Objective To investigate the effect of p15~(INK4B)(p15)gene transfection on the proliferation of pancreatic cancer cell line BxPC3.Methods In p15 transfection group.pCDNA3.1(+)p15 was transfected into BxPC3 cell by the vector of Lipofectamine2000.In empty plasmid transfection group, pCDNA3.1(+)neo was transfected into BxPC3 cell with the sa/ne method as a blank control group.In non-transfection group.the BxPC3 cell was not transfected as a negative control group.The p15 mRNA expressions were assayed by RT-PCR,and p15 protein expressions were assayed by Western blot.The proliferation was determined by MTY assay,ultra-structure changes were measured by transmission electron microscope.Cell cycle and apoptosis were measured by flow cytometry.Results In the pCDNA3.1(+)p15 transfeetion group,the expression of p15 mRNA and protein were resumed.Since the 2nd day of culture,the growth of pCDNA3.1(+)p15 transfeetion group was inhibited,till the 7th day,the inhibitory rate was 47.9%,G_0/G_1 Dhase cell accounted for(61.56±3.96)% of all the cells,which was significantly higher than(47.44±6.35)%ofthe black control groups and(49.22±7.23)%0f tlle negative control group(P<0.05).G_1apoptosis peak occurred and the apoptosis rate was(5.27±1.04)%in pCDNA3.1(+)p15 transfection group,which was significantly higher than(0.11±0.06)% of the black control groups and(0.09±0.07)% of the negative control group(P<0.05).Apoptosis was also observed by transmission electron microscope in the pCDNA3.1(+)-p15 transfection group cells.Conclusions After p15 gene transfection,BLPC3 cell proliferation could be significantly inhibited and apoptosis could be induced.

Objective To investigate the anti-fatigue activity of fermented grain-containing blueberry anthocyanins ( LANHE,LH) in mice.Methods Experiments were conducted in two phases .In the first phase , forty mice were randomly divided into four groups:control group(distilled water,dw) and three LH administration groups (8.75,17.5 and 35.08 ml/kg body mass).In the second phase, mice were randomly divided into two groups:high-dose LH group (35.08 ml/kg body mass) and control group (dw 35.08 ml/kg body mass).After four weeks, a forced swimming test was performed and the biochemical parameters related to fatigue were examined .Results and Conclusion The administration groups showed a significant increase of swimming time to exhaustion compared with the control group , especially the high-dose group ( P<0.001).There was no significant change in the blood lactate between the two groups , but at 20 min after swimming, the lactic acid(LA) contents of the high-dose group were lower than in the control group (P>0.05).LH could significantly increase the liver glycogen contents and decrease the serum contents (P<0.05).These data indicate that LH has anti-fatigue activity and can elevate the exercise tolerance in mice .

Objective To evaluate the necessity and safety of implanting temporary vena eava fihers to prevent pulmonary emboli in patients of lower extremity fractures concomitant with acute deep venous thrombosis(DVT). Methods A total of 782 patients with lower extremity fractures were complicated with DVT perioperatively. Among them, 91 received temporary vena cava filters implantation before orthopedic operations for the prevention of pulmonary embolism. All patients were followed up post-operation. Results Vena cava filters were successfully implanted in 89 patients. Mean implantation time was 27 days (range from 14 to 42 days). Thrombus trapped within the filters were found in 78 patients (87.6%) after the filters removal. Eight-two filters (92.1%) were retrived successfully at the first attempt as scheduled. Seven filters(7.9%) with big trapped thrombi were removed at the 2nd attempt after additional thrombolytic therapy. No patients needed a permanent filter. No fetal pulmonary embolism (PE) or other major complications were detected during the three to six months follow-ups period. Conclusion Temporary vena cava filter can reduce the incidence and mortality of pulmonary embolism as well as the occurrence of mid- or long-term complications in lower limb fracture patients complicated with DVT.

BACKGROUND:Little data have been available concerning the mechanism of drag resistance and anti-apoptosis in leukemic cells of leukemia children.The majority of studies focus on normal bone marrow mesenchymal stem cells (MSCs) and established stroma cells,but not interaction of MSCs and leukemic cells in leukemia children.OBJECTIVE:To explore the effect of MSCs in leukemia children on the proliferation and apoptosis of leukemic cell strain K562/AO_2.DESIGN,TIME AND SETTING:In vitro cytology experiment was performed at the laboratory of Department of Pediatrics,First Affiliated Hospital of Guangzhou Medical College from December 2007 to August 2008.MATERIALS:MSCs were provided by 30 leukemia children admitted to First Affiliated Hospital of Guangzhou Medical College,including 22 acute lymphoblastic leukemia and 8 acute myeloblastic leukemia.Written informed content was obtained from all families.K562/AO_2 was provided by Tianjin Institute of Hematopathy.METHODS:MSCs were isolated and cultured by Ficoll density gradient method.They were cultured in two conditions:the co-culture of MSCs and K562/AO_2 and K562/AO_2 suspension alone.In co-culture group,1×10~8 /L K562/AO_2 cells at log phase were added to confluent MSCs,and free floating K562/AO_2 cells were discarded after 24 hours.MAIN OUTCOME MEASURES:Effect of MSCs on the growth of K562/AO_2 cells was observed;effect of addamycln on K562/AO_2 cell apoptosis was detected by AnnexinV-FITC method.Cell cycle was determined by flow cytomtry,mdrl gene of K562/AO_2 cell was detected by Taqman-MGB probe real-time PCR.RESULTS:Compared with K562/AO_2 alone,the K562/AO_2 cell co-cultured with MSCs grew slower and the log phase of growth was not significant;the rate of apoptosis in earlier period was significantly decreased (P ＜ 0.05);co-cultured K562/AO_2 G_0-G_1 phase increased,but S phase decreased.No changes in mdr1 gene in cells were found between two culture conditions (P ＞ 0.05).CONCLUSION:In vitro cytology has demonstrated that leukemia children MSCs induce drug resistance of K562/AO_2 cells by changing K562/AO2 cell cycle through adhesion to avoid pro-apoptotic effect of drugs but not related with mdr1 gene.

ObjectiveTo analyze the risk factors related to perforation of lateral wall by lower cervical pedicle screw instrumentation in the treatment of cervical spinal disorders. MethodsA retrospective review was made to analyze 214 cervical vertebral injury patients ( 1 024 pedicle screws were used) treated with pedicle screw instrumentation at C3-7 from July 2004 to July 2009. Lateral walls of 18 patients were perforated (a total of 28 pedicle screws). The surgeons assessed the position and the angle of the screw in the pedicle, and the relation and the distance between the screw and the pedicle walls by carefully probing intraoperative walls of cervical pedicles and studying postoperative thin-slice computed tomography (CT) scan of the fixed vertebral segments of the cervical spine. The data of patients with lateral wall perforation was recorded and analyzed statistically. ResultsPerforation of the lateral wall occurred in 18 patients (28 pedicle screws). Backward step by step Logistic regression analysis was used and two variables were selected in the end. ConclusionsRatio variance between inner and lateral wall is the risk factor of perforation in the lateral wall associated with lower cervical pedicle screw instrumentation, while the angle variance between implanted screws and CT measurements is the protection factor of perforation in the lateral wall.

Objective: To evaluate the new compensation mechanism for primary healthcare institutions in Zhejiang province, in terms of fairness, performance, incentive mechanism and sustainability in pilot areas. Methods: Evaluation indicators were constructed based on stakeholder theory, fairness theory, expectation theory and sustainable development theory.Focus group interviews were conducted with stakeholders and quantitative data were collected through questionnaires. Meanwhile, the financial compensation, income and expenditure and work equivalent data were collected from such institutions of the four pilot areas, with quantitative data subject to descriptive analysis. Results: This study found the reform used reasonable proportion of funds allocated(the proportion of basic salary for employees was lower than 50%)and adjustment factors(1.0-1.8)of different primary healthcare institutions to guarantee the fairness of the reform; the increase of work equivalents(the per capita work equivalents of medical staff in pilot counties had increased from 38.435 million in the previous year to 42.590 million work equivalents)reflected the performance outcomes of the reform. The incentive and sustainability of the reforms were the weak parts. These were mainly due to the fact that the internal distribution system of primary healthcare institutions failed to make corresponding reforms. Conclusions The reform of the compensation mechanism based on the equivalent method has changed medical staff′s perception of the distribution of funds. The principle of" more pay for more work" and the use of information technology to capture work equivalents have improved the enthusiasm of primary medical staff and the operational efficiency of these institutions, thus, making reform generally scientific and reasonable.

This paper described the market reform′s influence and the healthcare reform on medical behaviors of both doctors and patients.It is held that the healthcare reform should ensure the public welfare nature of public hospitals and focus on medical quality and patient′s health,so as to promote the harmonious development of the doctor-patient relationship from the policy and practice aspect.

OBJECTIVE： To study the chemical constituents in the ethyl acetate extract of Balanophora involucrate， and to provide reference for further enriching chemical constituent of the plant and the development and utilization of B. involucrate. METHODS： The whole plant of B. involucrate was extracted with 75％ ethyl alcohol. The extraction was carried out by petroleum ether， dichloromethane， ethyl acetate and n-butyl alcohol in turn. The chemical compounds from ethyl acetate extract part were isolated and purified by silica gel column， gel column and semi-preparative HPLC. The structures were identified on the basis of spectral spectrum （mass spectrum， hydrogen spectrum and carbon spectrum） data and literature reports. RESULTS： Thirteen compounds were isolated from ethyl acetate extract part of B. involucrate， identified respectively as pyracanthoside （1）， 5，7，3′ ，5′ -tetrahydroxyflavanone （2）， naringenin （3）， homoeriodictyol （4）， hesperetin （5）， sakuranetin （6）， eriodyctiol （7）， aureusidin-6-O-β-D-glucopyranoside （8）， penicillic acid （9）， dihydropenicillic acid （10）， 2-methyl-3-foroic acid （11）， 5-hydroxymaltol （12） and 5， 7-dyhydroxy chromone （13）. Most of them were dihydroflavones. Compounds 2 to 13 are isolated from Balanophora genus for the first time. CONCLUSIONS： The study enriched the chemical constituents of the Balanophora genus and lays foundation for quality evaluation of B. involucrate.

This paper introduced the development of medical specialists alliance in Zhejiang University. With Jinhua Central Hospital as an example,merits and setbacks of the 3-level trusteeship model in practice were discussed,with improvement recommendations raised for progress of the hierarchical medical system.

Objective To investigate the difference of the expression of estrogen receptor,progesterone receptor,human epidermal growth factor receptor 2 and Ki-67 protein between primary lesions of breast cancer and its synchronous ipsilateral lymph node metastasis,as well as its clinical implications.Methods Retrospectively analyze invasive breast cancer patients treated in Peking University First Hospital from January 2012 to May 2016.The IHC expressions of estrogen receptor,progesterone receptor,human epidermal growth factor receptor 2 and Ki-67 protein in both the primary and lymph node metastatic lesions are compared and analyzed statistically.The count data were represented as n(％),and comparsion between groups were evaluated using the McNemar test.Results One hundred and fifty-six patients were included,of which on 2 cases (1.3％),estrogen receptor status of primary lesions is different from that of lymph node metastases(P =0.500);on 10 cases (6.4％),progesterone receptor status of primary lesions is different from that of lymph node metastases (P =0.344);on 28 cases (18.0％),Ki-67 protein status of primary lesions is different from that of lymph node metastases (P =0.000 18);on 3 cases (1.9％),human epidermal growth factor receptor 2 status of primary lesions is different from that of lymph node metastases (P =1.000).Conclusion There may be difference between primary lesions and lymph node metastases in the expression of estrogen receptor,progesterone receptor,human epidermal growth factor receptor 2 and Ki-67 protein,which can provide a reference for individualized treatment of breast cancer patients.