Objective: To examine and analyze the expression of two isoforms of livin (livin α/livin β), a novel inhibitor of apoptosis protein (IAP) family member, in tumor cell lines and malignant tumor tissues and analyze the relationship between livin expression and clinical pathological features of different tumors. Methods: Livin mBNA expression was detected by reverse transcription polymerase chain reaction (BT-PCR) in 12 malignant tumor cell lines, 50 different malignant tumor tissues, and 20 cases of normal tissues. Results: Livin mBNA was highly expressed in seven cell lines (Hela, SPEA-1, SBE-2, PC14, MKN45, LOVO, and HHCC) and poor expressed in A549 cells, It had negative expression in other four kinds of cell lines ( HL-7702, Hep-2, 7721 and K562). Livin mRNA was positively expressed in 32 of 50 malignant tumor cases (64.0%). The expression of livin mRNA was at low level in paracancerous tumors tissues (2%). Livin expression had no significant association with differentiation degree, lymph node metastasis and TNM staging of tumor patients. Conclusion: Livin expression in tumor cell lines and malignant tumor tissues provides stronger evidence for further investigating the relationship between livin and tumor initiation and development. It may serve as a new target for malignant tumor diagnosis and therapy.

Objeetive To investigate the clinical significance of combined detection of lipoprotein associated serum phospholipase A2 (Lp-PLa2),homocysteine (Hcy) and cystatin C (CysC) in the diagnosis of hypertensive disorders complicating pregnancy (HDCP).Methods From January 2013 to May 2016 in Changan Hospital,selected 113 cases of pregnancy induced hypertension patients as the observation group,and were divided into three group A,B and C (group A:55 cases of HDCP patients,group B:32 cases of mild preeclampsia and group C for patients with severe preeclampsia 26 cases).At the same period,selected 50 cases of normal college pregnancy as control group,serum Lp-PLa2 (enzyme-linked immunosorbent assay),Hcy (cyclophorase method) and CysC (particle enhanced turbidimetric method),the test results were analyzed and compared.Results Serum Lp-PLa2,Hcy and CysC test results in the control group,the observation group A,observation group B and observation group C increased significantly,in the observation group C increased most obviously.Compared with the control group,the serum levels of Lp-PLa2,Hcy and CysC in the observation group were significantly higher,the difference was statistically significant (F=8.102,7.231 and 6.926,all P＜0.05).Pearson correlation analysis showed that there was a positive correlation between serum Lp-PLa2,Hcy and CysC and blood pressure (r=0.71,0.69,0.63,all P＜0.05).The abnormal rate of serum Lp-PLa2,Hcy and CysC for three joint detection was higher than that of single detection,and the difference was statistically significant (x2 =6.725,P＜0.001).The abnormal rate of serum Lp-PLa2,Hcy and Cys single test results increased with the exacerbation of HDCP,and the difference was statistically significant (x2=9.351,P＜0.000).Conclusion Serum Lp-PLa2,Hey,CysC and pregnancy would be closely related to the occurrence and development of hypertension syndrome,so combined detection of HDCP can improve the detection rate of abnormal results,and it has important clinical significance for early diagnosis and prognosis of HDCP.

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.

OBJECTIVESTo test in vitro the spermatozocidine drug which can also prevent sex transmitting diseases (STD) pathogens.

METHODSChlorheridine diacetate and other three chemical compounds were applied in vitro spermatozocidine and sperm inhibitting tests.

RESULTSThe lowest concentrations of chlorheridine diacetate and p-nitrophenol which can inhibit human sperm in 20 seconds were 1.25 mg/ml. The minimal inhibitory concentration and minimal bactericidal concentration of chlorheridine diacetate and p-nitrophenol on Streptococcus albus Stemberg were 0.125 to 0.50 mg/ml and 0.25 to 1.00 mg/ml.

CONCLUSIONSChlorheridine diacetate and p-uitrophenol have strong spermatozocidine and antibacteria effects.

Acetates ; pharmacology ; therapeutic use ; Anti-Bacterial Agents ; pharmacology ; therapeutic use ; Humans ; Male ; Microbial Sensitivity Tests ; Neisseria gonorrhoeae ; drug effects ; Nitrophenols ; pharmacology ; therapeutic use ; Sexually Transmitted Diseases ; prevention & control ; Spermatocidal Agents ; pharmacology ; therapeutic use ; Spermatozoa ; drug effects ; Streptococcus ; drug effects

The distribution information of Lomatogonium rotatum. was collected by interview investigation and field survey, and 55 related environmental factors were collected, the habitat suitability study was conducted based on geographic information system (GIS) and maximum entropy model. The AUCs of ROC curve were both above 0.99, indicating that the predictive results with the maximum model were highly precise. The results showed that 13 major environmental factors have obvious influence on ecology suitability distributions of L. rotatum, including month average temperature of February et al., the suitable distribution areas are mainly concentrated in the east-central of Inner Mongolia, including Hexigten banner, Duolun county, Zhenglan banner et al., The zoning results basically coincide with the genuine producing areas, and further afford new suitable distribution areas, which can provide reference for L. rotatum's wild nursery and the siting of introduction and cultivation.
China ; Ecosystem ; Environment ; Gentianaceae ; growth & development ; Geographic Information Systems ; Rain ; Temperature

OBJECTIVETo investigate the effects of anluohuaqianwan on experimental hepatic fibrosis induced by dimethyl nitrosamine (DMN) in rats.

METHODS36 male SD rats were randomly dividied into three groups: model group, normal group, anluohuaqianwan group. The rats in the three groups were treated with DMN daily for 4 weeks. The liver function was detected using auto biochemistry analyzer, the serum HA, LN, IV-C, PIIIP were detected by immunoradiometry, the histopathology was observed in the left liver lobe after HE staining, the expression of matrix metalloproteinase-2 (MMP-2) in liver tissue were detected by immunohistochemistry.

RESULTSThe serum levels of ALT, AST, ALP, TP, ALB and the contents of HA, LN, IV-C in model group were significantly increased compared to these in the normal group (P less than 0.01). The serum levels of ALT, AST and the contents of HA in anluohuaqianwan group were significantly lower than those in the model group (P less than 0.01). The liver MMP-2 in the model group was significantly increased compared to that in the normal group (P less than 0.05). The expression of MMP-2 in liver tissue of model group was lower than that in the anluohuaqianwan group (P less than 0.05).

CONCLUSIONAnluohuaqianwan can inhibit liver fibrosis in rats induced by DMN.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Dimethylnitrosamine ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Hyaluronic Acid ; blood ; Hydroxyproline ; metabolism ; Immunohistochemistry ; Liver ; drug effects ; metabolism ; pathology ; Liver Cirrhosis, Experimental ; chemically induced ; drug therapy ; metabolism ; pathology ; Liver Function Tests ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley

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

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

OBJECTIVETo observe the effect of electroacupuncture on recovery of urinary bladder function after radical hysterectomy.

METHODSOne hundred and ten cases were randomly divided into an electroacupuncture (EA) group and a control group, 55 cases in each group. In the control group, the urinary tube was placed and kept with routine method and the urinary bladder was rinsed, and from the eighth day the abdomen was radiated with TDP, 30 min each day, for 5 days. In the EA group, on the basis of treatment in the control group EA was given at Sanyinjiao (SP 6), Zusanli (ST 36), Waiguan (TE 5), Shuidao (ST 28), Guilai (ST 29), etc. from the eighth day to twelfth day after operation. The recovery time of urinary bladder function after radical hysterectomy, urine dynamic indexes and hospitalization days were compared between the two groups.

RESULTSThe cases of the bladder function recovery, retention of urine, urinary incontinence were 51(51/55), 4(4/55), 0 on the 14 th day after operation and 53(53/55), 2(2/55), 0 on the 28 th day in the EA group, and 27(27/55), 25(25/55), 3(3/55) on the 14 th day and 43(43/55), 11(11/55), 1(1/55) on the 28th day in the control group, respectively, with a very significant difference between the two groups (P < 0.01); the EA group in residual urine volume, bladder volume, mean urinary flowing rate was better than the control group on the 14 th day after operation (P < 0.01 or P < 0.05); the hospitalization days after operation was (21.1 +/- 3.3) days in the EA group and (25.5 +/- 3.5) days in the control group, the former being shorter than the later (P < 0.01).

CONCLUSIONEA can promote recovery of bladder function, shorten the keeping time of urinary tube after radical hysterectomy, which is benefit to decreasing incidence rate of urinary system infection and shortening hospitalization days.

Adult ; Aged ; Electroacupuncture ; Female ; Humans ; Hysterectomy ; adverse effects ; Medicine, Chinese Traditional ; Middle Aged ; Postoperative Complications ; physiopathology ; therapy ; Urinary Bladder ; physiopathology ; Urinary Bladder Diseases ; physiopathology ; therapy

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