Objective To investigate the relationship between vascular endothelial cell nitric oxide synthase (eNOS) gene polymorphism and the pathogenesis of avascular femoral head necrosis (ANFH).Methods The eNOS full-length CDS fragments,containing 894G or 894T separately,was subcloned into lentiviral expression vector,and then infect the human umbilical vein endothelial cells (HUVEC).The contents of nitric oxide NO and cyclic guanosine monophosphate (cGMP) in cell culture supernatant were detected in a time-dependent manner.The luciferase-labeled reporter plasmid pNF-κB-luc was co-transfected with the reference control plasmid pRL-TK into HUVEC cells infected with LV-eNOS-894G,LV-eNOS-894T,and control lentivirus (LV-NC) for 48 h.The luciferase activity of each group was detected.The expression of nuclear factor (NF)-κB protein and eNOS protein in HUVEC cells were detected by Western blot assay.The HUVEC cells in each group were co-cultured with hFOB1.19 cells,and the concentration of alkaline phosphatase (ALP) or osteocalcin (OCN) in the supernatant were detected at different time points.Results The contents of NO and cGMP in the cell culture supernatant of the full-length lentivirus expressing eNOS gene (containing 894G or 894T) were significantly higher than that in the empty cell group and the empty vector group,and the contents of NO and cGMP in the cell culture supernatant of the 894G group were significantly higher than that of 894T group (P ＜ 0.01).Compared with blank cells,the expression levels of NF-κB/p65 protein and eNOS protein were significantly increased in cells expressing eNOS,and the expression of NF-κB/p65 protein in 894G group was significantly increased than 894T group,but there was no difference in eNOS protein expression between the 894G and 894T groups;each group of HUVEC cells were co-cultured with hFOB1.19 osteoblasts,and at each of the same time points,the concentrations of ALP and OCN in the cell culture supernatant expressing lentivirus were significantly higher than that in the empty cell group and the empty vector group,and the ALP and OCN concentrations in the cell culture supernatant of the 894G group were significantly higher than those in the 894T group.Conclusions The eNOS gene exon G894T mutation reduces the levels of nitric oxide and cGMP produced by vascular endothelial cells through the eNOS-NO pathway,affecting the expression levels of NF-κB/p65 protein and eNOS protein,and reducing osteoblast activity,and blood supply to blood vessels.It may be one of the pathogenic mechanisms leading to femoral head necrosis.

Objective To investigate the clinical effects and related complications of surgical and endovascular treatments of paraclinoid aneurysms. Methods Seventy-two consecutive patients with paraclinoid aneurysm admitted to the Department of Neurosurgery, the First Affiliated Hospital of Soochow University were enrolled retrospectively. They were confirmed by head and neck CT angiography and DSA examination,including 45 females and 27 males. According to the different treatment modalities, 72 patients were divided into endovascular embolization group ( n=38,stent-assisted coil embolization) and microsurgery group (n=34,including aneurysm clipping or intracranial and extracranial vascular bypass plus trapping of aneurysms). The baseline data and aneurysm diameter of both groups were recorded and analyzed. Baseline data included age,sex,cranial nerve injury,Hunt-Hess grade,etc. The new cranial nerve injury,aneurysm recurrence and Glasgow outcome scale ( GOS) scores were compared between the two groups,and the size of recurrent and non-recurrent aneurysms were analyzed. The age, aneurysm diameter,and prognostic score were compared in patients with Hunt-Hess grade Ⅲ-Ⅳ in both groups. Results (1 ) The age of patients in the endovascular embolization group was older than that in the microsurgery group (60 ±10 years vs. 54 ± 10 years,t= -2. 490),the diameter of aneurysms was smaller than that of microsurgery group (5[4,8] mm vs. 9[5,16] mm,Z=3. 026). There were significant differences between the two groups ( all P <0. 05 ) . There were no significant differences in gender, incidence of visual impairment and Hunt-Hess grading between the two groups (all P >0. 05). (2) The patients of both groups were followed up for 6 months. There were no significant differences in GOS score, improvement of postoperative visual acuity,and immediate visual impairment after surgery ( all P>0. 05). The recurrence rate of aneurysms in the endovascular embolization group was higher than that in the microsurgery group. The difference was statistically significant between the two groups (18. 4%[7/38] vs. 2. 9%[1/34], χ2=4. 350,P<0. 05). (3) In the endovascular embolization group, the diameter of aneurysm in recurrent patients was larger than that in non-recurrent patients,and the difference was statistically significant between the two groups (10[8,15] mm vs. 5[3,7] mm,Z = -2. 356, P<0. 05). (4) For aneurysm patients with Hunt-Hess grade Ⅲ-Ⅳ, the GOS score of the endovascular embolization group was lower than that of the microsurgery group,and the difference was statistically significant (4[3,4] vs. 5[4,5],Z= -2. 282,P<0. 05]. Conclusion For paraclinoid aneurysms,the recurrence rate of aneurysms with microsurgical treatment is lower than that of endovascular embolization,and is beneficial to the prognosis of patients with Hunt-Hess grade Ⅲ-Ⅳ.

Objective To explore the methods of improving diagnosis correctness between the patients with prostate cancer and benign prostatic hyperplasia.Methods Totally 87 patients with benign prostatic hyperplasia or prostate cancer which confirmed by MRI and prostate biopsy for the PSA significantly increased in our hospital from July 2013 to March 2016 were collected.By using the three methods of the PI-RADS V2 score,the T2WI+DWI/ADC+DCE-MRI+MRS and PI-RADS V2 score+MRS to diagnose,and comparing with the pathology results,the diagnostic consistency of the two physicians were analyzed.The sensitivity,accuracy and specificity of three ways were compared,and the correlation between PI-RADS V2 score and Gleason score were calculated.Results The diagnostic consistency of the two physicians:PI-RADS V2 score,K=0.951;T2WI+DWI/ADC+DCE-MRI+MRS score,K=0.838;PI-RADS V2+MRSI score,K=0.937.The correlation between PI-RADS V2 score and Gleason score,r=0.871,P=0.001 4;diagnostic sensitivity,specificity and accuracy of PI-RADS V2 score were 77.3％,74.4％,75.9％;diagnostic sensitivity,specificity and accuracy of T2WI+DWI/ADC+DCE-MRI+MRS were 88.6％,76.7％,82.8％;diagnostic sensitivity,specificity and accuracy of PI-RADS V2+MRSI score were 86.4％,81.4％,83.9％,respectively.Conclusion Compared with the traditional diagnostic methods,the combination of new prostate report and data system and MRSI can improve the diagnostic accuracy of prostate cancer and benign prostatic hyperplasia.The PI-RADS V2 score is more objective and accurate in the description of the lesion,but the low signal of benign hyperplastic nodules in transitional zone should be dialogued carefully through a variety of image parameters.

Objective To investigate the changes of airway resistance before and after Bronchial Diulation Test in patients with senile asthma and the effect of age on the degree of airway resistance. Methods A total of 29 (> 60 years)senile asthma patients and 21 younger(< 60 y)patients were enrolled in the study. The status of their pulmonary function was determined by MEFV(Maximum expiratory flow?volume curve)and an IOS(impulse oscillometry) system before and after Bronchial Diulation Test,The spirometric indexes such as forced vital capacity,total respiratory impedance,respiratory resistance and respiratory system reactance were collected and analyzed. Results Significant decreases of indexs were found after Bronchial Diulation Test compared with those before Bronchial Diulation Test in the study group(P<0.05,respectively). No significant difference in improvement of indexs was found between the group of patients with senile asthma(age > 60 years)and the group of healthy control(age < 60 years ). Each index of the airway resistance has a very good correlation with FEV1%,with the highest degree of relation is X5%. Conclusion The airway resistance of senile asthmatic patients was significantly improved after diuslation test and these indexes IOS would be valuable in evaluating the changes of airway resistance of senile asthma.

Objective To analyze the results from adopted evaluation of Composite Adaptation of Assistive Devices Program (CAADP) in Shanghai. Methods 1996 potential adaptation objects were evaluated by professional assessment panel with the questionnaire developed by our research group in the early. Results and Conclusion Physical disabilities were the majority. 97.69% of the objects needed different assistance from others or aids in their daily lives, with low quality of life. The assistive devices recommended were an average of 4.3 per person with complete species.

Objective To construct the recombinant modified vaccinia Ankara vaccine expressing HBsAg and investigate its antigenicity and immune effect by different strategies.Methods Target gene was cloned into plasmid pSC1l,the recombinant plasmid pSC11-S was transfected into BHK-21 cells that infected with MVA.Homologous recombination occurred between MVA and pSC1 1-S.The recombinant virus MVA-S was selected by several rounds of blue/clear plaques.Target gene was identified by PCR.The expression of target gene was analyzed by Western Blot.The BALB/c mice were immunized with single immunization and consecutive immunization strategy.The level of specific cellular response was measured by ELISpot assays.Results The recombinant virus expressing target gene was confirmed by PCR and Western Blot showed that MVA-S could elicit specific cellular response.There were no significant differences among two kinds of DNA vaccine and recombinant MVA-S.The prime/boost regimen could induce higher cellular response than single DNA vaccine immunization.High doses of MVA-S boost with the same DNA vaccine prime could induce highest response.There was no significant difference between different DNA vaccines (pVR-S,pcDNA-S) followed by same MVA-S boost.Conclusions MVA-S expressing target was constructed successfully,and it could induce specific cellular immune response which was higher with the strategy of DNA prime/MVA-S boost than that of single immunization.

Objective Cervical lymph node enlargement may be attributed to inflammation or tumors .This study was to analyze the pitfalls in fine-needle aspiration cytology ( FNAC) of cervical lymph nodes and the measures for avoiding misdiagnosis of cervical lymph node lesions . Methods We retrospectively analyzed the data about 435 cases of FNAC in comparison with the results of corre-sponding tissue biopsies in cervical lymph nodes . Results Among the 435 cases, 7 showed disagreement between the results of cytolog-ic and histologic diagnoses, which included 5 males and 2 females, at the age of 41 to 71 (58.4 ±8.9) years.Six of the cases presented with local lymph node enlargement and 1 with generalized lymphadenopathy, all with enlarged lymph nodes palpable 1-4 cm in diameter . Based on the results of FNAC, 1 case of malignant lymphoma was misdiagnosed as lowly differentiated adenocarcinoma, 1 case of lympho-ma misdiagnosed as poorly differentiated metastatic carcinoma, 2 cases of lymphoma diagnosed as lymphoproliferation and recommended for biopsy, 1 case suggestive of malignant tumor without further classification, and 2 cases microscopically characterized and recommended for lymph node biopsy.Compared with the results of the biopsy, FNAC achieved a 99.3%coincidence rate of qualitative diagnosis (432/425), with a misdiagnosis rate of 1.6%(7/435). Conclusion FNAC plays a very important role in the initial identification of the nature of lymph node lesions and the type of tumors.Practiced puncture skills and intimate knowledge about the histopathological features, diagnostic criteria, and differential diagnosis of the lymphatic system disorders are essential for improving the diagnostic accuracy of FNAC .

BACKGROUND:Young patients with cruciate ligament injury and high activity levels are prone to grade III medial colateral ligament injury in knee joint. Conventional construction methods such as direct restoration of injured ligaments and muscle-tendon transposition may induce the loosening of medial colateral ligament complex. OBJECTIVE:To explore the clinical effect of autologous tendon transplantation for grade III medial colateral ligament injury of the knee joint. METHODS: Forty-seven patients with grade III medial colateral ligament rupture were treated by operative repair and reconstruction. After repairing the anatomical structure of medial colateral ligaments, we liberated organic semitendinous tendon, bone-patela tendon-bone, and hamstring tendon, to reconstruct superficial medial colateral ligament. RESULTS AND CONCLUSION:Thirty-seven patients were folowed up for 48 months. Preoperative Lysholm score, IKDC score, Tegner score at the final folow-up, and the widened medial joint space on valgus stress X-Ray films were significantly improved compared with before reconstruction (P < 0.05). According to clinical efficacy scores, 12 cases achieved excelent reconstruction, 18 cases achieved good reconstruction, 4 cases achieved moderate reconstruction, and 3 cases achieved poor reconstruction. The excelent and good rate was 82%. Medial colateral ligament reconstruction based on repairing the anatomical structure is effective and satisfactory. Autologous bone-patela tendon-bone or semitendinous tendon is a good choice for medial colateral ligament reconstruction.

OBJECTIVE: To investigate the effect and mechanism of herpes simplex virus type 1 (HSV-1) based vector mediated interlukin-1 receptor antagonist (IL-1Rα) gene for knee osteoarthritis in rabbits. METHODS: HSV-1 vectors containing IL-1Rα genes were constructed and injected into the joint space of the osteoarthritis knee in rabbits for 4 weeks. The rabbits were sacrificed, and the knees were lavaged, dissected and the effect of transgene expression was analyzed. Levels of IL-1Rα and IL-1 expression in the recovered lavage fluids were measured with a cytokine ELISA kit. Cartilage from the lesion areas of medial femoral condyle and synovium were observed with hematoxylin and eosin (cartilage and synovium) and toluidine blue (cartilage). The blank control group was injected pHSV-LacZ vector into rabbit knees. RESULTS: Intra-articular delivery of pHSV-IL-1Rα-LacZ resulted in a significant inhibition of IL-1 level and cartilage degradation compared with those in the blank control group (P<0.05). CONCLUSION pHSV-LacZ is an ideal vector to mediate intra-articular gene delivery in the rabbit model of osteoarthritis. Continuous intra-articular expression of IL-1Rα can treat knee osteoarthritis by inhibiting IL-1.
Animals ; Arthritis, Experimental ; therapy ; Female ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; Herpesvirus 1, Human ; genetics ; Injections, Intra-Articular ; Interleukin 1 Receptor Antagonist Protein ; genetics ; Male ; Osteoarthritis, Knee ; therapy ; Rabbits

Objective To investigate the status and needs of professional training in the rehabilitation institutes associated to Shanghai Disabled Persons' Federation. Methods All the rehabilitation institutes associated to Shanghai Disabled Persons' Federation were surveyed with their theme, frequency and targeted participants of training in 2010. The intention of training of rehabilitation personnel were investigated and ranged as importance and urgency. Results There were some trainings in all the rehabilitation institutes, which varied with the property and level of the institutions. The theme of training did not meet the intention of the personnel. Conclusion A preliminary training system for the rehabilitation personnel has been established in Shanghai Disabled Persons' Federation system. It is needed to improve the needs assessment,curriculum, and teaching.