AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.

As a member of G protein coupled-receptors superfamily, free fatty acid receptor 1 (FFAR1), is also known as GPR40, has been shown to regulate numerous pathophysiological processes in a variety of tissues and organs. The activated FFAR1 has a variety of biological functions. For instance, it can not only regulate metabolism of fatty acids and glucose, but also play an important role in immune inflammatory response, it may be a potential drug target for the treatment of various chronic inflammatory diseases. In this review, we focus on the recent researches of FFAR1's action in the regulation of pathophysiological processes, its molecular mechanism and new agonists development. At the same time, this review will take the discovery of series FFAR1 agonists as examples, and display the applied prospects of FFAR1.

OBJECTIVETo investigate the effect of advanced maternal age on birth defects and postnatal complications of neonates.

METHODSAmong the 1 109 neonates who were born at The First People's Hospital of Yunnan Province between January 2014 and December 2015, 536 neonates whose mothers were aged ≥35 years were enrolled as advanced age group and 573 neonates whose mothers were aged <35 years were enrolled as appropriate-age group. The incidences of the comorbidities in pregnancy, fetal intrauterine distress, neonatal birth defects, and postnatal complications were compared between the two groups. A univariate logistic regression analysis was performed to analyze the effect of advanced maternal age on neonatal comorbidities during perinatal period.

RESULTSCompared with the appropriate-age group, the advanced age group had significantly higher rate of caesarean section and incidence rates of multiple birth, gestational diabetes, pregnancy-induced hypertension, in vitro fertilization, and fetal intrauterine distress (P<0.01). The neonates in the advanced age group had a significantly higher incidence rate of cleft lip and palate and a significantly lower rate of skeletal dysplasia than in the appropriate-age group (P<0.05). Advanced maternal age was the risk factor for fetal intrauterine distress (OR=2.27, 95%CI: 1.33-3.88, P=0.003), neonatal resuscitation (OR=1.66, 95%CI: 1.19-2.31, P=0.003), and intracranial hemorrhage (OR=2.70, 95%CI: 1.21-6.04, P=0.02).

CONCLUSIONSThe women of maternal advanced age have higher incidence rates of pregnancy comorbidities than those of appropriate age, and the neonates born to the mothers of advanced maternal age have a higher incidence rate of cleft lip and palate. Advanced maternal age may increase the risks of fetal intrauterine distress, neonatal resuscitation, and intracranial hemorrhage.

Adult ; Cerebral Hemorrhage ; etiology ; Cesarean Section ; Congenital Abnormalities ; etiology ; Female ; Humans ; Infant, Newborn ; Infant, Newborn, Diseases ; etiology ; Logistic Models ; Maternal Age ; Middle Aged ; Pregnancy ; Pregnancy Complications ; etiology

OBJECTIVE: To determine whether SC-435, a new ileal apical sodium-codependent bile acid transporter (IBAT) inhibitor, can alter the gastrointestinal motility in guinea pigs. METHODS: Sixty guinea pigs received regular diet or IBAT inhibitor (SC-435) diet for 2, 4, and 8 weeks, respectively. At the end of the feeding period, the gallbladder motility was assessed and then four bipolar silver electrodes were implanted on the antrum, duodenum, jejunum, and ileum. Seven days later, migrating motor complex (MMC) was recorded and the total bile acid pool size was measured according to the isotope dilution principle in the meantime. RESULTS: After feeding SC435, the gallbladder motility was declined in the 4-week group and the 8-week group. The bile acid pool size decreased by 17.11% (P <0.05) in the 4-week group and 48.35% (P < 0.05) in the 8-week group. The places of origin of MMC were changed where antral origins (37%) and duodenal origins (46%) decreased whereas jejunal origins (17%) increased. The MMC cycle period was prolonged in the duodenum (1.16 times in the 4-week group, P < 0.05; 1.38 times in the 8-week group, P < 0.05) whereas MMC amplitude fell in the duodenum (10.58% in the 4-week group, P <0.05; 49.17% in the 8-week group, P <0.05). There were not significant differences in all parameters of MMC between the control group and the 2-week group in guinea pigs. CONCLUSION The IBAT inhibitor (SC-435) reduces the bile acid pool size and inhibits the MMC cycle activity. MMC is related to the enterohepatic circulation of bile acids, which is consistent with the changes of the bile acid pool size in guinea pigs.
Animals ; Bile Acids and Salts ; Cyclic N-Oxides ; pharmacology ; Female ; Gallbladder ; physiology ; Gastrointestinal Motility ; drug effects ; physiology ; Guinea Pigs ; Myoelectric Complex, Migrating ; drug effects ; Organic Anion Transporters, Sodium-Dependent ; antagonists & inhibitors ; Random Allocation ; Symporters ; antagonists & inhibitors ; Tropanes ; pharmacology

OBJECTIVETo observe the inhibitory effect of targeted ribonuclease delivered via adenovirus against HBV replication in vitro.

METHODSThe shuttle plasmids pDC316 were constructed on the basis of the previous plasmids pcDNA3.1(-)/TRL, pcDNA3.1(-)/TR, pcDNA3.1(-)/TRmut, pcDNA3.1(-)/HBVc, and pcDNA3.1(-)/hEDN by subcloning the target gene sequences of TRL, TR, HBVc, and hEDN, respectively. HEK 293 cells were cotransfected with the pDC316 plasmids respectively in the presence of the rescue plasmid pBHGlox(delta)E1,3Cre to yield the recombinant adenoviral vectors which comprised the above genes. After transfection of HepG2.2.15 cells with the vectors, RAd/TRL expression was detected by indirect immunofluorescence staining and RT-PCR. Radioimmunoassay was used to analyse anti-HBV activity of RAd/TRL.

RESULTSRecombinant RAd vectors were prepared successfully. Effective expression of RAd/TRL in HepG2.2.15 cells resulted in a significant decrease of HBsAg and HBeAg concentration in comparison with the controls.

CONCLUSIONAdenoviral vector-mediated targeted ribonuclease can effectively inhibit HBV replication.

Adenoviridae ; genetics ; Cell Line ; Cell Line, Tumor ; Fluorescent Antibody Technique, Indirect ; Gene Expression ; Genetic Vectors ; genetics ; Hepatitis B Core Antigens ; genetics ; metabolism ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Plasmids ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Ribonucleases ; genetics ; metabolism ; Transfection ; Virus Replication ; genetics

Objective To observe the preventive value of recombinant human granulocyte colony stimulating factor(rhG-CSF)in cancer patients after chemotherapy.Methods In the open study,enrolled 52 patients with previously untreated cancer and with normal bone marrow function were randomly divided into 2 matched groups,A and B group.Each patient received one cycle of chemotherapy.In the study cycle,the pa- tients received a single subcutaneous injection of rhG-CSF 150 ?g before 24 hours of chemotherapy and in control cycle the patients only received chemotherapy.Efficacy and safety parameters were monitored.Results The incidence rates of leukopenia in the 26 valuable study cycles and 26 valuable control cycles were 19.23 % and 53.85 %,There were significant lower than those of group B(P

Objective To probe into the level and influencing factors of community stroke patients′health behaviors so as to provide evidence for health guidance of community stroke. Methods The health promoting lifestyle profile ( HPLPⅡ) and self-designed general information questionnaire were conducted in 200 stroke patients in the community in 2013 July to 2014 March, and the results of surveys had been analyzed. Results Stroke patients with HPLP Ⅱ score was (121. 43 ± 13. 71) score, and 74% patients had unhealthy lifestyle. The influencing factors of health behavior were gender, education background, income, BMI, smoking, regular diet and regular check-up. Conclusions Stroke patients were in poor health behavior level, so the medical care personnel should focus on patients with smoking, diet, not regular recheck and other bad habits according to different gender, education and income to promote their lifestyle.

OBJECTIVETo establish the reverse transcription loop-mediated isothermal amplification (RT-LAMP) methods for on-site HIV-1 detection.

METHODSAs for the real-time fluorescent RT-LAMP, we firstly tested the specificity and sensitivity, then explored its quantitative determination, and finally applied the method to the detection of 35 HIV-1 positive samples. For colorimetric judgment, after choosing different ameliorates to modify Hydroxynaphthol blue (HNB), we tested their real effects on coloration, and then picked out the modified dyes with obvious color change to test the sensitivity and the detection of the 35 HIV-1-positive samples.

RESULTSThe real-time fluorescent RT-LAMP showed great specificity of HIV-1, and the sensitivity to detect HIV-1 RNA was between 10 and 100 copies per reaction. On testing 35 HIV-1-positive samples, the method could reach 100 percent detection rate. However, for the quantitative determination, the quantitative relation was not observed regarding the HIV-1 RNA of below 10(3) copies per reaction. Three modified HNB dyes with clear color variation between the reaction tubes of the negative and the positive were got in the study, and their sensitivities equaled to the level of agarose gel electrophoresis. Similarly, 100% (35/35) detection rate was reached when the colorimetric RT-LAMP with the modified dyes was applied to detect 35 HIV-1-positive samples.

CONCLUSIONThe established real-time fluorescence method and the modified color judgment of RT-LAMP could be helpful for truly achieving rapid, accurate, and sensitive on-site detection of HIV-1.

HIV-1 ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo determine the frequency of mce gene in Leptospira interrogans, and to investigate the gene transcription levels of L. interrogans before and after infecting cells.

METHODSThe segments of entire mce genes from 13 L.interrogans strains and 1 L.biflexa strain were amplified by PCR and then sequenced after T-A cloning. A prokaryotic expression system of mce gene was constructed; the expression and output of the target recombinant protein rMce were examined by SDS-PAGE and Western Blot assay. Rabbits were intradermally immunized with rMce to prepare the antiserum, the titer of antiserum was measured by immunodiffusion test. The transcription levels of mce gene in L.interrogans serogroup Icterohaemorrhagiae serovar lai strain 56601 before and after infecting J774A.1 cells were monitored by real-time fluorescence quantitative RT-PCR.

RESULTmce gene was carried in all tested L.interrogans strains, but not in L.biflexa serogroup Semaranga serovar patoc strain Patoc I. The similarities of nucleotide and putative amino acid sequences of the cloned mce genes to the reported sequences (GenBank accession No: NP712236) were 99.02%-100% and 97.91%-100%, respectively. The constructed prokaryotic expression system of mce gene expressed rMce and the output of rMce was about 5% of the total bacterial proteins. The antiserum against whole cell of L.interrogans strain 56601 efficiently recognized rMce. After infecting J774A.1 cells, transcription levels of the mce gene in L.interrogans strain 56601 were remarkably up-regulated.

CONCLUSIONThe constructed prokaryotic expression system of mce gene and the prepared antiserum against rMce provide useful tools for further study of the gene function.

Amino Acid Sequence ; Animals ; Bacterial Proteins ; genetics ; metabolism ; DNA, Bacterial ; analysis ; Escherichia coli ; genetics ; metabolism ; Genes, Bacterial ; Leptospira interrogans ; classification ; genetics ; Molecular Sequence Data ; Rabbits ; Recombinant Proteins ; genetics ; metabolism ; Sequence Analysis, Protein ; Serotyping

OBJECTIVETo explore the interaction between SerpinB5 and MAFbx in gastric cancer cell and to identify the interaction sites.

METHODSThe interaction between SerpinB5 and MAFbx was screened and validated by yeast two-hybrid screening and co-immunoprecipitation. The expression of MAFbx was analyzed after SerpinB5 expression being modified by RNA interference and pGBKT7-SerpinB5 transfection. The impact of SerpinB5 on the expression of MAFbx was studied in gastric cancer cell line SUN-16. A model of MAFbx was constructed by homology modeling. The related residues for interaction were analyzed by Autodock4.0.

RESULTSThe interaction between SerpinB5 and MAFbx was validated. The expression of MAFbx changed along with SerpinB5 expression. Amino acids including PRO261, ASN361, and LYS362 were key residue in the interaction of SerpinB5 and MAFbx.

CONCLUSIONSerpinB5 interacts with MAFbx in gastric cancer cell. Amino acids including PRO261, ASN361, and LYS362 are potential binding sites.

Cell Line, Tumor ; Humans ; Immunoprecipitation ; Muscle Proteins ; genetics ; metabolism ; RNA Interference ; SKP Cullin F-Box Protein Ligases ; genetics ; metabolism ; Serpins ; genetics ; metabolism ; Stomach Neoplasms ; genetics ; metabolism ; Two-Hybrid System Techniques