BACKGROUND: Compared with the bone marrow mesenchymal stem cells, umbilical cord blood mesenchymal stem cells (UCB-MSCs) is an ideal source of tissue engineered seed cells, but the culture success rate is low.OBJECTIVE: To explore establish a stable reliable method to isolate and culture UCB-MSCs by optimizing medium choice,centrifugation speed and time, incubation density, choice of growth factor, first time of medium change.DESIGN, TIME AND SETTING: The cytological in vitro study was performed at the Department of Blood Transfusion, Bethune International Peace Hospital of Chinese PLA from January to October 2008.MATERIALS: A total of 20 samples of neonatal UCB by full-term uterine-incision delivery were supplied by Stem Cell Center,Bethune International Peace Hospital of Chinese PLA. Parturient and their family member signed the informed consent.METHODS: Under sterile conditions, UCB-MSCs were isolated by combination of density gradient centrifugation (1 500 r/min, totally 15 minutes) and different adherent time method. UCB-MSCs were incubated in DMEM/F12 medium, supplemented with 10% human UCB serum, 5 μg/L granulocyte-macrophage colony-stimulating factor (GM-CSF), 15 pg/L interleukin-3. MSCs at 1 ×1010/L were incubated in plastic flask coated with human UCB serum at 37℃ and 5% CO2 saturated humidity. The medium was changed following 3 days of culture. Non-adhered cells were removed. Subsequently, the medium was changed once every other 24 hours. When 80% confluence, UCB-MSCs were digested by the mixture of pancreatin-athylenediamine tetraacatic acid.MAIN OUTCOME MEASURES: Morphological changes of UCB-MSCs were observed by inverted phase contrast microscopy. Cell immunophenotypes were determined by flow cytometry.RESULTS: A small quantity of adherent round cells were determined after 24 hours, and adherent cells became more at 48 hours,with a few monopole spindle cells. Cell colonies were detected at day 7. Fibroblast-like cells arranged parallelly, presented whirlpool-shape and unclear boundary, with 80% 80% confluence 2-3 weeks following culture. At the second passage, these calls adhered at hour 12, and reached 80% 90% confluence at day 10. Flow cytometry showed that these calls were positive for CD29 and CD44, but negative for hematopoietic lineage marker CD34.CONCLUSION: MSCs can be successfully isolated from human UCB by using this modified method in vitro, with short culture cycle and high cell purification. Adherent cells have the same immunophenotype as bone marrow mesenchymal stem cells.

To study the therapeutic action of tuberculosis Ag85A DNA vaccines. Methods:BALB/c mice were infected by in-traperitoneal injection with M. tuberculosis H37Rv for 8 weeks, and then treated with the saline (A), plasmid vector (B), M. bovis BCG (C), M. vaccae (D), and Ag85A DNA vaccine (E). Ag85A-specific antibodies were determined by ELISA. The lungs, livers and spleens were taken and observed their pathological changes, weighted and performed mycobacterial cultures 2 or 5 monthes after treatment. Spleen cells were tested for proliferative responses.Results:Ag85A-specific antibodies increased in the mice of group E. At 2 months after immunothera-pies, the stimulation rates of spleen cells had no significant difference among each group. The numbers of viable bacteria in the lungs and spleens of therapeutic group were lower than those in the control group. The group C and B could be observed slight .lesion of lung. At 5 months after immunotherapies, the stimulation rates of spleen cells all increased significantly in the immunotherapeutic groups. The numbers of viable bacteria in the lung and spleen had no significant difference among each group. No lesions of lung could be observed in group E and D. Conclusion:The tuberculosis DNA vaccines seem to have some immunotherapeutic actions.

Background:Functional gastrointestinal disorders(FGIDs)is a common disease of digestive system which severely affects the quality of life. Recently,more and more attentions have been paid to the relationship between FGIDs and psychological factors. Aims:To investigate the relationship between FGIDs and psychological factors and related treatment. Methods:A total of 270 patients with FGIDs admitted from Sept. 2013 to Sept. 2014 at Anhui No. 2 Province People’s Hospital were enrolled. Hamilton Anxiety Scale(HAMA)and Hamilton Depression Scale(HAMD)were used to evaluate the mental state of patients,and symptom severity of digestive system was scored. Patients with psychological abnormalities received conventional therapy to relieve digestive symptoms,and those who failed from conventional therapy were assigned into refractory group and others were assigned into response group. Patients in refractory group were further divided into conventional therapy group and combined therapy group for receiving paroxetine in addition to conventional therapy. Results:The rates of anxiety and depression were 61. 5%(166 / 270)and 57. 0%(154 / 270),respectively. The scores of HAMA and HAMD were positively correlated with the score of digestive symptoms(r = 0. 585,P < 0. 01;r = 0. 584,P <0. 01)in patients with anxiety and depression in response group. The scores of HAMA,HAMD and digestive symptoms in response group after conventional therapy were significantly decreased(P < 0. 05),while the scores of HAMA and HAMD in refractory group after therapy were significantly higher than those in response group before therapy(P < 0. 05). The scores of HAMA,HAMD and digestive symptoms in combined therapy group were significantly lower than those in conventional therapy group(P < 0. 05). Conclusions:Relationship is found between FGIDs and psychological factors,and antidepressants might contribute to the amelioration of symptoms of FGIDs.

Objective: To observe the effect of Weiyanyin on Motilin (MOT) in blood plasma of experiment bile reflux gastritis rats. Methods: To make the model of experiment bile reflux gastritis by drinking reflux liquid, compared with Domperidone.The effect of Weiyanyin on Pathological histology and MOT in blood plasma of bile reflux gastritis rats was observed. Results: The mucosal of rats in model group was damaged, amotic, and a lot of inflammatory cell could be seen, glandular organ hyperplasy and intestinal metaplasia in most of rats; At the same time the content of the MOT was decreased; Compared with model group, Weiyanyin could improve pathological histology of experiment bile reflux gastritis rats, and increase the content of MOT(P

How to identify active constituents of traditional Chinese medicines (TCMs) and study their interactions are key problems in the development of TCMs. The inhibitory effect of six alkaloids from Rhizoma Coptidis (RC) on Shigella dysenteriae (S. dysenteria) growth had been investigated by microcalorimetry in this study. Main active constituents of RC were confirmed by comparing their contributions to the bacteriostatic effect, and the interactions among active constituents were further researched. According to the result, in 0.8 mg-mL-1 extract of RC, the contributions of six active alkaloids including berberine, coptisine, epiberberine, palmatine and the combination of jatrorrhizine and columbamine were 52.83%, 36.31%, 2.49%, 4.27% and 3.21%, respectively. Therefore, berberine and coptisine were the main active constituents of RC that inhibited the growth of S. dysenteria. The study of interactions among the six alkaloids indicated that, 1 there were some contstituents antagonizing the inhibitory effect of RC, 2 there was a synergy effect between berberine and coptisine, 3 there were additive effects between other four alkaloids and the main active constituents. These results may provide some useful references for the establishment of the quality standard for RC and the development of multi-component TCMs.

Objective To determine the influencing factors on informed consent associated with decline or delay of primary percutaneous coronary intervention (PCI) in patients with acute ST-segment elevation myocardial infarction(STEMI).Methods This study was conducted in Peking University People′s Hospital from 1 August, 2014 to 31 March, 2016, with 229 consecutive cases of acute STEMI enrolled in the study.Data were collected by reviewing medical records and STEMI treatment time records.180 patients with ischemic symptoms ≤12 hours were included in the final analysis.Patients were divided into the consent group and the refusal group according to the final decision.For patients who received primary PCI, they were further categorized into two groups based on the 30min cut-off time.Results Among the 180 STEMI patients reviewed, 139 patients agreed to primary PCI and the remaining 41 patients refused primary PCI.Multivariate logistic analysis showed that symptom relief (OR 5.532, 95% CI 1.165-26.278, P=0.031) and history of chronic kidney disease (OR 4.786, 95% CI 1.346-17.011, P=0.016) were predictors of dissent of primary PCI.Self-rated symptoms severity (OR 0.094, 95% CI 0.034-0.260, P<0.001)was predictor of consent to primary PCI.106 patients had complete time point records of informed consent in the consent group (n=139).Among these patients, the median informed consent delay was 24 min.64 patients made a decision within 30 minutes, and the other 42 patients had their decision made beyond 30 minutes.Symptom-to-door time ≥4 hours (OR 4.563,95% CI 1.682-12.385, P=0.003) was independent predictor of informed consent delay.Conclusions Patients with resolved symptoms, self-rated mild symptoms or renal insufficiency were more inclined to refuse primary PCI.For patients who eventually received primary PCI, symptom-to-door time ≥4 hours was the independent predictor of informed consent delay.

ObjectiveTo observe the clinical efficacy of electroacupuncture at Baihuanshu (BL30) and Huiyang (BL35) in treating chronic abacterial prostatitis.MethodSixty patients with chronic abacterial prostatitis were divided into a treatment group(n=30) and a control group (n=30), respectively to receive electroacupuncture plus Chinese herbal medicine and Chinese herbal medicine alone.ResultAfter intervention, the scores of National Institutes of Health-Chronic Prostatitis symptom index (NIH-CPS) dropped in both treatment and control groups, while the decrease was more significant in the treatment group (P<0.05); the pain or discomfort score from the NIH-CPS dropped in both groups, while the decrease was more marked in the treatment group (P<0.05).ConclusionElectroacupuncture and Chinese herbal medicine both are effective in treating chronic abacterial prostatitis, and electroacupuncture plus Chinese herbal medicine can produce a more significant efficacy than Chinese herbal medicine alone.

Objective To analyze the clinical feature,the endoscopic treatment,therapy of complication of colorectal polyps.Methods According to polyps'size,location and shape in 157 patients with colorectal polyps,corresponding endoscopic treatment method was selected and observed the complication of the endoscopic treatment and therapeutic efficacy.Results 219 colorectal polyps were found in 157 patients.Colorectal polyps occurred frequently in rectum and sigmoid colon.Moreover,inflammatory polyps and hyperplastic polyp were the majority,and a small portion of polyps may turn cancerous.By using endoscopic therapy,hemorrhage was found in 12 samples,and the bleeding ratio was only 5.5％.Conclusion A diversity of methods used in endoscopic therapy of colorectal polyps is safe,efficient,resulting in less complications.

To observe the cerebral activating effects of needling at Waiguan (SJ5) versus SJ5 plus Yanglingquan (GB34) points in young healthy volunteers based on the hypothesis of "needling effect of combined acupuncture points relates to the brain activation".