BACKGROUND: Tissue engineering development brings a hope for articular cartilage defect repair. Current studies have demonstrated that bone marrow mesenchymal stem cells (MSCs) are the best cell source to repair full-thickness cartilage defects.OBJECTIVE: To induce MSCs to differentiate into chondrocytes with Ad-hTGF-pi in pellet culture system in vitro and identify the differentiated cells.DESIGN, TIME AND SETTING: Repetitive cellular measurements were performed at the Central Laboratory of Xinqiao Hospital,Third Military Medical University of Chinese PLA from June 2007 to January 2008.MATERIALS: Japanese rabbits, 2 months old, were provided by the Laboratory Animal Center of Third Military Medical University of Chinese PLA.METHODS: The rabbit MSCs were isolated, cultured and expanded in vitro. After transacted with Ad-hTGF-β1, the cells were cultured in pellet culture system. Chondrogenic differentiation was evaluated by histological, immunohistochemical and RT-PCR techniques.MAIN OUTCOME MEASURES: Cell morphological changes were observed by histological staining; proteoglycan and type Ⅱ collagen expression was detected by immunohistochemical and RT-PCR techniques.RESULTS: The induced cells exhibited a chondrocyte-like morphology by histological staining. Immunohistochemical and RT-PCR results showed that proteoglycan and type Ⅱ collagen were expressed in the induced cells.CONCLUSION: Bone marrow MSCs cultured in pellet culture system can differentiate into chondrocytes under the induction of Ad-hTGF-β1.

Objective To compare the clinical effect of two hemostatic methods for patients with pelvic fracture combined with hemorrhagic shock in the early stage.Methods A retrospective analysis was done on clinical data of 90 patients with unstable pelvic fracture combined with hemorrhagic shock managed by damage control resuscitation from January 2008 to December 2012.Unstable blood pressure and abdominal distension were noted postoperatively.Forty patients in Group A received laparotomy and bilateral internal iliac artery ligation from January 2008 to January 2010.Fifty patients in Group B underwent internal iliac artery embolization from February 2010 to December 2012.Comparative measurement was made on parameters of mortality,24-hour blood transfusion volume,24-hour lactic acid value,postoperative systolic blood pressure,postoperative body temperature,and postoperative prothrombin time (PT).Results Following parameters differed significantly between Groups A and B (P ＜ 0.05):mortality rate (53％ vs 12％),24-hour blood transfusion volume[(3 865.5 ±451.3)ml vs (2 108.8 ±336.4)ml],24-hour lactic acid value[(13.0 ± 2.0)mmol/L vs (5.4 ± 1.2)mmol/L],postoperative systolic blood pressure [(80.50 ± 22.73) mmHg vs (113.23 ± 20.89) mmHg],postoperative body temperature [(32.4 ± 0.2)℃ vs (36.1 ±0.3)℃],postoperative PT [(24.5 ±3.6)s vs (18.4±2.1)s].Conclusion For pelvic fracture combined with hemorrhagic shock,if the indications of abdominal viscera rupture are unclear,the interventional embolization can gain advantage over laparotomy in improving treatment success rate and reducing mortality and complications.

According to the basic theory of traditional Chinese medicine (TCM), the pathogenetic factors such as platelet activation, adhesion, congregation and thrombosis fall into the category of blood stasis, while the pathological changes such as tissue necrosis, oxidative stress injury and inflammation, etc, are far beyond the etiological category of blood stasis. The toxin or the combination and transformation of toxin and blood stasis of TCM are involved in the pathogenesis of thrombotic cerebro-cardiovascular diseases. It is significant to recognize and stress the combination and transformation of toxin and stasis in pathogenicity so as to enrich TCM etiology and improve TCM clinical efficacy in the treatment of cerebro-cardiovascular and thrombotic diseases.

Objective To examine the levels of serum tissue inhibitor of metalloproteinase in severe sepsis patients with disseminated intra-vascular coagulation (DIC) and evaluate its diagnostic value in severe sepsis with DIC.Methods Sixty patients were divided into 3 groups,namely severe sepsis group (SS group,n =28),severe sepsis with DIC group (SSD group,n =12) and normal group (n =20).Clinical data including APACHE Ⅱ score and DIC score of these patients were collected.Serum levels of TIMP-1 and TIMP-2 of these patients were measured by using ELISA.Results The patients of SS group had higher levels of TIMP-1 (723.74 ± 96.27) and lower levels of TIMP-2 (68.08 ± 14.87) than healthy control subjects (TIMP-1:574.24 ± 79.99,TIMP-2:89.99 ± 18.45) (P ＜ 0.05).The patients of SSD group had higher levels of TIMP-1 (907.56 ± 200.20) and lower levels of TIMP-2 (44.84 ± 22.13) than patients of SS group (P ＜ 0.05).An association was found between TIMP-1 and fibrinogen (FIB) (r =-0.392,P ＜ 0.05),TIMP-1 and D-dimer (r =0.407,P ＜ 0.05),TIMP-2 and PLT (r =0.484,P ＜0.01),TIMP-2 and PCT (r=-0.523,P＜0.01),TIMP-2 and DIC score (r=-0.579,P＜0.01).The areas under the curves (AUC) for TIMP-1/TIMP-2 was 0.896 (95％ CI:0.843 ～ 0.950 (P ＜ 0.05).Conclusions Serum levels of TIMP-1 in patients with severe sepsis complicated with DIC increased and TIMP-2 decreased,suggesting they were valuable in diagnosis of severe sepsis complicated with DIC.

BACKGROUND:Tissue engineering development brings a hope for articular cartilage defect repair.Current studies have demonstrated that bone marrow mesenchymal stem cells (MSCs) are the best cell source to repair full-thickness cartilage defects.OBJECTIVE:To induce MSCs to differentiate into chondrocytes with Ad-hTGF-?1 in pellet culture system in vitro and identify the differentiated cells.DESIGN,TIME AND SETTING:Repetitive cellular measurements were performed at the Central Laboratory of Xinqiao Hospital,Third Military Medical University of Chinese PLA from June 2007 to January 2008.MATERIALS:Japanese rabbits,2 months old,were provided by the Laboratory Animal Center of Third Military Medical University of Chinese PLA.METHODS:The rabbit MSCs were isolated,cultured and expanded in vitro.After transfected with Ad-hTGF-?1,the cells were cultured in pellet culture system.Chondrogenic differentiation was evaluated by histological,immunohistochemical and RT-PCR techniques.MAIN OUTCOME MEASURES:Cell morphological changes were observed by histological staining;proteoglycan and type II collagen expression was detected by immunohistochemical and RT-PCR techniques.RESULTS:The induced cells exhibited a chondrocyte-like morphology by histological staining.Immunohistochemical and RT-PCR results showed that proteoglycan and type II collagen were expressed in the induced cells.CONCLUSION:Bone marrow MSCs cultured in pellet culture system can differentiate into chondrocytes under the induction of Ad-hTGF-?1.

Objective To examine the levels of plasma tissue factor pathway inhibitor (TFPI) in patients with acute pancreatitis (AP) to assess the clinical value of diagnosis for severe acute pancreatitis ( SAP).Methods Sixty-eight patients were divided into mild acute pancreatitis (MAP)group (n=36) and SAP group (n=32), and twenty volunteers were chosen into normal group ( n=20 ) .Clinical data of these patients were collected, including APACHEⅡscore and Ranson score.Plasma levels of TFPI were measured by ELISA.Results The plasma levels of TFPI in SAP group, MAP group and control group were (4274.25 ±639.83),(3026.81 ±465.76) and (2468.73 ±262.39)pg/ml, respectively(P<0.05).There were significant positive correlations between TFPI and WBC, AST, ALT, TBIL, Cr, PT, APTT, PCT, APACHEⅡscore and Ranson score (P<0.05).The area under the curve (AUC) of TFPI for SAP was 0.902(95%CI=0.845 -0.959, P<0.05 ) .The cutoff value was 4028.83 pg/ml for plasma TFPI with a sensitivity of 87% and a specificity of 78%.Conclusion Plasma levels of TFPI in patients with SAP are significantly increased, which maybe help diagnose SAP.

Objective To investigate the effect of berberine anti macrophages apoptosis, initially to determine the concentration of anti-apoptosis and to explore its mechanisms of treatment in atherosclerosis (AS). Methods Apoptosis of RAW264.7 macrophages is induced by 0.5 μmol/L Tg and 50 μg/ml Fuc. The cell survival rate and apoptosis rate was detected by MIT and flow cytometry, respectively. Results There was statistical significance between the berberine group of 800 μg/ml and normal group in the cell sur-vival rate(P<0.01). With the decline in concentration of berberine, the cell apoptosis rate showed a down-ward trend, berberine (0.02-0.63 μg/ml) effectively inhibited the cell apoptosis induced by Tg and Fuc. Conclusion Berberine group has little impact on the cell proliferation. Berberine can effectively inhibit the cell apoptosis induced by Tg and Fuc.

Objective To investigate the immunologic properties of osteogenic differentiated bone mesenchymal stem cells (BMSCs). Methods BMSCs were isolated from normal volunteers and induced in osteogenic medium for two weeks. Then,non-differentiated/osteogenic differentiated BMSCs were co-cultured with allogenic T cells and phytohemagglutinin (PHA).The proliferation of T cells was examined by MTT method.The concentrations of TGF-β1 in osteogenic differentiated BMSCs supernatants at week 2 and mixed lymphocytes reaction (MLR) supernatants at day 5 were determined by ELISA.Also,anti-TGF-β antibody was added into the MLR to detect the response of the mixed T cells. Results Non-differentiated and osteogenic differentiated BMSCs did not induce proliferation of the allogeneic T cells but both suppressed the proliferation of the T cells mediated by PHA.The TGF-β1 concentrations had significant elevation in the MLR.Anti-TGF-β antibody could counteract the immunosuppressive function of the osteogenic differentiated BMSCs. Conclusion Osteogenic differentiated BMSCs possess low immunogenicity and immunosuppressive property.

Objective To detect and explore the expression and clinical significance of dual specificity tyrosine phosphorylation regulated kinase1b (Dyrk1b) in the specimens and cells of cervical lesions. Methods (1)All the data were collected from 75 patients with cervical cancer and 52 cases with squamous intraepithelial lesion(SIL)admitted in the First Affiliated Hospital of Dalian Medical College during Jan. 2011 to Dec. 2013 and confirmed by pathological examination, included 60 cases of stageⅠand 15 cases of stageⅡ, 12 cases with low-grade squamous intraepithelial lesion(LSIL)and 40 cases with high-grade squamous intraepithelial lesion(HSIL). While, 28 cases with chronic cervicitis were chosen as the control group. The protein expression of Dyrk1b was detected by immunohistochemistry among the four groups.(2)The expression of Dyrk1b in HeLa and SiHa cells were detected by western blot method and the expression of Dyrk1b protein were also detected after treatment of AZ191 (5, 10 μmol/L) for 48 hours in HeLa and SiHa cells.(3)The cellular survival and proliferation of HeLa and SiHa cells treated by different concentrations of AZ191(2.5, 5, 10, 25, 50, 100 μmol/L)for 48 hours were detected by methyl thiazolyl tetrazolium (MTT) assay.(4)The rate of apoptosis of HeLa and SiHa cells was detected by flowcytometry after treatment of AZ191 (5, 10μmol/L) for 48 hours. Results (1)The positive rates of Dyrk1b protein in chronic cervicitis, LSIL, HSIL and cervical squamous cancer by immunohistochemistry were 11%(3/28), 1/12, 42%(17/40)and 71%(53/75), respectively. The expression of Dyrk1b in cervical squamous cancer and HISL were higher than those in LSIL and chronic cervicitis (P<0.01), there were significant difference between cervical squamous cancer and HSIL, or between HSIL and LSIL(all P<0.05), while there were not significant difference between LSIL and chronic cervicitis(P>0.05). Expression of Dyrk1b was correlated with stromal invasion depth of cervical cancer (P<0.05), but not with age, clinical stage, lymph node metastasis, and serum squamous cell carcinom antigen(SCC-Ag)levels (all P>0.05). (2) Dyrk1b protein was expressed in different levels in HeLa and SiHa cells, and the expression of Dyrk1b was decreased gradually as the increased of the concentration of AZ191 in both HeLa and SiHa cells by treatment of AZ191 for 48 hours. (3) Different concentration of AZ191 treated on cervical cancer cells could inhibit the cellular proliferation and induce cell apoptosis in a concentration-dependent manner(P<0.01), concomitant to the decreased cell survival rate. The apoptosis rate of HeLa and SiHa were increased significantly after 10μmol/L AZ191-treatment for 48 hours, but no any difference induced by 5 μmol/L AZ191-treatment compared to control group. Also,there was no any difference between Hela and SiHa cells in either inhibitory effect or apoptosis rate induced by AZ191. Conclusions Dyrk1b is over-expressed in either specimens or cells of cervical cancer. The expression of Dyrk1b protein in cervical lesions is increased as the progression of disease. Dyrk1b inhibitor AZ191 could inhibit cellular proliferation and induce apoptosis in a concentration-dependent manner in cervical cancer cells.

Objective:To construct a primary health care institutions performance evaluation index system from the perspective of health value orientation under the background of countywide medical alliances construction.Methods:From May 2021 to February 2022, preliminary screening was made on core performance evaluation indexes via literature review; purposive sampling was used to select the dean/vice dean, persons in charge of medical service, and those in charge of public health service responsible for performance evaluation at the community health service center. Then semi-structured interviews were made on the existing performance evaluation and assessment plans as well as existing problems of primary medical and health institutions. Based on the " input-process-output" performance evaluation model, the thematic framework analysis method was used to analyze the interview data, and combined with literature research results, a preliminary performance evaluation index system for primary medical and health institutions was built under the guidance of health value. From March to May 2022, the Delphi expert consultation method was used to evaluate the importance and operability of indexes. The threshold method was used to screen indexes, and analytic hierarchy process was used to calculate the weights of evaluation indexes.Results:The health value oriented performance evaluation index system for primary healthcare institutions included 3 first-level indexes, 9 second-level indexes, and 50 third-level indexes. The first-level indexes were output (0.377 3), input (0.336 3), and process (0.286 4) in descending order of weight. The top three weighted second-level indexes were health manpower(0.177 8), health literacy and health outcomes (0.157 6), as well as responsiveness and satisfaction (0.142 6). The third-level indexes included 17 medical indexes, 16 prevention indexes, and 17 medical prevention integration indexes. The top three weighted indexes for inpatient services were resident satisfaction with medical treatment (0.052 4), medical staff satisfaction (0.050 1), and responsiveness of residents seeking medical treatment (0.040 1); The top three weighted third-level indexes excluding inpatient services were resident satisfaction with medical treatment (0.052 4), medical staff satisfaction (0.050 1), and surplus funds used for personnel incentives (0.045 5).Conclusions:The performance evaluation index system of primary health care institutions built under the health value orientation is scientific, conducive to promoting the health-orientated transformation and improving the efficiency of primary health care services.