A large body of evidence demonstrates that dendritic cells (DC) play a pivotal role in the control of immunity by priming and tolerizing T cells. In multiple sclerosis (MS), autoreactive T cells are proposed to play a pathogenic role by secreting pro-inflammatory cytokines, but comparison studies on the effects of immature and mature dendritic cells on the cytokines profile of antigen-specific T cell lines are lacking. To evaluate the actions of dendritic cell maturation on T cell polarization, the effects of immature and mature dendritic cells derived from MS patients on in vitro proliferative responses, and cytokine production by glatiramer acetate (GA)- specific T cell lines (TCL) derived from MS patients were analyzed. The results demonstrated that it is easy to derive GA-specific TCL from MS patients with high specificity; lipopolysaccharide can efficiently induce DC maturation within 24 hours at a concentration of 5 micro g/ml; mature DC showed higher co-stimulatory capacity of GA-specific TCLs than immature DC. GA-specific TCLs produce dominantly IL-2, IL-4, IFN-gamma and IL-10, but low levels of IL-6. In contrast to immature DC, mature DC enhanced capacity to induce IL-6 and IL-10 secretion, but down-regulate IL-2, IL-4 and IFN-gamma production by GA- specific TCLs. It is concluded that DC maturation status modulating proliferation of TCL and production of cytokines may represent another focus for the study on both immuno-pathogenesis and immunotherapeutic interventions in MS.
Adult ; Cell Line ; Cytokines ; biosynthesis ; Dendritic Cells ; physiology ; Female ; Glatiramer Acetate ; Humans ; Lymphocyte Activation ; Male ; Middle Aged ; Multiple Sclerosis ; immunology ; Peptides ; immunology ; T-Lymphocytes ; immunology

To determine whether gamma irradiation influences phenotype and function of human dendritic cells (DC) in vitro, dendritic cells were induced from the peripheral blood mononuclear cells of multiple sclerosis patients with RPMI 1640 medium containing recombinant human GM-CSF (rhGM-CSF, 800 U/ml) and recombinant human IL-4 (rhIL-4, 500 U/ml). Phenotypic changes were monitored by light microscopy. Lipopolysaccharide at a concentration of 5 micro g/ml was added into the cultures after 6 days of growth for DC complete maturation, and the cells were cultured for another 24 hours. The harvested DC on day 7 were divided equally into several parts. One part was used as non-irradiated DC (naive DC) while the other parts were irradiated by gamma ray at a dose of 25 Gy and 30 Gy respectively. Cell surface molecules were analyzed by flow cytometry. The capability of DC to stimulated autologous T cell proliferation were determined. The results showed that gamma irradiation reduced expression of CD86, CD80 and HLA-DR molecules on dendritic cells, especially CD86 molecules. Dendritic cells effectively stimulated autologous T cells proliferation while irradiated DC in all groups showed profound decrease of capability to promote T cells proliferation. It is concluded that gamma irradiation of dendritic cells not only influenced phenotype of DC but also altered their function as stimulator cells in mixed lymphocyte reaction.
Antigens, CD ; analysis ; B7-1 Antigen ; analysis ; B7-2 Antigen ; Cell Division ; immunology ; Dendritic Cells ; drug effects ; immunology ; radiation effects ; Flow Cytometry ; Gamma Rays ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; HLA-DR Antigens ; analysis ; Humans ; Immunophenotyping ; Interleukin-4 ; pharmacology ; Membrane Glycoproteins ; analysis ; Multiple Sclerosis ; blood ; Recombinant Proteins ; pharmacology ; T-Lymphocytes ; cytology ; immunology

AIMTo investigate the toxic response in auditory cortex of guinea pigs caused by cis-platinum (DDP), and the protective role of melatonin in this effect.

METHODSCis-platinum and melatonin were injected peritoneally. LDH, MDA, NO in the auditory cortex were detected by spectrophotometeR.

RESULTSThe body weight of the guinea pigs was diminished by peritoneal injection of Cis-platinum for 7 days (P < 0.01). Peritoneal injection of Cis-platinum induced the increased leakage of LDH (P < 0.05 vs injection of normal saline). This effect was reduced by injection of MT (P < 0.05). The content of MDA in the auditory cortex was also increased because of injection of Cis-platinumv for 7 days (P < 0.01) and MT reduced this effect (P < 0.05). The change of NO in the auditory cortex was not statistically significant after injection of Cis-platinum or Cis-platinum with MT.

CONCLUSIONPeritoneal injection of Cis-platinum could destroy neurons in the auditory cortex. This effect could be reduced by melatonin by an anti-free radials mechanism.

Animals ; Auditory Cortex ; drug effects ; metabolism ; pathology ; Cisplatin ; antagonists & inhibitors ; toxicity ; Female ; Free Radical Scavengers ; pharmacology ; Guinea Pigs ; Male ; Malondialdehyde ; metabolism ; Melatonin ; pharmacology ; Neurons ; drug effects ; pathology ; Random Allocation

OBJECTIVETo clarify the impact of increased heme oxygenase-1 (HO-1) expression on cardiac function of diabetic rats with myocardial infarction and its mechanism.

METHODSSixty adult male Wistar rats were randomly divided into five groups (n = 12): sham operation group (sham), diabetes + sham operation group (DM + sham), diabetes + MI group (DM + MI) , diabetes + myocardial infarction + cobalt original porphyrin (CoPP) group (DM + MI + CoPP), diabetes + myocardial infarction + CoPP+ tin porphyrin (SnMP) group (DM + MI + CoPP + SnMP). CoPP 4.5 mg/kg or SnMP 15 mg/kg were administered at the day next to MI operation, for six weeks, once a week. At the 28th week post operation, the echocardiography, left heart via the carotid artery indoor intubation were used to observe the long-term influence of HO-1 inducer (cobalt protoporphyrin, CoPP) and activity of HO inhibitor (tin porphyrin, SnMP) on the indices of left ventricular remodeling and cardiac function after the intervention. Blood glucose (GLU), total cholesterol (TC), C-reactive protein (CRP), serum creatinine (Cr), aminotransferase (ALT) and other indicators were measured. ELISA was used to test interleukin-6 (IL-6), tumor necrosis factor (TNF), nitric oxide (NO), prostacyclin (PGI2), adiponectin, and ultra sensitive CRP (HsCRP) level.

RESULTSHO-1 inducer, CoPP, could ameliorate ± dp/dtmax, left ventricular ejection fraction and left ventricular shortening fraction in diabetic myocardial infarction rats. It could also decrease left ventricular end-diastolic diameter. The serum bilirubin, NO and PGI2 levels, myocardial phosphorylated endothelial nitric oxide synthasee(peNOS), phosphorylated activated protein kinase (pAkt), phosphorylated adenosine monophosphate-activated protein kinase (pAMPK) expression were also significantly elevated, and the serum hs-CRP and TNF levels were significantly inhibited. Compared to inducer group, cardiac function were worse in the inhibitor group.

CONCLUSIONUpregulated HO-1 level can improve the endothelial function, inhibite of the inflammatory response and enhance the antioxidant substances in serum bilirubin via peNOS-pAMPK pathway, which effectively inhibit ventricular remodeling and improve the long-term cardiac function after infarction in diabetic rats.

Animals ; Antioxidants ; metabolism ; Bilirubin ; blood ; Diabetes Mellitus, Experimental ; physiopathology ; Heme Oxygenase (Decyclizing) ; metabolism ; Male ; Myocardial Infarction ; enzymology ; Myocardium ; enzymology ; Rats ; Rats, Wistar ; Signal Transduction ; Up-Regulation ; Ventricular Function, Left ; Ventricular Remodeling ; drug effects

OBJECTIVETo establish a cytologic expressing system of rat glutathione S-transferase pi (GST-pi) cDNA for detecting the resistance of HeLa cells to anticancer drugs.

METHODSThe assessment was made with various anticancer drugs (adriamycin, mitomycin, cisplatinum and vincristine) that showed different cytotoxicities in transfectant HeLa cells with pSV-GT containing rat GST-pi cDNA (HeLa/pSV-GT) or control pSV-neo (HeLa/pSV-neo). Expression levels of GST-pi mRNA in HeLa/pSV-GT and HeLa/pSV-neo were measured by in situ hybridization using Digoxin-labelled cDNA probe.

RESULTSHeLa/pSV-GT expressed significantly high degree of GST-pi mRNA, whereas both HeLa/pSV-neo and HeLa cells had very low expression. Cytotoxicities of HeLa/pSV-GT and HeLa/pSV-neo with 4 anticancer drugs were measured by MTT assay. Drug concentrations for yielding 50% inhibition (IC50) in HeLa/pSV-GT by adriamycin, mitomycin and cisplatinum were 70.13 microg/mL, 10.95 microg/mL and 16.52 microg/mL, respectively. In contrast, IC50 in HeLa/pSV-neo was 10.34 microg/mL, 7.48 microg/mL and 13.70 microg/mL, respectively. The cytotoxicities of vincristine on both HeLa/pSV-GT and HeLa/pSV-neo were not significantly different.

CONCLUSIONSOur findings suggest that HeLa/pSV-GT containing rat GST-pi cDNA is resistant to some anticancer drugs due to overexpression of GST-pi. Also, HeLa/pSV-GT cell line could serve as a useful cytogenetic model for further research.

Animals ; Antineoplastic Agents ; pharmacology ; DNA, Complementary ; Drug Resistance ; Drug Screening Assays, Antitumor ; Glutathione Transferase ; biosynthesis ; pharmacology ; HeLa Cells ; Humans ; Rats ; Transfection

OBJECTIVETo develop human recombinant neutralizing IgG monoclonal antibodies to hepatitis A virus (HAV) by baculovirus expression system.

METHODSThe heavy and light chain genes of two human-derived neutralizing Fab antibodies to HAV were cloned into baculovirus expression vector Pac-kappa-Fc and Pac-L-Fc, and further expressed in insect cells as IgG antibodies. The IgG products were purified and well characterized.

RESULTSThe baculovirus expressed McAb HAFc16 fully retained the specificity of binding to hepatitis A virus and the competition with mouse anti-hepatitis A virus McAb using ELISA. The viral neutralization assay in vitro demonstrated the retention of antibody function after expression of the human antibody in insect cells. The other expressed antibody HAFc78 also has the neutralizing activity but it is directed against different epitopes of HAV when compared with HAFc16.

CONCLUSIONThe recombinant baculovirus/insect cells expressed human neutralizing IgG antibodies to hepatitis A virus retained all biological functions specific for hepatitis A virus. The results provided the possibility of using these antibodies to rapidly protect high risk or early exposure populations from hepatitis A virus infection.

Antibodies, Monoclonal ; biosynthesis ; immunology ; Baculoviridae ; genetics ; Hepatitis A virus ; immunology ; Hepatitis Antibodies ; biosynthesis ; immunology ; Humans ; Immunoglobulin Fab Fragments ; biosynthesis ; immunology ; Immunoglobulin G ; biosynthesis ; immunology ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Recombinant Proteins ; biosynthesis ; immunology

OBJECTIVETo compare enzyme-linked immunospot assay (ELISPOT) and tuberculin skin test (TST) and explore their roles in the auxiliary diagnosis of initial pulmonary tuberculosis.

METHODSTotally 123 patients with initial pulmonary tuberculosis (tuberculosis group) and 102 patients with non-tuberculosis pulmonary disease (control group) were enrolled. The peripheral blood mononuclear cells of all participants were co-cultured with early secretiny antigen target-6/culture filtrate protein-10 fusion protein (ESAT-6/CFP-10), and spot forming cells (SFCs) were enumerated by ELISPOT (ESAT-6/CFP-10-ELISPOT). TST was also performed simultaneously.

RESULTSESAT-6/CFP-10-ELISPOT showed significantly higher numbers of SFCs after stimulation in tuberculosis group than in control group (P = 0.000). The sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, positive predictive value, and negative predictive value of ESAT-6/CFP-10-ELISPOT were 91.1% (111/123), 81.4% (82/102), 4.60, 0.12, 0.85, and 0.87 respectively, while the above values of TST were 65.6% (59/90), 45.1% (46/102), 1.31, 0.76, 0.51, and 0.60, respectively. The sensitivity and specificity of ESAT-6/CFP-10-ELISPOT were significantly higher than those of TST (all P = 0.000). The number of SFCs were not significantly different between smear-positive tuberculosis subgroup and smear-negative tuberculosis subgroup (P = 0.166). The sensitivities were 91.8% (67/73) and 88.0% (44/50) in these two subgroups, respectively, (P = 0.448).

CONCLUSIONSESAT-6/CFP-10-ELISPOT may be a more accurate approach for the auxiliary diagnosis of initial pulmonary tuberculosis; meanwhile, it offers certain diagnostic evidences for smear-negative tuberculosis. However, its specificity may be affected by latent tuberculosis infection. On the contrary, TST has poor value in the auxiliary diagnosis of initial pulmonary tuberculosis.

Enzyme-Linked Immunospot Assay ; Humans ; Leukocytes, Mononuclear ; Sensitivity and Specificity ; Tuberculin Test ; Tuberculosis, Pulmonary ; diagnosis

OBJECTIVETo investigate the association between physical activity (PA) and the incidence of metabolic syndrome (MS) in Chinese adults.

METHODSData on PA and other variables were obtained at the baseline examination of China Multi-center study of Cardiovascular Epidemiology in 1998 and of International Collaborative study of Cardiovascular Disease in Asia(InterASIA) during 2000 - 2001. Follow-up study was conducted in 2007 - 2008. A total of 11 512 Chinese adults aged 35 - 74 years (5563 men and 5949 women) were included in the final data analysis. Information on demographics, PA, smoking and alcohol consumption were obtained and components of MS were examined. Participants were divided into four groups according to quartile of total metabolic equivalent (MET) values per day. In addition, subjects were grouped into the following categories according to occupational PA: inactive, light, moderate and vigorous. Binary logistic model was used to examine the association between PA and the incidence of MS.

RESULTSA total of 2527 cases with MS were documented during an average following up of 8.1 years. The annual incidence rate of MS was 2.71% (2527/93 178.68). After multivariate logistic regression analysis, compared with participants with total PA volume < 32.0 MET×h×d(-1) (annual incidence rate was 3.19% (697/21 830.74)), the RR (95%CI) value of participants with total PA volume during 32.00 - 37.85, 37.86 - 52.29, and ≥ 52.30 MET×h×d(-1) was 1.05(0.92 - 1.19), 0.98(0.86 - 1.12), and 0.68(0.59 - 0.80), respectively (χ(2)trend = 34.23, P < 0.05), with corresponding annual incidence rates of 2.82% (690/24 504.25), 2.73% (661/24 179.36) and 2.11% (479/22 664.33). In addition, compared to inactive occupational PA (annual incidence rate was 2.76% (402/14 588.33)), the corresponding RR (95%CI) value was 0.80 (0.69 - 0.92), 0.70 (0.59 - 0.82), and 0.54 (0.45 - 0.65) (χ(2)trend = 42.34, P < 0.05), and the annual incidence rates were 2.86% (648/22 663.41), 2.40% (455/18 956.14) and 1.89% (344/18 173.86) in participants with light, moderate and vigorous occupational PA, respectively.

CONCLUSIONBoth increased total PA volume and occupational PA intensity are significantly associated with decreased risk of incidence of MS.

Adult ; Aged ; China ; epidemiology ; Exercise ; Female ; Humans ; Incidence ; Male ; Metabolic Syndrome ; epidemiology ; Middle Aged ; Prospective Studies ; Risk Factors

OBJECTIVETo determine the efficacy and safety of combined L-carnitine and acetyl-L-carnitine therapy in infertile males with oligoasthenozoospermia.

METHODSOne hundred fifty patients with oligoasthenozoospermia were randomized selected into treatment and control groups. The treatment group with 90 patients were given L-carnitine (2 g/d) and acetyl-L-carnitine (1 g/d) orally, twice a day. The patients in control group were given Vitamin E 100 mg plus Vitamin C 100 mg, tid. The oral therapy lasted three months and patients accepted sperm analysis every one month. The L-carnitine level in seminal plasma was examined by high performance liquid chromatography (HPC). Side effects as well as pregnant rate were observed.

RESULTSIn the treatment group, 85 patients out of 90 finished the three month treatment. Female spouses of 10 patients (11.6%) achieved pregnancy. Moreover, their forward motile sperm per ejaculation, total motile sperm, as well as the concentration of L-carnitine in seminal plasma were increased significantly (P < 0.01). In control group, 53 patients out of 60 completed three months therapy. Two pregnancy (3.7%) was observed. Though some increase was seen in number of forward motile sperm and total motile sperm per ejaculation, the changes were not statistically significant (P > 0.05). The difference of the pregnant rate between two groups was statistically significant. No side effects were found.

CONCLUSIONCombined treatment with L-carnitine and acetyl-L-calmitine can be an effective and safe option for treating oligoasthenozoospermia by means of significantly improving forward motile sperm and total motile sperm per ejaculation, as well as increasing pregnant rates.

Acetylcarnitine ; administration & dosage ; adverse effects ; Administration, Oral ; Adult ; Carnitine ; administration & dosage ; adverse effects ; Double-Blind Method ; Drug Administration Schedule ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Oligospermia ; drug therapy ; Pregnancy ; Pregnancy Rate

OBJECTIVE: To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province. METHODS: The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017. RESULTS: The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01). CONCLUSIONS Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.
Anti-Bacterial Agents ; Birth Weight ; Humans ; Infant ; Infant, Extremely Low Birth Weight ; Infant, Newborn ; Intensive Care Units, Neonatal ; Surveys and Questionnaires