BACKGROUND: At present, there has been no definite experiment systemically evaluating adherent separation and density gradient centrifugation to isolate bone marrow mesenchymal stem cells (BMSCs). Whether the two methods produce different influences on BMSC induction and differentiation remains unclear.OBJECTIVE: This study was designed to verify difference of these two isolation methods in chondrogenic differentiation of BMSCs.DESIGN, TIME AND SETTING: A controlled observation was performed at the Shengjing Hospital Affiliated to China Medical University from March to September 2005. MATERIALS: Twenty Japanese big-ear rabbits, aged 2-3 months, weighing 1.2-2.0 kg, were included for this study. METHODS: BMSCs were isolated by adherent separation and density gradient centrifugation. Two groups of BMSCs were taken from the same passage and induced towards chondrogenic differentiation with transforming growth factor beta 1. MAIN OUTCOME MEASURES: Growth of BMSCs was observed under an inverted microscope to draw growth curves; Type II collagen expression was detected by immunohistochemistry. Type II collagen mRNA expression was determined by in situ hybridization. RESULTS: The growth curves demonstrated that cellular growth velocity of the two groups tended to be the same. Immunohistochemistry results showed that the efficiency of adherent separation and density gradient centrifugation for promoting chondrogenic differentiation of BMSCs was 76.1% and 77.7%, respectively, and in situ hybridization results showed that the efficiency was 70.3% and 71.0%, respectively. No significant difference in differentiation efficiency existed between the adherent separation and the density gradient centrifugation. CONCLUSION: Adherent separation and density gradient centrifugation had no different influences on BMSC growth and chondrogenic differentiation.

Objective To establish the model of hyperlipidemie acute pancreatitis(AP)to explore the mechanism of hyperlipidemia in aggravating AP.Methods Sprague-Dawley rats were divided into four groups at radmon:1)control group(C);2)hyperlipidemic group(TG);3)acute pancreatitis group(AP);4)hyperlipidemic acute pancreatitis group(HAP).After 8 weeks,blood,pancreas and peripancreas tissue were collected from each rats for determination of various parameters.Results The levels of hyperilidemic index were elevated in HAP group compared with those of other groups.The ADP/ATP ratio was obviously increased in HAP group.Caspase-3 and caspase-8 of HAP group presented in zymogen forms,while in AP group they were in activity forms.After the pancreas issue were stained bv HE and further analyzed through TUNEL assay,we found that there were more pancreatic acinar cells undergoing necrosis in HAP group.Conclusion Hyperilidemia may promote the course of apoptosis to necrosis of pancreatic acinar cells,which may be the mechanism of deterioration in hyperlipidemic acute pancreatitis.

Objective To investigate the level of nurses′preparedness and influencing factors of nurses with low seniority in Anhui province. Methods Totally 295 nurses with low seniority were recruited by convenience sampling method. They were investigated by a self-designed demographic data questionnaire and the Disaster Preparedness Evaluation Tool. Results The score of the disaster nursing ability of the nursing managers was 163.85 ± 32.41, the mean score was 3.64 ± 0.72. Regression analysis showed that the gender, the experience of disaster rescue,disaster training were the influencing factors of disaster preparedness. Conclusions The capacity of disaster preparedness for nurses is not ideal. There is still room for improvement among nurses in Anhui province to enhance disaster preparedness, and improved disaster nursing training system is needed to achieve this.

ObjectiveTo investigate fecal diversion in the management of pelvic fractures associated with perineal injuries.MethodsThis retrospective study involved 27 patients of pelvic fractures associated with perineal injuries during April 2005 to April 2011.There were 23 males and 4 females,and the mean age was 32.9 years old (range,16-62 years old).Fractures type according to Tile classification:4 cases were type A,10 cases were type B,and 13 cases were type C.The pelvic external fixator and fecal diversion were selectively used.Results Of the 27 patients of pelvic fractures associated with perineal injuries,24survived.The overall mortality of pelvic fractures associated with perineal injuries in the present study was 11％.The survived 24 patients were totally reviewed clinically.The mean follow-up time of these patients was 10.9 months (range,4-42 months).Of those patients who underwent early fecal diversion (＜ 48 h),none experienced infectious completions.However,of those underwent non-early (＞48 h) fecal diversion (including those who did not undergo fecal diversion),four patients experienced infectious completions.Fisher's exact test was used to compare the infection rate of these two groups.And the result of Fisher's exact test demonstrated that those patients who underwent early (＜ 48 h) fecal diversion and non-early (＞48 h) fecal diversion (including those who did not undergo fecal diversion).ConclusionStabilization of hemodynamic; selective fecal diversion; early stabilization of pelvic fracture are necessary for the emergency management of pelvic fractures associated with perineal injuries.Rectal injury and severe perineal injury without involvement of rectum should undergo fecal diversion.Early fecal diversion (＜ 48 h) could reduce the infection rate of pelvic fractures associated with perineal injuries.

ObjectiveTo detect mRNA and protein expression of steroidogenic factor-1 ( SF-1 ) and DAX-1 in human adrenocortical tumors and normal adrenal cortex,and to investigate the effect of SF-1 and DAX-1 on the steroidogenesis and development of adrenocortical tumors.Methods Total RNA and protein was extracted from angiotensin Ⅱ unresponsive aldoterone-producing adenomas ( A Ⅱ -U-APA,n =12 ),angiotensin Ⅱ responsive aldoterone-producing adenomas ( AⅡ -R-APA,n =5 ),cortisol-producing adenomas ( CPA,n =10 ),adrenal nonfunctional adenomas ( NFA,n =10 ),aldosterone-producing carcinoma ( APC,n =2 ) and normal adrenal cortex ( NAC,n =8).To analyze gene expression of SF-1,DAX-1,ACTH receptor(ACTHR),and β-actin by real-time quantitative PCR in different tissues.The protein expression of SF-1,DAX-1,and β-actin in the same tissues by Western blot.To study the relationship of ACTHR,SF-1,and DAX-1 with clinical data in adrenocortical tumors.ResultsThe expression of SF-1,DAX-1 mRNA and protein was different in NAC,AⅡ -U-APA,A Ⅱ -R- APA,APC,CPA,and NFA tissues [ relative expression of SF-1 mRNA:24.58±2.45,23.89±3.17,21.59±3.00,(38.75,44.16),14.17±2.80,and 36.38±3.50; DAX-1 mRNA:0.57±0.06,0.37±0.05,0.43±0.05,( 1.52,1.21 ),0.39 ±0.04,and 0.83 ±0.08 ; SF-1 protein:0.76 ±0.11,0.76 ±0.10,0.73 ±0.07,(1.24,1.40),0.55±0.04,and0.98±0.10; DAX-1 protein:0.65±0.14,0.39±0.13,0.43±0.14,(1.18,1.02),0.56±0.04,and 1.03±0.13 ; all P＜0.05 or P＜0.01 ].There was negative correlation by higher SF-1/DAX-1 ratio and tumor size in AⅡ -U-APA tissues.The mRNA and protein expression of SF-1 was lower in CPA and there was the positive correlation with tumor size.Conclusion SF-1 and DAX-1 might play a key role in the development of the adrenocortical tumorigenesis and steroidogenic tissues.

Objective:To ascertain whether the immune complexes (ICs) formed by Dengue virus 1 non-structure protein 1 (DENV1 NS1)and its IgG antibodies could mediate passive systemic anaphylaxis (PSA) and to explain the pathogenesis of Dengue hemorrhagic fever or Dengue shock syndrome (DHF/DSS).Methods:The monoclonal antibodies (mAbs) or mAb cocktails from 20 IgG mAbs of DENV1 NS1 prepared in this lab were screened to initiate PSA and passive cutaneous anaphylaxis (PCA) in mice.Meanwhile, the effects of GdCl3 and platelet activating factor ( PAF) antagonist CV-3988 on PSA induced by the NS1-IgG ICs were observed.Results:Two groups of monoclonal antibody cocktails with purified NS 1 were proved to be capable of provoking PCA and PSA in mice,whereas the other mAbs or mAb cocktails could be not .The murine PSA initiated by NS1-IgG(5D25+3B1) ICs could be sig-nificantly inhibited by in vivo treatment with GdCl3 or PAF antagonist CV-3988.Conclusion: The NS1-IgG ICs formed with DENV1 NS1 and IgG mAb cocktails can mediate PSA and PCA ,but not all of ICs formed by DENV 1 NS1 mAbs or mAb cocktails with DENV 1 NS1 can induce PSA ,indicating that it may be related to the special epitopes of DENV 1 NS1.The monocyte/macrophages and PAF may be as major effector cells and the major mediator for PSA induced by NS 1-IgG ICs,respectively.

Objective To evaluate the clinical effects of block calculus extraction catheter in the ureteroscopic lasertripsy. Methods Two hundred and thirty patients with ureter calculus were enrolled. Ninety-six patients were treated with block calculus extraction catheter (test group), and 134 patients were treated without block calculus extraction catheter (control group). The patients in 2 groups were treated with holmium laser ureteroscopic lithotripsy of Lumenis PowerSuite60W, and the treatment effectiveness was compared. Results The operation time and primary stone clearance rate in test group were significantly better than those in control group: (23.3 ± 7.4) min vs. (35.6 ± 11.1) min, 98.92%(92/93) vs. 88.89%(112/126), and there were significantly differences (P<0.01). The bleeding amounts and fever rate of 2 groups had no significant difference (P > 0.05). Conclusions The block calculus extraction catheter is very useful and safe in preventing the calculus shift, shortening the operation time and improving the calculus elimination rate .

Objective To compare the detection efficiency between multiplex RT-PCR method and liquichip technology for screening the viral etiological agents of diarrhea.Methods The development of the multiplex RT-PCR method.A total of 107 feces samples from patients who suffered from diarrhea and attended to Zhujiang Hospital of Southern University from September 2013 to February 2014 were collected and tested in parallel by both multiplex RT-PCR and xTAG Gastrointestinal Pathogen Panel ( xTAG GPP) for Adenovirus, Norovirus genogroupⅠandⅡ, as well as by both multiplex RT-PCR and monoplex RT-PCR for Astrovirus and Sapovirus.To evaluate the sensitivity and specificity of multiplex RT-PCR, xTAG GPP and monoplex RT-PCR were used as reference.Kappa coefficient test was used to evaluate the consistency among the methods.The detection limit and accuracy of multiplex RT-PCR were evaluated by detection of serial dilution of positive plasmids and products sequencing for the five viral agents.Results The multiplex RT-PCR showed high consistency with xTAG GPP and monoplex RT-PCR, in which Kappa value was 0.885 and 1.000 respectively( P=0.000 ).Compared to xTAG GPP, the sensitivity and specificity of the multiplex RT-PCR were at average of 80.8%( 21/26 ) and 100%( 295/295 ) respectively.The detection limit and accuracy of multiplex RT-PCR were 104 copies /μl-106 copies/μl.Conclusion The high consistency indicated that both the multiplex RT-PCR and xTAG GPP are useful as a special,sensitive, high throughput and rapid diagnostic tools for the detection of the major viral pathogens related to diarrhea in clinical laboratory.

ObjectiveTo evaluate the value of the measurement with a scale in arthroscopic anatomic anterior cruciate ligament (ACL) reconstruction.Methods Thirty-six patients with unilateral ACL-deficient knees were divided into single bundle group and double bundle group based on the length of the major axis of the native ACL footprint measured with a scale.Consistent with the International Knee Documentation Committee (IKDC) scoring systems,Lysholm knee scoring scale and Tegner activity rating were used to evaluate the clinical results,and data from 23 to 26 months follow-up were gathered and analyzed statistically.ResultsIn terms of IKDC evaluation,19 patients( 90.4％ ) in the double band group and 13 patients( 86.7％ )in the single band group were graded as normal or nearly normal ( Z =7.82,6.33 ; P ＜ 0.05 ).The mean Lysholm scores were 93.8 ± 9.9 and 94.2 ± 9.6 and the mean Tegner scores were 6.6 ± 1.9 and 6.5 ± 1.7 for the double and single band groups,respectively( t =2.31,5.60,3.55,7.09 ;P ＜0.05 ).ConclusionArthroscopic measurement with a scale combined with other traditional methods to determine the tunnel location is easy to conduct and intuitional to understand.It could be useful in anatomic ACL reconstruction.

Objective To elucidate the mutations of NPHS1 gene in children with sporadic steroid-resistant nephrotic syndrome (SRNS) in Southern Chinese Han ethnic group.Methods Peripheral blood samples were collected for genetic analysis from 40 patients with sporadic SRNS and 50 healthy volunteers as control.Genomic DNA was isolated from peripheral blood leucocytes.Twenty-nine exons and exon-intron boundaries of the NPHS1 gene were amplified by polymerase chain reaction.Mutational analysis was performed by DNA sequencing directly.Results Seven variants,928G＞A(D310N),2677A＞G (T893A),2869G＞C (V957L),IVS8+30C＞T,IVS21+14G＞A,IVS25-23C＞T and *142T＞C,of NPHS1 gene were found in 6 of 40 children with sporadic SRNS,whereas they were not found in 50 healthy controls.2677A ＞G,IVS8 +30C ＞T,IVS21 +14G＞A,IVS25-23C ＞T and *142T＞C were novel.Moreover,thirteen already reported NPHS1 polymorphisms,294C＞T,349G＞A,IVS3+15C＞T,IVS3+61A＞G,803G＞A,IVS8+68A＞G,1339G ＞A,1802G ＞C,2223C ＞T,2289C ＞T,IVS24 +36C ＞T,3315G＞A and IVS27 +45C ＞T,were detected in some patients and controls. Conclusions NPHS1 mutations in 6 of 40 children with sporadic SRNS in Southern Chinese Han ethnic group (15％) are detected.NPHS1 mutations are existed in Southern Chinese children,so it is necessary to perform the mutation analysis of NPHS1 gene in those children patients.