Objective To investigate the clinical significance of serum total bile acid(TBA)and cholyglycine(CG)detection in the early diagnosis of intrahepatic cholestasis of pregnancy(ICP)and perinatal adverse outcomes.Methods Chose 67 ca-ses of ICP pregnant women diagnosed and treated in Chang'an Hospital from June 2015 to June 2017 and they were selected as observation group.According to the 2015 edition of the diagnostic guidelines for the diagnosis and treatment of intrahe-patic cholestasis of pregnancy.The patients were divided into mild ICP group and severe ICP group,and 60 healthy pregnant women were selected as the control group.The serum TBA concentration was measured by fifth generation cyclic enzyme method and the concentration of serum CG was detected by latex enhanced turbidimetric immunoassay.The serum TBA,CG test results and the rate of abnormal test results,the incidence rate of perinatal adverse outcomes were compared between groups.Evaluation of serum TBA and CG detection of pregnancy early diagnosis of intrahepatic cholestasis and clinical value of perinatal adverse outcomes.Results The detection results of serum TBA and CG in the control group,mild ICP group and severe ICP group,there were significant differences between the three groups,the difference was statistically significant (P<0.01),the detection results in the CG group,serum TBA,ICP slightly higher than the control group,the difference was statistically significant(t=22.27,39.68,P<0.05).Weight of serum TBA and ICP group,the results of CG was higher than that of patients with mild ICP group,the difference was statistically significant(t=10.24,70.87,P<0.05).And in the con-trol group,mild ICP group,severe ICP group pregnant women serum TBA,CG test results increased with the aggravation of the disease.Serum TBA and CG abnormal results in 60 cases of the control group were not detected.In 67 cases of group ICP(mild ICP group and severe ICP group)were 63 cases and 61 cases,two groups of abnormal results rate comparison,and the difference was statistically significant(χ2=29.35,31.27,P<0.01).Perinatal premature labor,fetal distress,perinatal death and stillbirth incidence of adverse perinatal outcomes in the control group,mild ICP group and severe ICP group were significantly different between the three groups(χ2=39.17,56.31,13.02,6.92,P<0.01).Conclusion Intrahepatic chole-stasis of pregnancy,serum TBA and CG increased significantly,can be used as a sensitive indicator of ICP diagnosis,improve the detection rate of ICP,and effectively predict perinatal outcome.For intrahepatic cholestasis of pregnancy early detection and early diagnosis,it has important clinical significance.

OBJECTIVETo investigate the progression of visual field loss and to explore the prognosis of glaucomatous optic neuropathy in patients with chronic angle-closure glaucoma (CACG) after their intraocular pressures were well controlled under 21 mmHg.

METHODSForty-seven eyes of 29 patients in the Department of Ophthalmology in PUMC Hospital were included. All the patients had at least two separate tests of visual fields using the 24-2 program of the Humphery Visual Field Analyzer after their intraocular pressure were well controlled under 21 mmHg after sugery. The visual fields of patients were followed routinely for at least 1 year. In addition, all patients were divided into 2 groups according to follow-up period: 1-2 years group and over 2 years group. Visual field scores were calculated with the Advanced Glaucoma Intervention Study (AGIS) method. The visual fields were divided 5 sections and the sensitivity and defect depth of each section were calculated.

RESULTNo statistically significant differences were found in terms of AGIS scores, localized sensitivities and localized defects within the time interval of the observation.

CONCLUSIONGlaucomatous optic neuropathy is not likely to progressively deteriorate in CACG cases once their intraocular pressure are well controlled under 21 mmHg.

Aged ; Female ; Follow-Up Studies ; Glaucoma, Angle-Closure ; physiopathology ; surgery ; Humans ; Intraocular Pressure ; Male ; Middle Aged ; Optic Disk ; physiopathology ; Optic Nerve Diseases ; physiopathology ; Retrospective Studies ; Visual Fields

OBJECTIVETo explore a treatment program for increasing therapeutic effect on rheumatoid arthritis at active stage.

METHODSOne hundred and forty-six cases were randomly divided into treatment group (n = 74) and medicine control group (n = 72). The treatment group were treated by electroacupuncture at Quchi (LI 11), Hegu (LI 4), Yanglingquan (GB 34), etc. , combined with meloxicam, sulfasalazine and MTX. The control group treated by simple the Western medicines. Their therapeutic effects were compared.

RESULTSThe effective rate was 79.73% in the treatment group and 51.39% in the control group with a significant difference between the two groups (P< 0. 05).

CONCLUSIONElectroacupuncture combined with medicine has a better therapeutic effect than the simple medicine on rheumatoid arthritis at active stage.

Arthritis, Rheumatoid ; therapy ; Electroacupuncture ; Humans ; Medicine

OBJECTIVE To investigate the anti-metastatic effect of Alisol B on 4T1 breast cancer cells and explore its possi-ble mechanisms.METHODS 4T1 breast cancer cells were treated with Alisol B for 24 h.Cell viability was evaluated by MTT assay.Migration ability was evaluated by wound healing assay.Invasion ability was investigated by transwell invasion assay. The expressions of matrix metalloproteinases(MMP-2 and MMP-9)were detected by Western blot analysis.RESULTS Ali-sol B showed inhibitory effect on cell growth in 4T1 cells.Wound healing assay demonstrated that Alisol B inhibited the migra-tion of 4T1 cells after the treatment for 24 h.The migration rate was(52.66±8.48)%(P<0.01).Transwell invasion assay proved that Alisol B effectively suppressed the invasion of 4T1 cells.The invasion rate was(47.06 ± 9.32)%(P<0.01). Western blot results demonstrated that the expression of MMP-2 and MMP-9 were significantly down-regulated with the treat-ment of Alisol B.CONCLUSION Alisol B can suppress the metastasis of 4T1 breast cancer cells in vitro,which could be through down-regulation the expression of matrix metalloproteinases.

To explore the absorption mechanism of paeonol-beta-CD from various intestinal segments and offer biopharmaceutics data for paeonol new dosage form. The absorption kinetics and permeability rate consatants were investigated by the in situ perfusing method in rats. The absorption of the drug conforms to the firt-order kinetics and passive transport mechanism . The results indicate that paeonol-beta-CD absorption mechanism wasn't change.
Acetophenones ; pharmacokinetics ; Animals ; Intestinal Absorption ; physiology ; Intestines ; metabolism ; Kinetics ; Male ; Rats ; Rats, Sprague-Dawley

This study was aimed to construct the soluble HLA-A*0201-PR1 complex for preparation of HLA-A*0201-PR1 tetramer. The recombinant HLA-A*0201-BSP (BirA substrate peptide) fusion protein as heavy chain and beta(2)-microglobulin (beta(2) m) as light chain were expressed highly as insoluble aggregates in Escherichia coli and then purified with gel filtration, and the final purity reached above 90%. The two subunits were refolded to form an HLA-A*0201-peptide complex by dilution method in the presence of an antigenic peptide PR1, a HLA-A2-restricted peptide from proteinase 3 (aa 169 - 177, VLQELNVTV). Refolded HLA-A*0201-PR1 complex was biotinylated using a BirA enzyme and purified by anion exchange chromatography on a Q-Sepharose (fast flow) column. The extent of reconstitution of the HLA-A*0201-PR1 complex was analyzed by HPLC gel filtration. The refolded and biotinylated products were detected by Western blot and ELISA with monoclonal antibody BB7.2 that recognized the natural conformations of HLA-A2 and streptavidin. The results showed that the refolded complex was composed of HLA-A*0201-BSP aggregate, HLA-A*0201-PR1 complex and beta(2) m, and reconstitution yields of 18% with PR1 was obtained. Refolded HLA-A*0201-PR1 complex could be confirmed by practical immunological method and biotinylated efficiently. It is concluded that the refolding and biotinylation of HLA-A*0201-PR1 complex is successfully obtained. This work provides the basis for the preparation of HLA-A*0201-PR1 tetramer.
DNA Primers ; genetics ; Escherichia coli ; genetics ; HLA-A Antigens ; analysis ; biosynthesis ; genetics ; HLA-A2 Antigen ; Humans ; Oligopeptides ; genetics ; metabolism ; Protein Binding ; Protein Folding ; Recombinant Fusion Proteins ; analysis ; biosynthesis ; beta 2-Microglobulin ; biosynthesis ; chemistry ; genetics

Human beta(2)-microglobulin (beta(2)m) is the light chain of major histocompatibility complex (MHC) class I molecule. High-yield production of this protein is a prerequisite to the preparation of MHC class I tetramer. The present study was aimed to obtain recombinant human beta(2)m expressed in Escherichia coli (E. coli) for preparing MHC class I tetramers. For cloning of human beta(2)m gene, a pair of specific primers was designed based on the published sequence of this gene. A 300 bp specific DNA fragment corresponding to the encoding region of beta(2)m lack of the signal peptide sequence was obtained by RT-PCR from the total RNA of human leukocytes. The amplified cDNA was inserted into the IPTG-inducible expression plasmid pET-17b by Nde I and Bam H I sites and its sequence was confirmed by DNA sequence analysis. The recombinant plasmid pET-beta(2)m was transformed to the competent cells of E. coli BL21 (DE3). The results showed that beta(2)m was expressed in the form of inclusion body and amounted to over 32% of total cell proteins after IPTG induction. After washing with triton X-100 and urea, the inclusion body was dissolved with 4 mol/L urea and then purified with Sephacryl S-200 HR, and the final purity reached above 95%. The denatured protein was renatured by dilution method. Western blot assay indicated that the monoclonal antibody against human native beta(2)m could react specifically with the recombinant protein. In conclusion, the human beta(2)m gene was cloned successfully and expressed efficiently in E. coli BL21 (DE3). This work establishes a convenient approach for renaturation and purification of large quantity of recombinant beta(2)m. This provides the basis for the preparation of MHC tetramers.
Cloning, Molecular ; Escherichia coli ; genetics ; Gene Expression ; Histocompatibility Antigens Class I ; genetics ; Humans ; Recombinant Proteins ; biosynthesis ; beta 2-Microglobulin ; biosynthesis ; genetics

OBJECTIVETo evaluate the effect on pre-exposure prophylaxis (PrEP) to prevent HIV infection in high risk populations.

METHODSA computerized literature searching had been carried out in PubMed, EMbase, Ovid, Web of Science, Science Direct, Wanfang, Tsinghua Tongfang database and related websites to collect relevant papers (from establishment to June 2012) with the key words of pre-exposure prophylaxis, HIV, AIDS, high risk populations, relative risk, reduction. All randomized controlled trials (RCT) papers about using single or compound antiretroviral drugs (ARVs) orally or topically before HIV exposure or during HIV exposure in high risk populations were enrolled. Meta-analysis was conducted using Stata 10.0 to calculate the pooled RR value (95%CI). Consistency test was performed and publication bias was evaluated.

RESULTSFinally 5 RCT papers were enrolled, including 10 271 persons who were at high risk of HIV infection. The number of the experimental group was 5929, among which 116(1.96%) became infected. The number of the control group was 4342, among which 201(4.63%) became infected. Meta-analysis showed that the pooled relative risk (RR) and 95%CI was 0.49 (0.39 - 0.61), P < 0.05, indicating that the persons in experimental group had a 0.49 times lower risk of HIV infected, as compared with the control group. Publication bias analysis revealed a symmetry funnel plot. The fail-safe number was 825.

CONCLUSIONPrEP was an effective and safe protection measure to reduce HIV infection in high risk populations.

Anti-HIV Agents ; administration & dosage ; therapeutic use ; HIV Infections ; prevention & control ; Humans ; Randomized Controlled Trials as Topic ; Risk

High-yield production of HLA-A*0201 heavy chain is a prerequisite to the preparation of HLA-A2 tetramer. The present study was aimed to construct the expression vector of recombinant HLA-A*0201-BSP fusion gene for preparing HLA-A2 tetramers. The extracellular region HLA*0201 was cloned by RT-PCR from HLA-A2(+) donor, and a 15-amino acid biotin-protein ligase (BirA) substrate peptide (BSP) for BirA-dependent biotinylation was added to the COOH-terminus of HLA-A*0201 heavy chain. Then the fusion gene was cloned into pBV220 vector at EcoRI and Bam HI sites and its sequence was confirmed by DNA sequence analysis. The recombinant plasmid pBV220-HLA-A*0201-BSP was transformed to the competent cells of E.coli DH5alpha. The results showed that the HLA-A*0201-BSP fusion protein was successfully expressed in the form of inclusion body and amounted to over 28% of total cell proteins via induction at 42 degrees C. After washed with triton X-100 and urea, the inclusion body was dissolved with 8 mol/L urea and then purified with Sepharcyl S-300 HR, and the final purity reached above 90%. It is concluded that the HLA-A*0201-BSP fusion gene was cloned successfully and expressed efficiently in E.coli DH5alpha. This work establishes a convenient approach for purification of large quantity of recombinant HLA-A*0201-BSP. This provides the basis for the preparation of HLA-A2 tetramers.
Biotin ; biosynthesis ; genetics ; Carbon-Nitrogen Ligases ; biosynthesis ; genetics ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Escherichia coli Proteins ; biosynthesis ; genetics ; HLA-A Antigens ; biosynthesis ; genetics ; HLA-A2 Antigen ; Humans ; Ligases ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Repressor Proteins ; biosynthesis ; genetics ; Substrate Specificity ; Transcription Factors ; biosynthesis ; genetics

Objective To study the acceptability of pre-exposure prophylaxis (PrEP) to prevent the transmission of HIV among men who have sex with men (MSM) in Guangxi, China.Methods Snow-balling methods were used to recruit 650 MSM in Guangxi. Questionnaires and interview were administrated to these 650 men, using a self-designed questionnaire and face to face interviews to collect information on HIV-related risk behaviors, knowledge and acceptability of PrEP.effective, safe and free of charge', 597 (91.9%) of the 650 MSM claimed that they would accept it,who refused to use it, most of them said that were afraid of the side-effect and doubted on the effectiveness of PrEP. Data from logistic regression analysis showed that those who had found partners through friends (OR=6.21, P=0.020) and those who would advise his friend to use PrEP (OR=39.32, P=0.000) were more likely to accept PrEP. Those who thought they could protect themselves from HIV infection (OR=0.32, P=0.010) or not having sex with the ones who refused to use a condom (OR=0.34, P=0.010) were less likely to accept PrEP. Conclusion Effectiveness, safety and cost seemed to be the main influential factors related to the acceptability of PrEP. Peer education might improve the acceptability of PrEP.