Objective To investigate the effects of sex hormone-binding globulin (SHBG)gene in the apoptosis of human trophoblastic cells.Methods The siRNA specific-targeting SHBG gene was transfected into human trophoblastic cells and they were divided into six groups:trophoblasts without transfection in normal control groups(group Ⅰ);transfect liposome in blank control groups(group Ⅱ);transfect nonspecific siRNA in negative control groups(group Ⅲ);transfect SHBG siRNA-Ⅰ,SHBG siRNA-Ⅱ,SHBG siRNA-Ⅲ respectively in trans-fection group(group Ⅳ,Ⅴ,Ⅵ).Hoechst 33258 dying method was used to detect cell apoptosis.SHBG and Caspase-3 mRNA profiling and the level of SHBG and caspase-3 protein were detected by real-time PCR and Western blot.Results There was no statistical significant difference in the gene expression and protein level of SHBG and caspase-3 in group Ⅰ,Ⅱ and Ⅲ (P >0.05).In Ⅳ,Ⅴ and Ⅵ group,there was no statistical significant difference in the expression level of SHBG and caspase 3 (P >0.05).Compared with group Ⅰ,Ⅱ and Ⅲ,the a-mount of SHBG gene expression decreased obviously,the caspase-3 mRNA and protein level increased obviously and the trophoblast cell ap-optosis increased markedly (P <0.05).Conclusion Through siRNA interference technology can reduce SHBG gene expression in human trophoblastic cells,and it can lead to excessive apoptosis of human trophoblasts cells.

The study established a UPLC-MS/MS method that is used for simultaneous determination nine major bioactive compounds of Dachengqi Tang in rat plasma. Using Aglient C18 column (2.1 mm x 50 mm,1.7 microm) was chromatographed, using methanol-5 mmol x L(-1) ammonium formate mobile phase gradient, elution 0.3 mL x min(-1). In the plasma pre-treatment process, not only the method of methanol and acetonitrile protein precipitation was investigated, and different factors extraction solvent, the type of the scroll time, the number and the type of extraction solvent, the extraction volume of the extraction solution of liquid-liquid extraction is investigated. Finally, with ibuprofen as an internal standard, using ethyl acetate liquid-liquid extraction method pretreatment blood, N2 dry reconstituted supernatant after centrifugation UPLC-MS/MS analysis, in electrospray ionization (ESI) negative mode, using multiple reaction monitoring mode for testing. The linear range of emodin, rhein, aloe-emodin, chrysophanol, magnolol, honokiol, hesperidin and hesperitin is 0.33-660, 0.40-792, 0.41-827, 0.34-680, 0.45-907, 0.46-927, 0.43-867, 0.34-683, 0.39-787 microg x L(-1) respectively, good linear relationship; and extraction recovery were greater than 69.39%, days after the day of the RSD is less than 15%. This method can be used to study the rat gastric large bearing gas after Dachengqi Tang, the simultaneous determination of nine components in plasma for its pharmacokinetics and efficacy material base to provide a theoretical basis.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Female ; Male ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

OBJECTIVETo evaluate the effect of sustained-release alpha-lipoic acid tablets (SRLA) on blood lipid, glucose and insulin levels in hyperlipidemic New Zealand rabbits.

METHODSTwenty-four New Zealand rabbits were randomized into normal diet group, high-fat diet group, and high-fat diet + SRLA (300 mg/tablet) group with corresponding feed. At the beginning and 4 weeks after the feeding, the serum levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), blood glucose, and serum insulin were measured, and insulin sensitivity index (ISI) was calculated.

RESULTSFour weeks after feeding with high-fat diet, the insulin levels was elevated and the ISI lowered in the New Zealand rabbits, indicating successful establishment of the animal model of hyperlipidemia. Compared with the high-fat diet group, the serum levels of TG, TC, LDL-C and insulin were significantly reduced (P<0.05), and the ISI was significantly increased (P<0.05) in high fat diet + SRLA group. But no statistically significant difference was found in the blood glucose among the 3 groups.

CONCLUSIONSRLA can significantly correct blood lipid and insulin disorders in hyperlipidemic New Zealand rabbits and prevent the occurrence of insulin resistance and hyperlipidemia.

Animals ; Blood Glucose ; metabolism ; Delayed-Action Preparations ; Hyperlipidemias ; blood ; drug therapy ; metabolism ; Insulin ; metabolism ; Lipids ; blood ; Male ; Rabbits ; Tablets ; Thioctic Acid ; administration & dosage ; pharmacology ; therapeutic use

OBJECTIVETo prepare cryptotanshinone (CT)-cyclodextrin inclusion compound and improve dissolution of CT.

METHODInclusion ratio was determined by plotting the phase solubility curve of CT versus hydroxypropyl-beta-cyclodextrin (HPCD). CT-cyclodextrin inclusion compound was made by wet grinding method. Properties of the inclusion compound was investigated by in vitro dissolution test, DTA and IR spectrum.

RESULTInclusion ratio of CT versus HPCD was 1:1. Dissolution of CT-HPCD inclusion compound at 45 min was 21.6 times of material drug.

CONCLUSIONDissolution of CT was improved remarkably in CT-HPCD inclusion compound. The complexation force of the inclusion compound was hydrogen bond formed by carbonyl group of CT and hydroxyl group of HPCD.

2-Hydroxypropyl-beta-cyclodextrin ; Biological Availability ; Drug Carriers ; Drugs, Chinese Herbal ; chemistry ; Phenanthrenes ; chemistry ; isolation & purification ; Salvia miltiorrhiza ; chemistry ; Solubility ; Technology, Pharmaceutical ; methods ; Time Factors ; beta-Cyclodextrins ; chemistry

OBJECTIVETo establish a method for determining the content of alpha-lipoic acid in New Zealand rabbit plasma.

METHODSAlpha-lipoic acid in the plasma samples was purified by solid-phase extractor and analyzed on an HYPERSIL C18 column with isocratic mobile phase consisting of potassium dihydrogen phosphate-acetonitrile (50:50, v/v) at a flow rate of 1.0 ml/min and detection wavelength of 230 nm.

RESULTSThe standard curve was linear in the range of 5-100 microg/L (r=1) and the average recovery was 77.4%-82.1%. The relative standard deviations of intra-day and inter-day assay were within 1.5%-8.9%.

CONCLUSIONThe method is sensitive, accurate and simple for determining plasma alpha-lipoic acid levels in New Zealand rabbits.

Animals ; Chromatography, High Pressure Liquid ; methods ; Rabbits ; Thioctic Acid ; blood

AIMTo assess whether exposure to computers harms the semen quality of healthy young men.

METHODSA total of 178 subjects were recruited from two maternity and children healthcare centers in Shanghai, 91 with a history of exposure to computers (i.e., exposure for 20 h or more per week in the last 2 years) and 87 persons to act as control (no or little exposure to computers). Data on the history of exposure to computers and other characteristics were obtained by means of a structured questionnaire interview. Semen samples were collected by masturbation in the place where the semen samples were analyzed.

RESULTSNo differences in the distribution of the semen parameters (semen volume, sperm density, percentage of progressive sperm, sperm viability and percentage of normal form sperm) were found between the exposed group and the control group. Exposure to computers was not found to be a risk factor for inferior semen quality after adjusting for potential confounders, including abstinence days, testicle size, occupation, history of exposure to toxic substances.

CONCLUSIONThe present study did not find that healthy men exposed to computers had inferior semen quality.

Adult ; Case-Control Studies ; China ; Computers ; Electromagnetic Fields ; adverse effects ; Humans ; Male ; Semen ; Surveys and Questionnaires

OBJECTIVETo assess the effect of RNA interference-mediated gene silencing of plasma membrane-related Ca(2+)-ATPase-1 (PMR1) gene on the insulin secretion in islet beta cells NIT-1 in vitro.

METHODSA small interfering RNA duplex (siPMR1) corresponding to the nucleotides 337-357 of mouse PMR1 cDNA was introduced into NIT-1 cells via liposomes. The gene silencing effect was assessed by RT-PCR, and the total insulin level in the transfected cells was measured by radioimmunoassay.

RESULTSTransfection with siPMR1 resulted in obviously reduced PMR1 expression and increased insulin secretion in NIT-1 cells.

CONCLUSIONThe synthesized siPMR1 can significantly silence the expression of PMR1 and promote the secretion of insulin in the islet cells in vitro, which shed light on further studies of RNAi-based therapy of diabetes.

Animals ; Calcium-Transporting ATPases ; deficiency ; genetics ; Cell Line ; Gene Expression Regulation ; Insulin ; secretion ; Insulin-Secreting Cells ; metabolism ; secretion ; Mice ; RNA Interference ; RNA, Messenger ; genetics ; metabolism

AIMTo investigate the paclitaxel-loaded nanoparticles of poly(ethylene glycol)-b-poly(D,L-lactic acid) amphiphilic diblock copolymer (PMT).

METHODSPMT was prepared by solid dispersion technique. The average size and size distribution were determined by dynamic light scattering (DLS). The morphology was characterized by transmission electron microscopy (TEM) and 1HNMR. The influences of the copolymer molecular weight and the paclitaxel-fed amount on PMT were studied. Therapeutic effect of PMT was studied on Kunming mice liver cancer H22.

RESULTSPMT showed nanometer size and spherical morphology with core and shell. The sizes of PMT increased with increasing the molecular weight of the hydrophobic segment in PEDLLA or increasing the drug-loaded amount. The tumour inhibiting effect of PMT was similar with that of Taxol.

CONCLUSIONIt will provide an experiment basis for the development of new kind of intravenous administration of paclitaxel.

Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Lactic Acid ; Liver Neoplasms, Experimental ; pathology ; Mice ; Microspheres ; Nanotechnology ; Paclitaxel ; administration & dosage ; pharmacology ; Particle Size ; Polyethylene Glycols

This study was to investigate the protective effects of puerarin on myocardial ischemia/reperfusion (MI/R) injury and the underlying mechanism. The MI/R-model was established by ligating the left anterior descending artery (LAD) for 60 min followed by 24 h reperfusion, puerarin (10, 30, and 100 mg·kg^-1) was orally administered 20 min before reperfusion. Cardiac function, myocardial infarct index, cardiac damage markers, inflammatory cytokines, and apoptosis index were measured to evaluate the protective effects of puerarin on MI/R injury. The activation of Nod-like receptor protein 3 (NLRP3) inflammasome and Toll like receptor 4 (TLR4)/myeloid differentiation factor 88 (Myd88)/nuclear factor kappa B (NF-κB) pathway were determined by Western blot. All animal experimental procedures were approved by the ethics committee of the Institute of Materia Medica, Peking Union Medical College, Chinese Academy of Medical Sciences. The results showed that puerarin could significantly improve cardiac function, reduce myocardial infarct size, decease the levels of lactic dehydrogenase (LDH), aspartate transaminase (AST), creatine kinase-MB (CK-MB), and cardiac troponin T (cTnT) and suppress cardiomyocyte apoptosis. Meanwhile, puerarin could notably decrease the levels of inflammatory cytokines such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α). Western blot analysis revealed that puerarin could downregulate the expression of TLR4, Myd88, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cleaved-caspase 1, cleaved-gasdermin-D (GSDMD), IL-1β, and IL-18, as well as the phosphorylation levels of inhibitor of NF-κB α (IκBα), IκB kinase β (IKKβ), and NF-κB. These findings demonstrated that puerarin could alleviate MI/R injury by suppressing NLRP3 inflammasome activation, possibly via TLR4/Myd88/NF-κB pathway.

OBJECTIVETo observe the effects of Liuwei Dihuang Granule ([symbols; see text], LDG) for tonifying Kidney (Shen) on the outcomes of in vitro fertilization pre-embryo transfer (IVF-ET) of infertility women with Kidney-yin deficiency syndrome and to explore its mechanism by detecting the proteome expression in the follicular fluid.

METHODSSixty-six infertility patients of Kidney-yin deficiency syndrome who would undergo IVF-ET, were randomly assigned to a treatment group and a control group according to a random number table, 33 cases in each group. Another 33 cases of non-Kidney-yin deficiency syndrome was taken as a syndrome-control group. Besides Western routine therapy, LDG was given 3 menstrual cycles before IVF to the treatment group, and a placebo granule to the control and syndrome-control groups. The scores of Kidney-yin deficiency symptoms (sore waist and knees, dry vagina, dysphoria with feverish sensation in the chest, palms and soles, etc.) were assessed, the number of retrieved oocytes, rates of high quality oocytes and embryos, fertility rate and clinical pregnancy rate were recorded, and the follicular fluid was collected on the day when the ovum was picked up, the differential protein expression was detected using two-dimensional gel electrophoresis, and then, matrix assisted laser desorption ionization time-of flight mass spectrometry (MALDI-TOF-MS) was applied to identify the proteins.

RESULTSThe syndrome score in the treatment group decreased significantly from 16.09±2.58 to 8.67±2.13, while it changed insignificantly in the control group, with a significant difference in the lowering score between the two groups (P<0.05); the high quality rates of oocytes and embryos and clinical pregnancy rate were all superior in the treatment group to the control group (82.29% vs 78.08%, 76.76% vs 68.79%, 63.64% vs 36.36%, all P<0.05). The protein expression map from the follicular fluid showed that compared with the control group, 33 differential protein expressions were found in the syndrome-control group, among which 18 were down-regulated, and 15 up-regulated; in the treatment group 28 differential protein expressions were found, among which 15 were down-regulated, and 13 up-regulated. Through MALDI-TOF-MS, 14 proteins were identified (P<0.05).

CONCLUSIONSFor the infertility patients undergoing IVF, LDG could alleviate clinical symptoms, improve rates of high quality oocytes and embryos, so as to raise clinical pregnancy rate. The mechanism may be through regulating proteome expression in the follicular fluid to improve the developmental microenvironment for oocytes which would lead to a successful embryo implantation.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Embryo Transfer ; Female ; Fertilization in Vitro ; methods ; Follicular Fluid ; metabolism ; Holistic Health ; Humans ; Infertility, Female ; metabolism ; therapy ; Kidney ; metabolism ; Oocytes ; cytology ; metabolism ; Ovarian Hyperstimulation Syndrome ; metabolism ; Placebos ; Pregnancy ; Pregnancy Outcome ; Proteomics ; methods ; Yin Deficiency ; metabolism ; therapy