In this study, we applied in vivo electroporation(EP) for HBV DNA vaccine administration to improve the cell transfection rate of plasmid DNA and to enhance the immune response. In BALB/c mice (8 mice in each group), the luciferase activity (16170?12533RLU) was 4 digits higher than that of the non EP control (8 02?8 00RLU), the difference between them was very significant ( P

[Objective]To evaluate the inhibiting affection of Ad-BMP-7 to osteoporosis of ovariectomized rats.[Method]Mice model of osteoporosis was made by surgical ovariectomy,and divided into 4 groups:A.group of vertebral body intramedullary injection of normal saline;B.injection of Adovirus(Ad)-GFp;C.injection of Ad;D.injection of Ad+BMP(bone morphogenetic protein),the bone losing of whole body were monitored by measuring dry bone weight after 1 month,the osteoporosic condition of bone trabecular were observed by histology,and bone histomorphometry of vertebral bodies were carried to quatitatively measure the dynamic and static indexes of osteoporosic bone.[Result]The dry bone weight of AdBMP-7 rats werehigher than that of other groups.Under the microscope,the bone trabecula form of the Ad BMP-7 groups were more integrity,however,the bone trabecular of the other groups showed sharpen and ruptured.In the bone histomorphomitry,the numbers of the osteoblasts,the thickness of osteoid of group D were obviously higher than other groups.[Conclusion]Ad BMP-7 injected into the vertebral bodies of ovariectomized rats may secrete BMP-7,and inhibits the osteoporosis of ovariectomized rats in the early stage.The protective effect of this gene was not restricted to bones receiving intramedullary injection of the vector,but occurred in all bones that were evaluated.This proof of concept encourages further development of gene therapy approaches to the treatment of osteoporosis.

Objective To investigate the effect of pyrrolidine dithiocarbamate(PDTC) on monocyte chemotactic protein1 (MCP-1) in rejection of cardiac allograft and its mechanisms.Methods Heterotopic cervical heart transplantation was performed by cuff-technique.The SD rat recipients were randomly divided into 3 groups:AR group (Acute rejection,n =12),both the recipients and donors were without any treatment.CsA group(n =12),the recipients were treated with 10 mg/kg cyclosporine A after transplantation.PDTC group(n =12),the recipients were treated with 100 mg/kg PDTC after transplantation.All the cardiac allografts were harvested at different time post transplantation according to requirements.We studied allograft myocardial fibrosis wih the help of Masson stain,immuno-histo-chemistry and western blot also were used to detect the expression of MCP-1.Results The survival time of the cardiac allografts was significantly longer in PDTC group than in acute rejection group and CsA group(P ＜ 0.01),and myocardial fibrosis of cardiac allografts in PDTC group was significantly decreased (P ＜ 0.01).The IOD in PDTC group was markedly lower than in CsA group (P ＜ 0.01).Conclusion As the inhibitor of NF-κB,PDTC can significantly relieve rejection of cardiac allograft by inhibiting the expression of MCp-1.

Objective To investigate the potential roles of 14 -3 -3 in the pathogenesis of the cervical cancer.Methods Tissues from 107 patients with cervical cancer(cervical cancer group),50 cases of cervical intra-epithelial neoplasia (cervical intraepithelial neoplasia group)and 50 cases of chronic cervicitis (chronic cervicitis group)were obtained.Immunohistochemistry was performed to determine the expression of 14 -3 -3 protein in tissues of the three groups.Results The positive rate of 14 -3 -3 in cervical cancer group(75.70%)was signifi-cantly higher than that in cervical intraepithelial neoplasia group and chronic cervicitis group(42.00%,30.00%, respectively)(χ2 =17.00,29.96,all P <0.01),while there was no significant difference of 14 -3 -3 level between the cervical intraepithelial neoplasia group and chronic cervicitis group(χ2 =1.56,P >0.05).Moreover,in the cervi-cal cancer group,the positive rate of 14 -3 -3 in lymph node metastasis group was 88.89%,which was higher than that in negative lymph node metastasis group (63.16%)(χ2 =5.15,P <0.05).Conclusion The expression level of 14 -3 -3 protein in cervical cancer is increased,it may be correlated with cervical cancer cell malignant differenti-ation and metastasis.Perhaps,14 -3 -3 will play a key role as a novel target for tumor therapy.

Objective:To investigate the effect of Genistein combined with cyclosporine A on chemokine receptor CXCR3 in rejection of cardiac allograft in rats.Methods:Heterotopic cervical heart transplantation was performed from Wistar rats to SD rats by using cuff-technique.The SD rat recipients were randomly divided into 3 groups.No treatment was adopted in the acute rejection( AR) group.Rats in the cyclosporine A(CsA) group were treated with cyclosporine A after transplantion.Rats in CsA+Genistein(C+G) group were treated with Genistein combined with cyclosporine A after transplantion.All the cardiac allografts were harvested at 7th day, and made into tissue slices.The HE-staining was used to observe the pathology changes of the allograft myocardia.And the expression of CXCR3 was detected with immunehistochemistry and Western blot.Results:The expression of CXCR3 strongly positively expressed in AR group.The degree of myocardial inflammation in C+G group and the expression of CXCR3 were much lower than the other two groups.Conclusion:Genistein combined with cyclosporine A can significantly relieve rejection of cardiac allograft by inhibiting the ex-pression of CXCR3.

Obiective To set up a method of scaffold evaluation using human cell line as seed cells and screen appropriate scaffold for live tissue engineering, Methods HepG2 cells were plated onto biodegradable polymer scaffolds: PLGA, 3% chitosan (3%CS) and 2% silk fibroin (2%SF), and cultured in vitro. The growth, distribution and function of HepG2 cells in the scaffolds were evaluated using MTT assay, H.E. staining, and urea assay kit. Results HepG2 cells plated on the three scaffolds maintained a proliferative state. In contrast, the cells on the 2%SF proliferated strongly, while the cells on the PLGA and chitin proliferated poorly. Histological examination showed that HepG2 cells distributed evenly on the 2%SF scaffold with a high amount, while few cells could be found on the PLGA and ehitin at day 7. Cell function assay showed that HepG2 cells on the 2%SF and PLGA exhibited slower decrease of urea synthesis compared to HepG2 cells on the chitosan. Conclusion The three scaffolds have good biocompatibility. In contrast, 2%SF scaffold is more appropriate for liver tissue engineering. This method may be used for scale screening of scaffolds for liver tissue engineering.

Objective To explore the strategy and outcomes of surgical treatment of thoracic ossification of ligamentum flavum(OLF),especially combined with ossification of posterior longitudinal ligament,thoracic kyphosis and epidural adhesion.Methods Fifty-three cases of thoracic OLF from January 2003 to December 2009 were reviewed retrospectively.All patients were treated by the methods of en bloc resection of semi-facet and lamina.All patients were followed up for more than half an year,including 32 males and 21 females,aged from 43 to 73 years(average 54.7 years).The lesions located in upper thoracic for 18 patients,and in thoracolumbar for 35 patients.For multi-level or jumping OLF patients,the responsible levels were determined by combination of images and clinical symptoms.For multi-level OLF with ossification of posterior longitudinal ligament(OPLL)or thoracic kyphosis(＞50°),multi-level pedicle screw fixation and correction of kyphosis were performed.For dural adhesion patients,part of cerebrospinal fluid was released with a caudal incision of dural sac resulting in collapse and epidural arachnoid separation.Ossific and adhesion dura mater were removed with integrity of arachnoid.The surgical outcomes were evaluated with preoperative and postoperative thoracic Japanese Orthopaedic Association(JOA)score,Nurick grade and neurologic functional recovery ratio.Results Fifty-three cases were followed up for 6 months to 6 years,with an average of 18 months.The average preoperative JOA score was 4.3±2.3,which significantly increased to 8.3±1.8 after operation.Postoperative neurologic functional recovery rates were 11% to 80%(average 65.8%),including excellent in 18 cases,good in 20,fair in 10,and poor in 5.The excellent or good rate was 71.7%.The mean preoperative Nurick grade was 3.7(2-5 grade)and decreased to 2.3 grade after operation.Conclusion En bloc resection of semi-facet and lamina is a safe and effective method for treatment of thoracic OLF.For the patients with OPLL or kyphosis,pedicle screws fixation and kyphosis correction was beneficial for recovery of neurologic function of thoracic OLF patients.

The association of non-HDL-cholesterol and non-HDL-C-to-HDL-cholesterol ratio (non-HDL-C-to-HDL-C ratio) with early diabetic nephropathy in patients with type 2 diabetes mellitus was investigated.Non-HDL-C and non-HDL-C-to-HDL-C ratio were positively related with microalbuminuria (P＜0.05 or P＜0.01).Non-HDL-C-to-HDL-C ratio is an independent risk factor of early diabetic nephropathy in patients with type 2 diabetes mellitus.

Objective To compare the effectiveness of comprehensive rehabilitation therapy with that of surgical intervention for patients with recurrent lumbar disc herniation (LDH).Methods A total of 124 LDH patients were recruited antd randomly divided into an experimental group and a control group,each of 62.The experimental group was given comprehensive rehabilitation consisting of medicine,physical therapy,and spinal stabilization exercises,while the control group received lumbar spine surgery.The patients' back and leg pain scores and their Oswestry Disability Index (ODI) scores were compared at admission and discharge,as well as 3 and 12 months after discharge.Anxiety and depression were also assessed at admission and 3 months after discharge.Results At discharge and 3 and 12 months later the average back and leg pain scores and ODI ratings were significantly lower than that at admisshin for both groups.There were no significant inter-group differences.In terms of anxiety and depression,however,these had decreased signifieantly in both groups,but the experimental group's average rating was significantly better at diseharge.Conclusions Comprehensive rehabilitation is superior to surgical intervention in relieving the depression of patients with recurrent LDH.Comprehensive rehabilitation should be considered as the first ehoiee for patients with rccurrent LDH,and only supplemented by surgical intervention if it is not effective.

Objective To get minimally invasive and cosmetic effect,minimal incision was used in thyroid surgery.Methods By using MPBS series instruments,thyroid surgeries were performed through anterior cervical tiny incision approach.Results 164 cases of subtotal theroidectomy and 78 cases of thyroid individual lobe resection were performed.The length of incision were(2.0 ± 0.5)cm,and the duration of operation time were(40 ± 10)min.the volume of blood loss were(30 ± 10)ml.There was no conversion to traditional open operation.After 3 years follow-up,short and long term complications were not found.Conclusions Minimally incision thyroid surgery has advantages of less trauma,cosmetic effect and does not compromise the safety.The disadvantages of this method is limited operative field and inconvenience for operation,which need to be further improved.