Association of single nucleotide polymorphisms of CSPG2 and HSPG2 genes with intracranial aneurysm in ethnic Han Chinese population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association between CSPG2 and HSPG2 gene polymorphisms and intracranial aneurysm (IA) in ethnic Han Chinese population.</p><p><b>METHODS</b>A case-control study was carried out. A total of 537 IA patients and 1071 normal controls with matched age and gender were recruited. Peripheral blood samples were obtained from all subjects. Following extraction, target DNA was amplified with PCR and genotyped with a SNaPshot method. The association between 2 tag SNPs (rs251124 and rs3767137) of CSPG2 and HSPG2 genes and IA was assessed.</p><p><b>RESULTS</b>The genotype frequencies of rs251124 and rs3767137 were both in Hardy-Weinberg equilibrium. No significant difference has been found in the frequencies of rs251124 of CSPG2 between the two groups. Similarly, the frequency of rs3767137 (HSPG2) did not differ between the IA and control groups (P=0.22), albeit with an OR value of greater than 1 (OR=1.12, 95%CI=0.92-1.37). There were no significant difference in genotypic frequencies of the two SNPs between the two groups (P=0.46, 0.53).</p><p><b>CONCLUSION</b>No association has been found between polymorphisms of rs251124 and rs3767137 loci of CSPG2 and HSPG2 genes and IA in the selected population.</p>

Immunotherapy effect of dendritic cells pulsed by lysate of tumor cell Ar-He cryoablation on rat models of intracraniai gliomas / 中华神经医学杂志

Objective To investigate the role of C6 glioma cells mediated by rapid freezing and thawing ofAr-He cryoablation in the maturation of marrow-derived dendritic cells (BM-DCs) in Wistar rats,and the anti-tumor effect of these DCs on rat models of intracranial gliomas. Methods C6 glioma cells were routinely cultured in vitro; rapid freezing and thawing of Ar-He cryoablation was employed in C6 glioma cells of the experimental group, and C6 glioma cells of the negative control group were only performed insertion of the probe; blank control group (using rapid freezing and thawing of Ar-He cryoablation on the same amount of PBS) was also employed.Bone marrow-derived mononuclear cells (MNCs) were first prepared from tibia and femur bones of Wistar rats. These cells were cultured with such cytokines as recombinant granulocyte-macrophage colony-stimulating factor (rmGM-CSF),recombinant interleukin-4 (rmIL-4) and tumor necrosis factor-alpha (TNFα) to induce their maturation; BM-DCs were pulsed with or without tumor cell lysate obtained by rapid freezing and thawing of Ar-Hecryoablation at a ratio of (DC:tumor cells =1:3) 7 d after that.Morphological observation of BM-DCs was performed by light microscopy and the expression of DCs costimulatory molecules CD80 and CD86 were measured by flow cytometry 48 h after the addiction; the IL-12 level in the supematant of DCs was detected by ELISA. In order to determine whether or not vaccination with C6 TP DCs can induce the therapeutic potential in the established glioma-bearing models, the C6 cells cultured in vitro were stereotaxically implanted into the left caudate nucleus of Wistar rat brain; glioma-bearing rats were injected with vaccination with DCs,cells from the blank control group and negative control group on the 3rd and 10th d. Survival time was observed and determined using the method of Kaplan-Meier and Log-Rank analysis. Results DCs from rats' bone marrow cells cultured with cytokines and pulsed with tumor lysates showed the characters of typical mature DCs.Morphologically,these cells were large with oval or irregularly shaped nuclei and with many small dendrites. Phenotypically, they expressed high levels of CD80 and CD86 antigens (71.8 1％± 1.10％ and 74.66％± 1.48％ in experimental group,49.49％±1.08％ and 51.20％±2.06％ in negative control group, and 48.47％±1.09％ and 49.53％±1.89％ in blank control group); significant difference was noted between each 2 groups (P＜0.05).Functionally,the IL-12level in the supernatant of DCs showed obvious increment in the experimental group (245.99 ±3.20pg/mL) as compared with that in the negative control group (138.68±3.20 pg/mL) and blank control group (135.16±2.88 pg/mL,P＜0.05).These cells gained the capacity of mediating immunotherapy against intracranial gliomas in rats:the median survival in the experimental group (33 d) was significantly higher than that in the negative control and blank control groups (22 and 24 d, respectively, P＜0.05).Conclusion C6 glioma cells mediated by rapid freezing and thawing of Ar-He cryoablation can induce maturation of BM-DCs in Wistar rats; these BM-DCs pulsed with tumor lysates, as new therapeutic vaccines,can mediate immunotherapy against intracranial gliomas in rats.

Preparation of suspension of gastric mucous membrane single cell and expression of cyclins in cells / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore a method which can remove the gastric mucus in order to prepare mucous membrane single cell suspension for the research of cytomics.</p><p><b>METHODS</b>Enzymology was used to remove the mucus gel and to separate mucous layer from the normal fresh gastric tissue. The mucous layer was broken to prepare single cell suspension with machine method. Expression of major cyclins in mucous layer cells was examined by cytoimmunochemistry, flow cytometry(FCM) and confocal microscopy.</p><p><b>RESULTS</b>The 0.1% pepsin could dissolve the mucus gel and 1.2-2.4 U/L dispase could separate the mucous layer completely. The single mucous cell suspension was prepared successfully. FCM results from mucous single cell suspension revealed that expression of cyclin D(3), B(1) was obvious, that of cyclin D(2) was weak and that of cyclin D(1), A, E was the least. Similar results were found with confocal microscopy.</p><p><b>CONCLUSIONS</b>Single cell suspension from mucous layer can be easily prepared by pepsin and dispase. Cyclins schedule expression in vivo is different from cyclins schedule expression in vitro.</p>

Study on clinical effect and mechanism of jianpi qingre huayu recipe / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the effects of Jianpi Qingre Huayu Recipe in curing gastric ulcer and to preliminarily probe into its pathogenic mechanism.</p><p><b>METHODS</b>Fifty patients with gastric ulcer of Pi -insufficiency and stasis-heat syndrome type were assigned to the treated group (30 patients) and the control group (20 patients). They were treated respectively with JQH and Ranitidine. At the same time, another group consisting of 20 healthy persons was set up for normal control. The clinical effect on gastroscopic figure and traditional Chinese medicine (TCM) syndrome were observed. Changes of T-cell subsets and interleukin-8 (IL-8) in serum as well as IL-8 in mucosa around the gastric ulcer were determined before and after treatment by flow cytometry and ELISA.</p><p><b>RESULTS</b>Comparison of the total effective rate on gastroscopic figure in the treated group and the control group (86.7% vs 80.0%) showed insignificant difference, but the cure rate and markedly effective rate in the former (50.0% and 20.0%) was higher than that in the latter (40.0% and 15.0%) respectively. Comparison of the total effective rate on TCM syndrome in the treated group and in the control group (96.7% vs 70.0%) showed insignificant difference, but the cure rate and markedly effective rate in the former (63.3% and 23.3%) was higher than that in the latter (50.0% and 20.0%) respectively. Serum levels of CD3+, CD4+, CD8+ got restored to normal range in the treated group after treatment but it was not so in the control group. IL-8 level in gastric mucosa was improved in both groups but the improvement in the treated group was better.</p><p><b>CONCLUSION</b>JQH could effectively treat gastric ulcer and partly reduce its recurrence through improving patients' immune function.</p>

Cloning, ligation and expression of the variable region genes of the monoclonal antibody against human HnRNPA2/B1 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To clone the variable region genes of the monoclonal antibody (McAb) against human heterogeneous nuclear ribonucleoprotein A2/B1 (HnRNPA2/B1), ligate them to assemble single chain Fv (ScFv) gene and express in Escherichia coli.</p><p><b>METHODS</b>The specificity of the anti-HnRNPA2/B1 McAb 3E8 to synthetic HnRNPA2/B1 peptide, HnRNPA2/B1 protein in lung cancer cells were examined by dot-immunobinding assay, Western blot and immunohistochemistry. The variable region genes of heavy chain (VH) and light chain (VL) were amplified from hybridoma cell by reverse transcription-polymerase chain reaction(RT-PCR), and then were linked by a linker peptide using SOE-PCR (splicing by overlap extension-PCR) to construct recombination ScFv gene. The latter was cloned into the expression vector pET28 (a+) and expressed in E coli BL21. The expressed product was identified by SDS-PAGE and competitive ELISA inhibition test.</p><p><b>RESULTS</b>It was shown that the McAb combined specifically with synthetic HnRNPA2/B1 peptide and HnRNPA2/B1 protein in three lung cancer cells. The cloned VH gene and VL gene were 345 bp and 309 bp respectively and were linked successfully to obtain ScFv gene. The ScFv protein was expressed in the form of inclusion body, with molecular weight of 28,000 and immunoreactivity to HnRNPA2/B1.</p><p><b>CONCLUSION</b>VH gene, VL gene and ScFv gene of anti-HnRNPA2/B1 antibody were cloned, constructed and functionally expressed in E coli. These results provide the experimental basis for elucidating the role of HnRNPA2/B1 in lung cancer.</p>

Surveillance on hemorrhagic fever with renal syndrome in Guizhou during 1984-2000 / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the relations between different areas on hemorrhagic fever with renal syndrome (HFRS) in Guizhou.</p><p><b>METHODS</b>Various forms of infectious areas were surveyed continuously through epidemiologic surveillance system.</p><p><b>RESULTS</b>In fixed areas under surveillance system, the mean positive rate of HFRSV among Apodemus agrarius was 3.39%, comparing with Rattus norvegicus 1.61% in Apodemus infectious areas of Zunyi county, 3.19% in Rattus norvegicus, but no HFRSV of Apodemus agrarius was identified in Rattus infectious area of Shiqian county. Both Apodemus and Rattus infectious areas were relatively stabilized. In both banks of Luowang river, Kaiyang county, which had been identified as areas of infections for Apodemus in the eastern part, Rattus infectious area in the west, slow change was noticed. In 1983 - 1984 was not found in Apodemus agrarius HFRSV, however the infectious rate of HFRSV in Apodemus agrarius was 13.85% (Ag 1/65, Ab 8/65) in the western part of the province in 1995 - 1998.</p><p><b>CONCLUSION</b>Both Apodemus and Rattus infectious areas were stabilized but changed slowly. Mixed type and the result of mutual penetration were noticed.</p>