In this paper, published by COSOInternal Control-integrated framework Theory and the actual situation in China's state-owned hospitals, the hospital selected the elements of the control environment for the study, respectively, from the organizational structure, the values of the main staff, the hospital management and operation of the concept of style, the hospital audit department, as well as hospitals and other human resource policies in terms of five of the impact of the internal control system analysis, and made light of the actual current state-owned hospitals in the above-mentioned aspects of the prevailing problems and gives some solutions.

Objective To study the different mRNA expressions of big and small conductance calcium activated potassium channels (Bkca and Skca), and calcium activated chloride channels (Clca) in normal and instable detrusors. Methods Model of bladder outlet obstruction (BOO) of female Wistar rats was prepared by ligating the proximal urethra in the perineum. Detrusor instability (DI) was confirmed by conscious cystometry. The mRNA was extracted from the detrusors of normal and DI rats for the detection of the expressions of Bkca, Skca2, Skca3, and Clca by RT PCR. The different channel expression between normal and instable detrusors was identified by gel imaging. Results The incidence of DI in BOO rats was 76.17%. Bladder capacity and the maximal detrusor pressure increased significantly ( P

Objective To study the modulation alterations of calcium activated potassium channel (Kca) and calcium activated chloride channels (Clca) on detrusor instability(DI) and their roles in DI. Methods The detrusor instability (DI) animal model was established using female Wistar rats. Detrusor strips were obtained from normal and DI rats. The contraction frequency and amplitude of the strips were measured by an in vitro tension test. The actions of channel openers and blockers were studied. Results Spontaneous contraction frequency and amplitude of DI increased significantly. The block of big conductance calcium activated potassium channel(Bkca) by Iberiotoxin resulted in decrease of frequency but increase of amplitude in the control, but only increase of frequency in DI. The opener, NS1619, resulted in decreases of frequency and amplitude in the control, but only decreases of frequency in DI. The block of small conductance Kca (Skca) by apamin resulted in increases of frequency and amplitude in both control and DI groups, but the opener, Chlorzoxazone, resulted in decreases of frequency and amplitude in the control, but only decrease of frequency in DI. The contraction alterations of DI were significantly lower than those of the control no matter Kca was blocked or opened. NFA, a selective Clca blocker, resulted in no significant frequency and amplitude alteration in the control, but significant decrease in DI. Conclusion These results suggested that Kca and Clca play a critical role in the modulation of detrusor contraction, and the down regulation of Kca and up regulation of Clca suggests that disorder of calcium related regulation may play an important role in DI.

Objective To establish the co-culture model in vitro and induce bone marrow mesenchymal stem cells (MSCs) to differentiate into lung alveolar epithelial cells. Methods Each group had 6 samples, control group was MSCs alone; Group A was the MSCs cultured with the cells from normal lung; and Group B was the MSCs with the cells from injuried lung. Each group was cultured for 8 days and the two markers of lung alveolar epithelial cells including AQP5 and SP-C were tested by laser confocal microscopy and RT-PCR. Results Only AQP5 was detected in the control group and Group A, both AQP5 and SP-C were detected in Group B, the AQP5 mRNA expression in Group B was significantly increased compared with that in the control group(P<0.01). The AQP5 mRNA expression in Group B was also significantly increased compared with that in Group A (P<0.01). But there was no significant difference in AQP5 mRNA expression between Group A and control group. Conclusion We have successfully established the co-culture model in vitro to induce bone marrow mesenchymal stem cells to differentiate into lung epithelial cells.

Objective To construct HSP27 eukaryotic expression plasmids. Methods Full-length HSP27 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) from breast cancer cell line MCS-7 and cloned into eukaryotic expression vector pAAV-MCS. After the recombinant plasmids transfected into NIH3T3 cells, the expression of HSP27 protein in the host cells was characterized by ECL Western blotting. Results Full-length HSP27 gene was amplified by RT-PCR correctly. The correct cloning of HSP27 gene in pAAV-MCS was confirmed by restriction enzyme digestion and sequencing. ECL Western blotting results indicated that the target gene could express in the mammalian cell line NIH3T3. Conclusion Recombinant plasmid HSP27/pAAV-MCS had been cloned successfully, which would provide the foundation for investigating the role and the mechanism of HSP27 in the ischemia precondition of kidney.

Objective By using array-based single nucleotide polymorphisms comparative genomic hybridization (SNP-aCGH) to detect and fine mapping copy number variations (CNVs) in children with unexplained mental retardation/brain development delay(MR/BD),then the CNVs were analyzed to determine diagnosis and offer genetic counseling,finally to discuss the application of SNP-aCGH in genetic diagnosis of MR/BD with unknown causes.Methods Ninety-two children with unexplained MR/BD were recruited.SNP-aCGH was used to get CNVs from the whole genome-wide,and the correlation of CNVs and phenotype was analyzed to definit disease genes or pathogenic fragment.Statistics was performed to analyze the common phenotype between positive cases (case with CNVs) and negative cases.Results (1) The CNVs were detected in 10 cases with a detection rate of 10.86％,from which 8 cases showed subtelomeric aberration,5 cases without subtelomeric aberration,and the rate was 8.70％,5.40％,respectively.The CNVs related to MR/BD involved 10 different subtelomeric regions (9p,21q,3p,2p,15q,4p,12p,22q,16p,17p),and 7 different regions without subtelomeric (1p,4q,2p,14q,15q,12q,22q).The deletions involved 11 zones (size:1.05-8.80 Mb),and duplications referred to 8 zones (size:1.33-31.25 Mb).(2) One case was diagnosed as 9p duplication syndrome,for candidate genes:DOCK8,VLDLR.A case was detected with a gene fracture (CRBN).One case was diagnosed as Coffin-Sirrs syndrome combined with a deletion of 15q26.3-qter,for candidate genes:SOX11 and LINS1,respectively.One case referred to 12p13.3 deletion syndrome,for candidate genes:ELKS,ERC1.One case referred to 22q13.2-qter deletion,for candidate genes:SHANK3.Two cases were diagnosed as ATR-16 syndrome with 17p13.3 deletion syndrome,their candidate genes:HBA1,HBA2,SOX8 for the former,YWHAE,LIS1 for the latter.(3)There were statistically significant differences in comparison of positive cases to the negative ones for growth delay,internal organs deformity,low birth weight infant(LBW) and premature infant (all P ＜0.05).Conclusions (1) Besides MR/BD in different degrees in all the positive cases,they also showed growth delay,a portion of them with internal organs deformity,low birth weight infant and premature infant.(2) Subtelomeric aberrations are related to MR/BD,while the submicroscopic rearrangement in regions without subtelomeric is suspiciously pathogenic,and need to be further studied.(3) SNP-aCGH can fine mapping the region of CNVs by high resolution from the whole genome-wide,which does contribute to limit the zones for finding pathogenic region and candidate genes,as well as to offer a technology platform for investigating about the correlation of phenotype and genes or CNVs.

Objective To analyze the genotype-phenotype correlations of Angelman syndrome ( AS ) , and to discuss the advantage of applying array-based single nucleotide polymorphisms comparative genomic hybridization ( SNP aCGH) in diagnosis of AS. Methods Examination of electroencephalogram( EEG) and intelligence quotient( IQ) evaluation were done for 11 cases diagnosed as AS clinically. Gesell scares were chosen as the evaluation criterion of IQ. The screening techniques was methylation polymerase chain reaction( MS-PCR) ,then SNP aCGH was used to make genetic diagnosis. Results (1)Eleven cases of AS were confirmed:1 case had UPD(uniparental disomy),10 cases were type of deletion, from which 6 cases were deletion (Ⅱ) , 4 cases were deletion (Ⅰ) . ( 2 ) The copy number variations were detected in the region of 15q11-q13,which contained genes like MKRN3,MAGEL2,NDN,SNRPN, SNURF,GABRB3,GABRA5,GABRG3,UBE3A,OCA2,ATP10A. To search online Mendelian inheritance in man,genes above were correlated with AS manifestation. (3)All cases of deletion were 3-5 standard deviation(SD) in weight and height to normal children at the same age and with the same sex,while UPD was below 1. 5 SD. Gesell scares showed that the deletion(Ⅰ) was the most serious in mental retardation,deletion(Ⅱ) was moderate,and the UPD was mild. Eight cases were hypopigmentation,and one was the UPD. EEG revealed that 1 case of deletion(Ⅰ) and the UPD were spike occasionally,another one deletion(Ⅰ) was limit EEG. The rest cases displayed slow and spike waves paroxysmal-ly,with amplitude of medium or high,2. 5-3. 0 Hz. Conclusions Not only can SNP aCGH make a diagnosis of AS but discriminate the types of genetic pathology. Since different type contributes to a diverse of clinical features and the rate of recurrence is also different,it is significant for family genetic consultation. Moreover,the technology is advantageous for the study on the pathogenesis and gene function.

ObjectiveTo establish a mouse model for genital tract infection with different serovars of Chlamydia trachomatis(Ct),and to provide a basis for understanding the relationship between Ct serovars and virulence.MethodsSix-week-old female BALB/c mice were divided into 4 groups:study group (pretreated with progesterone and inoculated with 4.0 × 107 inclusion-forming units(IFU) of Ct serovar E,F,J and K,respectively),progesterone-control group(pretreated with progesterone and inoculated with culture medium of McCoy cells),non progesterone-control group(inoculated with 4.0 × 107 IFU of Ct serovar F),blank control group receiving no treatment.At 4 and 7 days after the inoculation,the appearance of vulva was observed,vaginal swab specimens were obtained and subjected to cell culture,direct immunofluorescence assay and real-time fluorescence-based PCR for the detection of Ct.ResultsA slight inflammation of the genital tract was observed in the mice 4 days after the inoculation with Ct serovar E,F,J or K in the study group.Cell culture,direct immunofluorescence assay and real-time fluorescence-based PCR all confirmed that Ct was present in vaginal specimens from the study group,but absent from the control groups.The longest duration of infection was 21 days in 1 out of 11 mice inoculated with Ct serovar K,followed by 17 days in 1 out of 11 mice inoculated with Ct serovar J,14 days in 5 out of 11 mice with Ct serovar F and in 2 out of 11 mice with Ct serovar K.Conclusion The model for genital tract infection with different serovars of Ct can be established in mice at a right age bypretreatment with progesterone and inoculation with a certain amount of Ct.

As the most common organ-specific autoimmune disease,autoimmune thyroiditis (AIT) is characterized by intrathyroidal lymphocyte infiltration and specificthyroid autoantibodies.Modern medicine is short of effective etiological treatment at present,and has no specific medicine foreuthyroidism of AIT patients,who have been followed-up passively.Chinese medicine treatmenthas the characteristics of multiple-targets,multiple-links and multiple-pathways,which plays a special superiorityrole in the prevention and treatment of AIT.The purpose of this article was to sort out and set forthmore Chinese medicine pharmacology on AIT recently,which provided evidence for further clinical apply.

Objective To assess the vitro susceptibility to 6 antimicrobial agents and genotypes of clinical isolates of Chlamydia trachomatis (Ct) from Guangzhou region. Methods Ct was isolated from clinical specimens by using McCoy cell culture and subjected to propagation. The minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of 6 antimicrobial agents (clarithromycin, roxithromycin, azithromycin, doxycycline, tetracycline, ofloxacin) against Ct isolates were determined in McCoy cell culture. Nested PCR was performed to amplify the outer membrane protein 1 (omp1) VS1-2 gene followed by sequencing. Results Seventy-six Ct strains were isolated from 346 urogenital specimens, and 40 strains met the require ments for susceptibility testing after serial propagation. The MIC50/MIC90 of clarithromycin, azithromycin, roxi thromycin, doxycycline, tetracycline and ofloxacin were as follows: 0.008/0.032, 0.080/0.160, 0.125/0.500, 0.032/0.064, 0.250/0.500 and 0.500/1.000 mg/L. Seven genotypes were observed. The most prevalent geno types in decreasing order were E (14, 35%), J (10, 25%)and F (6, 15%). The MIC50 was consistent for azithromycin among the 7 genotypes, but varied by 1 - 4 folds for doxycycline, ofloxacin and roxithromycin. Conclusions Clarithromycin, doxycycline and azithromycin exhibit an excellent activity against Ct, and the activity of azithromycin is consistent among the 7 genotypes of Ct.