Objective To discuss the protective effect of Tongsaimai pellets(TSMP) and butylphthalide(NBP) on PC12 cell in the hypoxia/hypoglycemia model.Methods Two ischemia models including hypoxia,hypoglycemia models were used to assay the anti-ischemic roles of TSMP and NBP in cultured PC12.Results TSMP and NBP possessed obvious protective effects on PC12 cells from two injured models.Both of them could increase the number of living cells and decrease LDH activity significantly,particularly in hypoglycemia injured model.Conclusion TSM and NBP have protective effect on PC12 cells from two injured models effectively in vitro.

To obtain the genome sequence of human bocavirus 2 (HBoV2), different regions of HBoV2 genome were amplified through PCR in fecal specimens which had been identified as single-positive for HBoV2 in 2010. A genome sequence of HBoV2 (HBoV2-NC, 5444 bp) was obtained after sequence assembly. The phylogenetic analysis showed that HBoV2-NC had the closest evolutionary relationship with HBoV2 Lanzhou strain. The predication of inverted terminal repeats of HBoV2-NC by DINAMelt showed that inverted terminal repeats were contained in HBoV2-NC 5' terminal, which had the typical stem-loop structure in other parvoviruses. Finally, some flanking sequences of HBoV2-NC were amplified by linker-PCR.
Base Sequence ; Gene Amplification ; Genome, Viral ; Human bocavirus ; chemistry ; classification ; genetics ; Humans ; Molecular Sequence Data ; Nucleic Acid Conformation ; Parvoviridae Infections ; virology ; Phylogeny ; RNA, Viral ; chemistry ; genetics ; Terminal Repeat Sequences

Human bocavirus 1 (HBoV1) is a novel virus that mainly causes respiratory tract infection, and it has the characteristic of genome of Parvovirus, containing three open reading frames that encode non-structural proteins NS1 and NP1 and structural proteins VP1 and VP2. Circular episome is present during the rolling circle replication of HBoV1, which provides the possibility of full genome amplification and infectious clone construction to save HBoV1. The recombination between HBoV1 and HBoV2-4 occurs frequently. With the three-dimensional culture, in vitro culture of HBoV1 provides a powerful tool for research on the pathogenesis of HBoV1. This review focuses on the molecular characteristics, association with diseases, in vitro culture, diagnosis and treatment of HBoV1.
Diarrhea ; virology ; Genomics ; Human bocavirus ; genetics ; isolation & purification ; physiology ; Humans ; Meningitis ; virology ; Respiratory Tract Diseases ; virology

Using sustained release tablets of gardenia extract as model drug and DPPH radical scavenging capacity as antioxidant index, the feasibility of using pharmacodynamics index was explored to evaluate sustained release tablets. Applying the established quantifiable method of DPPH radical scavenging to the dissolved liquid of model drug, release profiles and biological effects profiles were drawn, and their correlation was discussed. A good correlation was observed by linear regression and f2 actor, suggesting that the indicator could be used to evaluate sustained release tabletsofextracts of gardenia in which iridoids were mainly involved.
Antioxidants ; metabolism ; pharmacology ; Biphenyl Compounds ; metabolism ; Delayed-Action Preparations ; metabolism ; pharmacokinetics ; Free Radicals ; metabolism ; Gardenia ; chemistry ; Kinetics ; Linear Models ; Oxidation-Reduction ; drug effects ; Picrates ; metabolism ; Plant Extracts ; metabolism ; pharmacokinetics ; Tablets

BACKGROUND:Foreign scholars have researched hypoxia reperfusion in zebrafish embryos, but there is no research on c-fos gene expression and the mechanism during zebrafish cerebral hypoxia reperfusion.
OBJECTIVE:To observe the zebrafish embryonic brain cel apoptosis and expression of c-fos gene in brain tissues after hypoxia reperfusion.
METHODS:Zebrafish embryos were selected at 48 hours post fertilization. Neonatal hypoxia reperfusion injury was simulated by gradual y leading nitrogen (99.999%) into the device. After hypoxia treatment for 6, 12 and 24 hours, the embryos received reperfusion for 6 hours under normal oxygen concentration. The embryos in the control group received normoventilation (the dissolved oxygen concentration was about 7.0 mg/L). Acridine orange staining was performed to observe the effect of different hypoxia durations on the apoptosis of neurons in zebrafish, and then the c-fos gene expression was quantitative analyzed with real-time quantitative nucleic acid amplification detection system. And the expression level of c-fos gene was compared before and after hypoxia reperfusion.
RESULTS AND CONCLUSION:A smal amount of apoptotic brain cel s could be detected in the control group, and the c-fos gene expression level was decreased;in the experimental group, the number of apoptotic cel s was increased after hypoxia for 6, 12 and 24 hours, and the gene expression after hypoxia for 6 hours was increased distinctly. The results indicate that hypoxia can increase the c-fos gene expression in brain cel s of zebrafish embryos, which may be one of the mechanisms of brain cel apoptosis increasing after hypoxia.

Objective: in order to provide rapid and reliable method. Methods: Encoded Annexin Ⅴ cDNA was amplifyed from U937 cDNA libary by PCR and then subcloned into E coli expression vector. MS2-Annexin Ⅴ fusion protein could be overexpressed in E　coli. The MS2 bacteria protein could be removed by thrombin digestion.The mature Annexin Ⅴ was obtained by ion exchange chromatography and the FITC labled Annexin Ⅴ could be used in the detection of apoptosis. Results:Up to 37% of the total bacterial proteins was rhAnnexin Ⅴ as showed by SDS-PAGE. The purification of Annexin Ⅴ is over 99%. The FITC labled Annexin Ⅴ could efficiently detect apoptosis. Conclusion: We successfully established the technique procedure of obtaining a large quantity of Annexin Ⅴ and provided the basic routine for popularizing the detection of apoptosis' with high effciency.

AIM:To study the influence of material and configuration of microfiltration membranes on preparation of mono-ammonium glycyrrhizinate liposome(MGL) by membrane extrusion. METHODS: MGL were preparated by conventional rotary-evaporated film dispersion method and sonication,and MGL were extruding through 9 kinds of microfiltration membranes with 3 kinds of aperture specs of 0.1,0.2,0.45 ?m.The methods were evaluated by the filtration efficiency and the leakage rate of drug from liposomes. RESULTS: MGL were easily extruding through microfiltration membranes of NY6,PTFE,PVDF,PP and PC.MGL could not extruding through microfiltration membranes of PES,CN,CA and CA-CN.MGL become hardly to extrude through microfiltration membranes when increasing in the concentration of liposomes or the content of cholesterol.The leakage rate of drug from MGL was small when extruding through microfiltration membranes. CONCLUSION: The material and configuration of microfiltration membranes can remarkably influence the preparation of MGL by membrane extrusion method,and PC microfiltration membrane was selected.

Objective To investigate the Human papillomavirus(HPV)infection in different age groups of women in Shenyang, and explore its correlation with cervical biopsy diagnosis.Methods 7 311 women aged 13-85 did HPV test and thin-cytologic test (TCT)in the hospital.Some of them had biopsy detection under electronic colposcopy,and the pathological diagnosis was the golden standard for the diagnosis of cervical lesions.SPSS18.0 statistical software was used for all statistical analysis.Results The infection rate of <30 years old women was significantly higher than that of 30 - <40,40 - <50,≥50 years old women (P <0.05).The most prevalent high-risk HPV genotype in Shenyang were subtype 16,52,58,53,33,31 and 18,and the most prevalent low-risk HPV subtypes were 81,11 and 6.The former 4 subtypes of high-risk HPV infection accounted for 67.3% of all high-risk infection.As to the 4 subtypes with higher infection rate,the infection rate of ≥40 years old women was higher than that of <40 years old(χ2 =20.29,P =0.00).The top two low-risk HPV subtypes accounted for 74.8% of the infections.The mean age of the ICC patients were 48.3,which was statistically different from the other groups(P <0.05).Cervical lesions occured mostly in 40-49 years old,which accounted for 37.1% and was higher than the other agees(P <0.01).HPV16 infection rate increased with the severity of cervical lesions.Conclusion HPV DNA genotyping is a necessary methord for cervical cancer screen,an effective com-plement for precancerous lesions diagnosis which was missed in cytology test,and also an indispensable test for CIN treatment and follow-up after operation.

To study on the effects of Achyranthes bidentata on Tongsaimai pellets main active ingredients chlorogenic acid, isoliquiritin, harpagoside and glycyrrhizin in rats in vivo pharmacokinetic behaviors, a method for the simultaneous determination of chlorogenic acid, isoliquiritin, harpagoside and liquiritigenin in rat plasma was established by UPLC-MS/MS. The analysis was performed on a waters Acquity BEH C18 column (2.1 mm x 100 mm, 1.7 microm) with the mixture of acetonitrile and 0.1% formic acid/water as mobile phase, and the gradient elution at a flow rate of 0.3 mL x min(-1). The analytes were detected by tandem mass spectrometry with the electrospray ionization (ESI) source and in the multiple reaction monitoring (MRM) mode. It turned out that the analytes of Tongsaimai pellets groups C(max) and AUC(Q-infinity) values were higher than that with A. bidentata group, and the C(max) values of chlorogenic acid had significantly difference (P < 0.05), the AUC(0-infinity) values of chlorogenic acid and glycyrrhizin had significantly difference (P < 0.05); The T(max) and CL values of two groups had no significantly difference. Results showed that the established method was specific, rapid, accurate and sensitive for the studies of Tongsaimai pellets four main active ingredients in rat in vivo pharmacokinetic, and A. bidentata have varying degrees of effects on Tongsaimai pellets four main active ingredients in rat in vivo pharmacokinetic behaviors.
Achyranthes ; chemistry ; Animals ; Chalcone ; administration & dosage ; analogs & derivatives ; blood ; pharmacokinetics ; Chlorogenic Acid ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Glycosides ; administration & dosage ; blood ; pharmacokinetics ; Glycyrrhizic Acid ; administration & dosage ; pharmacokinetics ; Herb-Drug Interactions ; Male ; Pyrans ; administration & dosage ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

OBJECTIVETo study the absorption mechanism of Danshensu by using Caco-2 monolayer model .

METHODCaco-2 cell monolayer model was used to study the bi-direction transport of Danshensu. An LC-MS method was developed to measure the concentration of Danshensu in cell culture medium and calculate the apparent permeability coefficients (P(app)). The effects of time, drug concentration and inhibitor on the absorption of Danshensu were studied.

RESULTTransport of Danshensu was time and concentration dependent, and it was also effected by P-glycoprotein inhibitor. P(app) increased with time increase and tended to become saturated at some point. It, however,decreased while concentration of Danshensu increased. P(ratio) is larger than 1.5 . Verapamil can cause significantly effect on transport of Danshensu: P(app,A-B) increased and P(app,B-A) decreased .

CONCLUSIONThe absorption of Danshensu in Caco-2 cell model may be an active transportation mediated by P-glycoprotein transporter.

ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; Caco-2 Cells ; Cell Membrane Permeability ; drug effects ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Humans ; Lactates ; administration & dosage ; pharmacokinetics ; Plants, Medicinal ; chemistry ; Salvia miltiorrhiza ; chemistry ; Verapamil ; pharmacology