Analysis of Genetic Polymorphism and Population Genetic Structure of 57 Autosomal InDel Loci in Beichuan Qiang Population / 法医学杂志

OBJECTIVES@#To investigate the genetic information of 57 autosomal InDel loci (A-InDels) included in AGCU InDel 60 fluorescence detection kit in the Beichuan Qiang population of Sichuan Province and evaluate its application value in forensic medicine.@*METHODS@#A total of 200 unrelated healthy individuals from Beichuan Qiang population of Sichuan Province were typing detected by AGCU InDel 60 fluorescence detection kit. Allele frequencies and population genetic parameters of the 57 A-InDels were statistically analyzed and compared with the available data of 26 populations.@*RESULTS@#After Bonferroni correction, there was no linkage disequilibrium between the 57 A-InDels, and all loci were in Hardy-Weinberg equilibrium. Except for rs66595817 and rs72085595, the minor allele frequencies of 55 A-InDels were above 0.3. PIC ranged from 0.298 3 to 0.375 0, CDP was 1-2.974 8×10-24, CPEduo was 0.999 062 660, and CPEtrio was 0.999 999 999. The calculation of the genetic distance showed that Beichuan Qiang population had the closest genetic distances with Beijing Han and South China Han populations, but far away from African populations.@*CONCLUSIONS@#The 57 A-InDels in AGCU InDel 60 fluorescence detection kit have a good genetic polymorphism in Beichuan Qiang population of Sichuan Province, which can be used as effective supplemental for individual identification and paternity identification in forensic medicine.

Muscarinic receptor 3 agonist promotes epithelial-meschymal transition in human lung cancer A549 cells by activating PI3K/AKT signaling pathway / 中国病理生理杂志

AIM: To investigate the possible signaling pathway that promotes epithelial-mesenchymal transi-tion(EMT)of the lung cancer A549 cells stimulated with muscarinic receptor 3(M3R)agonist carbachol.METHODS:The lung cancer cells A549 were treated with 400 μmol/L carbachol.The morphological changes of the cells were observed under inverted phase contrast microscope.The migration and invasion abilites were measured by Wound healing and Tran-swell assays.qPCR was used to detect the mRNA level of vimentin and E-cadherin.The protein levels of p-AKT,vimentin and E-cadherin were determined by Western blot.RESULTS:After treatment with carbachol,the A549 cells showed loss of the close connection and the cell morphology was transformed from irregular polygon to spindle -like cells.The results of Wound healing and Transwell assays showed that the migration and invasion abilites of the A 549 cells were enhanced.Car-bachol increased the vimentin expression and decreased the E-cadherin expression at mRNA and protein level(P<0.05). The phosphorylation of AKT in the A549 cells was up-regulated(P<0.05).These changes was inhibited by M3R antago-nist 4-DAMP.CONCLUSION:Carbachol promotes EMT in the human lung cancer A 549 cells by activating PI3K/AKT signaling pathway.

Effects on the antibacterial activity of TiO(2) nanotubes with different diameters from ultraviolet ray-irradiation / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study the effects of ultraviolet ray (UV)-irradiation on the surface characteristic and antibacterial activity of TiO(2) nanotubes with different diameters.</p><p><b>METHODS</b>TiO(2) nanotubes with different diameters were fabricated on polished pure titanium (PT) samples by anodization at 5, 10 and 20 V with PT as control. The samples were exposed to UV-irradiation for 24 h, then the characteristic and antibacterial activity were analyzed and evaluated. The surface topograph was observed by field emission scanning electron microscope (FE-SEM). Contract angle measurements were carried out with three liquids. Staphylococcus aureus (Sa) were used to evaluate the antibacterial activity of samples with the film contact method. The bacterial morphology was observed by FE-SEM. The bacterial adhesion and cell membrane injury were evaluated by fluorescent staining analysis under laser scanning confocal microscope (LSCM).</p><p><b>RESULTS</b>After the TiO(2) nanotubes with different diameters were exposed to UV-irradiation, no change was observed in its surface topograph. With the increase of the diameters of nanotubes, each contract angle of nanotubes decreased, and bacterial FIt and dead/live ratio were also increased. We found 20 V FIt was the biggest (26.550 ± 2.940) and ranks the highest ratio of death/live (0.728 ± 0.091) among the others (P < 0.05).</p><p><b>CONCLUSIONS</b>The UV-irradiation can decrease the contract angle of TiO(2) nanotubes and promote the Sa adhesion on nanotubes. Meanwhile, the antibacterial activity of TiO(2) nanotubes with different diameters was remarkably enhanced by UV-irradiation. Nanotubes anodized at 20 V showed the best antibacterial activity.</p>

Lack of association between the TIGR gene mutation and the high myopia in Chinese children / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To screen TIGR/myocilin gene (MYOC) mutation in high myopic Chinese children with family history.METHODS: Gene sequencing was performed in exon 3 of the TIGR gene in high myopic Chinese Children. The coding sequence of TIGR exon 3 was screened by capillary electrophoresis sequencing. The sequence alterations were analyzed by bioinformatics.RESULTS: TIGR gene mutation was not found in high myopic patients and normal controls group.CONCLUSION: No identified gene mutation is found in TIGR gene in high myopic Chinese children.

Study on effect of sho-saiko-to compound on growth of nasopharyngeal carcinoma cells in CNE-bearing nude mice / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the role of sho-saiko-to compound (SSTC) on the growth of the well-differentiated squamous cell line 1 of nasopharyngeal carcinoma (CNE-1) and well-differentiated CNE-2 in tumor-bearing nude mouse, and try to supply scientific data for its clinical development.</p><p><b>METHOD</b>SSTC were prepared by concentration gradients, and the effect of SSTC on the growth and proliferation of the CNE-1 and CNE-2 were investigated by MT assay and soft-agar colony formation test. After setting up the subcutaneous tumor-bearing nude mouse model at the right lower back (0.2 mL CNE-2 cell suspension, 5 x 10(5)/mL), we randomly divided forty mice into 5 groups and gave high, middle and low concentration groups of SSTC (0.5, 0.25, 0.125 g X mL(-1) by intragastric administration. Positive and negative groups were set up for comparison. After constant administration for 15 days, the volume and weight of the tumor were measured for inhibition rate, so as to investigate the role of SSTC on the CNE-2 bearing tumor.</p><p><b>RESULT</b>In vitro, compared with negative control, SSTC at different gradient concentrations were cultured with the CNE-1 and CNE-2 for 24 h, 48 h and 72 h. It showed that the growth and proliferation of both cell lines were inhibited to some extent. The inhibition rate was increased as the concentration and culture time increasing. Both MTT assay and soft-agar colony formation test showed that the 50% inhibiting concentration (IC50) was about 2.5 g X L(-1). In vivo, compared with negative control, the SSTC could slow down the tumor growth in the SSTC treated groups. The tumor growth of the negative control group (0.76 +/- 0.28) g, (962.88 +/- 245.96) mm3 and the low concentration group of SSTC (0.88 +/- 0.40) g, (1239.66 +/- 421.93) mm3 were obviously faster than those of the high, middle concentration group of SSTC (0.22 +/- 0.14) g, (239.31 +/- 137.07) mm3; (0.20 +/- 0.16) g, (263.42 +/- 166.57) mm3 and CTX positive control group (0.20 +/- 0.10) g, (246.72 +/- 194.6) mm3 (P<0.05).</p><p><b>CONCLUSION</b>SSTC could efficiently inhibit the growth and proliferation of CNE-1 and CNE-2 in vitro, and slow down the tumor growth of the CNE-2 bearing nude mice. It may be a new compound of Chinese medicine for nasopharyngeal carcinoma therapy.</p>

Chemokine SR-PSOX/CXCL16 expression in peripheral blood of patients with acute coronary syndrome / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Scavenger receptor that binds phosphatidylserine and oxidized lipoprotein/CXC chemokine ligand 16 (SR-PSOX/CXCL16) promotes foam cell formation through the tumor necrosis factor (TNF)-alpha mediated mechanism. Because chemokine CXCL16 could be expressed in atherosclerotic lesions and induce smooth muscle cell (SMC) proliferation, we presume that the monocyte SR-PSOX/CXCL16 detection in the patients' peripheral blood will be important for early diagnosis and prognosis of atherosclerosis (AS).</p><p><b>METHODS</b>Enrolled in this study were 40 patients with acute coronary syndrome (ACS), including 20 patients with acute myocardial infarction (AMI) and 20 patients with unstable angina pectoris (UAP), and 20 normal controls. Monocytes in the peripheral blood were isolated, and the changes of expression of CXCL16/SR-PSOX mRNA were compared using reverse transcription-polymerase chain reaction (RT-PCR), with beta-actin as internal control. We compared the expression of CXCL16/SR-PSOX in the ACS subgroups, using Western-blot to analyze protein expression levels. Tissue sections were made from biopsy specimens taken from patients with infective endocarditis, liver cirrhosis, and lung cancer as well as normal controls. And the expression of CXCL16/SR-PSOX was analyzed with a confocal microscope.</p><p><b>RESULTS</b>The expression of CXCL16/SR-PSOX mRNA and protein in the monocytes of peripheral blood was significantly higher in ACS patients than in normal controls (P < 0.05); however, there was no significant difference in CXCL16/SR-PSOX expression between UAP group and AMI group (P > 0.05). Immunofluorescence showed that there were low expression of SR-PSOX in normal vascular endothelial cells and enhanced expression in every layer of the infected vessels, while spreading from endothelial cells to surrounding tissues as infection worsens. Confocal microscopy showed that the expression of SR-PSOX was enhanced in the infiltrated lymphocytes in liver cirrhosis, and that the expression level was proportionate to the degree of inflammation in the portal hepatis and folia.</p><p><b>CONCLUSIONS</b>The expression of CXCL16/SR-PSOX in the monocytes of peripheral blood was significantly higher in ACS patients than in the controls. CXCL16/SR-PSOX-mediated inflammation may contribute to the pathogenesis of ACS, and CXCL16 may play an important role in the pathogenesis and development of AS in humans.</p>

Inhibitory effects of immunotargeting of Chinese cobra cytotoxin and iodine-131 against nasopharyngeal carcinoma cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore a new specific therapy of nasopharyngeal carcinoma (NPC) using an anti-nasopharyngeal carcinoma (NPC) monoclonal antibody BAC5 conjugate with Chinese cobra (CT) and iodine-131(131I).</p><p><b>METHODS</b>BAC5 was labeled with 131I by chloramine-T method, CT was labeled with 125I using iodogen method, and BAC5 and 125I-CT were conjugated by SPDP method. The inhibitory effect of the conjugate on cultured human NPC CNE2 cells was observed using MTT assay.</p><p><b>RESULTS</b>The IC50 of 125I-CT-BAC5 conjugate was 9.17x10(-8) mol/L, and that of 131I-BAC5 was 5.83x10(8) Bq/L, and their combined administration showed obvious inhibitory effect on the NPC cells.</p><p><b>CONCLUSION</b>Both 125I-CT-BAC5 and 131I-BAC5 have obvious inhibition effects against NPC cells, but their combined use shows stronger effects.</p>

A reverse transcriptase-polymerase chain reaction method improving the specific amplification of caveolin-1 gene sequences / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To construct a reverse transcriptase-polymerase chain reaction (RT-PCR)approach that can improve the specificity of primers while dropping down the nonspecific amplification.</p><p><b>METHODS</b>In the recent study we reported a new RT-PCR assay which improved markedly the specificity. However its efficiency of regressing nonspecific amplification remains to be accurately checked and further documented. In primer design, we looked over again some sequences that showed differences at 5' or 3' ends between human CAV1 and mouse Cav1 genes. cDNAs and the diluted plasmids which harbored the sequence of human CAV1 or mouse Cav1 gene were chosen as the templates. The ordinary PCR compared with one, of which primers modified by phosphorothioate and combined with proofreading polymerase, for their efficiencies of nonspecific amplification inhibited.</p><p><b>RESULTS</b>Taq DNA polymerase without proofreading activity could efficiently catalyze the extension of primers with a single or multiple mismatched base pairs at the 3' terminus, but the kind of primer extension can be effectively blocked by phosphorothioate modified primers combined with proofreading polymerase. Compared with ordinary PCR reaction, this new PCR method can effectively regress the primer mismatched amplification of 50 ng DNA almost equaling to 2 x 10(4) unmatched template copies in a final volume of 50 microL.</p><p><b>CONCLUSION</b>Compared with the first generation of polymerases with or without proofreading activities mediating RT-PCR reaction, the introduction of nuclease-resistant 3' modified primers (3' phosphorothioate primer extension) can offer more simplicity, accuracy, and also decrease cost.</p>

Cloning, sequencing and expressing of microneme protein 1 partial gene in toxoplasma gondii ZS2 isolate / 中华预防医学杂志

<p><b>OBJECTIVE</b>To construct a recombinant prokaryotic expression vector (plasmid) containing microneme protein 1 (MIC1) partial gene in toxoplasma gondii (T. gondii) ZS2 isolate. The gene was expressed in varied Escherichia coli (E. coli) after sequencing.</p><p><b>METHODS</b>The gene fragment coding MIC 1 from the genomic DNA of T. gondii ZS2 isolate was amplified by polymerase chain reaction (PCR). The gene was inserted to a prokaryotic expression vector pWR450-1 by digesting with restriction enzymes and linking reaction. The positive clone was screened on LB plates containing ampicillin and identified by restrictive enzyme digestion, PCR amplification and sequence analysis. The recombinant plasmid was transferred into E. coli TG1, JM109 (DE3) and DH5 alpha, and was expressed under the induction of IPTG. The expression products were identified by SDS-PAGE. The MIC1 gene structure was analyzed and compared in homology with the gene sequence of RH isolate using computer software.</p><p><b>RESULTS</b>The recombinant plasmid pWR450-1/MIC1, after cloning from acquired 471 bp MIC1 gene fragment and amplified from the genome gene ZS2, was complete homologous to the sequence of RH isolate, reflecting its highly conservative. The gene could be expressed as fusion protein with 70,000 in varied E. coli.</p><p><b>CONCLUSION</b>Recombinant plasmid pWR450-MIC1 was successfully constructed and could be expressed in different strains of E. coli, laying a foundation for research on its structure and function.</p>