BACKGROUND:In mammalian cells,introduction of double-stranded small interfering RNA(19-25 bp)can cleave and destroy the cognate RNA,which can result in suppression of gene expression.OBJECTIVE:To construct siRNA expression plasmid for interference angiotensinogen(AGT),thereby,to resist AGT expression in adipose cells.METHODs:The mRNA sequence of AGT gene was searched from NCBI(NM000029).Utilize of GenScript siRNA technology,AGT-siRNA oliaonucletides were chemically synthesized and inserted into pRNAT-U6 1/Neo vector after annealing,then transformed into TOP10.The recombinant plasmid was identified by restriction endonuclease and DNA sequencing.RESULTS AND CONCLUS1ON:The recombinant plasmid psiRNAT-U6.1/Neo-AGT was obtained by connecting 19 bp segment containing AGT-mRNA sequence to pRNAT-U6.1/Neo After EcoR Ⅰ and Hind Ⅲ digestion.351 bp segment was obtained from empty vector.and 397 bp fragment band was obtained form recombinant plasmid,which was coincidence to the expectation.DNA sequencing showed Targeting siRNA oligonucleotides were correctly inserted into the eukaryotic expression vector pRNAT-U6.1/Neo without base mutation.The interference vector psiRNAT-U6.1/Neo-AGT was successfully constructed.

BACKGROUND:Studies have shown that methylation modification using CARM1-catalyzed histone H3R17/R26 can maintain the stemness of embryonic stem cel s. However, mechanism underlying CARM1 effect on the stemness of amniotic fluid-derived stem cel s is stil unclear.
OBJECTIVE:To investigate the function and underlying molecular mechanism of CARM1 to maintain stemness in the amniotic fluid-derived stem cel s.
METHODS:Amniotic fluid-derived stem cel s from term pregnancy were isolated and cultured. RT-PCR was used to identify the stem cel mark and CARM1 gene expression. CARM1 expression in amniotic fluid-derived stem cel s was knocked down by using two shRNA. RT-qPCR was used to detect the silencing efficiency, and western blot employed to examine the methylation level of Arginines 17 at N terminus of histone 3 (H3mR17). Moreover, the expression of embryonic stem cel markers, including OCT4, SOX2 and NANOG, were detected.
RESULTS AND CONCLUSION:Amniotic fluid-derived stem cel s from term pregnancy could express CARM1 and stem cel markers, including OCT4, SOX2, Nanog and KLF4. Both of the shRNAs could knock down the expression of CARM1 efficiently. When CARM1 was knocked down, the H3mR17 level was decreased and OCT4, SOX2 expression was also reduced, but NANOG expression had no change. Al these indicate that CARM1 is required for amniotic fluid-derived stem cel s to maintain stemness through regulating OCT4 and SOX2 expression.

OBJECTIVE:To observe the clinical efficacy and safety of Enalapril folic acid tablet combined with Shenkang injec-tion in the treatment of hypertensive nephropathy. METHODS:90 patients with hypertensive nephropathy were randomly divided in-to control group and observation group. All patients were given anti-infection,regulating electrolyte balance,supplying amino acids and other conventional treatment. Based on it,control group was orally given Maleic acid enalaprilat folic acid tablet 1 tablet in the morning,once a day;observation group was additionally given 100 ml Shenkang injection adding into 300 ml 10% Glucose injec-tion by intravenous infusion,20-30 drops/min,once a day. The treatment course for both groups was 4 weeks. The clinical effica-cy,and clearance rate of creatinine(Ccr),serum creatinine (Scr),morning urine test urine protein (Up)/urine creatinine (Ucr), serum malondialdehyde(MDA),superoxide dismutase(SOD),total antioxidant capacity(T-AOC)before and after treatment and incidence of adverse reactions in 2 groups were observed. RESULTS:The total effective rate in observation group was significantly higher than control group,the difference was statistically significant(P<0.05). After treatment,the Ccr,SOD and T-AOC in 2 groups were significantly higher than before,and observation group was higher than control group,Scr and MDA were significant-ly lower than before,and observation group was lower than control group,the differences were statistically significant(P<0.05). Morning urine test Up/Ucr,systolic blood pressure and diastolic blood pressure were significantly lower than before,and the differ-ences were statistically significant(P<0.05),however,there was no significant difference between 2 groups(P>0.05). And there was no significant difference in the incidence of adverse reactions(P>0.05). CONCLUSIONS:Based on the conventional treat-ment,Enalapril folic acid tablet combined with Shenkang injection has better efficacy than only Enalapril folic acid tablet in the treatment of hypertensive nephropathy,with similar safety.

Objective: To study the clinical effect of nursing after laparoscopic myomectomy after nursing intervention. Methods: Retrospective analysis of our hospital from 2009 May to 2012 October in our department, laparoscopic myomectomy in the treatment of 148 cases of uterine fibroids patients clinical data, were randomly divided into 2 groups, 74 cases in each group, the control group routine nursing care to patients, nursing intervention on the patients with study group, The quality of life of patients with depression and anxiety scores were compared between the two groups before and after the rate and nursing of exhaust and ambulation time, hospitalization time, complications. Results: Research group of exhaust, ambulation, time of hospitalization, the incidence of complications compared with the control group, showed significant difference; the study group before operation and nursing of patients with anxiety and depression and quality of life scores compared with the control group, there was no significant difference, nursing intervention, there were significant differences. Conclusion: Laparoscopic myomectomy nursing intervention, can promote the rehabilitation, and improve the living condition, patients with significant effect, should be popularized.

Biofilm formation makes bacteria adopt a multicellular lifestyle.Compared to planktonic cells, biofilm-grown cells express an increased resistance to antimicrobial agents.Bacterial biofilm formation is a crucial step in the pathogenesis of many subacute and chronic bacterial infections.In clinical setting, biofilm can cause bacterial biofilm disease and biomaterial associated infection.Understanding the mechanisms involved in biofilm-associated antimicrobial resistance is key to the development of new solutions.

Leukocytes from peripheral blood of 70 normal Chinese,40 males and 30 females, were cultured in vitro according to a modification of Moorhead's method.Chromosome studies were carried out.Table I lists the results obtained including age group,metaphase plates counted and analysed and the chromosome number of each cell.92.4 per cent of 8031 counted cells had the diploid number(2n=46).Chromosome measurements were made from camera lucida drawings.Measurements were estimated to the nearest 0.1 mm on a fine metric scale.The relative length,arm ratio and centromere index of each chromosome were calculated from measurements of 18 haploid chromosome sets of 11 metaphase plates in 6 males and 4 females.The averages,range of variation and 99% confidence limits are listed in Table Ⅲ.Systematic studies of individual chromosomes under the light microscope revealed the following morphological characteristics.The short arms of all 10 acrocentric chromosomes from groups 13—15 and 21—22 could be shown to possess satellites,but in general only 2—8 of these chromosomes at a time carried satellites.In many cells the satellite-carrying chromosomes tended to associate with each other through their satellites.The number of chromosomes involved in each association varied between 2 to 5,occasionally 6.It was not rare to find secondary constrictions to occur regularly in certain regions of chromosomes 1,4,9,13,and 16.Attachments of satellites to such regions of chromosomes 1,4,9,and 13 were also observed.Karyotypic analysis of 357 metaphase plates led us to conclude that in most cases the unequivocal identification of chromosome pairs 4—5,6—12+X,13—15,19—20,and 21—22 was im- possible.Accurate characterisation of these chromosome pairs will have to await further technical advances. The length of the Y chromosomes was systematically measured in order to deter- mine its variability and heritability.Among our sample of 40 male subjects 29 had a Y chromosome slightly larger than chromosome 21,8 slightly smaller or larger than chromo- some 18(long Y chromosome),and 3 smaller than chromosome 21(short Y chromo- some).The heritability of the size of the Y chromosome was established(Fig.Ⅳ). “Spontaneous”aberrations were discovered in 34 of 70 cases.Aberration rates varied from 0.88 to 13 per cent.The relation between aberrations and radiation injuries was briefly discussed.

OBJECTIVE To cultivate central supply unit member's custom of watching detail and dealing with everything carefully. To enhance central supply unit work's quality and efficiency,and decrease the phenomenon of article sterilized overdue and nipping tool's scathe. METHODS To organize study and enhance the cognition. To constitute regulations,systems and operation rule. To investigate the condition of medical instrument utilized,and pay attention to detail service for packaging sterilization. RESULTS The rate of overdue sterilized article decreased from 0.008% to 0.001%. The number of nipping tool's injuries decreased from 18 in 24 to 2 in 22 when the detail management was actualized. The rate of injury decreased from 75.0% to 9.09%. CONCLUSIONS In order to ensure the quality of asepsis of articles,we need to alter our conceptions,strengthen our responsibility,servicing more initiatively and carefulness.

Objectives To summarize the experience of nursing care to children with dermatomyositis complicated with interstitial pneumonia.Methods The nursing histories of 11 cases of children dermatomyositis complicated with interstitial pneumonia were retrospectively looked into for summarizing the nursing measures.Results The diseases were both effectively controlled in all the children,who were given maintenance therapy by oral taking of hormone after discharge.Conclusion Such nursing measures as close observation of disease,enhanced nursing and first aid of respiratory failure,nursing of infection and secondary infection,care after mesenchymal stem cells transplantation,strengthened disinfection and isolation,are of significance for controlling the disease.

Objective To study the clinical characteristics and treatment of functional articulation disorders. Methods 65 children withfunctional articulation disorders were reported. Results The main erroneous articulation pattern in these children was substitution, followedby omission and/or distortion. They were trained from oral function and articulation motion for 2~6 months. 58 children were corrected completely,and 7 were corrected partially. Conclusion The main erroneous articulation pattern of functional articulation disorder in children issubstitution. Speech training is effective on it.

Objective To investigate the relationship between Methotrexate (MTX) and its cognitive dysfunction,and to explore the possible mechanism of neurotoxicity induced by MTX.Methods Thirty healthy male SD rats weighting 180-220 g were divided into 3 groups using random number table:control group,60 mg/kg MTX (MTX60) group and 100 mg/kg MTX (MTXt00) group,with 10 rats in each group.The rats in MTX60 group,MTX100 group received 60 mg/kg MTX and 100 mg/kg MTX,respectively.The rats in control group accepted the same volume of 9 g/L saline injection as MTX group.Spatial memory of rats was evaluated by using Morris water maze test at different time points after pretreatment with MTX.After the Morris water maze test,the hippocampus were harvested and the expressions of C/EBP homologous protein (CHOP),cysteinyl aspartate specific proteinase 12(caspase-12) and cleave cysteinyl aspartate specific proteinase 3 (cleaved caspase-3) were detected by using Western blot.Meanwhile,cell apoptosis and pathological change of hippocampal neurons were detected by terminal dexynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay and HE staining respectively.Results In the Morris water maze test,the time in platform quadrant of rats in MTX60 group and MTX100 group was shorter than that of rats in control group during probe training [(27.30 ±3.98) s and (21.63 ±4.22) s vs (33.30 ±6.31) s,F =13.94,P ＜0.05],and the time in target quadrant of MTX100 group was shorter than that of MTX60 group (P ＜ 0.05).Compared with the control group,there were degenerated neurons in hippocampus cornu ammonis 1 (CA1) area in MTX60 group and MTX100 group.The number of TUNEL-positive cells of the hippocampus CA1 area increased significantly in MTX60 group and MTX100 group rats [(4.72 ±0.12)％ and (9.12±0.12)％ vs (1.11 ±0.49)％,F=95.272,P ＜0.05],and the TUNEL-positive cells of rats in MTX100 group were more than those of MTX60 group (P ＜ 0.05).The expressions of CHOP,caspase-12 and cleaved caspase-3 also increased compared with the control group (CHOP:2.98 ±0.31 and 4.15 ±0.61 vs 0.38 ±0.12,F =232.74,P ＜ 0.05;caspase-12:0.33 ±0.04 and 0.43 ±0.06 vs 0.14 ±0.02,F =120.70,P ＜ 0.05;cleaved caspase-3:0.35 ± 0.04 and 0.44 ± 0.06 vs 0.05 ± 0.03,F =198.64,P ＜ 0.05),and the protein expression levels of rats in MTX100 group were higher than MTX60 group (all P ＜ 0.05).Conclusions MTX can induce cognitive impairment in rats,and endoplasmic reticulum stress mediated hippocampal neurons apoptosis may play an important role in the mechanism of MTX-induced cognitive impairment in rats.