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Objective@#In this study we analyzed the genetic characteristics of echovirus 30 (E-30) VP1 gene sequences from Yunnan province isolated from viral meningitis (VM) cases in 2010-2013.@*Methods@#RT-PCR and VP1 gene sequencing were done for 9 E-30 strains isolated from VM cases in 2010-2013. VP1 gene sequences of E-30 reference strains were downloaded from the GenBank and their nucleotide (nt) and amino acid (aa) diversities were calculated by MEGA 5.1 software, the phylogenetic tree was constructed and the genetic characteristics and molecular epidemiology were analyzed.@*Results@#In 2010-2013, 9 strains of E-30 viruses were detected from 79 VM cases caused by echoviruses, accounting for 11.39%(9/79), the overall positive rate was 1.63%(9/553). Phylogenetic analysis revealed that E-30 strains can be divided into four genotypes (genotype A, B, C and D), and genotype D can be further divided into seven sub-genotypes. Nine Yunnan VM isolates were distributed in D7 sub-genotype, and can be further clustered into 3 branches: 5 strains isolated in 2010 were clustered in branch 1, it is evident that these viruses were responsible for an aseptic meningitis outbreak in Kunming in that year; one 2011 isolate, together with 2013 isolate and one isolate from healthy children in 2010 were clustered in branch 2, these two branches were Yunnan special branches, and two 2011 isolates had the highest homology with 2003 VM outbreaks′ strains isolated from Shandong, Jiangsu, and Zhejiang, showing that these strains may have the same evolutionary sources.@*Conclusions@#Nine Yunnan VM isolates were distributed in D7 sub-genotype, and these strains have different evolutionary sources, showing that at different times E-30 viruses in the same sub-genotypes branch might prevail in different areas.
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Objective To establish a mouse model of systemic C. albicans infection by oral inoculation of the pathogen and observe the proliferation and distribution of C. albicans in vivo tissues. Methods Male ICR mice(n=46) were used as the experiment group(n=40) and blank group (n=6). Cotton swabs with C. albicans were used to infect the mice (7 × 106 CFU/mL), and the blank group with saline. The mice of the experiment group were randomly divided into two groups:model group A for clinical assessment (n=20) and model group B for tissue fungal burden detection (n=20). Clinical score, survival and autopsy were carried out among the model group A. Five mice were randomly killed from the model group B at 3 d, 5 d and7 d after infection, respectively ( blank group killed 2 mice each time) . Microbial load tablet method was used to detect the tissue fungal burdens in different tissues, meanwhile samples of tongue, esophagus, stomach, liver, kidney, lung of infected mice were taken for pathological examination. Results White spot appeared on the surface of tongue since 3 d postinfection and increased with time and finally caused death. The mortality reached over 50% at 5 d. C. albicans was not only detected from the tongue (87?5%), stomach (87?5%), liver (54?5%), kidney (50?5%), lung (20%) and heart (4%), but also was microscopically seen mycelia proliferation in the tongue, stomach, liver, and kidney , yet not seen in the control group, showing that C. albicans caused disseminated systemic infection through mucosal infection in mice. Conclusions C. albicans can induce opportunistic systemic infection by breakthrough the mucosal immune barrier, so as to increase the infection to death.
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Excellent open video courses are network video courses and academic forum about science and cultural quality,which set college and university students as service principle and at the same time open freely to the public.Practice has proved that school support is the premise of setting up excellent open video courses,and how well teachers master curriculum contents and design diligent teaching is the basis.What's more,consideration should be given to higher shooting and production technique.Only the sufficient cooperation between the chief course instructor and production crew can guarantee the high quality of open video courses.
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Objective To construct and identify a recombinant Bifidobacteria(Bb) (pGEX-TSO45W-4B) vaccine of Taenia solium.Methods The TSO45W-4B gene of Taenia solium was synthesized.It was expected that the fragment length of the gene was 351 bp.The gene was cloned into Escherichia coli-Bifidobacteria(Bb) shutde vector pGEX-1λT to construct a recombinant plasmid pGEX-TSO45W-4B.The recombinant plasmid was electroporated into Bb to construct a recombinant Bb (pGEX-TSO45W-4B) vaccine.The vaccine was identified by restriction analysis,PCR and sequencing,which would demonstrate that the constructed recombinant Bb (pGEX-TSO45W-4B) vaccine of Taenia solium contained the gene of fragment length of 351 bp.Results The TSO45W-4B gene of 351 bp was synthesized.Restriction analysis,PCR and sequencing results showed that the recombinant Bb (pGEX-TSO45W-4B) vaccine of Taenia solium was successfully constructed and contained the gene fragment of 351 bp.Conclusion The recombinant Bb (pGEX-TSO45W-4B) vaccine of Taenia solium is successfully constructed,which lays a foundation for the study of expression and immunogenicity of the vaccine.
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Objective To study the influence of alprostadil on the serum adiponectin and leptin in patients with early diabetic nephropathy.Methods 98 patients with early diabetic nephropathy were randomly divided into two groups,the control group(n =48 cases) and the treatment group(n =50 cases).The patients of the control group were treated through the conventional treatment.However,the patients of the treatment group were treated plus alprostadil.They were treated for 21 days.The urinary albumin excretion rate (UAER),adiponectin,leptin and insulin resistance were detected before and after treatment.Results The UAER,adiponectin leptin and insulin resistance were improved after treatment,while those of the treatment group were more significantly improved than those of the control group(P ＜ 0.05 or P ＜ 0.01).Conclusion Alprostadil can improve the conditions of early diabetic nephropathy which is through improving the adiponectin and insulin resistance.
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p21 activated kinase 4 (PAK4) is a member of a family of serine/threonine kinase.PAK4plays an important role in a variety of cellular functions including cell cycle,cell proliferation,apoptosis,and cytoskeletal reorganization.Recently,PAK4 has been shown to overexpress in a variety of malignancies,and contribute to cancer cell migration and invasion through multiple signalling pathways.
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Objeetive To investigate the willingness to family care bed service and its influencing factors among the elderly with chronic diseases. Methods A three-stage random sampling method was adopted to carry out face-to-face questionnaire investigation in 652 community elderly patients with chron-ic diseases. Results The willingness to family care bed service was 57.2%.Its influencing factors includ-ed age,educational level,family economic income,activity of daily living,whether or not received treatment within 2 weeks and hospital stay within 1 year,whether or not understood the meaning of family care bed service or received it or similar at home service. Conclusions More than a half of the elderly with chronic diseases were willing to accept family care bed service.The community health service center or hos-pital should attaeh importance to and actively carry out family care bed service.They should enhance propanda work to induct the utilization of family care bed service by edlder people with chronic disease and tackle and satisfy their health requirement.
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Objective To study the relationship between the expression of p53 and its downstream effecter p21~(waf1)protein and multi-drug resistance(MDR) of Tca8113/BLM cell line.Methods The cDNA of wildtype p53 gene was introduced into the drug-resistance cell line Tca8113/BLM by Lipofectamine-mediated transfection.The expression of p53 mRNA in Tca8113/BLM and its parental counterpart Tca8113 cell lines were analyzed using molecular beacons.The expression of p53,p21~(waf1),P-gp,and MRP proteins in Tca8113/BLM cell line transfected by wt-p53 cDNA,the untransfected control Tca8113/BLM cell line,its parental counterpart Tca8113 were analyzed by Western blotting.MTT assay was used to determine the drug-sensitivity of different cell lines.The cell cycle distribution and apotosis of different cell lines were also observed by flow cytometery(FCM) and laser confocus microscope(LCM),respectively.Results The expression of p53 protein was significantly lower,and no p21~(waf1) protein was detected in Tca8113/BLM cells.The protein expression of P-gp and MRP were significantly higher in Tca8113/BLM cells than in Tca8113 cells.By Western Blotting,the expression of p53 and p21~(waf1)proteins was significantly increased,while the expression of P-gp and MRP proteins was significantly decreased in Tca8113/BLM/p53 cells.Comparing to Tca8113/BLM cells,the sensitivity of BLM treatment in Tca8113/BLM/p53 cells was 10.1 fold increased.Tca8113/BLM/p53 cells decreased at G1 and S phase,and increased at G2 phase.LCM results showed that the apoptosis of Tca8113/BLM/p53 cells was more obvious than Tca8113/BLM cells when treated with the drug BLM.Conclusion The multi-drug resistance in Tca8113/BLM cell line is involved in down-regulation of p53 and p21~(waf1) protein levels,and up-regulation of P-gp and MRP protein levels.
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Objective To study the correlation of replication efficiency and antigen expression of hepatitis B virus (HBV) strains in patient sera and transfected cells. Methods Quantification of five maternal serum HBV DNA was carried out using dot hybridization and polymerase chain reaction-enzyme linked immunosorbent assay (PCR-ELISA), hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) levels in the sera were determined by ELISA. Full-length genomes of HBV strains cloned from five pregnant women were separately used to transfect HepG2 cells. The levels of HBsAg and HBeAg in the supernatant of transfected cells were determined with Abbott EIA kits, the replication efficiency of intracelluar replicative intermediates and extracellular HBV DNA of viral particles was analyzed by Southern blot and hybridization. Results Intracellular and extracellular HBV replication and antigen expression of cloned HBV strains showed positive correlation tendency with the HBV DNA and antigen levels in serum samples. Conclusions Virus replication efficiency and expression of HBsAg and HBeAg by full-length HBV clones are in relative coincidence with the biological characteristics of HBV strains in patients. Cell transfection can be used to study the biological characteristics of individual isolates.
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Kunming mice were infected by feeding 150?5 larvae of Trichinella spiralis,established was also a normal control group.Blood was collected from the ophthalmic venous plexus respectively on 7 d,21 d,35 d and 49 d after infection and IL-12 in the serum was detected by ELISA.The level of IL-12 in serum decreased in groups of 7 d,21 d,and 35 d,with a significant difference to the control(P0.05),suggesting that the serum IL-12 of the Trichinella spiralis-infected mice significantly decreased at the earlier stage but approached to normal at a later stage.
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Objective To study the effects of extract from Narcissus tazetta var. chinensis on proliferation and apoptosis of human multiple myeloma cell line ARH-77. Methods The survival rate of cells were tested by MTT assay when cells were affected by 1.25, 2.5, 5, 10, 20 ?g/mL of the extract for 3 d. After being induced by 10 ?g/mL of the extract for 3 d, apoptotic cells were observed with fluorescence stain; changes of he csll cycle were analyzed by Flow Cytometry; the number and shape of nucleolar organizer region (NOR) were tested by AgNOR assay and submicroscopic structure were observed by transmission electron microscope. Results The extract from N. tazetta var. chinensis could significantly inhibit the proliferation and reduce the survival rate of ARH-77 cells (P
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Experimental acute serum sickness was produced in 20 rabbits By a sensitizing injection with bovine serum albumin (BSA) through the auricular vein. Five days after the sensitization, ten of the 20 animals were given a total body irradiation of 300 r from a 60Co sourse. No radiation was given to the other 10 animals which served as controls. Blood samples were taken from the rabbits of both groups before and 2 , 4 , 6 , 8 , 10, 12, 14, and 16 days after antigen injection. After the sera were seperated, the concentrations of the circulating immune complexes were determined with the PEG complement consumption test. It was found that the dynamic curves of the concentrations of the circulating immune complexes of the two groups were essentially similar. This result strongly suggests that total body irradiation of gamma rays given five days after the sensitization of an antigen exerts no influence on the formation of the circulating immune complexes though acute radiation sickness is well established.