Objective To observe the transfection efficiency of transferring plasmids pEGFP-N1 and pIRE-EGFP encoding enhanced green fluorescent protein(EGFP) to rabbit corneal endothelial cells(RCECs) by cationic polymer and the change of cell activity by detecting the activity of Na+-K+-ATPase.Methods RCEC was cultured by digestion and identification by neurone specific enolase(NSE) stain.Sofast TM mediated plasmid pEGFP-N1 and pIRE-EGFP to transfect RCECs and we compared the transfection efficiency at different time and in different groups by calculating the number of fluorescent cells.We observed the activity change of transfection and non-transfection RCEC by detecting the activity of Na+-K+-ATPase.Results After pEGFP-N1 and pIRE-EGFP transfected RCEC with Sofast TM by different ratio,RCEC expressed EGFP in 24-48 h.After transfection 48 h the group of SofastTM∶pEGFP-N1=3.2∶1 had the max transfection efficiency 3.6% and the max transfection efficiency of group SofastTM∶pIRE-EGFP =3.2∶1 was 3.5%,at 24 h after transfection.There were no obvious differences between the transfection and non-transfection groups of cell number and activity of Na+-K+-ATPase.Conclusion Sofast TM can mediate exogenous gene trasfering to RCEC effectively.pEGFP-N1 and pIRE-EGFP are the suitable non-virus vectors for gene transferring to RCECs.

Objective To study the pathogenesis of unstable bladder with partial bladder outlet obstruction through the experiment based research on connexins in both unstable bladder group and cell hypoxia group.Methods Unstable bladder rat models were established and smooth muscle cells were cultured in vitro.Then cell anoxia was induced.RT-PCR and Western blot were used to quantify the connexins.Results The mRNA levels of Cx40,Cx43 and the Cx43 protein expressed in the smooth muscle cells from the unstable bladder group and cell hypoxia groups were all higher than those in the control groups(the sham-operation group and normoxia groups)(P

SUMMARY Diffusion spectrum imaging ( DSI) , a newly developed MRI technique, affords the capacity to map complex fiber architectures in tissues with sufficient angular resolution by imaging the spectra of tissue water diffusion. By contrast, diffusion tensor imaging (DTI) , the currently widely used technique based on the 2nd order tensor model, obtains an approximation of the complex diffusion, and provides only one global maximal direction corresponding to the primary eigenvector for each voxel. As a generalized model-free diffusion imaging technique, firstly, DSI employs the probability density function to describe the diffusion process in each voxel; secondly, a sufficient dense signal sample derived from repeated applications of diffusion-weighed gradients ensures its capability to resolve the diffusion probability density function; thirdly, specific computer visualization techniques are used to extract the diffusion information and reconstruct the geometrical properties of tissue microstructure. The capacity to unravel complex tissue architecture, recent improvements in hardware and ongoing optimization of sequence design and algorithm enable a rapid growth of DSI for research use and future incorporation into clinical protocols. This paper introduces the basic principles of DSI and then compares the characteristics of DSI and DTI schemes. Finally, the typical applications of DSI to date are reviewed.

Objective:To investigate the curative effect of early enteral nutrition support in patients with COPD complicated with respiratory failure.Methods: When mechanical ventilation began,63 patients in the acute phase of COPD complicated with respiratory failure were divided into two groups: the early enteral nutrition group(EEN group,n=32) or the delayed enteral nutrition group(DEN group,n=31).After two weeks,the nutritional indexes,the times of mechanical ventilation,the incidence of complication and the mortality were observed.Results: Two weeks later,there was significant difference between EEN group and DEN group in total protein,albumin,prealbumin and hemoglobin(P

OBJECTIVE To explore the factors and effective preventive method of nosocomial lower respiratory(infection) after general anesthesia through trachea intubation.METHODS Using bacterial culture and identification technique to detect the samples collected from pipe and the sides of anesthetic machine,from disposable virus/(bacteri)a respiratory filter and buccal and bronchial secretion of patients with general anesthesia through trachea(intubation.) RESULTS Eight of 15 anesthetic machines without disinfection were with positive bacterial culture.All sides of virus/bacteria respiratory filter,and buccal and bronchial secretion of patients during operation had same(bacteria.) No bacteria growth was found in anesthetic machine and disposable virus/bacteria respiratory filter used after operation.The same bacteria as from buccal secretion were isolated from anesthetic machine in patients(without) using virus/bacteris respiratory filter.CONCLUSIONS The(results) showed the anesthetic machine is more easy to be contaminated by patients,and the reuse of anesthetic machine is an important factor for nosocomial(lower) respiratory infection after general anesthesia through trachea intubation.Intensifying periodic disinfection of(anesthetic) machine and using virus/bacteria respiratory filter can prevent contamination between anesthetic(machine) and patients and decrease nosocomial lower respiratory(infection) after operation.

Objective To explore the methylation status of hMLH1 and hMSH2 promoter region in the epithelial ovarian cancer and its role in oncogenesis.Methods Methylation status of hMLH1 and hMSH2 promoter region was assayed in 20 normal ovarian tissues,25 benign epithelial tumor,56 malignant epithelial tumor and cell lines SKOV3,3AO by methylation-specific PCR (MSP).SKOV3 and 3AO were analyzed before and after 5-aza-2′-deoxycytidine (5-Aza-CdR) treatment.In addition,an alterations of mRNA expression of hMLH1 and hMSH2 was observed by reverse transcription polymerase chain reaction (PT-PCR).Results No methylation of hMLH1 and hMSH2 promoter was found in normal ovarian tissues. CPG islands methylation of hMLH1 and hMSH2 was observed in 4% (1/25),8% (2/25) respectively in benign epithelial tumor,30.4% (17/56),51.8% (29/56) respectively in malignant epithelial tumor. Methylation status in promoter showed obvious correlation with pathological grade and lymph node metastasis (P

BACKGROUND:It is crucial for us to find an objective and sensitive means for the assessment ofthe severitv and development of dementia,so as to make instant diagnosis and examination of therapeutic effect on patientswith vascular dementias(VD).Event related potential endogenetic eomponent P300 is proved closely correlated with the cognitive management of examinees but not with stimulus property,thereby considered as one of the most valuable and objective index for brain superior psychological activity.OBJECTIVE:To study the role and significance of events related potential P300,providing objective basis for clinical diagnosis and therapeutic assessment for patients with VD.DESIGN:Non-randomized simultaneous controlled explorative study based on patients. SETTING:Neurological department and electromyography department in aprovincial hospital and neurological department in a medical college hospital.PARTICIPANTS:Between February 2000 and June 2001,64 patients were of VD at neurological clinic and ward ofthe Second Affiliated Hospital of Chongqing Medical College,including 38 males and 26 females with theaverage age of(66.0±4.6)years(VD group).Culturallevel:19 cases re-ceiyed over senior school education,31 cases of junior school education and14 cases of illiteracy.Meanwhile 60 inpatients without VD were enrolled(non-VD group)including 35 males and 25 females withthe average age of(68.0±3.7)years.Culturallevel:senior school or above,15 cases,junior school, 29 cases and illiteracy,16 cases.METHODS:Mental sate of both VD and non-VD groups were assessed with mini-mental state examination(MMSE),and tested with P300 potential observing the variability of latency and amplitude of P300 in patients.Both methods were compared between VD group and non-VD group, moreover relationship between P300 latency and MMSE scores was also explored.MAIN OUTCOME MEASURES:Variability of P300 latency and amplitude Relationship between P300 latency and MMSE scores.RESULTS:P300 latency was found prolonged in 74% and 7% of VD group and non-VD group respectively,with obvious significance between them,meanwhile prolonged P300 latency was proved closely correlated with the MMSE scores(r=0.609 6,P＜0.05).CONCLUSION:P300 latency can be considered as an obiective predictor for the earlier diagnosis and the assessment of therapeutic effects in patients with VD.

Objective To investigate the clinical effect of blocking nail in treatment of hypertrophic nonunion after interlocking intramedullary nailing of femoral shaft fractures.Methods A retrospective case series study was made on 11 patients with hypertrophic nonunion following interlocking intramedullary nailing of femoral shaft fractures from January 2006 to February 2016.There were 8 males and 3 females,aged (32.4 ± 6.8) years (range,19-48 years).Comminuted fractures were noted in three patients,long oblique or spiral fractures in two,multi-segment fractures in two,and transverse fractures in four.Time interval between this surgery and intramedullary nailing of femoral shaft fractures was (11.5 ±3.1) months (range,6-18 months).All blocking nails were inserted into pulp cavity of distal ends of the site of the bone nonunion.Fracture healing time,wound healing and postoperative complications were detected.Outcome was evaluated using the Visual Analogue Scale (VAS) and Oswestry Disability Index (ODI).Results All patients were followed up for (27.2 ± 4.6) months (range,24-36 months).Bone union was observed within (9.8 ± 3.4) months except that bone nonunion occurred in two cases requiring locking plate fixation and autologous bone grafting to provide union,with the healing rate of 91％.No wound infection,nail breakage and nail loosening occurred after operation.At postoperative 3 months,4 months,5 months,6 months,12 months and 24 months,VAS and ODI differed significant from the preoperative detections (P ＜ 0.05).Conclusion Blocking nail technique is associated with increased stability of intramedullary nails and fracture ends,low incidence of complications and high bone healing rate in treatment of hypertrophic nonunion after femoral interlocking intramedullary nailing.

Objective To investigate the changes of IL-6, IL-10 and TNF-αafter cholecystectomy.Methods 46 cases with cholecystolithiasis were selected and divided into 2 groups.23 in control group were treated with open cholecystectomy, experimental group were treated with laparoscopic cholecystectomy.The levels of IL-6, TNF-α, IL-10 and CD4 +/CD8 +T were compared in the two groups pre-and post-treatment.ResuIts Compared with pre-treatment, two groups of patients with IL-10, TNF-α, IL-6 and CD4 +T and CD4 +/CD8 +T increased (P<0.05), compared with control group, experimental group IL-10, TNF-α, IL-6, CD4 +/CD8 +T levels were higher (P<0.05).ConcIusion Laparoscopic cholecystectomy can significantly reduce the IL-6, TNF-αand IL-10 levels, reduce the body′s stress response to surgery, and reduce blood flow, shorten the operation time.

ABSTRACT:Nested‐PCR and loop‐mediated isothermal amplification (LAMP) were applied to identify the Borrelia burg‐dor f eri (B .burgdor f eri) in ticks in this study .A total of 112 adult ixodes were collected from vegetation in a forest area and farm animals in Xunhua County ,Qinghai Province and Xinbin County ,Liaoning Province .The ticks were examined for the presence of B .burgdorferi by nested‐PCR and LAMP .Results showed that 12 in 51 samples were found positive in Xunhua County (23 .53% ) .While positive rate in Xinbin County was 29 .51% with 18 samples positive in 61 samples .In total of 112 tick samples ,the PCR‐positive rate was 17 .86% with 20 positive samples identified ,whereas 15 positive samples were con‐firmed with positive rate of 13 .39 % by LAMP assay .There was no significant difference between the two assays (Х2 =0 .85 , P>0 .05) .Results suggest that both nested‐PCR and LAMP could be used in identifying B .burgdorferi in ticks .Combina‐tion of these two assays could improve the testing results .This is the first report of B .burgdorferi in ticks in Xunhua and Xinbin counties ,and helps to complete the database of the infection rate of B .burgdor f eri in ticks in the widely‐forested area of China .