Objective To develop a whole-genome DNA microarray based on the genomic sequences of Y. pestis CO92 and 91001 and its use in comparative genomic analysis. Methods A total number of 4 005 genes of Y. pestis were amplified by PCR and printed onto glass slides in duplicate. Fluorescently labeled probes were prepared by marking genomic DNAs with random hexamers and Klenow. Labeled DNAs were hybridized with the microarrays by the method of two-fluorescence comparative hybridization. Three sets of two-fluorescence hybridizations were performed to examine the absence/presence of each gene. Results The results agreed with those derived from the in silico genomic comparison. Conclusion The results demonstrate that the microarry can be a useful tool for comparative genomic analysis of Y. pestis.

Objective To identify and compare the genome differences among live plague vaccines prepared with different strains of the bacillus. Methods The whole-genome DNA microarray of Yersinia pestis was used as a tool to perform genomic comparison among live plague vaccines prepared with 19 different strains. Results Dozens of deletions and/or increased copies of the genomic fragments were identified in the studied vaccines of different strains. Conclusion The revealed genomic differences among the vaccines from different origins account for the variability of the immunogenic and protective potency of live plague vaccines. The whole-genome DNA microarray was also provesd to be an ideal tool for the pre-evaluation of a vaccine strain.

Objective To explore the surface electromyography (sEMG) assist for the rehabilitation training for children with cerebral palsy. Methods 60 children with spastic diplegia were divided into treatment group (n=30) and control group (n=30). All the children received comprehensive rehabilitation, while the rehabilitation program were designed with the assist of sEMG in the treatment group. They were assessed with Gross Motor Function Measure (GMFM) before and 3 months after rehabilitation. Results There was no statistically improvement in dimension A, B, and C of GMFM (P>0.05), but was in the total score and the dimension D and E (P<0.0001). There was no statistical difference in dimension A, B, and C between 2 groups (P>0.05), but was in the total score and the dimension D and E (P<0.01) after rehabilitation. Conclusion The sEMG might be helpful for the rehabilitation for children with cerebral palsy.

BACKGROUND: Diabetic cardiomyopathy, a serious complication of diabetes, is an important factor of increased mortality in patients with diabetes. Therefore, providing an effective experimental animal model is particularly important for studying the pathogenesis and treatment methods of diabetic cardiomyopathy. OBJECTIVE:To explore the method of establishing Wistar rat models of diabetic cardiomyopathy. METHODS:Forty rats were randomly divided into the control group (n=10) and diabetic cardiomyopathy group (n=30). The rats in the diabetic cardiomyopathy group were intraperitonealy injected with 60 mg/kg streptozotocin at a time to establish rat models of diabetic cardiomyopathy. The rats in the control group were given the same dosage of citric acid buffer by the same way. The rats in these two groups were al fed with non-fat high-sugar normal diet. RESULTS AND CONCLUSION:Compared with the control group, after 3 weeks of injection with streptozotocinin in rat models of diabetic cardiomyopathy, blood glucose level was significantly increased, myocardial cels arranged in disorder, the nuclei were of different sizes, colagen content in the myocardial tissue was significantly increased, and colagen fibers were thick and disordered. In addition, the mRNA and protein levels of transforming growth factorβ1 and type I colagen, two indices reflecting myocardial fibrosis, were markedly increased. These results indicate that intraperitonealy injecting large doses of streptozotocin (60 mg/kg) at a time and feeding with non-fat high-sugar normal diet could establish a stable rat model of type 1 diabetic cardiomyopathy. This method is safe and effective with high feasibility.

AIM:To investigate the effects of P34H gene silencing on the expression of P34H and activity of hyaluronidase (HYD) in mouse sperm.METHODS:The recombinant plasmid series of P34H targeted short hairpin RNA (shRNA) were constructed by GV248 plasmids vector.These recombinant plasmids were transformed into DH 5αcompetent cells, and the plasmids were taken from DNA sequencing analysis .The HEK293T cells were co-transfected with shRNA and lentiviral packaging plasmids .The 3 kinds of recombinant lentiviruses and negative control lentiviruses were used to in -ject into the mouse epididymis and the expression of P 34H at mRNA and protein levels was detected by real-time PCR and Western blot, respectively.The location of P34H protein on the mouse spermatozoa was determined by indirect immunofluo-rescent staining using P34H antibody.The positive rate and activity intensity of HYD was detected by modified sodium hya-luronate-gelatin membrane.RESULTS:DNA sequencing analysis confirmed that the 3 P34H-shRNA sequences were suc-cessfully inserted into the lentiviral vectors .P34H expression in epididymis tissue was significantly decreased at both mR-NA and protein levels compared with those of the non-transfected and normal control vectors (P<0.05).The GV-P34H-shRNA-1 played a significant role in reducing the percentage of P 34H positive rate and the activity of HYD in mouse sperm.The silencing effect did not significantly differ between the non-transfected and normal control vectors .CONCLU-SION:Silencing of P34H significantly inhibits the percentage of P 34H positive rate and the activity of hyaluronidase in mouse sperm.

Whether InfusaSleeve(IS) catheter can deliver antisense oligodeoxynucleotide (ODNs) following arterial denuation is unknown. We evaluate the feasibility of local endoluminal delivery of C-myc ODNs to the site of arterial denudation by using IS catheter and to determine the biological importance of these effects. IS catheter was introduced into right side of iliac artery of 21 rabbits after angioplasty of iliac artery. Animals were randomized to the control group (n＝6) receiving saline injection and the treated group receiving c-myc antisense (n＝15, 1 mg ODNs per vessel). In two weeks and 40 days following the operation, angiography was performed. Morphometric analyses were carried out in balloon-denuded iliac arteries. The expression of c-myc protein was detected by using a mouse monoclonal antibody to c-myc. Morphometric analyses carried out at 40 days after transcatheter c-myc antisense oligomer administration. The results showed that maximal neointimal area was reduced from 7.66±3.7(×105 μm2) in the control group (n＝6) to 4.04±1.02(×105 μm2) in the antisense treated group (n＝6, P＜0.05). These changes in vascular remodeling following denuding injury resulted in an increase in residual luman from 20～50% in the control group to 70～90% in the antisense-treated group. C-myc protein expression was virtually undetectable at baseline in locally ODNs-delivered arteries and detectable in control denuded arteries. The results show that: ①Single IS transcatheter administration allowed endoluminal delivery of ODNs to the site of arterial injury; ② c-myc antisense oligomer reduced the formation of neointime in denuded arteries, implying a therapeutic potential of this approach.

This paper summarized the experience of caring a patient with pancreatic cancer-related depression treated with pancreaticoduodenectomy and postoperative delayed gastric emptying.The nursing included several key points.On the base of collaboration of muhi-disciplinary teams,to strengthen supportive psychological intervention and safety management after admission;to use multimodal analgesia combined with cognitive behavioral therapy to reduce postoperative pain.After the patient was complicated with gastric emptying obstacles,solution-focus mode,sham feeding and nutritional support programs were implemented.Long-term follow-up with physician-nurse collaboration mode was implemented to enhance quality of life of the patient.

ObjectiveTo make sure the factors of influencing supply and utilization rehabilitation service of hospitals in Beijing.MethodsFive grade A hospitals in different districts in Beijing were surveyed to realize the condition and factors of influencing supply and utilization of rehabilitation service in polyclinics in Beijing and make sure the developing situation of departments of rehabilitation medicine, and its restriction factors in their hospitals.ResultsThe departments of rehabilitation medicine in the five hospitals had not reached to requirement of administer criterion to comprehensive hospitals, issued by Ministry of Health. The factors of influencing supply and utilization of rehabilitation service in polyclinics in Beijing included: discordant cooperation between rehabilitation section and other sections in hospitals; leaders of hospitals and clinical personnel not understanding the importance of rehabilitation early intervention much; patients and their family not understanding the special curative effect to prevent and treat disability of rehabilitation; the application for reimbursement limitation to the fee for patients to accept rehabilitation treatment and/or stay in rehabilitation section in hospitalization insurance and plan as a whole, and so on.ConclusionThe factors above limit the further development of rehabilitation service of polyclinics in Beijing. Effective measures should be taken and correlative policies should be established in order to promote the rehabilitation service in Beijing.

Objective To establish the genetic test technique of trisomy 21 concurrently conducts with prenatal diagnosis for hereditary hearing loss. Methods Fifty-four pregnant women who underwent prenatal diagnosis for hearing loss of their fetuses in Chinese People's Liberation Army General Hospital from March 2009 to May 2010 were enrolled in this study. All probands from the deaf families have confirmed the causative mutation for hearing loss in Genetic Testing Center in Chinese People's Liberation Army General Hospital. The mean age of 54 pregnant women is 31 years at pregnancy of 18 - 26 weeks, 5 cases ＞ pregnancy of 23 weeks, 9 cases ≥ 35 years. All subjects did not conduct the serologic tests for trisomy 21before. Fifteen to twenty ml amniotic fluid was drawn from 49 cases at pregnancy of 18 - 23 weeks and 5 cases ＞ pregnancy of 23 weeks. One to two ml umbilical blood was drawn from 5 cases ＞ pregnancy of 23 weeks. For 9 cases ≥ 35 years, amniotic fluid cell culture and karyotyping analysis were conducted concurrently. A multiple quantitative fluorescent ( QF) PCR and six microsatellite markers were applied to as trisomy 21. Results (1) Fifty-four fetuses were successfully conducted prenatal genetic diagnosis for hearing loss (included GJB2 and SLC26A4). Ten fetuses copied the exactly same genotypes as the probands. The other 44 cases fetuses did not copy the same genotypes as the probands and won't develop hearing loss. The hearing test showed normal hearing for the neonates. (2) All the 54 fetuses were excluded of trisomy 21 by QF-PCR and were verified after birth. Five fetuses with advanced maternal age were performed karyotyping analysis and showed normal. The diagnostic results of QF-PCR can be obtained in 1 - 3 days without misdiagnosed. Conclusions QF-PCR is an efficient, rapid and accurate technique for detection of trisomy 21 without increasing sample amount. It can be used for fetuses who were undertaken hearing loss gene test or other prenatal gene test.

Objective To investigate the infection status quo of 21 kinds of genotypes of human papillomavirus (HPV)and their distribution rule among the women in Hengyang region.Methods A total of 8 032 women voluntarily accepting cervical cancer screening in the Hengyang Municipal Maternal and Child Health Care Hospital fromg April 2012 to May 2013 were selected as the research subjects.The exfoliated cells samples of cervical tissue were collected for detecting 21 kinds of HPV genotypes by Hybri-Max.The HPV infectious rate and the HPV gene distribution were analyzed by using SSPS13.0.Results Of 8 032 women,1 664 cases were detected out 1 kind or more than 2 kinds of HPV,the HPV infectious rate was 20.72%(1 664/8 032).The top 6 geno-types of HPV were HPV16,52,58,81,53 and 18.Among 1 664 female cases of HPV infection,the single HPV infection rate was 76.44%,the multiple infection rate was 23.56%,which was dominated by the double infection;the total positive rate in the various age groups of HPV infection presented theU-type distribution with the age increase,however the total positive rate had no statis-tically significant differences among all age groups(P >0.05 ).Conclusion The HPV female infection rate in Hengyang region is relatively higher,moreover the high risk HPV is predominant.HPV 16 occupies the top ranking.