OBJECTIVETo study the active constituents from Alternanthera philoxeroides.

METHODThe constituents were isolated with silica gel and Toyopearl HW-40C gel column chromatography and purified by HPLC. Their structures were elucidated by spectroscopy.

RESULTNine compounds were isolated and identified as phaeophytin a (1), pheophytin a' (2), oleanoic acid (3), beta-sitosterol (4), 3beta-hydroxystigmast-5-en-7-one (5), alpha-spinasterol (6), 24-methylenecycloartanol (7), cycloeucalenol (8), phytol (9).

CONCLUSIONCompounds 1,2,5,7-9 were isolated from this plant for the first time.

Amaranthaceae ; chemistry ; Chlorophyll ; chemistry ; isolation & purification ; Phytol ; chemistry ; isolation & purification ; Phytosterols ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

Objective To investigate the bioequivalence of gliclazide sustained-release tablets in Chinese healthy subjects.Methods The study was designed using a single-center,open,randomized,single-dose,two-cycle,two-sequence administration method;subjects were orally administered the test/reference preparation 30 mg on an fasting or fed conditions,with self-cross-dosing.The concentration of gliclazide in human plasma was determined by liquid chromatography tandem mass spectrometry(LC-MS/MS)method.The main pharmacokinetic parameters of gliclazide(Cmax,AUC0-t and AUC0-∞)were analyzed by non-atrioventricular model of WinNonlin.Result In the fasting study,24 subjects were recruited and 22 completed the study.The main pharmacokinetic parameters of gliclazide sustained-release tablets test preparation and reference preparation in the fasting group were as follows:Cmax were(862.48±294.48)and(902.96±259.09)ng·mL-1;AUC0-t were(2.60 × 104±8 930.46)and(2.50 ×104±7 573.42)h·ng-1·mL-1;AUC0-∞ were(3.00 × 104±1.43 × 104)and(2.68 × 104±7 085.99)h·ng·mL-1.In the fed study,twenty-four subjects were enrolled and 23 completed the study.The main pharmacokinetic parameters of gliclazide sustained-release tablets test preparation and reference preparation in fed group:Cmax were(1 531.74±273.49)and(1 510.87±241.08)ng·mL-1;AUC0-t were(2.78 ×104±9 565.89)and(2.76 ×104±9 821.43)h·ng·mL-1;AUC0-∞ were(3.02 ×104±1.24 ×104)and(3.02 × 104±1.30 × 104)h·ng·mL-1 h·ng·mL-1.The 90％confidence intervals of the geometric mean ratios of Cmax,AUC0-t,AUC0-∞ for the test preparation and reference preparation gliclazide sustained-release tablets were all between 80％and 125％.Conclusion The test and the reference preparation of gliclazide sustained-release tablets are bioequivalent in Chinese healthy subjects.

Objective To find out the status of HIV infection,sexual behaviors with both men and women as well as condom use among currently married men who have sex with men (MSM) so as to document for HIV intervention targeting this sub-population,in Beijing,Chengdu,Harbin and Zhengzhou cities.Methods “Snowballing” sampling method was used to recruit subjects in four cities.Participants were investigated by anonymous questionnaire containing demographics,sexual orientation,both homosexual and heterosexual behaviors and condom use during the past 6 months.Blood samples from participants were used to test HIV antibodies.Results Of the 858 participants for four cities,the average age was 38.3 (SD=9.1) years.36.7％ of them had completed the junior high school or under.The percentage of participants who identified themselves as homosexual,bisexual,and heterosexual or undecided were 40.9％,54.3％ and 4.8％,respectively.The overall HIV prevalence was 8.0％,and among four cities the HIV prevalence in Chengdu was the highest ( 13.3％ ).Results from the Multiple logistic regression analysis showed that never used condoms when having anal sex with men during the past 6 months among participants who did not have a city residential card,identifying himself as having homosexual orientation,living in Harbin,having first anal sex experience before 18 years of age the range of OR value was 1.5-2.1.In addition,the faetors associated with never using condoms in vaginal sex with women during the past 6 months were as follows:being over 46 years old,not having a city residential card,living in Beijing,identifying himself as heterosexual orientated or with unknown sexual orientation,never using condoms in anal sex with men during the past 6 months,the range of OR value was 1.7-5.9.Conclusion The prevalence of HIV infection seemed to be quite high among those currently married MSM.High rates of unprotected homosexual and heterosexual behaviors might accelerate the spreading of HIV from currently married MSM to their wives.

Objective To explore the clinical effects of sequential flap of thigh perforator for closure of the anterolateral thigh free flap donor site.Methods From March,2014 to May,2015,8 consecutive patients underwent elective anterolateral thigh free flap reconstruction for limb soft tissue defect.To design a sequential flap of thigh perforator to achieve primary closure of the anterolateral thigh free flap donor site.The area of anterolateral thigh free flap ranged from 9.0 cm×6.0 cm to 20.0 cm×1 1.0 cm,respectively.The area of thigh perforator flap ranged from 7.0 cm×3.0 cm to 12.0 cm×7.0 cm.Results All flaps healed uneventfully.Distal skin flap necrosis appeared in 1 flap which was healed after local transposition flap.All the patients were followed up for 1 to 6 months,with mean time of 4 months.The color,texture and appearance of flaps were satisfactory.Conclusion Sequential flap of thigh perforator,with reliable blood supply and constant position,represents an ideal model to close the anterolateral thigh free flap donor site of small defect.

Objective Coronary arterial plaque rupture and secondary thrombosis are the major pathogenesis of acute coronary syndrome ( ACS) . Metalloprotease ( MMPs) secreted by monocyte/ macrophage was the main predisposing factor of the plaque rupture and peroxisotne proliferator-activated receptor-γ (PPAR-γ) is involved in a variety of inflammatory cytokine gene transcriptional regulations. We explored the possible role of PPAR-γ in the regulation of MMP-9 and TIMP-1 expressed by peripheral monocyte-derived macrophages (MDMs) from patients with ACS. Methods Peripheral blood mononuclear cells were isolated from 48 patients with ACS and 28 healthy controls and stimulated by macrophage colony-stimulating factor (0. 1 μg/ml for 24 hours) to form MDMs. MDMs were then incubated under various concentrations of rosiglitazone (0, 1, 10, 20 μmol/L) for 48 hours. The concentrations of MMP-9 and TIMP-1 in the supernatant were measured by enzyme linked immunosorbent assay, and the mRNA expression of PPAR-γ, MMP-9 by RT-PCR and nuclear factor-KB P65 ( NF-kB P65 ) expression by immunohistochemistry. Results PPAR-γ mRNA expression was significantly lower while NF-kB P65 and MMP-9 expression as well as MMP-9 and TIMP-1 concentrations in supernatant were significantly higher in ACS group than those in control group (all P <0. 05). After rosiglitazone intervention, PPAR-γ mRNA expression was significantly upregulated in both ACS and control groups in a dose-dependent manner. Both the MMP-9 concentration in the supernatant and MMP-9 mRNA expression were reduced post intervention with rosiglitazone in both groups. The TIMP-1 mRNA expression and concentration in supernatant were not affected by rosiglitazone in both groups. Rosiglitazone induced significant downregulation of NF-kB P65 expression in both groups. Conclusion Rosiglitazone intervention may downregulate MMP-9 expression by upregulating PPAR-γ expression, and by downregulaiting NF-kB expression in MDMs isolated from patients with ACS.

OBJECTIVETo investigate HIV and treponema pallidum infection status, risky sexual behavior among men who have sex with men (MSM) and its impact factors in China.

METHODSSnowball sampling was used to recruit subjects from April to August in 2008 in Beijing, Harbin, Zhengzhou and Chengdu city. Serological test of HIV and treponema pallidum were conducted and a questionnaire survey was undertaken among subjects. The questionnaire included social demographics, characteristics of sexual partners and sexual behaviors.

RESULTSA total of 1693 subjects was enrolled in present study, which included 1390 MSM/M (82.1%) and 303 MSMW (17.9%). The infection rate of HIV among MSM/M and MSMW subjects were 7.0% (97/1390) and 6.6% (20/303), respectively. The infection rate of treponema pallidum among MSM/M and MSMW subjects were 11.9% (166/1390) and 13.2% (40/303), respectively. The proportions of MSM/M subjects who never used or sometimes used condoms when having same-sex anal intercourse in recent 6 months were 8.6% (120/1390), 45.3% (630/1390), respectively. The according proportions among MSMW subjects were 10.2% (31/303), 44.6% (135/303), respectively. Among MSM/M subjects, the risk factors of risk sexual behaviors included having less than 6 sexual partners (OR = 6.03, 95%CI: 2.54 - 14.28), no same-sex regular sexual partner (OR = 2.18, 95%CI: 1.30 - 3.65), no same-sex casual sexual partner (OR = 2.90, 95%CI: 1.79 - 4.71), T-pattern only during anal intercourse (OR = 1.64, 95%CI: 1.13 - 2.37) or P-pattern only (OR = 1.58, 95%CI: 1.04 - 2.41). Among MSMW subjects, the risk factors of same-sex risk sexual behaviors included having less than 6 sexual partners (OR = 12.95, 95%CI: 2.38 - 70.52), no same-sex regular sexual partners (OR = 0.42, 95%CI: 0.21 - 0.85), never used condom during heterosexual intercourse (OR = 3.53, 95%CI: 1.48 - 8.42).

CONCLUSIONThe infection rate of HIV and treponema pallidum among MSM subjects were quite high, and the same-sex risk sexual behaviors among MSM subjects were ubiquity, whose risk factors including having less than six of sexual partners, no same-sex casual sexual partner, T-pattern or P-pattern only during anal intercourse and never used condemn during heterosexual intercourse among MSMW subjects no same-sex regular partner was a risk factor in MSMS population, while having same-sex regular partners was a risk factor in MSMW population.

Adolescent ; Adult ; China ; epidemiology ; HIV Infections ; epidemiology ; Homosexuality, Male ; Humans ; Logistic Models ; Male ; Risk Factors ; Risk-Taking ; Sexual Behavior ; Surveys and Questionnaires ; Syphilis ; epidemiology ; Young Adult

To evaluate in vitro release and transdermal behaviors of Zhitong cataplasm, modified Franz diffusion cell method was applied to investigate in vitro transdermal absorption of Zhitong cataplasm and the content of tetrahydropalmatine was determined by HPLC. In 24 hours, accumulative release rate of tetrahydropalmatine was 81. 9%, transmission rate was 2.26 microg x cm(-2) x h(-1). In 48 hours, accumulative transdermal rate and transmission rate of tetrahydropalmatine were 20.31%, 0.22 pg x cm(-2) x h(-1). So Zhitong cataplasm had a good release and transdermal properties and transdermal actions were consistent with zero-order kinetics process.
Administration, Cutaneous ; Animals ; Berberine Alkaloids ; administration & dosage ; chemistry ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacokinetics ; Male ; Mice ; Plant Extracts ; administration & dosage ; chemistry ; pharmacokinetics ; Skin ; metabolism ; Skin Absorption

OBJECTIVETo study effects of Yuquan pills on the pharmacokinetics process of metformin hydrochloride in diabetic rats.

METHODAfter administration Yuquan pills 7 day to the diabetic rats, the metformin hydrochloride was orally administrated, then the blood samples were collected at different time. The concentrations of metformin hydrochloride in plasma were determined by HPLC method and the pharmacokinetic parameters were calculated.

RESULTThe pharmacokinetic parameter Cmax of the controlling group and the testing group were respectively, 18.95, 21.76 mg x L(-1); t1/2 were 1,069.8, 1,767.4 min, respectively; CL/F were 0.013, 0.008 L x min(-1) x kg(-1); AUC were 10,042.1, 10,712.2 mg z L(-1) x min(-1) respectively.

CONCLUSIONThe pharmacokinetics process of metformin hydrochloride in diabetic rats fits one-compartment model. Yuquan pills has a significant effect on the pharmacokinetics of metformin hydrochloride in diabetic rats.

Animals ; Chromatography, High Pressure Liquid ; Diabetes Mellitus, Experimental ; blood ; drug therapy ; metabolism ; Drug Interactions ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Metformin ; blood ; pharmacokinetics ; Rats ; Rats, Wistar

OBJECTIVETo construct inducible lentiviral vector containing human Notch1 intracellular domain (NICD) gene and enhanced green fluorescent protein (EGFP), and to study its expression in PC12 cells.

METHODSNICD cDNA was amplified by RT-PCR from human placenta tissue. EGFP gene was amplified by PCR from pEGFP-C1. Both NICD and EGFP were cloned into pcDNA 3.1 (+) plasmid to form pcDNA3.1-Notch1-EGFP. Then the Notch1-EGFP fragment was separated and cloned into pLVX-Tight-puro to form pLVX-Notch1-EGFP. The lentivirus were packaged and harvested, which were used to infect PC12 cells. After antibody selection for 2 weeks, the PC12 cells were induced by doxycycline (Dox). The expression of Notch1-EGFP was detected by fluorescence microscope and flow cytometry.

RESULTSThe recombinant inducible lentiviral vectors (pLVX-Notch1-EGFP) were success fully constructed. The EGFP positive cell percentage was over 90% in transfected PC12 cells after 500 ng/ml Dox induction for 36 h. The expression of Notch1 was posited correlated to the Dox concentration. The expression of Notch1 increased with the duration of Dox induction, which got the peak at 36 h after Dox induction.

CONCLUSIONThe recombinant inducible lentiviral vectors containing Notch1 and EGFP gene are successfully constructed, which provides an effective and simple method to regulate the expression of Notch1 in PC12 cells.

Animals ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Lentivirus ; genetics ; PC12 Cells ; Plasmids ; Rats ; Receptor, Notch1 ; genetics ; Transfection