OBJECTIVE:To simultaneously determine the contents of CIPRO and MNZ in compound ciprofloxacin ear dro_ ps by nodical polyploid UV-spectrophotometry.METHODS:CIPRO and MNZ were nodical absorption at wavelength of350.0nm and this was just the sum of isoabsorptive point of two components while DXM did not absorb UV ray at this point.Therefore350.0nm was adopted as the detecting wavelength for these two components.RESULTS:The average recovery and relative standard deviation of CIPRO were99.7%and1.4%respectively.The average recovery and relative standard de?viation of MNZ were100.7%and1.6%respectively.CONCLUSION:The method is accurate,rapid,simple and sensitive.This method can be used to control the quality of this preparation.

To study the influence of 5?-reductase on the pathogenesis of human benign prostatic hyperplasia (BPH), the activity of this enzyme was measured in mechanically separated stroma and epithelium from 7 normal and 16 hyperplastic prostates. Samples were incubated in the presence of tritium labelled testosterone. The yield of DHT was used to estimate the enzyme activity. The results showed that the specific activity of the enzyme (pmol DHT / mg protein/30 rain) was91.4?18,1 and 28.6?7.4 in stroma (S) and epithelium (E) of BPH, 44.7?8.9 and 23.9?6.8 in S and E of normal prostates respectively. It indicated that the enzyme is predominantly localized in the stroma and is elevated ia BPH, the primary abnormality of BPH is in the stroma and the increase of 5?-reductase may have some contribution to the pathogenesis of BPH.

Objective To conduct the clinical and pathological evaluation on the conservative surgery in tubal pregnancy . Methods 513 cases of tubal pregnancy in this hospital from January 2006 to December 2012 were divide into the conservative sur-gery group(A ,314 cases) ,samlpintectomy group(B ,43 cases) and the medication conservative treatment group (C ,156 cases) .The hospitalization days ,HCG negative-conversion time ,cure rate ,re-pregnancy outcome ,pregnancy during 1-year follow-up were com-pared between the group A and C .In group B ,43 cases of tubal pregnancy and accomplishing fertility were firstly performed the tubal linear incision in the pregnant site ,then the biopsy was conducted after stopping bleeding and finally the salpingectomy in af-fected side was performed .The tubal electric injury degree by the unipolar or bipolar electrocoagulation and the pathological changes under light microscope were observed .Results The hospitalization days ,symptom relief rate ,HCG negative-conversion time ,cure rate and re-pregnancy outcome had statistical differences between the group A and C (P<0 .05) .Conclusion The tubal conserva-tive surgery has short hospitalization time ,high symptom relief rate ,high cure rate and few complications .The pathological observa-tion shows the localized electrocoagulation injury .The pregnancy rate after surgery is high .

The effects of microRNA-34a (miR-34a)-regulated Notch1 gene on the proliferation and apoptosis of the human glioma cell line U87 were investigated in this study. The U87 cells were divided into miR-34a mimics, negative control, mock transfection and blank control groups in terms of different treatments. In miR-34a mimics group, human U87 glioma cells were transfected with miR-34a mimics by using lipofectamine 2000. The cells transfected with nonsense microRNA were set up as negative control group. Those treated with lipofectamine 2000 only were designated to the mock tranfection group. In the blank control group, the cells were cultured routinely and no treatment was given. The expression of miR-34a and Notch1 was detected by using real-time RT-PCR. Western blotting was employed to monitor the change in Notch1 protein. Cell proliferation and apoptosis were measured by CCK-8 and flow cytometry. The results showed that the proliferative ability of U87 cells was significantly reduced and the apoptotic cells increased in miR-34a mimics group relative to control groups. The expression of miR-34a was significantly up-regulated in mimics group as compared with control groups (P<0.05). Furthermore, Notch1 protein levels were significantly decreased in miR-34a mimics group when compared with control groups (P<0.05), but the mRNA expression of Notch1 showed no significant difference among these groups. It was concluded that miR-34a may suppress the proliferation and induce apoptosis of U87 cells by decreasing the expression of target gene Notch1, suggesting that miR-34a may become a promising gene therapeutic target for brain glioma.

Objective: To investigate the prevalence and influencing factors of repeated HIV antibody tests among men who have sex with men (MSM) in Wuhan City, so as to provide insights into the development of interventions against AIDS among MSM. Methods: MSM with self-reported homosexual behaviors in a community studio in Wuhan City were recruited using a convenient sampling method from January to April 2019, and participants' demographic features, sexual behaviors, HIV antibody tests and recognition of HIV antibody tests were collected using questionnaires. Factors affecting repeated HIV antibody tests were identified using a multivariable logistic regression model. Results: Totally 300 valid questionnaires were recovered, with a mean age of (31.06±10.24) years. Among all the participants, 196 participants had the first homosexual behaviors over 18 years of age (65.33%), 107 participants were insertive partners (35.67%), and 125 participants received sex-transmitted diseases (STDs) tests in the past six months (41.67%). The mean score for recognizing the risk of HIV infection was 12.41±3.09, for HIV antibody test self-efficiency was 17.07±2.12, and for perceived social supports was 17.42±2.41. A total of 287 respondents received HIV antibody tests (95.67%), including 192 participants receiving repeated HIV antibody tests (64.00%). Multivariable logistic regression analysis showed that age of >18 years for the first homosexual sex behavior (OR=0.404, 95%CI: 0.223-0.734), receiving STDs tests in the past six months (OR=3.896, 95%CI: 2.145-7.076), sex role as receptive partners or both receptive and insertive partners (OR=0.502, 95%CI: 0.275-0.917), satisfying with HIV antibody test services (OR=2.955, 95%CI: 1.311-6.660), and high score for HIV antibody test self-efficiency (OR=1.149, 95%CI: 1.005-1.314) were factors affecting repeated HIV antibody tests among MSM. Conclusions The detection of repeated HIV antibody tests was 64.00% among MSM in Wuhan City in 2019, and age for the first homosexual behavior, STDs tests, sex role, evaluation of HIV antibody test services and self-efficiency of HIV antibody tests may be factors affecting repeated HIV antibody tests among MSM.

Chemical constituents of 95% ethanol extract of the dried persistent calyx of Physalis pubescens were investigated. By chromatography on a silica gel column and reverse-phase preparative HPLC, 10 compounds were isolated from the dichloromethane fraction. Based on the MS and 1D/2D NMR data, these compounds were identified as 5-O-(E-feruloyl) blumenol (1), isovanillin (2), (E) -ethyl 3-(4-hydroxyphenyl) acrylate (3), 4-hydroxybenzaldehyde(4), 4-methylphenol (5), (E) -methyl cinnamate (6), 7,3',4' trimethoxyquercetin (7), 5,3', 5'-trihydroxy-3,7,4'-trimethoxyflavone(8), danielone (9), and 5,5'-diisobutoxy-2,2'-bifuran (10).
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Molecular Structure ; Physalis ; chemistry ; Spectrometry, Mass, Electrospray Ionization

BACKGROUND: Previous studies have confirmed that embryonic stem cells call be induced to differentiate into insulin-producing cells, but the induction process takes a long time. Most of the processes take about one month.OBJECTIVE: Activin A, all-trans retinoic acid (ATRA), basic fibroblast growth factor (bFGF) and nicotinamide were applied in vitro in combination to observe whether mouse embryonic stem cells could be induct to differentiate into insulin-producing cell in a relatively short time.DESIGN: Cell observation experiment.SETTING: Shanghai Institute of Endocrine and Metabolic Diseases, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.MATERIALS: This study was performed at Shanghai Institute of Endocrine and Metabolic Diseases from October 2004 to February 2006. Two mice of clean grade and of 12.5-14.5 days of gestational age were provided by Shanghai SLAC Laboratory Animal Co., Ltd (Permission No. 2004A034). The protocol was performed in accordance with ethical guidelines for the use and care of animals. Mouse embryonic stem cell lines were supplied by Dr Changxian Zhang (CNRS UMR5641, France). Activin A was the product of the R & D Corporation. ATRA and nicotinamide were supplied by the Sigma Corporation, USA. BFGF was supplied by Gibco Corporaion. METHODS: Head and viscera were removed from embryos of the pregnant mouse. The remaining tissues were cut into pieces and digested with trypsin. Cell suspension was centrifuged and inoculated at 3×108L-1. The cells could be used as mouse feeder layer after 2-3 times of passage. The mouse embryonic stem cells (ESCs) were inoculated onto the feeder layer in knockout Dulbecco's modified Eagle medium (DMEM) supplemented with leukemia inhibitory factors (LIF). ESCs were passaged at 1:3-1:6 after 2-3 days of culture. Culture medium with serum was added into the culture dishes to terminate the digestion. Cell fluid was centrifuged and supernatant was discarded. The sediments were prepared into suspension and inoculated at 2.5×104 with LIF-free culture medium. After 24-48 hours, embryonic bodies (EBs) were collected and replated in 1% Matrigel-coated dishes. When began to adhere to the dishes, EBs were cultured in 10% FBS/DMEM supplemented with 100μg/L activin A for 24 hours. Then EBs were switched to 10% FBS/DMEM for 6-8 hours as an interval. After this interval. EBs differentiated were cultured in 10% FBS/DMEM with 10<-6mol/L RA for another 24 hours followed by culture in 10% FBS/DMEM supplemented with 10μg/L bFGFs for 3-5 days. Finally, EBs differentiated were cultured in DMEM/F12 supplemented with N2 supplement, B27 supplement, 1μg/L laminin, 10μg/L bFGFs, and 10mmol/L nicotinamide for 3-5 days. Dithizone (DTZ) staining, inununofluorescent staining and reverse transcription-polymerase chain reaction (RT-PCR) were applied to detect insulin expression in the differentiated cells.MAIN OUTCOME MEASURES: Induction of ESCs, DTZ staining and immunofluorescent staining as wel as RT-PCR detection.RESULTS: Mouse ESCs growing on a feeder layer formed many colonies with clear boundary and dense structure. However, there was no obvious outer limit between these ESCs. EBs began to adhere to the dishes, which were coated with matrigel, on the 2nd day. After activin A and ATRA interval induction, EBs spread, and most of the living cells were epithelial cell-like when cultured in 10% FBS/DMEM supplemented with 10μg/L bFGFs. After culturing in DMEM/F12 supplemented with N2, B27, nicotinamide, bFGFs and laminin, the cells formed small clusters. The insulin-producing cells were stained dark red with DTZ, and the cells stained with primary antibody to insulin were insulin-positive. After 2 weeks of induction of activin A, ATRA, bFGFs and nicotinamide, the insulin-producing cells expressed insulin 2, Pdxl, Nkx6.1, Nkx2.2, PP, IAPP, Glut2, Somastatin, Hnf3β and Neuro D mRNA but did not express insulin 1 mRNA.CONCLUSION: Mouse ESCs call be induced to differentiate into insulin-producing cells by activin A, ATRA, bFGFs and nicotinamide in vitro. Induction time call be shortened to 2 weeks.

Objective To over-express human trefoil factor 2 (hTFF2) by Escherichia coli system and an-alyze its activities in promoting migration and anchorage-independent growth in SW480 colonic cancer cells. Meth-ods hTFF2 gene encoding mature peptide was obtained by RT-PCR, and the recombinant expression vector pET32a-hTFF2 was constructed. Then pET32a-hTFF2 was transformed into E. coli BL21-32a and TrxA-hTFF2 fu-sion protein was induced to over-express. The expressed product was isolated by Ni-NTA affinity chromatography, purified by dialysis and identified by Western blotting. The activities of the recombinant hTFF2 in promoting SW480 cells migration and anchorage-independent growth were analyzed by MicroChemotaxis Chamber migration assay and Soft-agar assay,respectively. Results The TrxA-hTFF2 fusion protein was expressed to 220 mg/L at high purity. In vitro model demonstrated that recombinant hTFF2 obviously enhanced SW480 cell migration activity and anchor-age-independent growth. Conclusion The recombinant hTFF2 can be expressed in E. coli with high production, purity and biological activities. And its roles in cell migration and anchorage-independent growth suggest that up-regulation of TFF2 in colonic cancer might be involved in cancer invasion and metastases.

Objective To explore the expression of microRNA in Eaf2 knockout mice and the effect of Eaf2 on the apoptosis of human lens epithe?lial cells and the expression of microRNA in lens. Methods pEGFP?C1?Eaf2 was transfected into SRA01/04 cells using Lipofectamine 2000 to over express Eaf2 gene,and then the flow cytometry was used to detect cell apoptosis rate. And real time q?PCR was used to measure the expression of microRNA both in human lens epithelial cells and Eaf2 knockout mice. Results Compared with controls,the apoptosis rate of cells transfected with pEGFP?C1?Eaf2 was reduced,the expression of miR?125b and let?7a was significantly increased and miR?204 was decreased in cells transfect?ed with pEGFP?C1?Eaf2. Compared with controls,the expression of miR?125b and let?7a was lower and miR?204 was higher in Eaf2 knockout mice. Each result was statistically significant(P<0.01). Conclusion Eaf2 might inhibit apoptosis of human lens epithelial cells via regulating the expression of microRNA. Eaf2 may have a protective effect for the lens in the genesis of cataract.