultaneous determination of geniposide, genipin 1-O-beta-D-gentiobioside and geniposidic acid in rat brains and study the brain pharmacokinetics of the three iridoid glycosides in stroke rat after the oral administration of Xingnaojing. In this experiment, brain samples were precipitated with protein for twice. Acquity BEH C18 column was adopted, with acetonitrile-0.1% formic acid-water as the mobile phase for gradient elution. ESI source was adopted for mass spectra; multiple reaction monitoring (MRM) was conducted to detect negative ions. The time for sample analysis was 3.5 min. the results showed good linear relations among the three iridoid glycosides, with the extraction recovery between 99.6% and 114.3%, good intra- and inter-day precisions and accuracies and stability in line with the requirements. The t1/2 and MRT in the three components were similar in brains of stroke rats. Geniposide and genipin 1-O-beta-D-gentiobioside showed double peaks; where as geniposidic acid showed a single peak. In conclusion, the method is so specific, sensitive, accurate and reliable that it can be used to study the brain pharmacokinetics of Xingnaojing oral preparation.
Animals ; Brain ; metabolism ; Brain Chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Iridoids ; chemistry ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Objective To observe the effects of 75 gram glucose oral tolerance test (75 g OGTT) and standard mixed meal test (SMMT) on insulin secretion function of the islets of Langerhans and plasma free fatty acid (FFA) in patients with type 2 diabetes mellitus.Methods Seventy-six patients with type 2 diabetes without using insulin and with no obvious complications were recruited for 75 g OGTT following overnight fasting on the first day and SMMT (bread 50 g,egg 50 g and milk 250 ml) on the 7th day.Blood specimens were collected from each patients before the tests and 30 min,60 min,120 min and 180 min after glucose or meal load to measure their levels of plasma glucose,serum insulin,C peptide,FFA and lipids (total cholesterol,triglyceride,high-density and low-density lipoprotein cholesterol).Results No difference in fasting plasma glucose,serum insulin,C peptide,FFA and lipids between 75 g OGTT and SMMT was found.Postprandial plasma glucose 30 min,60 min,120 min and 180 min after 75 g OGTT was significantly higher than that after SMMT,with (15.3?3.5) vs (9.9?3.4) mmol/L,(18.2?4.8) vs (12.8?4.0) mmol/L,(16.3?5.8) vs (12.2?4.9) mmol/L and (10.6?5.4) vs (9.5?4.5) mmol/L (F=28.1,P

The filtrate of Staphylococcus aureus culture has been used in an ampule form named as staphylococcal enterotoxin C injection for cancer therapy in clinic for ten years in China and proved to be effective. The active constituent of three kinds of injections is claimed to be staphylococcal enterotoxin C2 (SEC2), and the content of SEC2 is used as quality control. However, the correct content of SEC2 was not known and the relative amount of SEC2 was very low because of the complicated components of the filtrate. In this research, we established a proper ELISA system for the detection of SEC2 in staphylococcal enterotoxin C injection, which will improve the quality control of the injection. We produced and identified polyclonal and monoclonal antibodies of SEC2 and established BA-ELISA method based on the method of sandwich ELISA. It was found that the BA-ELISA method had good specificity, sensitivity and reproducibility, and being able to detect SEC2 at concentration from 2 to 20 ng x mL(-1), with an average CV value of 5.08%. The SEC2 content in staphylococcal enterotoxin C injection was calculated. There is some difference between the actual and labeled contents in the injections.
Animals ; Antibodies, Bacterial ; biosynthesis ; immunology ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antineoplastic Agents ; administration & dosage ; analysis ; immunology ; Enterotoxins ; administration & dosage ; analysis ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Hybridomas ; secretion ; Injections ; Mice ; Mice, Inbred BALB C ; Quality Control ; Rabbits ; Staphylococcus aureus ; chemistry

This study is to clone the gene of staphylococcal enterotoxins O, obtain recombinant protein (rSEO) and investigate its activity on mice lymphocyte. Staphylococcus aureus O gene is cloned into GST gene fusion vector pGEX-4T-1. The resultant plasmid pGEX-4T-SEO was used to transform E. coLi BL21, where the GST-SEO fusion protein was expressed efficiently. Then SEO was purified by Glutathione Sepharose 4B affinity column and digested with thrombin. The bioactivity of SEO was analyzed by MTT assay on mice lymphocyte and tumor cells. The nucleotide sequence was confirmed to code for the protein correctly, and soluble SEO was expressed efficiently in E. coli BL21 with pGEX-4T-SEO. The protein purified by affinity chromatography resulted to be one single band by SDS-PAGE detection. The MTT assay of the purified rSEO demonstrated that its abilities of stimulating T cells and inhibiting the proliferation of K562, K562-ADM and B16 cells were equivalent to that of SEC in vitro. The expression plasmid pGEX-4T-SEO was constructed and the recombinant superantigen was expressed successfully, which may provide a foundation for the further research of the anticancer activity of SEO.
Animals ; Cell Proliferation ; Cloning, Molecular ; Enterotoxins ; genetics ; immunology ; metabolism ; Escherichia coli ; genetics ; metabolism ; Female ; Genetic Vectors ; Humans ; K562 Cells ; Lymphocytes ; cytology ; Male ; Melanoma, Experimental ; pathology ; Mice ; Mice, Inbred ICR ; Plasmids ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Spleen ; cytology ; Staphylococcus aureus ; genetics ; Superantigens ; genetics ; immunology ; metabolism