Purpose　The aim is to invest the mechanism and activity of kringle1～3 gene of angiostatin. Methods　Kringle1～3 gene of angiostatin was abtained from hepatocyte of normal human by RT-PCR and inserted into expression vector pPIC9K in Pichia expression system. The recombinant protein was induced secrete , purified and test the activity by MTT.Results　　Krigle1～3 protein could inhibit vascular endothelial cell proliferative activity.Conclusion　The reaction similar apoptosis induced by kringle1～3 recombinant protein of angiostatin inhibit cell proliferation.

The comparative analysis of the biological characterization and the genetic background study of EV71 circulating strains is commonly recognized as basic work necessary for development of an effective EV71 vaccine. In this study, we sequenced five EV71 circulating strains, isolated from Fuyang, Hefei, Kunming and Shenzhen city of China and named them FY-23, FY-22, H44, K9 and S1 respectively. The sequence alignment demonstrated their genotypes be C4. The genetic distance of the VP1 gene from these isolates suggested that they were highly co-related with genetic identity similar to other previously reported EV71 strains in China. Additionally, these strains were identified to display some obvious proliferation dynamics and plaque morphology when propagated in Vero cells. However, a distinctive difference in pathogenic ability in neonatal mice was found. Some differences in cross neutralization test & immunogenic analysis were also found. All these results are related to the biological characterization of circulating EV71 strains in China and aid in the development of an EV71 vaccine in the future.

Objective To investigate the dynamic changes of three types of anti-poliovirus neutralizing antibodies and anti-hepatitis A virus (HAV) IgG antibody in children who were immunized with inactivated enterovirus 71 (EV71) vaccine (human diploid cell).Methods Serum samples were collected from the subjects immunized with inactivated EV71 vaccine.Neutralizing antibodies against EV71 and poliovirus were detected by micro-cytopathic effect neutralization test.Enzyme linked immunosorbent assay (ELISA) was used to detect IgG antibody against HAV.Results The geometric mean titers (GMTs) of anti-EV71 neutralizing antibody increased to 4.85 following the first-dose injection of inactivated EV71 vaccine.A significant increase of GMTs (up to 64.37) could be observed 28 days after the second-dose vaccination.Meanwhile, results of the dynamic monitor showed that there were slight fluctuations in the neutralizing antibodies against three types of poliovirus on day 28 (28 days after the first-dose vaccination) compared with those on day 0 (before vaccination) (P<0.05);types Ⅰ and Ⅲ anti-poliovirus neutralizing antibodies on day 56 (28 days after the second-dose vaccination) remained slightly different from those on day 0 (P<0.05), but type Ⅱ anti-poliovirus neutralizing antibody on day 56 had restored to normal level (P>0.05).The level of anti-HAV IgG antibody was stable and no significant difference was found during the observation period (P>0.05).Conclusion This study shows that inactivated EV71 vaccine has no impact on anti-HAV IgG antibody in Children during the two-dose vaccination and in anti-EV71 antibody-producing period, but has slight influence on the anti-poliovirus antibodies.In general, changes in antibody profile do not affect the clinical efficacy of immune response.

Objective To compare the simple cases of hand-foot-mouth disease(HFMD) with HFMD patients complicated with encephalitis and HFMD cases complicated with pulmonary edema (PE). To explore predictor factors of disease progression and unfavorable prognosis. Methods Forty-one EV71-infected children admitted to the Fuyang First People's Hospital in Anhui Province from March to May in 2008 were investigated in the research, who were classified as encephalitis-complicated cases ( encephalitis group, n = 15 ), PE-complicated cases ( PE group, n = 15 ) and simple cases (simple group, n= 11 ). Their clinical manifestation, laboratory findings, and immunophenotypes of peripheral blood lymphocyte were analyzed to find predictors associated with disease progression and unfavorable outcomes. Results The mortality rate in PE group was 66.7%, which was significantly higher than that in encephalitis group. Ninty-three point three percent cases in PE group and encephalitis group were younger than 3 years old, with statistic difference compared to simple group. Patients in PE group had higher total blood white cell (WBC) counts and higher absolute neutrophil counts and tended to have higher breathing rate, heart i'ate and glucose level than encephalitis group. The percentages of T cells and natural killer (NK) cells were significantly lower among patients complicated with encephalitis than simple HFMD patients.Conclusions PE is one predictor for poor prognosis. Factors correlated with unfavorable outcome include high WBC, high absolute neutrophil counts; elevated breathing rate, heart rate and glucose level. The immunophenotypes of peripheral blood lymphocytes can also predict the disease progression.

PURPOSE: This study investigated the role of natriuretic peptide receptor 2 (NPR2) on cell proliferation and testosterone secretion in mouse Leydig cells. MATERIALS AND METHODS: Mouse testis of different postnatal stages was isolated to detect the expression C-type natriuretic peptide (CNP) and its receptor NPR2 by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Leydig cells isolated from mouse testis were cultured and treated with shNPR2 lentiviruses or CNP. And then the cyclic guanosine monophosphate production, testosterone secretion, cell proliferation, cell cycle and cell apoptosis in mouse Leydig cells were analyzed by ELISA, RT-qPCR, Cell Counting Kit-8, and flow cytometry. Moreover, the expression of NPR2, cell cycle, apoptosis proliferation and cell cycle related gene were detected by RT-qPCR and Western blot. RESULTS: Knockdown of NPR2 by RNAi resulted in S phase cell cycle arrest, cell apoptosis, and decreased testosterone secretion in mouse Leydig cells. CONCLUSIONS: Our study provides more evidences to better understand the function of CNP/NPR2 pathway in male reproduction, which may help us to treat male infertility.
Animals ; Apoptosis ; Blotting, Western ; Cell Count ; Cell Cycle ; Cell Cycle Checkpoints ; Cell Proliferation ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Germ Cells ; Guanosine Monophosphate ; Humans ; Infertility, Male ; Lentivirus ; Leydig Cells ; Male ; Mice ; Natriuretic Peptide, C-Type ; Polymerase Chain Reaction ; Receptors, Peptide ; Reproduction ; Reverse Transcription ; RNA Interference ; S Phase ; Testicular Diseases ; Testis ; Testosterone