4 were analyzed before, during and after dripping acid. Results The time of onset of pain in patients with empty stomach was 4.67?0.04, which was earlier than that in control group (9.69?1.32min, P4 after dripping acid in dismal part of duodenal bulb was less than that in the control group (P

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 investigate the feasibility of laparoscopic radical hysterectomy and abdominopelvic lymphadenectomy in the treatment of malignant uterine tumors.Methods Laparoscopic radical hysterectomy and abdominopelvic lymphadenectomy was performed in 62 cases of biopsy-confirmed early-stage malignant uterine tumors from February 2003 to August 2005.There were 26 cases of endometrial cancer and 36 cases of cervical cancer.Pelvic lymphadenectomy was conducted in all the cases;while selective lymphadenectomy of peripheral lymph nodes of the abdominal aorta was performed in 5 cases,followed by laparoscope-assisted vaginal hysterectomy(LAVH).Results The operation was completed under laparoscope in 61 cases,and a conversion to open surgery because of venous injuries was encountered in 1 case.The operation time was 165～265 min(mean,217 min);the intraoperative hemorrhage volume was 150～1200 ml(mean,260 ml);the number of excised lymph nodes was 13～23(mean,17).Bladder injury happened in 1 case and was successfully repaired under laparoscope.Postoperatively,urine retention developed in 4 cases and lymphatic cyst occurred in 5.Follow-up checkups for 1~28 months in 61 cases showed 1 case of recurrence at 3 postoperative month(stage Ⅱ_B cervical adeno-squamous carcinoma,withdrew from the treatment and died 4 months later).Conclusions Laparoscopic radical hysterectomy and abdominopelvic lymphadenectomy is safe,feasible,effective,and reliable.

Objective To investigate the clinical value of laparoscopic myomectomy.Methods A retrospective analysis was made on clinical data of 58 cases of laparoscopic myomectomy(Laparoscope Group) and 52 cases of open myomectomy(Open Group) from October 2002 to February 2004 in this hospital.Results The laparoscopic operation was all accomplished.The Laparoscope Group had significantly shorter operation time(58.0?11.2 min) than the Open Group(69.4?10.3 min)(t=(-5.535),(P=0.000)),less intraoperative blood loss(71.6?34.8 ml) than the Open Group(149.1?38.9 ml)(t=-11.029,P=0.000),lower postoperative pyrexia rate(21/58) than the Open Group(39/52)(?~2=16.642,P=0.000),shorter hospital stay(4.5?1.6 d) than the Open Group(7.6?2.1 d)(t=-8.760,P=0.000),higher total hospitalization costs (6 511.3?566.7 yuan) than the Open Group(6 286.8?387.5 yuan)(t=2.398,P=0.018),higher costs for anesthesia and operation(1 566.7?154.7 yuan) than the Open Group(946.6?156.6 yuan)(t=20.868,P=0.000),and lower costs for postoperative medication(703.5?140.2 yuan) than the Open Group(1 278.4?237.6 yuan)(t=-15.643,P=0.000).Follow-up observations in 56 cases in the Laparoscope Group and 50 cases in the Open Group for 18~34 months(mean,26 months) revealed no statistical differences in recurrence rate and pregnancy rate between the two groups.Conclusions Laparoscopic myomectomy has advantages of little hemorrhage,fast recovery,short hospital stay,and low complication rate,being one of ideal methods for the treatment of hysteromyoma.

BACKGROUND:Existing evidence has confirmed that umbilical cord blood mesenchymal stem cel s have an effect on functional recovery of a variety of damaged cel s.
OBJECTIVE:To explore the effect of umbilical cord blood mesenchymal stem cel transplantation on ovarian cancer chemotherapy.
METHODS:Sixty healthy adult female Wistar rats were randomly divided into normal control group, damage group and treatment group (n=20/group). There was no treatment in the control group, and a rat model of ovarian cancer chemotherapy damage was made in the damage group and treatment group. After successful modeling, rats in the control group were given normal saline injection via the tail vein, and those in the damage and treatment groups were given paclitaxel chemotherapy and pacligaxel chemotherapy plus umbilical cord blood mesenchymal stem cel transplantation, respectively. After transplantation of 2 weeks, mRNA and protein expressions of XAF1 and Survivin in ovarian tumor tissues were detected by RT-PCR and western blot assay, respectively. Apoptosis in ovarian cancer cel s were detected using TUNEL method.
RESULTS AND CONCLUSION:Compared with the damage group, a significant up-regulation of XAF1 mRNA and protein but a remarkable down-regulation of Survivin mRNA and protein were obtained in the treatment group (P<0.05). A severe damage to the ovarian tissues was visible in the damage group, presenting with large hemorrhage and necrosis area. This damage was markedly reduced in the treatment group. Additional y, the apoptotic rate of ovarian cancer cel s was significantly higher in the treatment group than the damage group (P<0.05). Al these findings indicate that umbilical cord blood mesenchymal stem cel transplantation aids in ovarian cancer chemotherapy to promote ovarian tissue repair in rats, and XAF1 and Survivin cannot be ignored in tumor angiogenesis and ovarian cancer cel apoptosis.

Objective To investigate the expression of IL-10 and IL-6 in tissue and plasma of subtypes lymphoma and its significance. Methods Immunohistochemical technique and enzyme-linked immunosorbent assay (ELISA) were used to assess the expression of IL-10 and IL-6 in 97 cases paraffin tissue and plasma samples from different subtypes of lymphoma.Results Expression of IL-10 and IL-6 was positive in all lymphoma tissue samples, but there is no significant difference among different subtypes lymphoma (x2 =0.815, x2 =0.542, P ＞ 0.05). Tumor samples were found positive IL-10 and IL-6 expression in macrophages and vascular endothelial cells by immunohistochemistry respectively. Expression of IL-10 and IL-6 in plasma of lymphoma were higher than that in control respectively [(232.57±191.59) pg/ml vs (59.12±68.35) pg/ml,(80.70±89.68) pg/ml vs (45.68±33.66) pg/ml],with significant difference (t =6.968,t =2.896,P ＜0.05).IL-10 and IL-6 plasma levels increased with enhanced expression in lymphoma (x2 =0.815,x2 =0.542,P ＜ 0.05).Which was found to be correlated with each other (rs =0.394,P ＜ 0.05). Conclusion The expression of IL-10 and IL-6 had been found at various degrees in tissues and plasma of lymphoma.IL-10 and IL-6 in plasma of lymphoma were higher than that in controls. These results showed that IL-10 and IL-6 may cooperate in vivo with lymphoma cell proliferation and infiltrate. IL-6 and IL-10 maybe served as useful indicators for diagnosis of lymphoma.

Objective To investigate the feasibility,reliability,and complications of laparoscopic second-look exploration for ovary epithelial cancer.Methods The operation was performed under general anaesthesia.The 4-port laparoscopy was carried out.Physiological saline was used to wash the abdominal and pelvic cavity for cytological examination.Then the exploration of abdominal and pelvic cavity was performed,adhesions were dissected,and multiple punch biopsies(at more than 20 sites) were conducted in the abdominal and pelvic cavity.Results All the operations were completed successfully.The operative time was 61.3?16.7 min,and the hemorrhage volume was 98.7?32.1 ml.Positive findings were encountered in 4 cases(28.6%),3 of which(21.4%) were identified by the naked eye and 1 of which was identified by microscope,and negative findings,10 cases(71.4%).Of the 4 cases of positive findings,the tumor was located at the pelvic peritoneum or the paracolic sulci peritoneum.No abdominal organ injuries occurred,and no conversion to open surgery was needed.Delayed healing of umbilical incision happened in 1 case.Of the 10 cases of negative findings,recurrence of tumor was observed in 1 case(10.0%) at 48 months after operation.Conclusions Laparoscopic second-look exploration for ovary epithelial cancer is feasible,with advantages of minimal invasion,fast recovery,and good patient's tolerance.

Objective To explore the influence of stereoscopic vision on the development of posterior capsule opacification (PCO) after intraocular lens implantation. The efficacy of Nd: YAG laser was studied. Design Case-controlled study. Participants 57 consecutive patients with POC(59 eyes with phacoemulsification and IOL implantation). Methods A prospective observational study was conducted comprising 57 patients (59 eyes) who had postoperative PCO. Neodymium: YAG (Nd: YAG) laser posterior capsulatomy was performed on all patients following a standardizes procedure. Preoperative and postoperative evaluation included LogMAR visual acuity, best-corrected visual acuity (BCVA), contrast sensitivity and stereoscopic vision. Main Outcome Measures LogMAR visual acuity, BCVA, contrast sensitivity and stereoscopic vision. Results The average LogMAR eyesight median of sick eye before laser treatment was (0.22?0.31) and the contrast sensitive degree of each frequency area was relatively lower than the opposite eye; After the treatment, there was evident improvement of the sick eye, and the LogMAR eyesight median of that was 0.00; The ratio of reaching the central spatial eyesight and the average sharpness degree both improve much than ahead of treatment. Those whose spatial eyesight sharpness don't get normal after the low best remedy eyesight. Irregular dipodic and previous eye disease. Conclusions LogMAR visual acuity, BCVA, contrast sensitivity and stereoscopic vision were damaged markedly with PCO. Our results indicate that Nd: YAG capsulotomy may affect the reconstruction of stereoscopic vision. Nd:YAG laser posterior capsulotomy is a valuable and clinically relevant method for clinical treatment of PCO.

Objective To screen a cell line which stably suppresses DAPK expression and to observe the growth characters.Methods Four pairs of shRNA were designed,synthesized and inserted into the pDsRed1-N1-U6 vector.The recombinant plasmids were purified and transfected into PC12 cell.Meanwhile,a pDsRed1-N1-U6 vector was transfected as control.The cell clones were screened by G418,and the stable PC12 cell line was established.DAPK expression was detected by Western blot.MTT method and Flow Cytometry(FCM) assay were used to assess the growth characters of the cell line.Results The shRNAs were transfected into PC12 cell and the cell clones were successfully screened out.Of the four recombinant plasmids,the F2 was the best interfering shRNA.Beyond our expectation,the F1 recombinant plasmid had an enhanced effect on DAPK expression.Conclusion A stable PC12 cell line with stable inhibition of DAPK expression by was established using siRNA expression vectors.