Background p16INK4a gene plays an important role during the aging and senility.So it was well known to be a leading gene associated with aging.Corneal endothelial cells(CECs) always get trapped in the G1 phase due to the lack of proliferative ability.Whether it is relative with cell senescence is unclear.Objective This study was to investigate the expression ofp16INK4a gene and Bmi1 gene in human CECs from different aged donors ex vivo.Methods The corneal rims,the residual of corneal tissue preserved in DX solution after penetrating keratoplasty,was used in the present study.Parameters were recorded for the donor,including the age,death to preservation interval and preservation to surgery interval.Corneal endothelium survival rate and endothelial cell density were evaluated by trypan blue-alizarin red dying immediately after penetrating keratoplasty.Routine haematoxylin and eosin staining was also performed to proof the normal structure of the cornea.Sections of corneas from different aged donors were classified into ＜30 years group,30-50 years group and ＞50 years group and were immunostained to assess the expressions of p16INK4aprotein,Bmi1 protein and Ki67 protein in CECs.Total RNA was extracted from independent corneal sample for the evaluation of p16INK4a mRNA,Bmi1 mRNA and Ki67 mRNA expression in CECs by quantitative real-time PCR(qRT-PCR).Results The endothelial cell density of each group was (3069 ±172),(2748±64),(2444 ±178)cells/mm2,respectively.Haematoxylin and eosin staining showed the normal structure of corneal epithelium,stroma and endothelium.qRT-PCR examination revealed an age-related increase in p16INK4a mRNA expression in the CECs(F =5.703,P =0.014) and a decrease in Bmi1 mRNA and Ki67 mRNA expression (F =3.950,P =0.042;F=548.500,P =0.000).The further comparison verified a significant elevation in the expression of p16INK4a mRNA in the CECs of the ＞50 years group compared with ＜30 years group and significant decline in Bmil mRNA and Ki67 mRNA the expression (P =0.006,0.013,0.000).Immunohistochemistry in situ confirmed the expression and nuclear localization of p16INK4a protein in CECs,and the expressing intensities of Bmi1 and Ki67 proteins in the elder donors were weaker than those of the younger donors.The immunofluorescence exhibited that the expressing intensity of p16INK4a protein in CECs of 58 years old donor was higher than that of 23 years old donor,showing a consistent result with that of qRT-PCR.Conclusions Expression of p16INK4a gene increases and that of Bmi1 gene decreases upon age.These results suggest that p16INK4a gene is associated with senescence of human CECs.

Objective To compare safety,completeness and long-term results between lobectomy by video-assisted thoracoscopic surgery(VATS) and thoracotomy for clinical stage Ⅰ / Ⅱ non-small cell lung cancer(NSCLC).Methods Between July 2010 and December 2011,673 patients with clinical stage Ⅰ / Ⅱ NSCLC from 11 hospitals underwent pulmonary lobectomy(425 VATS,248 thoracotomy).Patients were matched by propensity score matoh method to create two comparable groups,402 patients remained eligible for analysis,with 201 patients in each group.Perioperative variables and follow-up results were compared between the two groups.Results Patients characteristics were comparable between the two groups,except for a slight lower preoperative FEV1,FEV1％ and FEV1/FVC in the VATS group (P ＜ 0.05).There was one postoperative death in each group.There was no significant difference in operative time,mediastinal lymph nodes stations/numbers dissected and postoperative drainage time between the two groups.The operative blood loss was lower in the VATS group(P ＜ 0.001),the mean hospital stay was shorter in the VATS group(P ＜ 0.001).Postoperative pneumonia/pulmonary atelectasis rate was lower in the VATS group than in the thoracotomy group (5.1％ vs.10.4％,P ＜ 0.05).Median follow-up time was 20 months in both groups.One-year and two-year recurrence-free survival was 90％ vs.88％ and 80％ vs.70％,respectively(P =0.163).One-year and two-year overall survival were 96％ vs.95％ and 88％ vs.85％,respectively(P =0.226).Conclusion For clinical stage Ⅰ / Ⅱ NSCLC,VATS lobectomy is a feasible procedure with shorter hospital stay and less complications,while its surgical and oncologic outcome is comparable to that of thoracotomy lobectomy.

Objective To investigate the relaxant effect of salanic acid A isopropyl ester(isopropyl 3-(3,4-dihydroxyphenyl)-2-hydroxypropanoate)and its mechanism.Methods Isolated rat pulmonary artery was perfused and the tension of the vessel was measured.To observe the relaxant effect of salianic acid A isopropyl ester on the pulmonary artery precontracted by noradrenaline(NE)and the role of endothelial cell and vascular smooth muscle cell on vasodilation.Results Salianic acid A isopropyl ester relaxed the endothelium-intact artery precontracted by NE in a concentration-dependent manner,and the effect was depressed with denuded endothelium.Salianic acid A isopropyl ester right-shifted the concentration-response curve of KCl and suppressed the contractions induced by NE and CaCl2 in the Ca2+-free-K-H solution.The relaxant effect of salianic acid A isopropyl ester was inhibited by N?-nitro-L-arginine methyl ester hydrochloride(L-NAME),Indometacin,CsCl,Tetraethylammonium(TEA),Glibenclamide,and BaCl2,but not that by Propranolol,Metoprolol,and Salbutamol.Conclusion Salranic acid A isopropyl ester could significantly relax the pulmonary artery of rats,which may derive in part from endothelium.The relaxant effect of salianic acid A ispropyl ester may relate to NO pathway,prostacyclin(PGI2)pathway,calcium channel,and potassium channel.

Objective:To investigate the effects of autophagy-targeted DNA vaccine against Japanese encephalitis virus (JEV) on the immune-related functions of dendritic cells (DCs) in BALB/c mice.Methods:Healthy female BALB/c mice were randomly divided into 5 groups and injected with pcDNA3.1(+ ) empty vector (negative control), pJME plasmid, recombinant pJME-LC3 plasmid muscle, sterile PBS (blank control group) and live attenuated JEV vaccine (positive control group), respectively. The mice were immunized three times with an interval of two weeks. Splenic DCs were isolated two weeks after the last immunization. Immunofluorescence assay was used to observe the expression of the recombinant plasmid in dendritic cells. Expression of major histocompatibility complex Ⅱ (MHC Ⅱ) on DCs and the uptake of FITC-Dextran by DCs were observed by flow cytometry. CCK8 assay was used to detect the effects of DCs on the proliferation of spleen mononuclear cells from allogeneic mice.Results:The expression of plasmid-encoded protein in the DCs of the pJME-LC3 group was significantly higher than that of the pJME, pJME-LC3+ 3-MA and pcDNA3.1(+ ) empty vector groups. The uptake of FITC-Dextran by DCs was significantly enhanced in the pJME-LC3 group than in the other groups ( P<0.05), and the expression of MHC Ⅱ moleculars on DCs was increased in the pJME-LC3 group as well ( P<0.05). The splenic DCs from the mice in the pJME-LC3 group had a stronger effect on the proliferation of spleen mononuclear cells from allogeneic mice that those from other groups ( P<0.05). Conclusions:The recombinant plasmid pJME-LC3 could promote the ability of mouse splenic DCs to present antigen and to stimulate lymphocyte proliferation after immunization, suggesting that the autophagy-targeted recombinant DNA vaccine against JEV could enhance immune responses by affecting the function of DCs in BALB/c mice.

Objective To evaluate safety and validity of 3D video-assisted thoracoscopic technology in cardiac surgery.Methods Between October 2013 and July 2014,50 patients underwent 3 D Video-assisted thoracoscopic cardiac surgery including:5 atrial septal defect closure,1 ventricular septal defect closure,repairment of 1 unroofed coronary sinus,1 partial atrioventricular septal defect and 1 cor triatriatum,28 mitral valve replacement (1 re-MVR),1 tricuspid valve replacement,9 mitral valvuloplasty,2 left atrial myoxoma resection,and 1 left internal mammary artery harvest + minimally invasive direct coronary artery bypass.All procedures were performed under extracorporeal circulation,using 30° thoracoscope and 3D high resolution monitor.Results There was no perioperative death.One patient was converted to median sternotomy because of pericardial adhesions.Mean operative time was(232.8 ± 54.7) min,mean cardiopulmonary bypass time was(128.0 ±42.5) min,mean aortic cross-clamp time was (74.7 ± 25.1) min.Mean ICU stay was (30.4 ± 22.0) h,mean duration of mechanical ventilation was(13.7 ± 11.9) h.Mean postoperative drainage time was (3.34 ± 1.91) d,mean hospital stay was (6.52 ± 3.06) days.There were 11 (22％) operative complications,including 4 intrathoracic bleeding,1 wound infection,3 hemothorax,1 brachial plexus injury,1 intractable hiccup and 1 mild paravalvular leakage.Compared with 20 mitral valve replacement by 2D thoracoscopy,the 3 D group was slightly less time consuming(P ＞ 0.05).Conclusion 3 D video-assisted thoracoscopic cardiac surgery is feasible and safe,and it' s worthwhile to promote this promising technology.

Objective To analyze the causes and outcome of conversion from tacrolimus (FK506) to cyclosporine-A (CsA) in recipients after liver transplantation.Methods 317 consecutive liver transplantation recipients in our department received anti-CD25 monoclonal antibody,FK506, mycophenolate mofetil and corticoid for prophylaxis of cellular rejection.The blood FK506 trough level was (10-15)?g/L within the first 30 days,(8-12)?g/L within next 60 days,and (5-8)?g/L was kept during 90 to 180 days after transplantation.All recipients reveived a follow-up of 6 months. Remits Sixteen out of 317 recipients (5.05%) required conversion from FK506 to CsA.The clinical indications for conversion included:neurological adverse effect of FK506 in 5 cases (31.25%),hema- tological adverse effect in 2 cases (12.5%),gastrointestinal effect in one case (6.25%),not capable of reaching therapeutic window concentration in 3 cases (18.75%),refractory hyperglycemia in 2 ca- ses (12.5%),and economic factor in 3 cases (18.75%).The majority of recipients demonstrated clinical improvement after the switch,except 2 of 16 patients (12.5%) had to be reconverted to FK506 due to renal disadvantage.No dead recipient and adverse effect correlated to immunosuppres- sive agent conversion were seen.Conclusion If necessary,conversion from FK506 to CsA in patients undergoing liver transplantation is safe and effective.

Objective To analyze the characteristics of erectile dysfunction(ED)in aged men.Methods Erectile dysfunction(ED)was diagnosed according to the International Index of Erectile Function(IIEF).78 aged men(average 65.9 yrs)as the study group,and 82 young patients(average 36.0 yrs)as the control group,all with ED,Were compared in complicating diseases,self-rating SDS scores,penile-brachial indexes(PBI),the time of achieving erection,and ejaculatory latency time.Results The main complicating diseases in study group were cardiovascular diseases(42 cases,53.9%),lower urinary tract symptoms(LUTS)(31 cases,39.7%)and diabetes(26 cases,33.3%).In the control group,the main complicating diseases were chronic prostatitis(52 cages,63.4%),premature ejaculation(PE)(32 cases,39.0%)and depression(12 csses,14.6%).SDS scores of study group and control group were(29.13±5.63)and(39.59±13.31),PBI were(0.78±0.12)and(0.91±0.06),the time of achieving erection were(13.85±5.75)min and(3.61±4.29)min,ejaculatory latency time were(7.03±5.35)min and(3.81±5.53)min.All with significant difference(P＜0.01).Conclusion Most of the ED old men were complicated with organic diseases,such as the time of achieving erection,PBI low scores and longer ejaculatory latency time.

BACKGROUND: Tissue engineering plays an important role in treating rotator cuff injury. Promoting the healing of injured rotator cuff and preventing adhesion by regulating growth factor expression are main functions. OBJECTIVE:To compare the effects of corticosteroids and hyaluronic acid on the expressions of epidermal growth factor (EGF),platelet-derived growth factor (PDGF),transforming growth factor ? (TGF-?) and basic fibroblast growth factor (bFGF) in injured rotator cuff. DESIGN,TIME AND SETTING: The randomized controlled animal experiment was performed at the Animal Experimental Center,School of Medicine,Shanghai Jiao Tong University from July to September 2007. MATERIALS: Totally thirty-six Sprague-Dawley (SD) rats were randomly assigned into four groups: normal group (n=6),rotator cuff injury group (n=6),hyaluronic acid group (n=12) and corticosteroids group (n=12). METHODS: Full-thickness defects across 50% of the total width (5 mm) of the bilateral infraspinatus tendon were excised. Models of rotator cuff injury were created at the synovium. 0.05 mL hyaluronic acid was injected into the subacromial bursa mucosa in the hyaluronic acid group,whereas 0.05 mL corticosteroids in the corticosteroids group. MAIN OUTCOME MEASURES: At three and six weeks after injury,the infraspinatus were harvested to detect EGF,PDGF,TGF-? and BFGF by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: By using corticosteroids treatment,the expression of EGF remained greater at the six weeks than twofold at the three weeks; the expression of EGF after hyaluronic acid treatment greatly decreased at six weeks (P

Objective To study the rehabilitation effect of Bellidifolin for injuried sciatic nerve,and to explore whether ciliary neurotro-phic factor ( CNTF) is involved in this mechanism. Methods The right sciatic nerver of 225 male wistar rats was cut and sewed under mi-croscopy. Rats were devided into 5 groups,as control group,Bellidifolin 25 mg group,50 mg group、75 mg group and Mecobalamin group. The control group were injected sodium chloride,other groups were injected different dose of Bellidifolin and Mecobalamin. 1,3 and 5 weeks later, the motor nerve conduction velocity( MNVC) and gastrocnemius muscle cross-sectional area were detected,CNTF positive area were analysed by immunohistochemical method. Results There were differences among bellidifolin groups,control group and mecobalamin group in Nerve conduction velocity. Within Bellidifolin groups,50 mg group compared with 25 mg and 75 mg groups,there were statistically differences( P=0. 025). Three weeks after operation,gastrocnemius muscle cross-sectional area of control group,mecobalaming grop and Bellidifolin 25 mg group,50 mg group,and 75 mg group were(455. 06 ± 29. 38),(679. 03 ± 81. 48),(465. 31 ± 71. 55),(670. 24 ± 91. 26) and (669. 28 ± 78. 54) respectively,compared with control group and Bellidifolin 25 mg group,others had a significant difference(P<0. 05). CNTF expres-sion showed billidifolin 50 mg group are higher than others(P<0. 05). Conclusion Bellidifolin can improve the rehabilitation of injured sciatic nerve. CNTF is involved in this mechnism.

OBJECTIVETo explore the theory of "Fei and Dachang being interior-exteriorly related" and the pathogenesis of lung injury by observing changes of inflammatory cytokines and oxygen free radicals in ulcerative colitis (UC) rats.

METHODSTotally 50 healthy male Wistar rats were randomly divided into two groups, the normal control group and the model group, 25 rats in each group. The UC model was established by allergizing colon mucosa combined with TNBS-alcohol (50%) enema. Another 25 rats were recruited as the normal control group. At week 2 and 4 after modeling, the pathomorphological changes of the lung were observed. Furthermore, the contents of tumor necrosis factor alpha (TNF-alpha) and IL-1beta were determined by ELISA. The activities of superoxide dismutase (SOD) and malondialdehyde (MDA) were evaluated with colorimetry.

RESULTSCompared with the normal control group, the pathomorphology of the lung tissue in the model group appeared abnormal at week 2 and 4. Compared with the normal control group, levels of TNF-alpha, IL-1beta, and MDA in the lung tissue significantly increased in the model group (P < 0. 01) and the activities of SOD significantly decreased (P < 0.05, P < 0.01).

CONCLUSIONTNF-alpha, IL-1beta, SOD, and MDA might be common material bases for the large intestine involved in lung disease of UC patients, thus providing a modern scientific basis for the theory of Fei and Dachang being interior-exteriorly related.

Animals ; Colitis, Ulcerative ; diagnosis ; metabolism ; pathology ; Cytokines ; metabolism ; Interleukin-1beta ; metabolism ; Lung ; metabolism ; Male ; Malondialdehyde ; metabolism ; Medicine, Chinese Traditional ; Rats ; Rats, Wistar ; Reactive Oxygen Species ; metabolism ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism