Objective To provide an experimental evidence for the clinical applications of glyceryl trinatrate(GTN)and isosorbide dinitrate(ISDN)against Coxsackievirus B(CVB)-related myocarditis.Methods Coxsackievirus B3 was propagated in HeLa cells.Virus yields were determinded by 50% tissue culture infective dosage(TCID50).BALB/c mice were attacked with 5000 TCID50 of CVB3,meanwhile,the mice were administrated with GTN and ISDN.All mice were killed at the 14th day.The myocardial tissues were harvested for histologic evaluation.Results The infection plaques in the myocardial tissues obtained from CVB3-infected BALB/c mice treated with GTN were siginificantly reduced(0.89?0.18 in GTN group and 1.25?0.22 in ISDN group)compared with that of the untreated CVB3-infected mice(P

Objective To construct a ciaR gene-knockout (ΔciaR) mutant of Streptococcus pneu-moniae ( S. pneumoniae) and to investigate the effects of CiaR in CiaH/CiaR, a streptococcal two-component signal-transducing system, on the expression of genes encoding penicillin-binding proteins ( pbps genes) and cia-dependent small RNAs (csRNAs). Methods Electrophoretic mobility shift assay (ESMA) was per-formed to detect the recombinant CiaR (rCiaR)-binding pbps genes. A suicide plasmid pEVP3ciaR for ciaR gene knockout was constructed and then aΔciaR mutant was obtained through homologous recombination and insertion inactivation of the suicide plasmid, and screening with chloromycin. The mutant was identified using PCR and sequencing analysis. E-test was used to detect the minimal inhibitory concentrations ( MIC) of penicillin ( PCN) and cefotaxime ( CTX) against S. pneumoniae strains. Changes and differences in the expression of pbps genes and csRNAs in theΔciaR mutant and its wild-type strain before and after treatment with 1/4 MIC of PCN or CTX were detected using real-time quantitative RT-PCR. Results The rCiaR could bind to the promoter regions in pbp1a, pbp1b and pbp2b genes of S. pneumoniae. The ciaR gene in ΔciaR mutant was inactivated by insertion according to the results of PCR and sequencing analysis. After treatment with 1/4 MIC of PCN or CTX, the expression of pbps genes at mRNA level ( pbps-mRNAs) in theΔciaR mu-tant was significantly increased (P<0. 05), but the levels of csRNAs were significantly decreased (P<0. 05);whereas a significantly decreased pbps-mRNAs (P<0. 05) and increased csRNAs (P<0. 05) were observed in its wild-type strain. The result of E-test showed that the MICs of PCN and CTX against ΔciaR mutant were increased by 250-fold as compared with those against its wild-type strain. Conclusion The CiaR can enhance the drug resistance of S. pneumoniae to PCN and CTX through down-regulating the expres-sion of PBP1a, PBP1b and PBP2b and up-regulating the expression of csRNAs to inhibit the expression of PBPs.

Objective:To investigate the KRAS gene mutation features in lung and gastric cancers and their relationship with clinicopathologic characteristics. Methods:A total of 128 lung cancer and 115 gastric cancer patients were included. Polymerase chain reaction amplification and DNA sequencing were conducted to detect mutations in exon 2 of the KRAS gene. Results:The mutation frequency of KRAS was different in lung and gastric cancers;however, it did not show any statistical significance (6.3%vs. 4.3%, P>0.05). The KRAS codon 12 gene mutation ranks the first in both types of cancer. No significant correlation was observed between the prevalence of KRAS mutations and patient's age and gender. KRAS gene mutation rate was higher in lung adenocarcinoma than in non-adenocarcinoma, such as squamous cancer (10.7%vs. 0%, P<0.05). Conclusion:No correlation was found between the KRAS gene mutation and the sex and age of lung and gastric cancer patients in Jiangsu Province. The rate of KRAS mutation was low. KRAS gene mutation rate was relatively higher in lung adenocarcinoma patients;thus, the mutation status of the KRAS gene should be evaluated be-fore undergoing EGFR-TKI therapy.

microRNAs is a group of small non-coding RNAs that play a negative regulation role in expression of target genes at post-transcriptional level by inhibition or degradation of target mRNAs after combination of the seed sequence (SS) in microRNAs with the SS-binding sequences usually located at 5'ends of target mRNAs.microRNAs was firstly found in Caenorhabditis elegans.Subsequently,many different microRNAs in eukaryocytes were revealed.In eukaryocytes,microRNA precursors are transcribed at first and then become functional microRNAs with 21-23 nt in size after splice.Most of eukaryocytic microRNAs combime with the sequences at 3'end of target mRNAs that cause the translation inhibition or degradation of the mRNAs.In the recent years,many different prokaryocytes,such as bacteria,have been confirmed to possess microRNAs.However,the microRNAs in prokaryotes such as bacteria are 50-400 nt in size and have the biological activity without splice.Moreover,the characteristics,action sites and mechanisms of the prokaryotic microRNAs have some certain diversity compared to the eukaryotic microRNAs.Our review briefly introduce the major regulation mechanisms of gene expression as well as the general characteristics of microRNAs and their regulation mechanisms of gene expression in prokaryocytes and eukaryocytes,which will provide a basis for further and profound study on the gene expression regulation and pathogenic mechanisms of prokaryotic microbial pathogens.

Objective To investigate the correlation between Streptococcus pneumoniae (S.pneumoniae) StkP kinase and drug resistance and to analyze the binding ability of StkP extracellular region (EC-StkP) to β-lactam antibiotics.Methods A stkP gene knockout (ΔstkP) mutant was constructed from S.pneumoniae strain ATCC6306 by insertional inactivation method.E-test was performed to detect the minimum inhibitory concentrations (MIC) of penicillin (PCN) and cefotaxime (CTX) against ΔstkP mutant and its wild-type strain.Bioinformatic softwares were used to predict the EC-StkP of S.pneumonia strain ATCC6306,to generate the three-dimensional structure model of EC-StkP and to analyze the correlation between the structure and functions of EC-StkP.PCR was performed to amplify the extracellular segment of stkP (EC-stkP) gene and the product of it was sequenced after T-A cloning.A prokaryotic expression system of EC-stkP gene was constructed.SDS-PAGE in combination with a gel image analysis system was used to detect the expression of the recombinant EC-StkP (EC-rStkP).The expressed EC-rStkP was extracted by Ni-NTA affinity chromatography.The binding abilities of EC-rStkP to PCN and CTX were detected by isothermal titration calorimetry (VT-ITC) and surface plasmon resonance (Biacore).Results S.pneumonia strain ATCC6306 was sensitive to PCN (MIC=0.06 μg/ml) and CTX (MIC=0.12 μg/ml),but its ΔstkP mutant was resistant to the two antibiotics (PCN MIC=16 μg/ml,CTX MIC=32 μg/ml).The 295 aa segment was predicted as the extracellular region at C-end of StkP of S.pneumoniae strain ATCC6306,containing four penicillin-binding proteins and Ser/Thr kinase-associated (PASTA) domains.The cloned EC-stkP segment and the EC-stkP segment in GenBank shared 99.6% similarity in nucleotide sequence and 100% in amino acid sequence.The constructed prokaryotic expression system for EC-stkP gene expressed EC-rStkP in soluble form.Both PCN and CTX could bind to EC-rStkP and CTX was better than PCN in term of binding ability.Conclusion The stkP gene of S.pneumonia is closely related to drug resistance and the encoded protein,Ser/Thr kinase StkP,can recognize and bind to β-lactam antibiotics.

Abstract The integration of health education into physical education in colleges is an important carrier to achieve the goal of school health education, and an inevitable requirement for improving health literacy of college students. Exploration and construction of one curriculum content system integrating health knowledge and sports skills, mental health and sports spirit, basic theory of physiology, sports injury and disease prevention. The integration of health education into physical education includes curriculum and material design, classroom based teaching process, as well as teaching assessment inside and outside the classroom, developing a multi faceted, multi level and multi angle optimized implementation approach, aiming to explore the joint connection of health education and physical education, the implementation of Lide Shuren can better achieve the cultivation of students health awareness and healthy lifestyle, and lay a solid foundation for lifelong health.

colon cancer and to find out treatment and prevention by using different gene changes.

Objective To explore the relationship between the abnormal expression of anti-oncogene PTEN in gastric carcinoma and the clinicopathological characteristics. Methods Mythylation specific PCR was applied to detect the expression of PTEN methylation in gastric carcinoma and their normal tissues from 45 patients. Results The methylation took place in 40.0 percent in gastric carcinoma and 2.2 percent in their normal tissues, and the difference was significant(P<0.05). The methylation rate in poorly differentiated adenocarcinoma was 60.0 percent ,while 15.0 percent in highly-moderately differentiated group, the difference was significant (P<0.05). In the 24 cases with lymph node metastasis, mythylation was observed in 13 cases, and the difference was significant (P<0.05). Conclusion The methylation of PTEN gene was associated with gastric carcinoma, it may play an important role in the development of the disease.

Objective To explore the expression and effect of MP0 and TGF-β1 in ulcerative colitis tissues.Methods The expressions of MPO and TGF-Bl in colonal tissue specimens were determined by immunohistochemistry(two step method)in 37 cases of ulcerative colitis,10 crohng disease and 20 normal mucosa.Results The positive rates of MPO and TGF.B1 were 10％and 20.0％in the normal mucosa,55.0％and 35.O％in crohn's disease,and 89.2％and 86.5％in ulcerative colitis.Expression of MPO and TGF-β1 in the ulcerative colitis were higIIer than that in crohn's disease and normal mucosa(P<0.05),there was correlation between MPO and TGF-β1(r=O.51,P<0.05).Conclusion MPO and FGF-β1 are involved in the pathology of UC,and detection of MPO and TGFβ1 can provide some value to evaluate the conditions of UC.

Objective To analyze the correlation of traditional Chinese medical constitution types withcardiovascular risk factors in community hypertension patients. Methods We defined the constitution types of 1108 community hypertension patients, created personal health archives and provided routine examination for the subjects. After that, we input and analyzed the relevant data and results by computer. Results In 1108 cases of hypertension patients, 229 (20.67%) were defined as harmony physique, 879 (79.33%) as biased physique. In patients with biased physique, phlegm-dampness physique accounted for 25.54% (283 cases) , and then came yin deficiency physique and Qi deficiency physique, which accounted for 15.16% (168 cases) and 13.18% (146 cases) , respectively. The biased physique was dominated by single biased physique with a total of 590 cases ( 67.12%) , but the complex physique was also in large quantity, with a total of 289 cases ( 32.88%) . The complex physique consisting of two kinds of single physique was commonly seen in the patients with complex physique, accounting for 15.70%. The complex physique consisting of more kinds of single physique had less percentage. Phlegm-dampness physique was a kind of physique involving the most cardiovascular risk factors. Patients with phlegm-dampness physique had higher incidence of overweight, abnormal lipid level, family history of cardiovascular disease, lack of physical exercise and electrocardiographic abnormality, and the difference was statistically significant compared with other physique types ( P<0.05 or P<0.01) . Conclusion Phlegm-dampness, yin deficiency and Qi deficiency physique are the three commonly-seen constitution types in hypertension patients. Phlegm-dampness physique is the most commonly-seen biased physique and is also a kind of physique involving the most cardiovascular risk factors. Prevention and management in consideration with the constitution characteristics and cardiovascular risk factors will be a new prospective method for the treatment of hypertension patients in community.