BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) are in advantages of easy collection and amplification in vitro, but the culture and induction in vitro still need to be modified. OBJECTIVE: To investigate the differentiated potency of BMSCs into ostcoblasts by modified in vitro culture methods and inductor matching. DESIGN: Observational study. SETTING: Department of Medical Laboratory, General Hospital of Guangzhou Military Area Command of Chinese PLA. MATERIALS: This study was performed in the Stem Cell Tissue Engineering Laboratory, Department of Medical Laboratory, Genera Hospital of Guangzhou Military Area Command of Chinese PLA from July 2004 to June 2006. Twenty adult SD rats of clean grade, irrespective of gender, weighing 140-180 g were provided by Animal Experimental Center, General Hospital of Guangzhou Military Area Command of Chinese PLA. The experimental animals were disposed according to ethical criteria. Β-sodium glycerophosphate, dexamethasone, and vitamin C were provided by Sigma Company, USA; goat-anti-rat osteocalcin antibody by DSL Company, USA; S-P immunohistochemical kit by Maixin Biotechnology Developing Co., Ltd., Fujian. METHODS: Cells were cultured and induced into osteoblasts by modified methods. Separation and culture of BMSCs: By anesthesia, bone marrow of bilateral femur and tibia was separated to prepare single cell suspension; subsequently, the suspension was inoculated in culture bottle, and the culture media was semi-quantitatively changed after 48 and 96 hours in order to get rid of non-adherent hematopoietic cells. The liquid was changed every three days to further get rid of non-adherent cells. At 80% cell confluence, the medium was digested with 0.25% trypsin and cells were subcultured in the ratio of 1:2. MSCs in the second generation were inoculated in 6-well culture plate and glass fiat plate; after 48 hours, basic culture fluid was removed. Differentiation of BMSCs into osteoblasts: Subcultured BMSCs differentiated into osteoblasts induced by dexamethasone (10 mmol/L), β-sodium glycerophosphate (10-7 mol/L), and vitamin C (50 mg/L). MAIN OUTCOME MEASURES: Ten days after differentiation by modified culture methods and inductor matching, alkaline phosphatase activity was measured with Ca-Co technique. Twelve days after culture, osteocalcin secretion was detected with immunohistochemical method. Two weeks after culture, cell mineralization was detected with Von kossa staining. RESULTS: Alkaline phosphatase activity: Alkaline phosphatase staining of cells was apparent; gray-black particles or massive precipitations were observed in cytoplasm after positive reaction; regions expressing alkaline phosphatase activity were brown-black. Osteocalcin secretion: Osteocalcin was apparently positive; nucleus was blue; cytoplasm was brown. Cell mineralization: Induced cells grew layer by layer, and adiaphanous nodus was observed at the same time. Black mineralized nodus granules in various sizes were observed after Von kossa staining, and this suggested that mineralized apposition occurred. CONCLUSION: BMSCs may be successfully cultured and induced into osteoblasts by modified culture method.

BACKGROUND:With the potential of multi-directional differentiation and high proliferation, bone marrow mesenchymal stem cells (BMSCs) have wide application prospect in tissue engineering. OBJECTIVE:To investigate changes in cells and bioactivity in the differentiation from BMSCs into osteoblasts. DESIGN, TIME AND SETTING:A randomized control experiment was performed at the Laboratory of Stem Cells and Tissue Engineering, Department of Medical Experiment, General Hospital of Guangzhou Military Area Command of Chinese PLA from July 2004 to June 2006. MATERIALS:Twenty adult SD rats of both genders were used for bone marrow collection. METHODS:Rats were equally and randomly divided into an experimental group and a control group. BMSCs were isolated from adult rats by modified bone marrow culture method. BMSCs were inoculated in basic medium containing 10% fetal bovine serum, high glucose DMEM, 100 U/L penicillin, 100 U/L streptomycin and 2 mmol/L glutamine in the control group. BMSCs were inoculated in conditioned medium (basic medium, 10 mmol/L β-glycerophosphoric sodium, 10-7 mol/L dexamethasone, 50 mg/L vitamin C). Cell slide was prepared. MAIN OUTCOME MEASURES:Inverted microscope and transmission electron microscope were used to observe cell appearance. Gomori modified calcium-cobalt method was applied to do alkaline phosphatase staining. Immunocytochemistry was employed to measure osteocalcin expression. RESULTS:Forty-eight hours after inoculation, a mass of BMSCs adhered to a wall. Seventy-two hours later, BMSCs proliferated and well grew. Seven to nine days later, cells were grown to 80% confluency. Transmission electron microscope showed BMSCs with big cell nucleus and immature appearance. After in vitro osteoblast induction, BMSCs proliferated, aggregated, had node and mineralized. One week later, alkaline phosphatase activities were expressed in BMSCs. Two weeks later, osteocalcin expression was detected. CONCLUSION:After one week of in vitro osteogenic induction, BMSCs enter the process of osteoblast transformation, remain proliferative activities, and can be used as seed cells in bone tissue engineering

Objective To discuss the significance of platelet rich plasma in promoting bFGF and VEGF expression in would healing of rabbit buns .Methods The 24 rabbits were randomly divided into control group and experimental group .The sulfadiazine silver paint to the wound for control group and the platelet rich plasma gel evenly spread to the wound for experimental group .At 7th ,10th ,14th day ,4 rabbits of each group were randomly selected to sacrificed after anesthesiaed ,wound healing rate was compare in 2 groups ,HE staining and testimmunohistochemistry were conducted in each group .Results At 7th ,10th ,14th day ,the wound healing rate of experimental group were higher than those of control group .At 7th d ,the focal granulation ,cells and vessels inten-sive were more significant in experimental group than in control group ,at 10th day ,the wound fibroblast cells and capillary number are more significant in experimental group ,and at 14th day ,the most of fibroblasts translate into fiber cell and the capillary number decreased ,the fibroblasts proliferation was still active and fiber cell was less in the control group .At 7th ,10th day ,the expression of bFGF and VEGF were both higher ,but experimental group is obvious higher than control group(P<0 .05) ,but at 14th day ,the ex-pression of bFGF and VEGF were gradually declined and was not obvious statistics differences in the two group (P>0 .05) .Conclu-sion The platelet rich plasma could promote the big white rabbit scald wound healing ,while the main mechanism maybe the ex-pression of bFGF and VEGF increased in the early .

Objective To study the efficacy of CPT-11 TACE in the treatment of unresectable HCC.Methods A retrospective review was undertaken on unresectable HCC patients receiving doxorubicin transarterial chemoembolization (59 cases) and irinotecan(CPT-11) in 24 cases from May 2003 to November 2011.Survival analysis was performed with Kaplan-Meier statistics.Differences in survival between the two groups were assessed for statistical significance with the log-rank test.Results Overall survival time was significantly longer in patients treated with CPT-11 compared with doxorubicin treated group (21.7 vs 14.5 months,P =0.042).There was no significant difference in time to progression between the two groups,but time to progression was longer in CPT-11 group than doxorubicin treated group (11.42 vs 9.46 months,P =0.091).Subgroup analysis showed that for intermediate-stage HCC,CPT-11 resulted in a significantly longer time to progression and overall survival compared with doxorubicin treated group (P =0.029 and P =0.014,respectively).There were no significant differences in adverse events among the two groups (P ＞ 0.05).Conclusions Chemotherapeutic agent CPT-11 in the form of TACE significantly improved overall survival when compared with doxorubicin for the treatment of unresectable HCC.

Objective To evaluate the efficacy of chemoradiotherapy in patients with unresectable pancreatic cancer who were previously treated with PTCD.Methods From September 2005 to December 2012,47 unresectable pancreatic cancer patients with obstructive jaundice were enrolled in this study.They were divided into two groups.21 patients received after PTCD chemotherapy or radiation,or chemoradiotherapy.26 patients in support care group received only nutrition,analgesia and other related support treatment.Survival analysis was performed with Kaplan-Meier statistics.Differences in survival between the groups were assessed for statistical significance with the log-rank test.Results The median overall survival time of patients after PTCD was 7.19 months.The median overall survival time of chemoradiation group was 9.07 months,which was higher than that of support care group (5.52 months),P=0.017.12 patients received single therapy (either chemo or radiation),and 9 patients received chemoradiotherapy.The median overall survival times were 8.31 months and 11.15 months,respectively (P =0.325).Conclusions Post PTCD chemoradiotherapy helps prolong the survival time in unresectable pancreatic cancer patients.

OBJECTIVE:To promote effective development of services in the intravenous drugs allocation centre and to bring the pharmacists'function into full play in the hospital pharmacy.METHODS:Information concerning services in the intravenous drugs allocation centre of our hospital and experiences from which was introduced,some problems and difficulties in the pharmacy services were analyzed.RESULTS&CONCLUSION:The centralized allocation and management of intra-venous drugs have ensured the safety and effectiveness of the clinical intravenous drug use and which have become the essential part of the pharmaceutical care with a core of rational drug use.

Objective:To evaluate the relationship between breast reconstruction and postoperative complications by meta-analysis.Methods:Through a defined search strategy, related literature was collected in databases from PubMed, MEDLINE, EMBASE, Cochrane, CNKI, Wanfang Database and VIP Database, from January 1990 to November 2017. Data were extracted and each merged data was analyzed using RevMan 5.3 software. The postoperative complications between transverse rectus abdominis musculocutaneous-flap (TRAM) and deep inferior epigastric perforatorflap (DIEP), TRAM and latissimus dorsiflap (LDF), LDF and LDF+ prosthesis, LDF+ prosthesis and simple prosthesis implantation were compared.Results:Twenty-nine papers met inclusion criteria of our study. The Meta-analysis results showed that the risk of local flap necrosis, seroma, infection, fat liquefaction, abdominal wall hernia and abdominal bulging in TRAM group were higher than those in DIEP group, and the differences were significant. There was no significant difference in total flap necrosis, wound dehiscence, venous congestion between the two groups; the risk of total flap necrosis, wound dehiscence and infection in TRAM group were higher than those in LDF group, and the differences were significant. The risk of seroma in TRAM group was lower than that in LDF group, and the differences were significant. There was no significant difference in fat liquefaction between the two groups; there was no significant difference in the postoperative complications between LDF group and LDF+ prosthesis group; the risk of seroma in LDF+ prosthetic group was higher than that in prosthetic group, and the differences was significant. The risk of prosthetic capsular contracture and prosthesis displacement in LDF+ prosthetic group were lower than those in prosthetic group, and the differences were significant. There was no significant difference in prosthesis exposure and infection between the two groups.Conclusions:DIEP has most of the advantages of TRAM and fewer complications. It plays an important role in breast reconstruction in the future. We should make a choice of breast reconstruction methods according to the patient＇s conditions as far as possible in clinical practice.

Objective To detect and analyze thyroid tumor by two-dimensional sonogram and Doppler parameter, and evaluate the value of high-frequency ultrasound in diagnosing thyroid benign and malignant tumors. Methods The ultrasonic images of 104 thyroid tumor from 80 patients with typical features were collected. Thyroid tumor was classified into benign and malignant nodules, based on the shape, border, or the rear wall echo, echo attenuation loss, internal echo, and microcalcifications in two-dimensional sonogram and systolic blood peak velocity (Vmax) and resistant index (RI) in Doppler examination. The expected results of high frequency ultrasound were compared with pathological results on consistency and error rate. Results Prediction of benign tumor by high frequency ultrasound was 66, and pathology 61, consistency rate of the two was 92.4%. Prediction of malignant lesions was 38, and pathological examination 32, consistency rate of the two was 84.2%. The total coincidence rate was 89.4%(93/104) and the error rate was 11.6%( 11/104). Conclusions The typical features of thyroid tumor on high-frequency ultrasound are helpful in diagnosis of benign or malignant nodules, which is valuable in guiding clinical treatment.

In this experiment, Hca-F or L615 elemene combo-TCV (H-TCV and L-TCV), H-TCV lysates, corynebacterium parvum (CP) were used to immunize 615 and Balb/c inbred mice, and their splenic DCs were prepared and pulsed in vitro with tumor cell lysates (H or L) and TCV lysates (TH or TL). The capacities of DCs to stimulate the proliferation of syngeneic nonadherent spleenic cells were tested with MTT assay. The results showed that the splenic DCs from normal mice in vitro pulsed with TH or TL could induced syngeneic noradherent splenic cells to proliferate (P＜0.01), while the H or L pulsed DCs could not. The splenic DCs from H-TCV or L-TCV or TH immunized mice re-pulsed in vitro with TH or TL exhibited stronger stimulating effects than the DCs from normal mice pulsed in vitro for the firth time pulsed with TH or TL (P＜0.01 or P＜0.05); The capacities of DCs to induce proliferation of syngeneic nonadherent splenic cells could be further enhanced by CP immunization, especially when were pulsed with TH (P＜0.01). Normal inbred 615 mice were transferred with DCs pulsed with lysates of elemene TCV (TDCs) or pulsed with lysates of Hca-F tumor cells (HDCs) on day 7 before challenged with lethal dose of live Hca-F cells, significant adoptive immunoprotective effects were seen, with 61.6% tumor inhibition rate and 25% survival in TDC adoptive transfer group. This study indicated that DCs might play a role in the mechanisms of active immunization and pulsing DCs with lysate of elemene combo TCV and isolating DCs from elemene combo TCV immunized mice were useful methods for DCs vaccine preparation.

Objective To assess the causes of sildenafil failure and the feasibility of successfully rechallenging non-responding patients. Methods A total of 126 consecutive erectile dysfunction (ED) patients from Jan 2009 to Dec 2009 who claimed poor response to sildenafil (sildenafil 100 mg on demand, at lease 4 epiodes) were enrolled into the study. All patients received sexual reeducation and were treated with sildenafil, taken on a daily dose of 50 mg for 4 weeks. The International Index of Erectile Function-5 (IIEF-5), Rigiscan, serum testosterone or penile Doppler were used to evaluate ED and the cause of on-demand sildenafil failure. End point efficacy of rechallenging was evaluated using the IIEF-5 and the sexual encounter profile (SEP) 'Were you able to insert your penis into your partner's vagina?' and 'Did your erection last long enough to achieve successful intercourse?'. Results The recruited patients comprised of 41 cases with psychological ED, 39 cases with hypogonadism ED, 28 cases with diabetes mellitus ED and 18 cases with vascular ED. Compared with pretreatment and on-demand sildenafil baseline, daily administration of sildenafil significantly enhanced all efficacy outcome variables. The IIEF-5 was significantly improved after daily sildenafil (12.3 ± 2.9 vs18. 8±4.4, P＜0.01), 78 patients responded to daily sildenafil. The overall salvage rate was 61.9%(78/126). Conclusions Sexual reeducation and daily administration of sildenafil may be able to salvage many patients with ED who were sildenafil non-responders.