Objective To investigate p16 promoter methylation in premature rats with chronic lung disease induced by hyperoxia. Methods Eighty premature Wistar rats were randomly divided into two groups: hyperoxia group (fraction of inspiratory oxygen) 0. 90 and control group (fraction of inspiratory oxygen 0. 21), 40 rats for each group. Semi-nested methylation specific polymerase chain reaction and methylation specific polymerase chain reaction were applied respectively to detect p16 promoter methylation in lung tissues. Additionally, p16 mRNA and protein expressions in lung tissue were detected by reverse transcription- polymerase chain reaction, Western blot and immunohistochemistry method. Results The methylation was not found in control group by seminested methylation specific polymerase chain reaction and methylation specific polymerase chain reaction, while was found in different aged rats of the hyperoxia group. The methylation detection rate was higher by using the semi-nested methylation-specific polymerase chain reaction (52.5%, 21/40) than that by methylation specific polymerase chain reaction (42.5%, 17/40) in the hyperoxia group,but there was no statistically significant difference between the two methods. The p16 mRNA in the hyperoxia group were significantly lower than in the control group at day 7, 14 and 21(1.73 ± 0.40 vs 2.11±0. 37,1.29±0. 19 vs 1.60±0. 27,0. 95±0.25 vs 1.72±0. 34, t=2.19, 2.95 and 10. 43,P＜0. 05). The p16 protein expressions by western blot in the hyperoxia group were significantly lower than in the control group at day 7, 14 and 21 also (88. 1±8. 7 vs 95.0±4.1,65.7±4.5 vs 83. 5±13.6 and 50.4±4.9 vs 86.7±11.9, t=2.27,3.95 and 13.40,P＜0.05). The expression of p16 mRNA (1.06±0.61) and protein (62.32±25.65) in lung tissues of rats with methylation was lower than that without methylation (1.63±0.62 and 94.93±22.21, respectively) (t=2.95, OR=0. 86;t=4.28, OR=0. 85,P＜0.01, respectively). Conclusions Exposure to hyperoxia might induce p16 promoter methylation in lung tissues in premature rats. Methylation risk increases as exposure time extends. p16 promoter methylation induced by hyperoxia might participate in the mechanism of lowering p16 mRNA and protein expression, but might not result in p16 gene silence.

Inositol,also called cyclohexanehexol,is a water-soluble Vitamin (Vitamin B8),which is widely distributed in plants and animals.It is an essential and indispensable nutrient for human and animals to maintain normal physiological functions with multiple bioactivities.Inositol can promote the maturity of a variety of ingredients in pulmonary surfactants,participating in the regulation of cell growth and surviving,biofilm formation,as well as the information transmission processes of a variety of signaling molecules.Therefore,inositol plays a key role in the growth and development of children and is closely related to pediatric developmental diseases.In order to provide a new way of thinking for prevention and treatment of inositol and its molecular mechanisms in pediatric developmental diseases,this article reviews the relationship between inositol and pediatric clinical diseases and its research progresses combined with the results of our laboratory studies.

Objective To investigate the frequency distribution of proprotein convertase subtilisin/kexin 9 (PCSK9) gene I474V polymorphisms and their relationship with patients with ischemic stroke (IS)of Uygur and Han ethnic groups in Xinjiang Uygur Autonomous Region.Methods The I474V polymorphism was identified by polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) in 407 patients with IS(including 219 Hans and 188 Uygurs)and 425 health controls (including 255 Hans and 170 Uygurs),and some specimens were sequenced.Results (1) Between IS group and control group,the genotypes Ⅱ and Ⅳ had no statistically significant differences in the levels of triglycerides (TG),high-density lipoprotein cholesterol (HDL-C) ; Total cholesterol (TC),low-density lipoprotein cholesterol (LDL-C) levels had statistically significant differences; LDL-C levels had also statistically significant differences.Between IS and control groups,TC,LDL,HDL-C levels of genotype Ⅱ showed statistically significant difference.In the IS group,TC,LDL-C levels of Ⅳ genotype were significantly higher than the control group,the difference being statistically significant.(2) There was statistically significant difference in the genotype distribution between IS and control groups (9.5％ (77/814) vs 4.5％ (38/850),x2 =16.09,P =0.000).And the distribution of allele frequency was statistically different (18.9％ (77/407) vs 8.9％ (38/425),x2 =17.38,P =0.000).(3) The differences of I474V loci Ⅳ genotype frequency distribution in Xinjiang Uygurs and Hans were statistically significant (27.7％ (52/188) vs 11.4％ (25/219),x2 =17.40,P =0.000; 12.9％ (22/170) vs 6.3％ (16/255),x2 =5.57,P =0.018) ; So did the Ⅴ allele frenquency distribution (13.8％ (52/376) vs 5.7％ (25/438),x2 =15.58,P =0.000; 6.5％ (22/340) vs 3.1％ (16/510),x2 =10.44,P =0.001).(4) There was statistically significant difference in the genotype distribution and allele frenquency distribution between IS group and control group in the Xinjiang Uygurs (27.7％ (52/188) vs 12.9％ (22/170),x2 =11.79,P =0.001 ; 13.8％ (52/376) vs 6.5％ (22/340),x2 =10.44,P =0.001) ; But there was no statistically significant difference in the Hans.Conclusions Ⅱ and Ⅳ genotypes are dominant in the I474V polymorphism loci of PCSK9 gene.The genotype of PCSK9 gene I474V polymorphism is correlated with increasing serum levels of TC and LDL-C.I474V polymorphism is associated with cerebral IS course in Xinjiang region.There is statistically significant difference in the genotype I474V distribution between Uygur and Han groups.I474V polymorphism has a relationship with the occurrence of IS in Xinjiang Uygurs.Ⅳ may be a susceptible genotype and Ⅴ may be a genetic susceptible allele of the Xinjiang Uygurs.

Objective To evaluate the therapeutic effect of intravenously use of ambroxol in treatment of mechanical ventilator patients.Methods Meta-analysis was used.The Cochrane Library,PubMed,Elsevier (ScienceDirect),OVID series medical databases and domestic main databases were searched,the quality of included studies was evaluated by certain standards.The Review Manager 5.0 software was used for analysis.Results 14 studies were included,1 184 mechanical ventilation patients were involved.The Meta-analysis showed that there were significant differences between two groups in blood gas analysis,airway resistance and pressure and lung compliance.However,there were no differences between two groups in expectorant effect and VAP incidence.Conclusions Intravenously use of ambroxol in treatment of mechanical ventilator patients was effective.A large multi-center sample of high-quality clinical studies is needed to confirm the results.

Objective: To construct a phage display library of human single-chain Fv antibodies against blood group Rh(D) substance. Methods: Combining phage display library techniques, isolated total RNA from B lymphoblastoid cell lines secreting anti-Rh(D) antibodies was used for the synthesis of the first strand of cDNA, V_ H and V_ L genes were amplified by 2nd PCR and linked together by splicing overlap extension (SOE) with the use of a (Gly_ 4Ser)_ 3 linker. The resulted scFv genes were then cloned into pCANTAB5E vectors and displayed on the phage. Phage clones were selected using intact red cells as a source of antigen. After 4 rounds of "binding-elution-enrichment", each clone was assayed for specificity by Dot ELISA. Results: A phage antibody library, with the sink size being 1.2?107, was obtained. The percentage of full-length scFv gene inserted into phage DNA was 0.80. Rescued by helper phage, a phage scFv library with titer of 3?108 pfu/ml was established. Specific phages with scFv were acquired after 4 rounds of panning, one clone exhibiting specific binding to Rh+ cell was identified by Dot ELISA. Conclusion: A strategy for construction phage antibody library by means of phage display technique was practicable, which would be useful in screening engineered antibodies against human Rh (D) blood group substances.

OBJECTIVE:To provide reference for the medication safety in elderly patients with chronic diseases. METHODS:Elderly inpatients with chronic diseases and nursing staff in affiliated hospital of shaoyang medical college,from Mar. 2014 to May 2015 were randomly selected for a questionnaire survey to analyze the influential factors of drug safety and health education de-mand,and the difficulty sources of nursing staff in developing health education was explored. RESULTS:Totally 500 questionnaire was sent out to elderly patients with chronic diseases,500 were effectively received with effective recovery of 100%;and totally 200 were sent out to nursing staff,200 were effectively received with effective recovery of 100%. In the 500 surveyed patients, 193 had ADR,which was affected by age,educational background,monthly income,disease course,whether received health edu-cation,etc.(P<0.05),patients with older age,lower educational background and monthly income,longer disease course and no receiving health education showed higher incidence of ADR;the influential factors for health education demands included education-al background,occupational status,disease course,quality evaluation of nursing staff,etc.(P<0.05),patients with higher educa-tional background,lower disease course,higher occupational status and quality evaluation of nursing staff showed stronger health education demands. The difficulty sources of surveyed nursing staff in developing health education were busy work,being afraid of misunderstanding,embarrassing,cognitive and skill deficits. CONCLUSIONS:The drug safety of elderly patients with chronic dis-eases is related to many influential factors,as well as the health education demands. Nursing staff should continuously improve their ability and quality,combined with the above influential factors,carry out health education with physicians and pharmacists to ensure the medication safety.

Objective: To develop Epstein-Barr virus(EBV)-transformed human peripheral blood B cell lines from healthy volunteers of type B.Methods: B lymphocytes from healthy volunteers of type B capable of producing anti-A antibodies were transformed by EBV,while Cyclosporine A(CsA),as an immunosuppressive agent that could selectively inhibit T-lymphocytes and protect B-lymphocytes from regression,was used in the experiment.Then,the supernanant of the cell culture medium was tested with red blood cells of type A,B and O by agglutinin assay.Results: Twelve of the15 EBV-transformed B lymphocyte cell lines were established,with a success rate of 80%,while 9 human B lymphocyte lines secreted anti-A antibodies.Conclusion: Human B lymphocyte lines secreting antibodies to A antigens were successfully developed,which helps further studies of human blood specific monoclonal antibodies.

目的探讨系统功能训练对老年人工全膝关节置换(TKR)术后康复的效果观察。方法接受TKR老年患者21例36膝,入院后随机分A、B两组,A组实施系统功能训练,B组采用骨科常规护理。采用美国特种外科医院(HSS)膝关节评分标准在术前及术后3周末对A、B两组进行膝关节评分。结果两组患者TKR术后膝关节评分有显著差异(P<0.05)。结论系统功能训练有利于促进老年TKR术后康复。

Objective To investigate the pattern of the cerebral white matter lesions in patients with mild and moderate Alzheimer's disease(AD)and healthy controls using proton magnetic resonance spectroscopy(~1H-MRS)and diffusion tensor imaging(DTI).Methods Twenty AD patients and Twenty healthy controls were recrnited.All subjects underwent clinical examination,neuropsychological assessment.The quantitative analysis of N-acetylaspartate(NAA),myoinositol(mI),Chotine(Cho)and Creatine(Cr)resonance signals in region of interests(ROIs)located in the paraventricular white matter region bilaterally were measured.Ratios of NAA/Cr,mI/Cr and Cho/Cr were calculated in two groups.In addition,conventional MRI and DTI scanning were received,fractional anisotropy(FA)and mean diffusivity(MD)values of white matter in the same regions were measured respectively.Results No significant difference between two groups were observed in NAA/Cr ratio(P＞0.05).A significantly increased mI/Cr and Cho/Cr were found in AD patients than in controls(P＜0.05).FA and MD values in AD patients were 0.470±0.082 and 0.771±0.099,and in controls were 0.539±0.068 and 0.691±0.064,respectively.FA value decreased significantly in AD patients(P＜0.05),M D value increased significantly in AD patients(P＜0.05).After controlling for age-related,partial correlation analysis revealed a negtive correlation between mI/Cr and FA value in the patients with AD(P＜0.05).No correlation between mI/Cr and MD was found(P＞0.05).Conclusion The results suggest that not only the gray matter is injured,but also the white matter is abnormal in AD patients.Combining ~1H-MRS with DTI alterations could provide the valuable informations about white matter lesions in AD patients.

BACKGROUND: An ideal marked molecule has not been found to detarmine bone marrow mesenchymal stem cells (MSCs) so as to make sure the homogenicity.OBJECTIVE: To verify the in vitro differentiation from BrdU-treced MSCs into osteoblasts.DESIGN, TIME AND SETTING: A cytological observation/n vitro was performed in Shanxi Medical University from January to October 2008.MATERIALS: Wistar rats aging 4 weeks old were provided by Experimental Animal Center of Shanxi Medical University.METHODS: MSCs were isolated and cultured by using density gradient cantrifugation combined with attachment culture method.At about 80% confluence, trypsin was used for passage and amplification. MSCs at density of 5×1010/L were inoculated in a 25-mm culture dish with L-DMDM culture medium containing dexamethasone, β -phosphoglycarol, vitamin C, and 10% fetal bovine serum. The third-passaged MSCs were labeled in vitro with 10 μmol/L BrdU tracer. Thereafter, 10 visual fields were randomly selected to calculate numbers of positive and negative ceils so as to obtain BrdU tracing rate under a fluorescence microscope (×200).MAIN OUTCOME MEASURES: Inverted microscope was used to observe cell morphology; flow cytometry was used to detect cell surface antigen, differentiation into osteoblasts, and BrdU tracing rate in vitro.RESULTS: The purified MSCs which were like fibroblasts were adherent and fusiform. The third-passaged cells were changed equidirectionally and whirlpool-arranged, and the survival rate was more than 95%. The seventh-passaged cells still grew rapidly.CD44, CD71, and CD105 expressions were positive, but CD45 expression was negative. Black particles were visualized in MSCs after Von kossa staining. BrdU tracing rate was more than 90%.CONCLUSION: Density gradient centrifugation combined with attachment culture method can effectively isolate and purify rat MSCs which are cultured in vitro for a long period and differentiated into osteoblasts. BrdU tracer is safe, effective, and convenient to successfully label MSCs.