Objective To study the effects of suicide gene system mediated by adenovirus containing the CD-TK fusion gene controlled by human vascular endothelial growth factor(VEGF)promoter on apoptosis of human hepatocellular carcinoma cells HepG2 in vitro.Methods The VEGF-expressing HepG2 cells were infected by adenovirus vector containing CD-TK fusion gene controlled by the VEGF promoter(Ad-VEGFP-CDglyTK).The infection efficiencies in HepG2 cells were observed under a fluorescence microscope.The toxic effect of 5-fluorocytosine(5-FC)and ganciclovic(GCV)on infected cells was determined by light microscopy,electron-microscopy and flow cytometry(FCM).Results The transfection efficiency in HepG2 cells increased with the increasing adenoviral titer.The pro-drug(5-FC and GCV)could induce apoptosis of HepG2 cells in certain range of dosage(the doses of GCV+5-FC:1mg/L + 20mg/L,10mg/L + 40mg/L,100mg/L + 60mg/L)at the multiplicity of infection(MOI)of 100.The effect showed a time-dependent manner.HepG2 cells showed typical morphologic changes of apoptosis after administration of the pro-drug(GCV:10mg/L,5-FC:40mg/L)for 72 hours:chromatin condensation and disposition along nuclear membrane.Karyopyknosis and karyoklasis were seen under electron microscopy(?10 000).Apoptotic peak was also shown in HepG2 cells treated with the pro-drug(5-FC and GCV)by flow cytometry.The cell apoptosis rate was increased accordingly as the concentration of pro-drug(5-FC and GCV)increased.The apoptosis was increased obviously in comparison with the negative control group(P

Objective To observe the therapeutic effects and comphcations of treatment by using transcatheter arterial chemombolization(TACE) combined with functional splenic embolization(FSE) on patients with primary hepatocelluar carcinoma (HCC) combinded with hypersplenism.Methods Eighty HCC patients with hypersplenism were randomly divided into two groups by computer program.Observation group was treated with TACE and FSE,while control group was treated by only TACE.The iodine oil deposition,blood routine parameters and related complications were observed.Results (1) After one-month' s treatment,abdominal CT scan showed that 38 cases(79.17％) of patients in the observation group and 27 cases (84.38％) of patients in control group had iodine oil deposition in over 50％ of the tumor area.(2) The white blood cell counting in observation group were (7.65 ± 2.3) × 109/L,(5.89 ± 1.9) × 109/L,(5.02 ± 1.7) × 109/L in the follow-up examination 1 week,1month and 3 month after splenic embolization,respectively,which were significantly increased from the preoperative level of (2.21 ± 1.1) × 109/L(P ＜ 0.05).Platelets counting were (93 ± 29)× 109/L,(127 ± 32) × 109/L and (119 ± 26) × 109/L in observation group at 1 week,1 month and 3 month after splenic embolization,respectively,which were significantly higher than the preoperative level of (39 ± 20)× 109/L (P ＜ 0.05).In the control group,there were no statistically significant difference between the preoperative and postoperative levels (P ＞ 0.05).(3) Both of the the two groups showed no serious complications.Conclusion TACE combined with FSE is a safe and an effective method to treat hypersplenism combined in HCC patients.

Objective To explore the application effect of Sandwich teaching method in clinical teaching in the department of neurosurgery.Methods Totally 144 clinical medicine students were divided into 2 groups:Sandwich teaching group (n=72) and traditional teaching group (n=72).The analysis of test and the questionnaire of the students were conducted after the course.Comparison between the two groups was made using independent sample t test and measurement data were expressed as as ± s.P＜0.05 signifies statistically significant differences.Rank sum test was used to compare the distribution of the test scores.Results The test score was statistically higher in Sandwich teaching group than control group [(76.38 ± 10.84) vs.(69.67 ± 10.50)].And the main distribution of the test scores was 70～90 in Sandwich teaching group while 60～80 in control group.Questionnaire showed that the scores of Sandwich teaching group were statistically higher than those ofthe control group in five aspects such as enhancing self-learning ability,improving the ability to analyze and solve problems,and so on.Conclusions Sandwich teaching method achieves good teaching effect and it is worth recommending in clinical teachifig.

BACKGROUND:Erythropoietin and bone marrow mesenchymal stem cells have been shown to affect myocardial apoptosis. However, few studies concerned their combined application to sepsis-related myocardial injury. OBJECTIVE:To observe the effects of the combination of erythropoietin and bone marrow mesenchymal stem cells on pathology and apoptosis of sepsis rat cardiomyocytes. METHODS:A total of 50 Sprague-Dawley rats were randomly selected and assigned to five groups (n=10). Sepsis models were established by cecal ligation perforation method. Rat models in the bone marrow mesenchymal stem cellgroup, erythropoietin group and erythropoietin+bone marrow mesenchymal stem cellgroup were respectively treated with bone marrow mesenchymal stem cells, erythropoietin and erythropoietin+bone marrow mesenchymal stem cells immediately after model induction via intraperitoneal injection or caudal vein. Model group received cecum ligation and puncture. Control group did not undergo any treatment after the abdomen was opened. Model and control groups were infused with an equal volume of physiological saline via caudal vein. At 24 hours, experimental animals were sacrificed by anesthesia. Myocardial specimens were col ected. Myocardial appearance was observed using hematoxylin-eosin staining. Bax, Caspase-3 and Bcl-2 were tested by western blot assay. RESULTS AND CONCLUSION:Hematoxylin-eosin staining:cardiomyocytes were regularly arranged, showing integrated structure in the control group. Extensive myocardial fiber breakage, disordered arrangement, cardiomyocyte swel ing or shrinkage, and vacuolar degeneration were observed in the model group. Moreover, myocardial interstitial vascular congestion, edema, and inflammatory cellinfiltration were visible. Myocardial tissue was similar between erythropoietin and bone marrow mesenchymal stem cellgroups, with the presence of mild inflammatory cellinfiltration and scattered normal cardiomyocytes. In the erythropoietin+bone marrow mesenchymal stem cellgroup, cardiomyocytes were slightly damaged. Interstitial vascular congestion was not apparent, and a few inflammatory cells infiltrated. Western blot assay results demonstrated that Bcl-2 protein expression was significantly higher (P<0.01), but Bax and Caspase-3 protein expression was lower (P<0.05) in the erythropoietin+bone marrow mesenchymal stem cellgroup than in the erythropoietin, model and control groups. The combination of erythropoietin and bone marrow mesenchymal stem cells in treatment of sepsis-related myocardial injury could lessen myocardial pathological changes, and inhibit cardiomyocyte apoptosis. The mechanisms maybe exert by upregulating anti-apoptotic protein expression and downregulating apoptotic protein expression.

Objective To explore the effect and mechanism of Kruppel-associated box(KRAB) type zinc-finger protein Apak on the transcription of ribosomal RNA(rRNA) under hypoxic conditions.Methods Real-time quantitative PCR(qRT-PCR) was used to detect the level of rRNA and the effect of Apak on the transcription of rRNA.The expression and phosphorylation level of proteins were examined by Western blotting.Immunofluorescence assay was used to observe the location of Apak.Results Under hypoxia (0.3%O2),the transcription level of rRNA increased first and then decreased in Apak wild type cells.In Apak knock-out cells,the transcription level of rRNA kept decreasing under hypoxic conditions.The repression of Apak on rRNA synthesis and the nucleolus location of Apak disappeared,and the expression and phosphorylation level of Apak were down-regulated under hypoxic conditions for 3 h.Conclusion Under hypoxic conditions,the disapperance of Apak repression on rRNA transcription could lead to a temporary increase in rRNA.

AIM: To explore the therapeutic effect of malleolus radialis implantation with vascularized periosteal flap for disunion of carpal navicular fracture, and investigate the characteristics of tissue implantation. METHODS: Thirty-two patients with disunion in carpal navicular fracture were selected from Jiangyin People's Hospital from January 2000 to January 2006, with the informed consents of all the patients. They were treated with implantation of malleolus radialis with vascularized periosteal flap, fixed with plaster cast for 8-10 weeks. Therapeutic effect was evaluated as: ①excellent: normal activity of carpal joints, without soreness sense, fracture line disappeared by X ray;②fine: uncomfortable activity of carpal joints, with slight limitations, fracture line disappeared by X ray. RESULTS: ①All 32 cases of the disunion in carpal navicular fracture were synostosis displayed by X ray.②There were 23 cases followed up for 3-12 months, 7 cases for 13-24 months, and 2 cases for 25-36 months. Carpal joints function achieved the standard of the uninjured side, without pain or affection on daily life.③The therapeutic effect was excellent in 22 cases, fine in 10 cases, and good rate was 100%. CONCLUSION: Implantation of malleolus radialis with vascularized periosteal flap is an effective way to treat the disunion of carpal navicular fracture.

Objective To investigate the evaluation method of processing pieces' colour and quantificationally analyse the relactionship between the pieces' colour and the internal quality. Methods Taking different processing techniques on Rhizoma Alismatis as research objects,using a camera and Adobe Photoshop software to acquire the elementary information of processing pieces' colour and determine the contents of the active principle in Rhizoma Alismatis by HPLC. The active principle in Rhizoma Alismatis and the colour of processing pieces are analyzed quantificationally by SPSS 13.0 statistical software and data mining software Clementine 8.0. Results The method for acquiring and handling colour information of processing pieces in Rhizoma Alismatis could be used to quantificationally analyze the correlation of the colour of processing pieces. There is a significant correlation between processing pieces' colour difference and the internal quality. Conclusion The colour difference of processing pieces can be one of target for the quality assessment. The method can be popularized to other precessing pieces control of techniques.

Objective To evaluate CT findings and dynamic changes of post-traumatic intrapulmonary hematoma.Methods CT data of 43 patients with traumatic intrapulmonary hematomas were analyzed retrospectively.Results A total of 52 lesions were found in 43 patients with intrapulmonary hematomas.The majority of hematomas were located in the peripheral lung fields close to the pleura,single or multiple in number.CT showed nodular,mass,fusiform,ribbon and irregular shadows with uniform and high density in lung.There are two types of evolution: (1) The hematomas gradually reduced in size and finally disappeared;(2)The hematomas evolved into gas-liquid cyst cavity or gas cyst cavity, and then was absorbed gradually.Conclusion CT examination is helpful in the early diagnosis of post-traumatic intrapulmonary hematomas, and can monitor the morphology changes over time.

Cultivation of Medical Humanism Spirit has special value in the teaching of psychiatry,but which was ignored in the teaching. This paper explored the way to integrate humanistic spirit education into the teaching process of psychiatry.

Recombinant Fl-V (rFl-V) fusion protein is the main ingredient of the current candidate vaccine against Yersinia pestis infection, which has been under investigation in clinical trial in USA. We investigated the soluble expression conditions of rF1-V in Escherichia coli BL21 (DE3) that we constructed before. After scale-up and optimization of fermentation processes, we got the optimized fermentation process parameters: the culture was induced at the middle exponential phase with 50 µmol/L of IPTG at 25 °C for 5 h. Soluble rFl-V protein was isolated to 99% purity by ammonium sulfate precipitation, ion exchange chromatography, hydrophobic chromatography and gel filter chromatography. The protein recovery was above 20%. Protein identity and primary structure were verified by mass spectrometry and Edman sequencing. Results of purity, quality and western blotting analysis indicated that the target protein is a consistent and properly folded product. Furthermore, the immunogenicity of various antigens formulated with aluminum hydroxide adjuvant was evaluated in mice. Serum antibody titers of 4 groups including 20 µg rFl, rV and rFl-V and 10 µg rFl+10 µg rV, were assayed by ELISA after 2 doses. The antibody titers of anti-Fl with 20 µg rFl-V were obviously higher than titers with other groups; meanwhile there were no significant difference of anti-V antibody titers among them. These findings confirm that rFl-V would be the active pharmaceutical ingredient of the plague subunit vaccine.
Adjuvants, Immunologic ; Animals ; Antibodies, Bacterial ; blood ; Antibody Formation ; Antigens, Bacterial ; immunology ; Blotting, Western ; Chromatography, Ion Exchange ; Enzyme-Linked Immunosorbent Assay ; Mice ; Plague ; prevention & control ; Plague Vaccine ; immunology ; Recombinant Fusion Proteins ; immunology ; Vaccines, Subunit ; immunology ; Yersinia pestis