Objective To investigate the therapeutic time window and the dose response effects of epirubicin on the expression of c-FLIP in breast cancer.Methods MCF-7and MDA-MB-231 breast cancer cells were divided into two groups: epirubicin groups were treated with 4.0,2.0,1.0,0.5 and 0.25mg/L of epirubicin,and control groups were treated with 0.9% sodium chloride solution at the same dose.After treatment for 24,48 and 72 h,the incubated cells were collected for the measurement of c-FLIP by RT-PCR,and for examination of percent of apoptosis cells with flow cytometry.ResultsA dose-time-dependent pattern was observed.The expression of c-FLIP in MCF-7and MDA-MB-231 breast cancer cell lines declined gradually as the epirubicin concentration increased and treatment time was prolonged.Percentage of apoptosis breast cancer cells increased gradually as the epirubicin concentration was increased and treatment time was prolonged,and percentage of apoptosis cells was the highest when breast cancer cells were treated with 2 mg/L epirubicin for 72 h.ConclusionsEpirubicin can promote apoptosis of breast cancer cells by inhibiting the expression of c-FLIP,and its inhibitory effect is most pronounced when breast cancer cells are treated with 2 mg/L epirubicin for 72 h.

OBJECTIVE To identify the possible risk factors for the development of surgical site infection (SSI). METHODS A total of 218 consecutive patients who received open cholecystectomy due to gallbladder disease and stone of common bile duct during from the Jun to Dec in 2007 were included in the study. The potential risk factors including clinical features,biochemical data,operative types and incision types were analyzed by univariate analysis. RESULTS The overall incidence of SSI was 5.04%.The incidence of SSI in cholecystectomy alone group was lower than in cholecystectomy with exploration of common bile duct group (10.9% vs 3.1%,P=0.022).The incidence of SSI in emergency group was higher than that in selective operation group (12.5% vs 3.8%,P=0.037). The incidence of SSI among patients with white blood cell count more than 10.0?109 befove surgery was higher (12.2% vs 3.0%,P=0.025). The incidence was 1.5%,6.1% and 26.3%,respectively,for patients with Ⅱ,Ⅲ and Ⅳ types incision (P=0.000). CONCLUSIONS These findings indicate that risk factors for the development of SSI after open cholecystectomy include operation manner,operation type,incision type and preoperative leucocyte count.

OBJECTIVE: To explore the effects of Shenfu Injection on prostacyclin, thromboxane A2 and the activities of ATPases in rats exposed to hepatic ischemia-reperfusion injury. METHODS: Twenty-four male Wistar rats weighing 200-250 g were randomly divided into two groups: Shenfu Injection (SF)-treated group (rats were treated with Shenfu Injection of 10 ml/kg through intraperitoneal injection) and untreated group (rats were administered with normal saline at the same dose and served as a control group). Hepatic ischemia was caused by Pringle's maneuver and lasted for fifteen minutes, and then one-hour or three-hour reperfusion was performed. Venous blood samples for the measurement of thromboxane B(2) (TXB(2)) and 6-keto-prostaglandin F(1 alpha)(6-keto-PGF(1 alpha)) were collected three hours after reperfusion. Liver tissue samples were collected one hour or three hours after reperfusion for the measurement of Na(+)-K(+)-ATPase and Ca(+)-Mg(+)-ATPase and for morphological studies. RESULTS: Plasma TXB(2) was lower in the SF-treated group than that in the untreated group after three-hour reperfusion (P>0.05), while 6-keto-PGF(1 alpha) was higher in the SF-treated group than that in the untreated group (P>0.05). The ratio of TXB(2) and 6-keto-PGF(1 alpha) was significantly lower in the SF-treated group than that in the untreated group (P<0.05). The activities of Na(+)-K(+)-ATPase and Ca(+)-Mg(+)-ATPase in the SF-treated group were improved obviously. A three-hour reperfusion after fifteen-minute ischemia caused important hepatic histological alterations. Marked structural abnormalities were observed in the untreated group, such as massive hepatocyte swelling, necrosis, mitochondria edema and vacuolar changes. In the SF-treated group, hepatic tissue injury was reduced significantly. CONCLUSION: Shenfu Injection protects hepatic tissue from ischemia-reperfusion injury, and such protective effects are achieved by decreasing the ratio of thromboxane A(2) and prostacyclin, and increasing the activities of Na(+)-K(+)-ATPase and Ca(+)-Mg(+)- ATPase.

Objective To study the clinical significance of liver failure staging and MELD in predicting the short term prognosis of HBV-acute-on-chronic liver failure (HBV-ACLF).Methods One hundred and ten HBV-ACLF patients admitted to our hospital from July 2013 to July 2015 were included into this study.They were divided into early stage group (n=18),middle stage group (n=48) and end stage group (n=44),the fatality rate in each group was evaluated.According to the MELD score at baseline,they were divided into four groups,MELD＜20 (n =24),20≤ MELD＜30 (n=54),30≤MELD＜40(n =28),40≤MELD (n =4).The fatality rate in each group was evaluated.In the middle stage group,they were be divided into two groups,/△MELD＜0 and △MELD＞0(△MELD=MELD1w-MELDbaseline).The fatality rate in each group was evaluated.Results The fatality rate of the 3 groups(Early,Middle and End stage group) at 3th month was 0,50％,95 ％ respectively(P＜ 0.05).The fatality rate of the 4 groups (MELD＜20,20≤MELD＜30,30≤MELD＜40,40≤MELD)was 31.58％,66.67％,85.71％ and 100％ respectively (P＜ 0.05).In the middle stage group,the fatality rate of the two groups was (△MELD＜0 and △MELD＞ 0)41.18％ and 85.71％ (P=0.001).Conclusion It can be shown that the survival probability of early stage group was high,the probability of death in end stage group and middle stage group with△MELD＞0 was high.

BACKGROUND: Bone marrow stromal cells(BMSCs) are the ideal gene target cells and will have a bright future in the gene therapy of spinal cord injury.OBJECTIVE :To detect the expression of glial cell line - derived neurotrophic factor(GDNF) gene after BMSCs were infected by adenovirus-medialed GDNF (Adv-GDNF) in vitro and to explore its biological activity.DESIGN: A randomized controlled trial study.SETTING: Laboratory of Orthopedic DepartmentMATERIALS: The experiment was completed in the Laboratory of Orthopedic Department, Affiliated Tongji Hospital of Tong ji Meidcal College,Huazhong University of Science and Technology. Twenty-four SD rats of either gender, weighing (180 ± 20) g.INTERVENTIONS: BMSCs were infected by Adv-GDNF in vitro and then cocultured with spinal cord dorsal root ganglion. The three methods, immunofluorescent chemistry, reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay(ELISA) were used to evaluate GDNF expression in the BMSCs. The biological activity of GDNF was observed by a phase contrast microscope.MAIN OUTCOME MEASURES:Primary outcomes:①RT-PCR;②results of immunofluorescent chemical examination;③biological activity of GDNF in vitro. Secondary outcomes:①culturing and identification of BMSCs②time-effect relationship of GDNF expression revealed by ELISA.RESULTS: Immunofluorescence displayed expression of GDNF in BMSCs 48hours after Adv-GNDF infection. RT-PCR analysis demonstrated expression of GDNF mRNA 24 hours after Adv-GNDF infection. ELISA confirmed the presence of GDNF in the liquid supernatant of BMSCs 24 hours after Adv-GDNF infectionn and showed that GDNF was secreted. The supernatant can promote the neurite outgrowth in the rat dorsal root ganglion(DRG).CONCLUSION: It is demonstrated that BMSCs infected by Adv-GDNF can express GDNF steadily and the expressed GDNF has the activity of promoting neurite outgrowth, which lays a foundation of the GDNF gene therapy for spinal cord injury.

With the development of cell separation technique, hepatocyte transplantation becomes a hot topic; however, the application is limited by donor deficiency and immunological rejection. Microencapsulated hepatocytes contribute to the promotion and application for liver cell transplantation, for which provide a large amount of high activity and good function of liver cells, in this paper, liver cell microencapsulation technology and its progress in applications were reviewed, providing prospective way for large-scale and high-active culture in vitro and long-term cryopreservation.

OBJECTIVE: To analyze the research literatures related to bioartificial liver, and to make a conclusion concerning the development of bio-artificial liver.DATA SOURCES: Using bioartificial liver, liver cell, hepatocyte culture and bioreactor as search terms, searching Ovid, Springer Link database, and China National Knowledge Infrastructure, Vip Information database and Wanfang Date (1990.09-2008.09). Literatures search was limited to English and Chinese languages.DATA SELECTION: Researches regarding liver cells of bioartificial liver, reactors and auxiliary equipment was included, and the studies about immune and animal infection studies of bioartificial liver were excluded.MAIN OUTCOME MEASURES: ①The source, quantity and culturing of bio-artificial liver hepatocytes. ②Bioreactor type, nature and type of films. ③Composition of oxygen and temperature control devices of bioartificial liver.RESULTS: Totally 3898 documents seized initially in the searching by computer, according to inclusion and exclusion criteria, 29 were analyzed. Bioartificial liver was a hybrid device in which can culture hepatocytes in vitro, when the patient's blood flows through the device, material exchange with the cultured hepatocytes through semi-permeable membrane or direct contacting can take place, which can perform the same roles of detoxification, synthesis, biological transformation and other functions as real liver cells, so as to achieve the purpose of support and treatment. Bioartificial liver can also be involved in metabolism of the three major nutritive substances, as well as secretion of hepatocyte growth promo ting substances. So it is an effective alternative to the real liver as the function of detoxification and synthesis, and can fills the essential gap between the transplantation and acute liver failure.CONCLUSION: Although the bioartificial liver research has made significant progress, it still faces the problems such as limited liver cells sources, long-term maintenance of liver cell activity and function, and further optimization of the reactor design.

By culturing bone marrow mesenchymal stem cells of rabbits with fibrin glue in vitro, the biocompatibility of fibrin glue was investigated to study whether this material can be used as scaffolds in bone tissue engineering. After 2-months old New Zealand rabbits had been anesthetized, about 4-6 ml of bone marrow were aspirated from rabbit femoral trochanter. The monocytes suspension was aspirated after bone marrow was centrifuged with lymphocyte separating medium and cultured primarily. Then the cells were divided into two groups: one was cultured with complete medium and the other with induced medium. The cells of the two groups were collected and inoculated to the culture plate containing fibrin glue. In the control group, cells were inoculated without fibrin glue. The implanted cells and materials were observed at different stages under a phase-contrast microscope and scanning electron microscope. MTT and alkaline phosphatase (ALP) were measured. Bone marrow mesenchymal stem cells grew on the surface of fibrin glue and adhered to it gradually. Cells light absorption value (A value) and the ALP content showed no significant difference. Fibrin glue had no inhibitory effect on cell morphology, growth, proliferation and differentiation. It has good biocompatibility and can be used as scaffold materials for bone marrow mesenchymal stem cells in bone tissue engineering.
Biocompatible Materials/*pharmacology ; Bone Marrow Cells/*cytology ; Cells, Cultured ; Coculture Techniques ; Fibrin Tissue Adhesive/*pharmacology ; Mesenchymal Stem Cells/*cytology ; Tissue Engineering

In order to study the effects of electromagnetic fields (EMFs) on proliferation, differentiation and intercellular cyclic AMP (cAMP) in mouse bone marrow mesenchymal stem cells (MSCs) in vitro, the mouse bone MSCs were isolated and cultured in vitro. The third passage MSCs were divided into 4 groups and stimulated with EMFs. The cellular proliferation (MTT), the cellular differentiation (alkaline phosphatase activity, ALP), and the intercellular cAMP level were investigated at different time points. The results showed that EMF (50Hz pulse burst 2 mT peak) inhibited the cellular proliferation (P < 0.05), enhanced the cellular differentiation (P < 0. 05), and increased the intercellular cAMP level (P < 0.01) in the early time of the stimulation (1-3 days), but the intercellular cAMP level did not increased further in the later days. We are led to conclude that the cAMP may be involved in the mediation of the growth inhibitory and differentiation-inducing signals of specific EMFs in vitro.
Alkaline Phosphatase/metabolism ; Bone Marrow Cells/*cytology ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Cyclic AMP/metabolism ; Electromagnetic Fields ; Mesenchymal Stem Cells/*cytology