To provide the basis for preparation of diagnostic kits and vaccines in Toxoplasma gondii infection, the gene coding for the qualified recombinant p30 protein (SAG1) of this parasite was amplified by PCR, and the amplified gene was cloned into prokaryotic expression vector pET-30a(+) to construct the recombinant plasmid, and then transformed to E.coli DH5α. The positive recombinant plasmid was screened by PCR and double enzymes digestion, and the nucleotide sequence of p30 gene was determined by automated DNA sequencing. Meanwhile, the identified recombinant plasmid was transformed to E.coli BL21(DE3) with the expression of p30 on bacteria induced by IPTG and the expressed protein was identified by SDS-PAGE. The protein obtained was then further purified and refolded, and its biological activity was checked by Western blotting. It was shown that the size of the amplified gene was 750 bp with molecular weight of 30 ku, and this protein could specifically react with monoclonal antibody against p30 protein.

OBJECTIVETo explore the risk factors and clinical characteristics of non-erosive reflux disease (NERD) based on a prospective single disease database of functional gastrointestinal disease.

METHODSUsing a customized case report form, we collected the personal and clinical data of all study participants in an online database for further analysis. High-resolution manometry and multichannel intraluminal impedance-pH monitoring were performed in some cases.

RESULTSA total of 504 NERD cases and 152 control cases were included in our database. The NERD patients consisted of 266 (52.8%) female patients and 238 (47.2%) male patients; 32.7% of the patients were from rural areas and 67.3% from urban areas; 23.1% of the patients worked in the line of business, 19.6% were civil servants, 19.2% were unemployed, and 17.1% were workers; the mean disease duration of the patients was 27.88∓16.33 month. Our analysis showed that adverse events in life (P=0.045, OR=1.954), frequent drinking (P=0.040, OR=3.957), snoring (P=0.002, OR=2.334), late meals (P=0.002, OR=2.752), and anxiety or depression (P=0.003, OR=2.723) were all independent risk factors for NERD. Of these patients, 60.81% had varying degrees of ineffective contraction of the esophageal body. The total liquid reflux events differed significantly between NERD patients with hiatal hernia and those without (P<0.05).

CONCLUSIONUnhealthy eating habits and lifestyle, history of adverse events, anxiety and depression, snoring, poor esophageal motor function and hiatal hernia are significant factors contributing to NERD, which is related with occupation and living areas and occurs most commonly at 30-50 years of age.

Databases, Factual ; Female ; Gastroesophageal Reflux ; diagnosis ; physiopathology ; Humans ; Male ; Manometry ; Prospective Studies ; Risk Factors

Objective To study the protective effect of ROP2 nuclei acid vaccine in mice.Methods Forty-two BALB/c mice were divided into three groups.Each mouse in experiment group was injected with 50 ?g recombinant plasmid pc-DNA3-ROP2 through musculus quadriceps fexoris.In control groups,each mouse was injected with 50 ?g blank plasmid pc-DNA3 and with 50 ?l PBS respectively.All mice were immunized for three times with an interval of three weeks.The volume was doubled for the final injection in the two plasmid groups.Blood,spleens and lymph nodes of 4 mice in each group were taken for the detection of CD4+,CD8+ T cells and cytokines 2 weeks after the final immunization.The rest mice in 3 groups were challenged with 500 tachyzoites of Toxoplasm gondii RH strain for further observation.Results The vaccine induced strong cellular and humoral immune response.The titer of antibody in serum was high after inoculation and recognized ROP2 protein antigen expressed in vitro.The lymphocyte phenotype was analyzed.CD4+ T cells proliferated sharply(69.5?3.4)%,and the ratio of CD4+/CD8+ increased considerably by(4.69?1.32)%(P

Objective To explore the risk factors of cardiovascular disease in adolescents with a family history of hypertension.Methods A cross-sectional study was conducted in 3874 adolescents ages 13-18 years,with normal BP in 3724 people.Based on family history of hypertension (FH),the cohort of adolescents were dichoto- mized as postive family history (FH~+,n=1145) and negative (FH~-,n=2579).Height,weight,waist circum- ference,hip circumference,blood pressure and fasting plasma glucose(FPG),total cholesterol(TC),triglyceride (TG) and high-density lipoprotein cholesterol(HDL-C) were determined.Results FH~+ adolescents had signifi- cantly higher levels of body mass index(BMI),waist circumference,WHR,FPG,TC and LDL-C(P

OBJECTIVETo observe the effects of different ways in repairing scrotum of pigs with full-thickness burn on spermatogenesis of testis.

METHODSTwenty male Guizhou miniature pigs were divided into normal control (NC), natural-healing (NH), flap-repairing (FR), and skin-grafting (SG) groups according to the random number table, with 5 pigs in each group. Pigs in NC group were not subjected to any injury. Scrotum of pigs in the latter three groups were inflicted with full-thickness burn. Wounds in NH group healed naturally. Wounds in FR group were repaired with inguinal region flap, and those in SG group with full-thickness skin from lower abdomen. Appearance of scrotum in the latter three groups was observed right after injury, and three months post injury or surgery (PIM or PSM). Specimens of testes of pigs in the latter three groups were obtained in PIM or PSM 3 to detect apoptosis of spermatogenic cells with TUNEL, and bcl-2 protein expression with immunohistochemistry. The same indexes were observed and determined in pigs of NC group. Data were processed with one-way analysis of variance and LSD test.

RESULTS(1) Scrotum of pigs in NC group had skin folds with contraction function. Scrotum of pigs became hard with a leathery appearance right after burn in the other three groups. In PIM or PSM 3, wounds of pigs in NH group healed with scar, and the testes were squeezed into inguinal region. Scrotal skin of pigs in FR group was thick with testes in the scrotum, and that of pigs in SG group was thin with testes in the scrotum. (2) Spermatogenic cells in each level in NC group were arranged regularly, with few apoptotic spermatocytes and spermatoblasts. In NH, FR, and SG groups, seminiferous epithelium was thinner with most of the spermatogenic cells showing apoptosis, and they were mainly spermatogonia and spermatocytes. Apoptotic index of spermatogenic cells in NH, FR, SG, and NC groups was respectively (46.3 ± 3.3)%, (40.9 ± 3.5)%, (20.6 ± 2.3)%, (7.5 ± 1.9)%, and the difference among them was statistically significant (F = 405.65, P < 0.01). There were significant statistical differences among the former three groups (with P values below 0.01). (3) bcl-2 protein expression in NH, FR, SG, and NC groups was respectively (52 ± 5)%, (53 ± 4)%, (64 ± 5)%, (75 ± 5)%, and the difference among them was statistically significant (F = 56.63, P < 0.01). There was no significant statistical difference in bcl-2 expression between NH group and FR group (P = 0.66), and it was lower in both groups as compared with SG group (with P values below 0.01).

CONCLUSIONSEither scar healing, flap transplantation, or SG in repairing scrotum with full-thickness burn in pigs inhibits spermatogenesis, but repair with SG produces less deleterious effect on the testis.

Animals ; Apoptosis ; Burns ; metabolism ; pathology ; surgery ; Male ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Reconstructive Surgical Procedures ; methods ; Scrotum ; injuries ; metabolism ; Skin Transplantation ; Spermatogenesis ; Swine

OBJECTIVETo understand the influencing factors on the death of infants born to HIV infected mothers in areas with high prevalence of HIV/AIDS in China.

METHODSBased on the follow-up cohort study targeting at HIV/AIDS infected pregnant women and their babies initiated in 2004, a survey on the death status and influencing factors on the infants born to HIV/AIDS infected mothers enrolled in this cohort from Jan.2004 to Nov.2007 was carried out during Aug.to Nov.2008 in seven counties of four provinces in China. A total of 498 pairs of HIV-infected mothers and their infants were enrolled and their related information was collected. Single factor and multiple factors Cox model methods were adopted for data analysis.

RESULTSThe total observed person-years of 498 infants was 406.22, among which, 45 infants died, and the mortality density was 110.78 per 1000 child-year. A single factor Cox model showed, the pregnancy in pre-period of HIV/AIDS and HIV/AIDS period (RR = 1.971, 95%CI: 1.143 - 3.396), living status of the pregnancy (RR = 3.062, 95%CI: 1.097 - 8.550), multipara women (RR = 0.517, 95%CI: 0.278 - 0.961), natural childbirth (RR = 0.561, 95%CI: 0.345 - 0.910), premature labor (RR = 5.302, 95%CI: 2.944 - 9.547), low birth weight (RR = 4.920, 95%CI: 2.691 - 8.994), mother-child pairs taking antiretroviral drugs (RR = 0.227, 95%CI: 0.121 - 0.428) and infants infected HIV (RR = 5.870, 95%CI: 3.232 - 10.660) could affect the infants death. The death of HIV-exposed infants was influenced by various factors. The death risk of infants born to HIV infected mothers who were in the danger of pre-period of HIV/AIDS and HIV/AIDS period was greater than the infants delivered by HIV infected mothers who were in preclinical period of HIV/AIDS (RR = 6.99, 95%CI: 1.92 - 25.64). The death risks were greater in the group that the women whose CD4(+)TLC count number lower than 200 cells/microl (RR = 2.05, 95%CI: 1.01 - 4.15). The infants whose mothers had no ARV treatment had higher possibility to die than the others (RR = 6.17, 95%CI: 1.62 - 23.26). The death risk of premature delivered infants was 2.87 times of mature delivered infants (95%CI: 1.12 - 7.35). The death risk of HIV/AIDS infected infants was 9.87 times of the HIV/AIDS uninfected infants (95%CI: 3.81 - 25.62).

CONCLUSIONSome measurements including improving HIV-infected pregnant women's immunity, reducing mother to child transmission of HIV and premature birth, low birth weight are beneficial to reducing infant mortality.

Acquired Immunodeficiency Syndrome ; epidemiology ; mortality ; transmission ; Cause of Death ; China ; Female ; Follow-Up Studies ; Humans ; Infant ; Infant Mortality ; Infant, Newborn ; Mothers ; Pregnancy ; Pregnancy Complications, Infectious ; epidemiology ; Proportional Hazards Models

OBJECTIVETo investigate the oxidative damage to lung tissue and peripherial blood in PM2.5-treated rats.

METHODSPM2.5 samples were collected using an auto-sampling instrument in summer and winter. Treated samples were endotracheally instilled into rats. Activity of reduced glutathione peroxidase (GSH-Px) and concentration of malondialdehyde (MDA) were used as oxidative damage biomarkers of lung tissue and peripheral blood detected with the biochemical method. DNA migration length (microm) and rate of tail were used as DNA damage biomarkers of lung tissue and peripheral blood detected with the biochemical method.

RESULTSThe activity of GSH-Px and the concentration of MDA in lung tissue significantly decreased after exposure to PM2.5 for 7-14 days. In peripheral blood, the concentration of MDA decreased, but the activity of GSH-Px increased 7 and 14 days after experiments. The two indicators had a dose-effect relation and similar changing tendency in lung tissue and peripheral blood. The DNA migration length (microm) and rate of tail in lung tissue and peripheral blood significantly increased 7 and 14 days after exposure to PM2.5. The two indicators had a dose-effect relation and similar changing tendency in lung tissue and peripheral blood.

CONCLUSIONPM2.5 has a definite oxidative effect on lung tissue and peripheral blood. The activity of GSH-Px and the concentration of MDA are valuable biomarkers of oxidative lung tissue damage induced by PM2.5. The DNA migration length (microm) and rate of tail are simple and valuable biomarkers of PM2.5-induced DNA damage in lung tissues and peripheral blood. The degree of DNA damage in peripheral blood can predict the degree of DNA damage in lung tissue.

Animals ; DNA Damage ; drug effects ; Drug Administration Routes ; Drug Administration Schedule ; Lung ; drug effects ; pathology ; Lung Diseases ; blood ; chemically induced ; pathology ; Male ; Oxidative Stress ; Particle Size ; Particulate Matter ; administration & dosage ; toxicity ; Rats ; Rats, Wistar ; Seasons

BACKGROUNDLiver injury is one of the most important adverse effects of antiretroviral therapy, leading to therapy changing or discontinuation. Data on liver injury in human immunodeficiency virus-1-infected patients receiving antiretroviral therapy are limited in China. The purpose of this study was to investigate the features of liver injury in human immunodeficiency virus type 1-infected patients receiving non-nucleosides reverse transcriptase inhibitors-based antiretroviral therapy in China.

METHODSSeventy-five patients on antiretroviral therapy containing non-nucleosides reverse transcriptase inhibitors were retrospectively studied. The patients were divided into 2 groups: group 1 (with liver injury, n = 45) and group 2 (without liver injury, n = 30). The features of liver injury were analyzed. The sex, age, baseline CD4 counts, hepatitis B virus (HBV) and/or hepatitis C virus (HCV) co-infection, hepatotoxic drug use and nevirapine or efavirenz use were compared between two groups.

RESULTSForty-five patients (60.0%), 31 (68.9%) males and 14 (31.1%) females, aged 12 to 52 years (averaged (39 ± 9) years), experienced at least one episode of liver injury. Forty (53.3%) patients were co-infected with HBV and/or HCV, 42 (56%) patients had concomitant use of antituberculosis drugs or cotrimoxazole, 46 (61.3%) and 29 (38.7%) patients received regimen containing nevirapine and efavirenz, respectively. Grade 1 liver injuries were observed in 26 (57.8%) patients, grade 2 in 16 (35.6%), grade 3 in 2 (4.0%) and grade 4 in 1 (2.2%). Three (6.7%) patients discontinued highly active antiretroviral therapy (HAART) due to liver injury. In group 1, there were 29 (64.4%) patients co-infected with HBV and/or HCV, 32 (71.1%) patients received regimen containing nevirapine, and 30 (66.7%) patients had concomitant use of anti-tuberculosis drugs or cotrimoxazole, respectively, significantly higher than those in group 2 (11 (36.7%), 14 (46.7%) and 12 (40%), respectively; P = 0.018, 0.033, 0.023, respectively). The sex, age, baseline CD4 counts and disease stage were not factors associated with liver injury.

CONCLUSIONSLiver injury associated with HAART containing non-nucleosides reverse transcriptase inhibitors was mild to moderate and those who were co-infected with HBV and/or HCV, had concomitant use of antituberculosis drugs or cotrimoxazole and received a regimen containing nevirapine were prone to liver injury while receiving HAART.

Acquired Immunodeficiency Syndrome ; drug therapy ; Adolescent ; Adult ; Antiretroviral Therapy, Highly Active ; adverse effects ; Chemical and Drug Induced Liver Injury ; etiology ; Child ; Female ; HIV-1 ; Humans ; Male ; Middle Aged ; Nevirapine ; adverse effects ; Retrospective Studies ; Reverse Transcriptase Inhibitors ; adverse effects

HPV16 L1 gene was amplified from HPV16 positive vaginal secretion sample by PCR, and inserted into pTO-T7 to obtain the recombinant expression vector pTO-T7-HPV16-L1. Then, the pTO-T7-HPV16-L1 was transformed into E. coil strain ER2566 and the recombinant protein HPV16 L1 was expressed in soluble form. After purification by ammonium sulfate precipitation, ion-exchange chromatography, and hydrophobic interaction chromatography, the recombinant protein HPV16 L1 had a purity of more than 98%. By removing DTT, purified HPV16 L1 proteins self-assembled in vitro into VLPs with the diameter of 50 nm. The vaccination experiments on experimental animals showed the VLPs could elicit high titer of neutralizing antibodies against HPV 16. HPV16 VLPs with high immunogenicity and high purity can be produced easily and effectively from an E. coli expression system in the study, and thus can be used in structure investigation and HPV16 vaccine development.
Animals ; Antibodies, Viral ; immunology ; ultrastructure ; Capsid Proteins ; genetics ; immunology ; isolation & purification ; Goats ; Human papillomavirus 16 ; genetics ; immunology ; ultrastructure ; Humans ; Male ; Oncogene Proteins, Viral ; genetics ; immunology ; isolation & purification ; Papillomavirus Infections ; immunology ; virology ; Rabbits ; Recombinant Proteins ; genetics ; immunology ; Vaccination ; Virion ; genetics ; immunology

OBJECTIVETo investigate and analyze the therapeutic level and the existing problems on acute myocardial infarction (AMI) management in Beijing.

METHODSWe collected clinic data of 1242 AMI patients from 12 hospitals in Beijing, from January 2000 to March 2001, using a uniformed questionnaire, and evaluated the status of the diagnosis and treatment of AMI according to the Chinese guidelines issued on Decmeber 2001. Corresponding factors which influencing the mortality were also analyzed by one-way factor and multiple factors analysis methodologies.

RESULTSThe mean age of the 1242 AMI patients was 63.0 years old and about one third of them were under 55 years old. In hospitals, the total mortality was 9.10%. 37.9% of the patients had received therapy of the intravenous thrombolysis and emergency PCI with a total rate of reperfusion therapy as 56.0%. The in-hospital rates of drug use were as follows: Nitrates 90.0%, Aspirin 87.8%, heparin 88.7%, beta-blockers 73.4%, angiotensin converting enzyme inhibitors(ACEI) 77.6%, lipid regulating agents 43.6%. The rate of intravenous therapy of TCM by promoting the blood circulation and supplementing the vital energy was 30.5%. Results from multiple factors analysis showed that the compositive factors which could lower the mortality were reperfusion therapy,lipid regulating agents, intravenous therapy of TCM, beta-blockers, ACEI, lower molecule heparin and digitalis.

CONCLUSIONData from this study showed that there still existed a gap between clinical management on AMI and the guideline in Beijing. To set up a straightway passage of reperfusion therapy, to become more standardized to follow the guideline in undertaking the medical treatment practice, and to go deep into discuss the status of TCM on AMI management seemed the important tasks we are facing.

China ; Cities ; Female ; Guideline Adherence ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; therapy ; Quality of Health Care