Objective To prepare quality control samples for St.Louis encephalitis virus(SLEV)molecular detection by constructing pseudovirus containing target sequences of SLEV.Methods According to the principles of armored RNA technique, the prM gene sequence of SLEV was cloned into the prokaryotic expression vector to generate recombinant plasmid pSE380-MS2-SLEV.Then, recombinant E.coli transformed with the corresponding plasmid was induced with IPTG to produce recombinant pseudovirus particles.The particles were purified by chloroform and further characterized by double enzyme digestion and transmission electron microscopy.The temperature sensitivity experiments and quantitative RT-PCR were performed to validate the potential of these pseudovirus particles as quality control samples.Results PCR amplification and sequencing analysis confirmed that the prM gene sequence of SLEV was cloned into vector pSE380-MS2.Transmission electron microscopy showed that homogenous spherical particles with a diameter of about 25 nm were produced upon IPTG induction.The SLEV genomic RNA within the pseudovirus particles was resistant to DNaseⅠand RNase A digestion, and remained stable for 20 days at 37℃.These samples were validated with quantitative RT-PCR for SLEV.Conclusion The RNase-resistant and stable pseudovirus particles containing prM fragment of SLEV are constructed successfully, which can be used as positive quality control samples for RNA extraction and molecular detection.

Objective To systematically assess the relation between angiotensin-I converting enzyme(ACE) gene insertion/deletion (I/D) variation and type 2 diabetic nephropathy (T2DN) onset risk among Chinese population.Methods The related literatures were retrieved from the China National Knowledge Infrastructure (CNKI) and Wanfang Data until June 1st,2016.The RevMan 5.0 was used to conduct the statistical analysis.The merge OR value and corresponding 95% confidence interval(95%CI) were used to assess ACE gene I/D polymorphism and T2DN onset risk.Results Totally 29 papers with 4 357 subjects were included according to the inclusion and exclusion standard,including 2 208 cases of DN and 2 149 cases of T2DM without DN.Meta analysis showed that compared with ACE gene I/D polymorphism I allele,D allele could significantly increase the risk of T2DM patients suffering from DN,the OR value and corresponding 95%CI were 1.44(1.25,1.66);the gene analysis showed that ACE gene I/D polymorphism loci were significantly correlated with DN onset risk in the Asian population.The corresponding relative onset risk OR and 95%CI were 1.42(1.15,1.76) and 1.75(1.46,2.10) in the dominant and recessive genetic model.The Begg′s test showed that the included data had no obvious publication bias existence.Conclusion ACE gene I/D polymorphism is closely correlated with the onset risk of T2DN,and D allele might be a risk genetic factor for DN occurrence in the patients with T2DM.

Objective To study expressions of heme oxygenase-1 mRNA and protein in rat hippocampus after hypoxic-ischemia brain damage(HIBD) as well as the relationship with apoptosis in brain.Methods Seven-day-old SD rats were randomly divided into hypoxic-ischemia brain damage group and sham control group.Expressions of HO-1 protein and mRNA as welll as the relationship with apoptosis after HIBD in neonatal rat were determined by immunohisochemistry and in situs hybridizaion as well as terminal deoxynucleotidy transferase mediated UTP-biotin nick end labeling(TUNEL).Results 1.In the right hippocampus,expression of HO-1 gene increased sharply at 4 h (P

Objective To study changes of phosphorylated cyclic adenosine monophosphate(c-AMP) response element binding protein in hippocampus(CA1) of neonatal rats after cerebral hypoxic-ischemia(HI)and effects of gangliosides (GM1)on the p-CREB.Methods An animal model of neonatal hypoxic-ischemia brain damage was established. Changes of p-CREB in hippocampal CA1 was detected with immunohistochemical methods.Results The p-CREB levels in the CA1 of HI and GM1 groups increased transiently and then decreased quickly, but there was no significant difference between HI and GM1 group.Conclusion The p-CREB levels in the CA1 of HI group increase transiently and then decrease quickly after HI ;GM1 has little effect on p-CREB in the CA1 after HI.

Most cases of prostate cancer become hormone refractory after 12 to 18 months of androgen deprivation therapy. The etiology of the disease is thought to be multifactorial, associated with genetic, dietary, and environmental factors. The article reviews the current situation of researches at home and abroad on the molecule mechanism of hormone refractory. It expounds the influence of the androgen receptor and its genetic mutation, apoptosis and the gene changes of p53, p21, EphB2 on prostate cancer. It is hoped to be of some directive value for the studies of prostate cancer.
Androgens ; pharmacology ; Animals ; Apoptosis ; Genes, p53 ; genetics ; Humans ; Male ; Mutation ; Oncogene Protein p21(ras) ; genetics ; Prostatic Neoplasms ; drug therapy ; genetics ; pathology ; Rats ; Receptor, EphB2 ; genetics ; Receptors, Androgen ; genetics

OBJECTIVETo investigate the significance of human papillomavirus test in triage of patients with atypical squamous cell of undetermined significance (ASCUS) diagnosed by cervical cytology.

METHODSHuman papillomavirus test,colposcope and cervical biopsy were performed in 184 patients with a referral diagnosis of ASCUS by cervical cytology.

RESULTSConfirmed by pathological diagnosis of cervical biopsy, 112 cases were chronic inflammation (60.87%), 33 CIN I (17.93%), 17 CIN II (9.24%), 8 CIN III (4.35%), 4 cervical squamous carcinoma (2.17%) and 10 condyloma (5.43%). Of the 184 women with cytological ASCUS, 124 (67.39%) cases were positive in high-risk HPV test among which 66 cases were histologically confirmed as chronic inflammation (53.23%), 22 as CIN I (17.74%), 16 as CIN II (12.90%), 8 as CIN III (6.45%), 4 as cervical squamous carcinoma (3.23%) and 8 as condyloma (6.45%). The positive rate of HPV in groups of ASCUS were higher than those with negative HPV (P < 0.003).

CONCLUSIONWomen with ASCUS should be tested for HPV. Cervical biopsy under colposcopy is recommended for women with HR-HPV infection.

Adult ; Aged ; Alphapapillomavirus ; isolation & purification ; Carcinoma, Squamous Cell ; diagnosis ; pathology ; virology ; Female ; Humans ; Middle Aged ; Papillomavirus Infections ; diagnosis ; pathology ; virology ; Uterine Cervical Neoplasms ; diagnosis ; pathology ; virology ; Vaginal Smears ; Young Adult

OBJECTIVETo observe the central nervous system symptoms and alterations in the blood indicators in rats within a short term after benzene poisoning.

METHODTwenty-four female SD rats were randomized into 4 equal groups to receive intraperitoneal injection of low-, medium- or high-dose benzene (39.05, 78.11, and 234.33 mg/kg, respectively) or peanut oil. Blood samples were taken from the rats via the femoral artery 24 h after the injections for routine blood test and liver and kidney function test.

RESULTSIntraperitoneal injection of benzene at a high dose, but not at a low or medium dose, caused obvious symptoms in the central nervous system. Benzene either at a low or medium dose did not produce obvious changes in routine blood test or liver and kidney function test as compared with the control group, but a high dose resulted in significant changes in WBC, PLT, ALT and AST (P<0.05). Abnormalities in the renal function were found in none of the groups (P>0.05).

CONCLUSIONExposure to high-dose benzene can result in abnormalities in the central nervous system, routine blood indicators and liver function, but does not obviously affect the kidney function in rats.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Benzene ; toxicity ; Blood Cell Count ; Central Nervous System Diseases ; chemically induced ; Female ; Kidney ; drug effects ; Liver ; drug effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the clinic value of combination of high-risk human papillomavirus test and cervical cytology test in diagnosis of cervical lesions.

METHODSPatients underwent physical examination at our department were checked by high-risk human papillomavirus test, cervical cytology test and colposcope from October 2004 to December 2006. Abnormal patients with cervical abnormalities were asked for pathological test.The diagnostic value of cervical lesions among these different methods were compared.

RESULTSBased on the criteria of histopathology, the sensitivity, specificity, positive-predictive value and negative-predictive value of high-risk human papillomavirus test for detecting all cases of CIN II and CIN III were 94.83%, 31.06%, 55.22% and 87.02% respectively, and those of the cervical cytology were 92.10%, 31.06%, 54.50% and 81.43% respectively.Those values changed to 99.65%, 18.55%, 61.46% and 97.62% respectively if two methods were combined.

CONCLUSIONSHuman papillomavirus test and cervical cytology test combined with pathological test can improve the detective rate of cervical lesions and facilitate the treatment.

Adolescent ; Adult ; Aged ; Cervical Intraepithelial Neoplasia ; diagnosis ; virology ; Cytodiagnosis ; Female ; Humans ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; diagnosis ; Pregnancy ; Reagent Kits, Diagnostic ; Uterine Cervical Diseases ; diagnosis ; Uterine Cervical Neoplasms ; diagnosis ; Vaginal Smears ; Young Adult

Objective: To investigate the abnormalities of the blood system in landfill workers. Methods: A cohort study was conducted for 224 landfill workers who were followed up for 6 consecutive years with abnormal routine blood test results and a low platelet count as the outcome events. The life-table method was used to analyze the incidence rates of these two outcome events, and the incidence rates were compared between first-and second-line workers. Results: A total of 71 workers had abnormal routine blood test results, among whom 29 had abnormal leukocyte count, 14 had abnormal erythrocyte count, 40 had abnormal platelet count, 17 had abnormal hemoglobin, and 29 had a reduction in platelet count. For these landfill workers, the 6-year abnormal rate of routine blood test results was 43.2%, and the incidence rate of low platelet count within 6 years was 13.5%. The first-line workers had a significantly lower abnormal rate of routine blood test results than the second-line workers (P=0.020) , and the relative risk of abnormal routine blood test results in the first-and second-line workers was 0.592. There was no significant difference in the incidence rate of low platelet count between the two groups of workers (P≥0.05) . Conclusion The landfill has an adverse effect on the blood system of landfill workers, and the second-line workers have greater impairment than the first-line workers.

OBJECTIVETo screen for genetic mutations in families featuring non-syndromic hearing loss.

METHODSSixteen families with non-syndromic hearing loss were interviewed to identify medical histories by a questionnaire. Audiological and neurological examinations were conducted for all families. Coding regions of GJB2 and 12S rRNA genes were amplified and sequenced.

RESULTSOf the 17 patients with sensorineural hearing loss, 3 were homozygous mutation for GJB2 235 delC, 1 was 235 delC heterozygous mutation, 1 was 235 delC+299_300 delAT compound heterozygous mutation, and 6 were 79G>A+341G>A heterozygosis in cis mutation. No 1555A>G mutation of mitochondrial DNA (mtDNA) was found in the 16 families.

CONCLUSIONThe incidence of mtDNA 12S rRNA 1555A>G mutation in Jiangsu province may be lower than the average across China. Mutations of GJB2 genes may account for as much as 64.7% of non-syndromic hearing loss in this study. Screening for such mutations and genetic counseling may play an important role in the prevention of hereditary hearing loss.

Adolescent ; Base Sequence ; Child ; Child, Preschool ; Connexin 26 ; Connexins ; genetics ; DNA, Mitochondrial ; genetics ; Female ; Genetic Predisposition to Disease ; Hearing Loss ; genetics ; Heterozygote ; Homozygote ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; RNA, Ribosomal ; genetics