To explore the role and application of Meta-regression and subgroup analyses to recognize and control the heterogeneity in Meta-analysis, Meta-regression models were established by secondary data to screen the factors resulting heterogeneity,and subgroup analyses were used to compare the change of heterogeneity before and after.The heterogeneity was found in the Meta-analysis(Q=44.71,df=27,P=0.017).Sample size and region were selected(P=0.012 and P=0.091,respectively)by Meta-regression from many possible factors such as sample size,year,region and case/contml ratio.The Q values were lowered from 44.71 to 32.11 after subgroup analyses.Thus,Metaregression method was convenient and reliable to screen the affected factors of heterogeneity,and subgroup analyses based on the hypothesis that could significantly lower the heterogeneity.It was recommended to a combined use when an obvious heterogeneity existed but was in need to get an overall result in Metaanalysis.We could correctly judge and lower the heterogeneity to increase the robustness and rationality of results from Meta-analysis.

OBJECTIVETo explore the influence of the lentiviral vector mediated mouse genetic engineering regulatory T cells (Treg) infusion on post-allogeneic bone marrow transplantation (allo-BMT) acute graft-versus-host disease (GVHD) in mice.

METHODSLentivirus-mediated Forkhead box P3 (Foxp3) gene was transduced into BALB/c mice CD4(+)CD25(-) T cells (Treg) to construct engineered Tregs in vitro. An allo-BMT model of BALB/c to C57BL/6 mice was established. After irradiation, the recipients were injected with donor cells plus the genetic engineering Tregs. Survival time, clinical GVHD score, histopathological findings, activation of donor T cells or serum levels of inflammatory cytokines were observed after allo-BMT.

RESULTSThe mean survival times for radiation alone group (Gp I), transplantation control group (Gp II), engineering Treg infusion group (Gp III) and empty vector control group (Gp IV) were (8.8 ± 0.6) d, (36.7 ± 2.5) d, (51.6 ± 4.0) d and (34.1 ± 2.3) d, respectively. The survival time was significantly longer in Gp III than in other groups (P < 0.05). Histopathological finding in several target organs (skin, liver and small intestine) confirmed the presence of severe GVHD in Gp II and Gp IV, while no histological signs of GVHD were observed in long survival recipients in Gp III, and clinical GVHD scores in Gp III were significantly lower than that in Gps II and IV. The numbers of donor T cells and the percentage of IFN-producing donor T cells in the spleen of recipients in Gp III were significant lower than those in Gps II and IV at days 3 and 4, and at day 3 after transplantation, respectively (P < 0.05). The serum levels of IFN-γ, IL-2 and TNF-α were increased at day 21 to 28 after transplantation in all groups. The peak concentrations of IFN-γ, IL-2 and TNF-α in Gp III were significantly lower than those in Gps II and IV control groups at day 21 (P < 0.05).

CONCLUSIONCo-injection of genetic engineering Treg can efficiently prevent recipients from lethal GVHD after allo-BMT in mice by inhibiting the early activation and expansion of donor T cells and reducing the serum levels of inflammatory cytokines.

Animals ; Bone Marrow Transplantation ; adverse effects ; Cytokines ; blood ; Female ; Forkhead Transcription Factors ; genetics ; Genetic Engineering ; Genetic Vectors ; Graft vs Host Disease ; immunology ; Lentivirus ; Lymphocyte Activation ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; T-Lymphocytes, Regulatory ; cytology ; immunology ; Transduction, Genetic ; Transplantation, Homologous

OBJECTIVETo prepare the genetic engineering regulatory T cells (Treg) via the self-inactivating (SIN) lentiviral vectors carrying Foxp3 gene, and assay the phenotype and abilities of its proliferation and immunosuppression.

METHODSThe bicistronic SIN lentiviral transfer plasmid containing Foxp3 gene and internal ribosomal entry site-green fluorescent protein gene (IRES-GFP) was constructed. Human embryonic kidney 293T cells were co-transfected using liposome by lentiviral packing system, which included the packaging plasmid Delta NRF, the transfer plasmid and the envelope plasmid VSVG. The efficiency of gene transduction and the expressions of Foxp3, CD25, GITR, CTLA-4 of CD4(+)CD25(-) T cells, which were isolated by magnetic beads from the spleen, and then co-cultured with 293T cells, were detected by flow cytometry (FCM). The proliferative and suppressive capacities of transduced T cells were estimated by Cell Count Kit-8 (CCK-8) and the cytokine production was performed by ELISA.

RESULTSThe lentiviral transfer plasmid pXZ208-Foxp3-IRES-GFP was successfully constructed, the virus titers were above 10(6) IU/ml in the supernatant. pXZ208-IRES-GFP was used as control group. After cocultured, the CD4(+)CD25(-) T cells expressed significantly higher Foxp3, CD25, GITR and CTLA-4 in experimental group than in control group. Upon stimulation with anti-CD3 epsilon and APCs, the proliferative capacity of Foxp3-transduced T cells and the production of IL-2, IL-4, IL-10, IFN-gamma were significantly lower than those in control group (P < 0.01); Foxp3-transduced T cells also significantly inhibited the proliferation of CD4(+)CD25(-) T cells.

CONCLUSIONSThe genetic engineering Treg mediated by SIN lentiviral vectors are successfully constructed and their phenotype and function are similar to natural CD4(+)CD25(+) Treg.

Animals ; Cell Proliferation ; Cells, Cultured ; Forkhead Transcription Factors ; genetics ; metabolism ; Genetic Engineering ; Genetic Vectors ; HEK293 Cells ; Humans ; Lentivirus ; genetics ; Mice ; Mice, Inbred BALB C ; Phenotype ; T-Lymphocytes, Regulatory ; cytology ; immunology ; metabolism ; Transfection

OBJECTIVETo study the clinical feature and more reasonable diagnostic typing criteria for patients with liver failure.

METHODS13/21 cases of ALF, SALF with no past liver disease, 49/72 cases of with chronic hepatitis, and 23/73 cases ALF, SALF with liver cirrhosis, were analyzed respectively.

RESULTS1 ALF patients (1). There exist significant statistic differences in ALB, ALT, CHE in three ALF groups.(2). It had statistic differences in those patients with hepatic encephalopathy.(3). The prognosis of the patients with chronic hepatitis group (42.85 percent) was best than that of chronic cirrhosis (26.09 percent) and no past liver disease (15.38 percent). (2) In SALF patients (1). There exist significant statistic differences in ALB, GLO, ALT, AST, BDIL, GLU and CHE in three SALF groups.(2). It had statistic differences in those patients with hepatic encephalopathy in three SALF groups.(3). The prognosis of the patients with chronic hepatitis group (51.39 percent) was best than that of chronic cirrhosis (36.85 percent) and no past liver disease (33.33 percent).

CONCLUSIONThere are different clinic feature and prognosis in three ALF or SALF groups, so we suggest that it were clinic practicability and science in classify of liver failure at present.

Humans ; Liver Failure ; classification ; Liver Failure, Acute ; classification ; Prognosis

The aim of this study was to detect the nerve growth factor (NGF) level in serum and NGF low affinity acceptor CD271 expression on bone marrow leukemic cells in acute B lymphoid leukemia (B-ALL) patients and to analyze their clinical significance. The NGF level in serum and CD271 expression on leukemic cells in bone marrow were detected by enzyme linked immunosorbent assay and flow cytometry in B-ALL patients respectively. The results indicated that compared with control group, the NGF level in serum of patient group significantly increased (t = 4.191, p < 0.05), but CD271 expression on leukemic cells in bone marrow decreased significantly (t = 4.898, p < 0.05). The complete remission (CR) rate of 25 B-ALL patients was 64% (16/25) after one course of CVAD chemotherapy. There were statistically significant differences of NGF level and CD271 expression in non-remission (NR) group and control group (t = 3.976, p < 0.05 vs t = 5.052, p < 0.05), but there were no statistically difference of NGF level and CD271 expression in CR group (t = 1.102, p > 0.05 vs t = 1.150, p > 0.05) as compared with control group. The CD271 expression before and after chemotherapy between CR and NR groups showed statistically significant differences (t = 3.889, p < 0.05; t = 3.751, p < 0.05 and t = 4.678, p < 0.05 respectively), but NGF level before and after chemotherapy showed no statistical difference between these 2 groups (t = 0.476, p > 0.05). 50% (8/16) patients relapsed during following up, and of their NGF level [(168.00 ± 61.66) pg/ml] and CD271 expression [(52.29 ± 13.00)%] showed the significantly differences, compared with those in control group (t = 5.284, p < 0.05 vs. t = 6.073, p < 0.05), but the NGF level [(81.13 ± 25.32) pg/ml] and CD271 expression [(78.45 ± 7.12)%] of other 8 patients showed no statistical difference as compared with control group (t = 1.228, p > 0.05 vs t = 1.144, p > 0.05). Compared with low NGF level and CD271 low expression groups, the survival time of B-ALL patients with high NGF level and CD271 expression was not changed significantly (p = 0.750 vs p = 0.170). It is concluded that the increased NGF level in serum and decreased CD271 expression on bone marrow leukemic cells in B-ALL patients are related with leukemia development and may be the useful indexes to evaluate curative effect and prognosis.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; metabolism ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Nerve Growth Factor ; blood ; Nerve Tissue Proteins ; metabolism ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; metabolism ; Receptors, Nerve Growth Factor ; metabolism ; Young Adult

This study was aimed to propagate and identify the prdm1 gene-knockout mice, so as to lay the foundation for studying Blimp-1 protein. Two kinds of transgenic homozygous mice with B6.prdm1(flox/flox) and B6.Lck-Cre were feed and propagated; after successful propagating, the first passage mice were obtained; after the first passage mice were copulated once again, the genotypes were obtained as follows: B6. prdm1(wild/wild). Lck-Cre, B6. prdm1(wild/wild), B6.prdm1(flox/flox). Lck-Cre, B6.prdm1(flox/wild). Lck-Cre, B6.prdm1(flox/flox), B6. prdm1(flox/wild). The genomic DNA of second passage mice was extracted, the Cre and loxp gene fragments were amplified by PCR, then the size of Cre and loxp genomic DNA were detected by agarose gel electrophoresis. The mice with B6.prdm1(flow/flox). Lek-Cre were used as conditionally prdm1-knockout mice, B6.prdm1(flox/wild). Lck-Cre mice, B6.prdm1(flox/flox) and B6 mice were used as controls. The spleen T lymphocytes and B lymphocytes were sorted by using magnetic beads, the blimp-1 target protein was identified by Western blot. The results showed that the two transgenic homozygous mice had the ability to reproduce, and the separation ratio of second passage mice generated from propagation of their offspring cach other meet Mendelian laws, and the prdm1 gene-knockout mice also could successfully obtained. It is concluded that the application of Cre-loxp system may successfully obtain plentiful prdm1 gene-knockout mice.
Animals ; Genotype ; Mice ; Mice, Inbred C57BL ; genetics ; Mice, Knockout ; genetics ; Reproduction ; Transcription Factors ; genetics

Objective To investigate personality characteristics, coping styles and influential factors on quality of life (QOL) of patients with cancer.Methods Patients in three groups (patients with cancer,hepatitis patients and health control) were assessed by using Eysenck personality questionnaire (EPQ), coping style questionnaires and WHO quality of life questionnaire-brief (WHOQOL-BREF).Results Compared with the hepatitis patients and healthy persons, E and N scores of cancer patients were lower (P <0.01), resolvingproblem score and general score of WHOQOL were also lower (P < 0.01); However, self-reproach and evasive scores were higher (P <0.01).The quality of life of patients with cancer had positive correlation with resolvingproblem factor, rationalization, and course of disease.Conclusions Cancer patients more frequently had passive coping styles and introvert personality; The quality of life of patients with cancer was poorer and had positive correlation with active coping styles.

The aim of this study was to investigate the expression of Th17 cells and regulatory T (Treg) cells in peripheral blood of patients with immune thrombocytopenia (ITP) and to clarify the role of the Th17/Treg cell ratio imbalance in pathogenesis of ITP. Patients were divided into the pre-treatment group (active group) (n = 38) and post-treatment group (remission group) according to the platelet count and curative effect. Post-treatment group was further divided into remission group (n = 24), partial remission group (n = 10), and non-remission group (n = 4). 30 healthy subjects were enrolled in control group. Flow cytometry was used to detect the percentages of peripheral blood Th17 cells and Treg cells in CD4(+) T cells from ITP patients and controls respectively. The results showed that the percentages of Th17 cells in active group and non-remission group were significantly higher than those in control group (p < 0.05). The percentages of Th17 cells in remission group, partial-remission group were also higher than those in control group, but there were no statistically significant differences between these groups. The percentage of Th17 cells in remission group was lower than that in active group, but there was also no statistically difference between two groups. The percentages of Treg cells in active group, partial-remission group and non-remission group significantly decreased, compared with in control group (p < 0.01). The percentage of Treg cells in remission group was lower than that in control group, but there was no statistically significant difference. The ratio of peripheral blood Th17/Treg cells in active group, partial-remission group and non-remission group was higher, as compared with in control group. The ratio of peripheral blood Th17/Treg cells in remission group was higher than that in control group, but there was no statistically difference between two groups. It is concluded the percentage of Th17 cells and the ratio of Th17/Treg cells are higher in active group. The percentage of Treg cells is low in active group, partial remission and non-remission groups. The imbalance of Th17/Treg ratio may play a critical role in ITP pathogenesis.
Adolescent ; Adult ; Aged ; Blood Cell Count ; Case-Control Studies ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Purpura, Thrombocytopenic ; blood ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology ; Young Adult

OBJECTIVETo study the effects of 1-bromopropane (1-BP) on the functions of learning-memory and the central cholinergic system in rats.

METHODSForty male Wistar rats were randomly divided into four groups: low 1-BP group (200 mg/kg), middle 1-BP group (400 mg/kg), high 1-BP group (800 mg/kg) and control group, and the exposure time was 7 days. The Morris water maze (MWM) test was applied to evaluate the learning-memory function in rats. After the MWM test, the rats were sacrificed, the cerebral cortex and hippocampus were quickly dissected and homogenized in ice bath. The activity of acetylcholine esterase (AChE) and choline acetyltransferase (ChAT) in supernatant of homogenate were detected.

RESULTSThe latency and swim path-length of rats in middle and high 1-BP groups prolonged significantly in place navigation test and the efficiency of searching strategy obviously decreased, as compared with control group (P < 0.05 or P < 0.01). In spatial probe test, the number of crossing platform in three 1-BP groups decreased significantly, as compared with control group (P < 0.05 or P < 0.01). The cortical AChE activity of rats in middle and high 1-BP groups was significantly higher than that of control and low 1-BP group (P < 0.05 or P < 0.01). The AChE activity in rat hippocampus of high 1-BP group obviously increased, as compared with control group as compared with control group (P < 0.05). There was no significant difference of cortical ChAT activity between three 1-BP groups and control group (P > 0.05). In the hippocampus, there was no difference of ChAT activity among the groups (P > 0.05).

CONCLUSION1-BP exposure could significantly influence the learning-memory function in rats due to the increase of AChE activity.

Acetylcholinesterase ; metabolism ; Animals ; Cerebral Cortex ; drug effects ; enzymology ; Choline O-Acetyltransferase ; metabolism ; Hippocampus ; drug effects ; enzymology ; Hydrocarbons, Brominated ; toxicity ; Male ; Maze Learning ; drug effects ; Rats ; Rats, Wistar

Objective To understand the epidemiological characteristics and the influencing factors of Japanese encephalitis (Abbreviated JE) at different periods in Hainan province, and to provide evidence for JE control strategies. Methods Data on JE had been collected since 1952 and related incidence, mortality, fatality rates of JE and the rate of JE vaccination were calculated for four periods: the natural epidemic period (1952-1978), period for inactivated JE vaccine promotion (1979-1986), national immunization program period (1987-1990) and the promotion of live-attenuated JE vaccine period (1991-2007). History on the immunity of JE and data from the monitoring programs on cases and insect vectors were also analyzed. Results The average annual incidence in natural epidemic period was 7.12/100 000, which was also the highest among the 4 periods with the mortality rate as 0.89/100 000. The average annual incidence was 0.11/100 000 which was the lowest while the mortality rate was 0.01/100 000 in the period when live-attenuated JE vaccination was under promotion. Since the establishment of JE surveillance system was set up in Hainan province in 2002, the specimen collection rate of JE reported suspected cases had reached 92.8%, while the incidence of confirmed JE cases fluctuated between 0.05/100 000 and 0.2/100 000. Of the confirmed cases, 86% did not have clear history of JE vaccination. The annual average protective rate reached 95.69% in 2004-2007. The density of insect vectors was high in Hainan province. The infection rate of JE in host animal swine, kept rising from April to October, and in some surveillance points, the rate reached 99.3%. The density of insect vector plotted in trans-curve with the infection of host animals. Conclusion The application of live-attenuated JE vaccine was found to have had significant protective effect on JE in susceptible children. It helped reducing the incidence and controlling the epidemics of JE. It appears feasible to include live-attenuated JE vaccine into the expanded immunization program.