Objective To investigate the impacts of long cold ischemic preservation time on the ultrastructural changes of donor heart and to provide the data for expanding the heart donor pool. Methods Heart was obtained from brain dead donor and preserved in the the University of Wisconsin (UW) solution at 4℃. Cardiac tissues were harvested at different time points of cold ischemic preservation (6, 8, 10, 12, 14 h) and observed under electron miscroscope. Results Donor heart did not have significant pathologied and ultrastructural changes when cold ischemic preservation time was 6 h. After that, time related impairment of myocardia and endothelium of coronary artery was seen.When ischemic time was longer than 12 h, focal myocardial necrosis and complete loss of the endothelium were detected. Conclusions Myocardial ultrastructure is an important index to evaluate the donor heart quality. Heart, which underwent 10 h of cold ischemia preservation time, causes no significant irreversible and pathological ultrastructural changes, and could be used for heart transplantation. When ischemia time was over 10 h, the donor heart presented with irreversible change and was nolonger unsuitable for transplantation.

Objective To analyze diagnosis and management of traumatic atlantal fractures complicated with disruption of the transverse atlantal ligament. Methods Twenty-six patients with traumatic atlantal fractures complicated with disruption of the transverse atlantal ligament, 20 acute ones and six old ones, were managed in our department from March 1995 to March 2005. All the patients had the symptom of neck pain and a certain extent of neurological deficits. The radiographic examination showed that there were fractures in the anterior arch or/and lateral mass of the atlas and that the atlanto-dental interval (ADI) in these patients was 4 to 7 mm. Twenty-one cases were operated on with occipitocervical or atlantoaxial fusion immediately after injury. Five cases received conservative treatment. Results All were followed up for 15 (from 6 to 24) months on average. The 21 cases treated operatively reported relieved symptoms. Four of the five conservative treatments succeeded, but one of them underwent a successful atlantoaxial fusion because the original conservative treatment had failed. Conclusions The severity of clinical symptoms mostly depends on the degrees of atlantal displacement and cord compression. Diagnosis should be made on the basis of the ADI change as well as clinical presentations. For patients who have potential risk of atlantoaxial instability, occipitocervical or atlantoaxial,usion should be performed early.

Objective To summarize the outcomes and clinical experience of orthotopic heart transplantations in Shanghai Zhongshan Hospital.Methods From May 2000 through October 2005,141 patients,101 males and 40 females,diagnosed as having dilated cardiomyopathy in 118,hypertrophic cardiomyopathy in 2,restrictive cardiomyopathy in 2,end-stage ischemic heart disease in 9, primary malignant cardiac tumor in 4,valvular heart disease in 3 and others in 3,underwent orthotopic heart(transplantations) at our center.The operative procedures included 120 bicaval anastomotic cardiac transplantations,19 conventional Stanford orthotopic cardiac transplantations and 2 total heart transplantations.The immunosuppressive therapy strategies included Cyclosporine A or tacrolimus,corticosteroids and mycophenolate mofetil.For the latest 28 patients,induction therapy with Daclizumab was applied.Results There were 3 operative deaths with an operative survival of 97.9 %.During the follow-up from 1 month to 65 months,the actuarial survival rate was 90.8 % after 1 year,84.6 % after 3 year and 81.4 % after 5 year.Acute rejection,infections,graft failure and metastasis of primary cardiac tumor were the main causes of death in the 1st postoperative year,while acute rejection and graft coronary vasculopathy were the leading causes of death thereafter.Postoperative complications included acute rejection,infections,renal dysfunction and graft failure.Conclusions Orthotopic heart transplantation was proved to be a reliable choice for endstage heart disease with excellent outcomes.More attention should be paid to surveillance and management of acute rejection, infections and graft coronary vasculopathy in the long-term follow-up.

Objective To investigate the effect of paclitaxel on the intimal proliferation in rat aortic allografts and the possible mechanism on preventing graft arteriosclerosis.Methods Thirty-two inbred(Wista)r rats and 16 SD rats were divided into three groups:the isografts control group(Wistar/Wistar),the(allografts) control group ((Wistar)/SD) and test group(Wistar/SD) randomly(16 rats each groups).The rat abdominal aortic allograft model was used.The rats in test group were treated with paclitaxel after operation and those in control group with(0.9 %) normal saline.The grafts were removed and measured by means of pathology and immunohistochemistry 30 days later.Results The results showed that the thickness of the(aortic) intima,the degree of inflammatory cells infiltration in adventitia,stenosis ratio and the expression of(PCNA) were decreased in test group as compared with the allografts control group.Conclusions Paclitaxel can inhibit intimal proliferation in aortic allografts and prevent the graft arteriosclerosis.The mechanism is related to inhibition of vascular smooth(muscle) cell proliferation and alleviation of aortic allografts rejection.

Objective To discuss appropriate strategy about treatment of rotatory fixation or dislocation of atlantoaxial articulation in children. Methods 32 cases of rotatory fixation or dislocation of atlantoaxial articulation in children were analyzed retrospectively from January 2000 to March 2005, 24 males and 8 females; patients' age was from 3 to 12 years old with an average of 5.5 years old. The mean course of diseases was 6.5 weeks (range: 2 days to 50 weeks). According to Fielding-Hawkins clinical classification, typeⅠ17 cases(1 case with os odontoideum), type Ⅱ 12 cases(1 case with os odontoideum and 1 case with congenital absence of the arch of posterior atlas), type Ⅲ 3 cases. 26 cases received conservational treatments which were recommended as 2-3 weeks of cervical traction, traction weight should be controlled to 1.0-1.5 kg. After a successful reduction a proper external fixation was required to maintain reduction. And another 6 serious cases received posterior atlantoaxial or occipitocervical autograft fusion. Indications for posterior atlantoaxial or occipitocervical atuograft fusion includes: 1) have difficulty in reduction even under proper traction; 2) with obvious neurological symptoms; 3) combined with compensatory deformity of atlantoccipital articulation or other occiptocervical deformities. Results All of 32 cases were followed up from 3 to 50 months with an average of 32 months. 26 cases who received cervical traction and external fixation resulted in satisfactory outcome in which all the torticollis were rectified, bilateral masses were symmetrical on AP position, and ADI was less than 4 mm in dynamic extension and flexation lateral view. ROM in rotation and extension and flexion were completely recovered. All the 6 cases surgically treated obtained sound bony fusion and neural symptoms were improved obviously and torticollis were rectified completely in 4 cases and others (2 cases) stopped progressing after operation. Conclusion Conservative treatment including Glission traction, cranial traction, collar, cast or cervicothoracic external fixation has been proved to be very effective in most of rotatory fixation or dislocation of atlantoaxial articulation in children; however, operative treatment should be considered in the following situations: patients with difficulty in reduction, with neural involvement and compensatory deformity of atlantoccipital articulation or other occipitocervical deformities.

Objective To investigate changes in the number of endothelial progenitor cells (EPC) from peripheral blood and pathological feature in the development of transplant arteriosclerosis in mouse abdominal aortic allografts, and discuss their correlations. Methods A segment of abdominal aorta was transplanted orthotopically from C57BL/6 to Balb/c mice. The grafts were harvested at 3rd day, 2nd week, 4th week and 6th week after the operation and studied by light and electronic microscopy. Regional changes in the lumen and intima were measured with computer imaging analysis system. EPC from peripheral blood were quantified by flow cytometry. Results Endothelium injury and inflammatory cells infiltration were seen in the aortic allografts at 3rd day after transplantation.Neointimal lesions and acute rejection were observed as early as 2nd week after surgery. The lumen of allografts was significantly narrowed due to neointima hyperplasia and had progressed at 4th and 6th week postoperatively. The number of circulation EPC was increased from 1 st day after operation and reached the peak at 3rd day. Thereafter the number of EPC was decreased rapidly and significantly less at 14th and 28th day postoperation than that pre-operation. Conclusion Abdominal aortic transplantation from C57BL/6 to Balb/c mice presents typical pathological feature of transplant arteriosclerosis. The number of EPC from peripheral blood is related to the process of injured endothelial repair and neointima formation of aortic grafts. EPC count may be considered a novel biological marker and therapeutic intervention for transplant arteriosclerosis.

objective To observe the cyclooxygenase-2(COX-2)expression and the cell proliferation and apoptosis of gastric adenocarcinoma cells after transiently transfecting gastric cancer cells using specific COX-2 small interfering RNA(siRNA)and discuss the role of COX-2 in gastric tumorigenesis and the effect of RNA interference(RNAi)as anti-cancer gene therapy.Methods Three groups were included in the study,i.e.a COX-2 siRNA group,a non-sense siRNA group and a control group.Gastric adenocarcinoma cells SGC7901 were cuhured at 37℃ in an atmosphere containing 5%CO2.72 hours after transfecting SGC7901 cells with specific COX-2 siRNA or non-sense siRNA,RT-PCR was used to detect COX-2 mRNA,immunohistochemistry and Western blot were taken to detect the expression of COX-2 protein and flow cytometry was taken to detect the cell cycle and apoptosis.MTT method was used to detect the proliferation and activity of the cells every day for one week after transfection.Results The expression of COX-2 mRNA and protein in the COX-2 siRNA group was obviously suppressed as compared with non-sense siRNA group and control group.No change was found between the non-sense siRNA group and the control group.The reduced expression of COX-2 could inhibit SGC7901 cells proliferation and induce cell apoptosis,but had no effect on cancer cell cycle.Conclusions The experimental results suggest that effectively inhibiting the expression of COX-2 can suppress the proliferation of gastric adenocarcinoma cell and promote the process of cancer cell apoptosis.so RNAi is a powerful tool of gene therapeutic method.

Objective This research investigated the coagulation of critically ill patients for predicting the prognosis of 28 day in a university hospital emergency room.Methods A prospective investigation was done in the emergency room of Beijing Chao-Yang Hospital,Capital Medical University from June 2015 to May 2016,and 28-day mortality was recorded.Whole blood cell analysis,blood gas analysis and clotting test were done and repeated after patients in hospital.Results A total of 1 992 patients were enrolled,and divided into two groups:survival (n =1 522) and dead (n =470).No significant difference of age,gender,body mass index and disease composition were found between the two groups (P ＞0.05).APACHE Ⅱ of the survival and dead groups were (12.11 ±4.12) and (21.15 ± 5.55) respectively.D-dimer and platelet account of the dead group were M (Qr) 265 (0,718) μg/L and (208.16±89.87) × 109/L-1 respectively,significant differences were found between the two groups (P ＜ 0.05).Coagulation was found deteriorated progressively in the dead group,whereas improved in the survival group.The risk factors of poor prognosis,which were the increased APACHE Ⅱ and D-dimer,were detected by Logistic analysis and ROC curve,especially the D-dimer.Conclusions Coagulation abnormalities were found in the critically ill patients of emergency room.The increasing of D-dimer is one of the risk factors of poor prognosis.

Objective Ascending aortic dissection(AAD),for which the pathogenesis remains unknown,is life-threatening.Matrix metalloproteinase-9(MMP-9)and the pathological changes of vascular smooth muscle cells(VSMCs)have been reported to have roles the pathogenesis.The study examined the expression of matrix metalloproteinase-9(MMP-9)and the pathological changes of,VSMCs in patients with AAD.Methods AAD samples were taken from 35 patients(disease group)in acute phase during aortic replacement operation for AAD and control samples were corresponding part of ascending aorta(control group,n=21)collected from the donor hearts for transplantation.Transmission electron microscepe,hematoxylin-eosin(H-E)staining.Mallory staining were used for observing the pathological changes of VSMCs and matrix in the affected aortic wall.The immunohistochemicai staining of MMP-9 was carried out in both groups and semi-quantified by staining intensity analysis.The affected patients were further grouped according to the diameter of dissected aorta as with a AAD of <55 mm or with a AAD of≥55 mm.The associations of clinical factors,such as smoking status,hypertensive disease and aneurysm diameter,with the expression of MMP-9 were analyzed.Results Increased synthetic function of VSMCs with decreased density,disrupted elastic fibers and fibrosis in the dissected aortic wall were observed in the disease group,but not in the control group.MMP-9 was scarcely expressed in the aortic wall of the patients in the control group,though it was notably expressed in the VSMCs of disease group.Both subgroups presented more MMP-9 than the control group(both P<0.001).In the disease group,sub-group with a AAD diameter of ≥55 mm presented more MMP-9 than that with a diameter of <55 mm(P<0.05).MMP-9 expression was positively correlated with a history of hypertension(P<0.01)or a great aneurysm diameter(P<0.05).MMP-9 expression was not associated with age,smoking status or other clinical factors.Conclusion Increased secretion of VSMCs and the expression of MMP-9 induced by elevated blood pressure may lead to the destruction of matrix proteins.The resulting fibrosis of the aortic wall would decrease the tensile strength of the wall.When the fibrotic aortic wall dilated further,the increased expression of MMP-9 would aggravate the damage to the wall.It can be speculated that acute AAD would occur as a result of partial tearing of the aortic intima.

Objective To construct a short hairpin RNA (shRNA) adenovirus vector targeting P85 and protein kinase B1 (PKB1/Akt1) and study its effects on the growth of SGC-7901 human gastric adenocareinoma cells. Methods P85 and Aktl shRNA expression frames were subcloned to pGSadeno adenovirus vector with homologous recombination technology to construct pGSadeno-P85 + Akt1 (rAd5-P + A) vector. After screening and amplification, the recombinant adenovirus vector was digested with PacI and transfected into SGC-7901 cells and then its titer and transfection efficiency were detected with fluorescent microscope. P85 and Akt1 mRNA protein expression was identified with real-time PCR and Western blot. The proliferative activity of tumor cells was evaluated with MTr assay and flow cytometry in vitro, rAd5-HK and rAd5-P + A mediated by adenovirus were injected into the established subcutancous SGC-7901 gastric adenocarcinoma in nude mice. During the observation period of 21 days, tumor volume was measured every 3 days to further testify the anti-tumor effect of rAd5-P + A on the SGC-7901 gastric adenocarcinoma cells and cell in situ apoptosis was detected with TUNEL assay. Results The adenovirus vector rAd5-P + A was successfully constructed and it dramatically downregulated P85 and Akt1 mRNA expression in SGC-7901 gastric adenocarcinoma cells. Compared with a control group of SGC-7901 cells and cells transfected with general adenovirus rAd5-HK as control, P85 and Akt1 protein expression 48 h and 72 h after rAd5-P + A transfection was decreased by 57.5% and 63. 7%, 67. 8% and 75.6% with statistical significance(P = 0. 005, P = 0. 003). Cell proliferative activity in rAd5-P + A transfected cells was suppressed from the second day (P <0. 001) and the decreased P85 and Akt1 expression was accompanied by 5.9% -7. 1% decrease of S phase fraction and 12. 1% - 13.7% increase of G0/G1 phase. The tumor volume of rAd5-P + A treated group was smaller than that of the control and rAd.5-HK group with statistical significance (F = 9. 871, P = 0. 025) . Moreover, rAd5-P + A could induce cell in situ apoptosis. Conclusions Adenovirus-mediated targeting P85 and Akt1 shRNA can inhibit the growth of SGC-7901 human gastric adenocarcinoma cells and this may provide a new strategy of combination gene therapy in gastric adenocarcinoma.