Changes in T helper lymphocytes and their subsets in children with tic disorders / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the changes in T helper lymphocytes and their subsets in children with tic disorders (TD) and their clinical significance.</p><p><b>METHODS</b>Flow cytometry was used to measure the percentages of T helper lymphocytes and their subsets in the peripheral blood of children with TD and healthy children (controls).</p><p><b>RESULTS</b>The percentage of T helper lymphocytes was significantly lower in the TD group than in the control group (P<0.001). The abnormal rate of T helper lymphocytes in the TD group was significantly higher than that in the control group (68.7% vs 18.8%; P<0.001). The percentage of T helper lymphocytes was negatively correlated with Yale Global Tic Severity Scale score (r=-0.3945, P<0.001). As for the subsets of T helper lymphocytes, the TD group had a significantly higher percentage of Th1 cells and a significantly lower percentage of Th2 cells compared with the control group (P<0.001).</p><p><b>CONCLUSIONS</b>The abnormality of T helper lymphocytes and the imbalance of their subsets may be associated with the pathogenesis of TD in children. The percentage of T helper lymphocytes can be used as an indicator for assessing the severity of TD.</p>

Inhibitory effect of saikosaponin-d on the proliferation and migration of human breast cancer MDA-MB-231 cells / 国际药学研究杂志

Objective To investigate the effect and related mechanism of saikosaponin-d (SS-d) on the proliferation and migration of breast cancer MDA-MB-231 cells. Methods The MDA-MB-231 cells were treated with varying concentrations of SS-d.CCK-8 and colony formation analysis were used to evaluate the proliferation ability of MDA-MB-231 cells;3D cell culture, wound healing assay and Transwell migration and invasion assays were used to analyze the ability of cell migration and invasion;flow cytometry was utilized to analyze the cell cycle and apoptosis;semi-quantitative RT (qRT)-PCR and Western blot were used to explore the action mechanism of SS-d on the cell migration and invation. Results SS-d inhibited the proliferation and colony formation ability of MDA-MB-231 cells, both dose-and time-dependently;migration and invasion abilities of the MB-231 cells were also significantly inhibited by the treatment with SS-d (P＜0.01);the treatment with SS-d caused the significant cell cycle arrest at G1 phase and also significant apoptosis-induction in MDA-MB-231 cells;SS-d could up-regulate the expression of P21, P27 E-cadherin (P＜0.01), but down-regulate the expression of Bcl-2, vimentin and N-cadherin (P＜0.01), all detected in both the m RNA and protein levels. Conclusion SS-d was likely to be a potential antitumor drug for the breast cancer therapy to inhibit the proliferation and metastasis of the cancer cells.

Synthesis of DNA Encoding Botulinum Neurotoxin Receptor syt II and Its Fusion Expression in E.coli / 中国生物工程杂志

In order to construct a prokaryotic expression vector of human receptor syt II N-fragment and to express recombinant MBP-Syt fusion protein in E.coli and to purify and identify its activity. According to codon preference of E.coli, a DNA fragment encoding human syt II N-fragment was synthesized, and then cloned into prokaryotic vector pMAL-c2x for sequencing. Then the recombinant plasmid pMAL-Syt was introduced into E.coli ER2566 by transformation for expression and the obtained engineered bacteria were induced by IPTG. The fusion protein was purified by amylose resin affinity chromatography and identified by SDS-PAGE and Western blot. The binding activity of the protein was determined by ELISA. It is concluded that MBP-Syt protein is of good binding activity.

Preparation of alpha-toxin's protective antigen of clostridium perfringens type A and research for its primary immunological protective function / 生物工程学报

Induced by 42 degrees C, the recombinant engineering bacterial pBV/cpa408 was highly expressed. After having been pelleted by 80% (NH4)2 SO4 and dialysised, the expressed protein was isolated and purified by the gel filtration choromatography. Then according to an amount of 1.0 mg/kg, the Kunming Mice (body weighted 18g) were immuned with the purified protein by intraperitoneal inoculation. One week after the first enhanced immunization, the Kunming Mice were attacked with an amount of 1.0MLD alpha-toxin, in which the eight mice immuned all survive and the control group all died. During the period of immunization, the titre of the mouse's serum antibody was measured by ELISA. One week after the first immunization, the titre of the mice's serum antibody was 1:800, but that of one week after the first enhanced immunization reached to 1:6400.

Analysis on mutation of adrenoleukodystrophy gene in exon 1 and exon 5 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To elucidate the molecular mechanism of X-linked adrenoleukodystrophy(ALD) in Chinese.</p><p><b>METHODS</b>Polymerase chain reaction in exon 1, exon 5 and their flanking sequences and direct DNA sequencing of ALD gene were performed in four patients, their mothers and twenty normal individuals as controls.</p><p><b>RESULTS</b>A splice mutation was identified in the interface of exon 5 and intron 5 (1875 G-->A). This splice mutation in 5' end of intron 5 might lead to abnormal splice in exon 5 and exon 6 and bring about unstable and abnormal ALD protein; the lignoceryl CoA ligase could not transport very long chain fatty acids (VLCFA) into peroxisome and could not function normally; consequently, defective beta-oxidation of VLCFA in peroxisome could result in an accumulation of VLCFAS in the central nervous system, adrenal gland and blood.</p><p><b>CONCLUSION</b>The splice mutation in 5' end of intron 5 leading to abnormal splice in exon 5 and exon 6 appears to be one of the causes of X-linked recessive adrenoleukodystrophy.</p>

STUDIES ON HARMFUL MICROBES IN RECIRCULATING WATER SYSTEM OF HEATING PIPELINE IN TAIYUAN / 微生物学通报

The microbial counts, type, as well as relationship between microbial counts and the temperature of water in reticulating water system of heating pipeline in Taiyuan were studied, which the main biofouling harmful microbes included slimeforming heterotrophic bacteria, sulfate reducing bacteria, iron bacteria and fungi, respectively. The results showed that the harmful microbes in water system were lower than that of control guideline during heating period, whereas the microbes were higher than that of control guideline, which would result in biofouling of water tube during non- heating period.

Construction and Screening of a Phage Display Library of Repertoire Single Chain Fv Antibody from Mouse Immunized with BoNTB/Hc / 微生物学通报

To produce antibodies capable of neutralizing botulinum neurotoxin type B(BoNT/B),We cloned the carboxy-terminal end of Hc containing the major determinants responsible for specific toxin,induced and purifed.The heavy-chain and kappa light-chain variable region gene repertoire of immunoglobulin were amplified individually from the spleen cell mRNA by RT-PCR and joined as a single-chain Fv(scFv)DNA fragment.These fragment were cloned into the phagemid pCANTAB5E and the phage display library was constructed.Results showed that the high affinity scFv was obtained after 4 rounds of panning,with its DNA sequence conforming to that of mouse antibody.

High-level Secretion Expression of Human ScFv Against Botulinum Neurotoxin A in Pichia pastoris* / 微生物学通报

The specific ScFv gene against botulinum neurotoxin A (BoNTa)was cloned into pPIC9k. Positive integrators were screened by increasing the dose of G418 in culture and expressed in Pichia pastoris GS115. As a result, engineered recombinant clone were obtained. 26 kD product of interest was seen easily in SDS-PAGE. Expression of human ScFv got the highest level 15% of total secreted proteins during 72～84 h after 1% methanol inducing. Purification of ScFv was finished by two steps: gel filter and ion exchange. Competing ELISA showed that recombinant ScFv could compete with antiserum to specific bind BoNTa.