Objective To investigate the diagnosis and treatment of secondary sclerosing cholangitis after billary tract operation.Methods The clinical clata of 7 cases with secondary sclerosing cholangitis after billary tract operation treated in our hospital from January 2000 to June 2008 were retrospectively analyzed.Results Recurrent episodes of bacterial cholangitis were present in all patients,and they all had history of biliary tract operation.After definite diagnosis,2 cases underwent operation and 5 cases underwent conservative treatment.Among the latter,1 case underwent endoscopic sphincterotomy and 1 case had liver transplantation.Of all the patients,2 cases recovered(2/7),2 cases died(2/7) and 3 cases are still undergoing treatment.Conclusions The prognosis of secondary sclerosing cholangitis after billary tract operation is relatively poor.The key measure is prevention,and early diagnosis together with rational treatment can have certain therapeutic effect.

Objective To investigate the clinical effect of splanchnicectomy for pain palliation with advanced pancreatic carcinoma.Methods Altogether 27 cases were treated from January 2005 to February 2008,retrospective study was used to analyze the clinical data of them.All the patients had diverse extent pain,among these 16 cases who had underwent splanchnicectomy (therapy group),others were in control group.Results Pain extent was scored according to visual analogue scale (VAS),preoperative score of therapy group (6.06±1.93) scores,postoperative score 3 months(2.67±1.68) scores,6 months(2.69±1.75) scores,the discrepancy had statistical significance (P<0.01),preoperative score of control group (5.91±1.87) scores,postoperative score 3 months(5.11±2.03) scores,6 months(5.33±2.25) scores,the discrepancy had no statistical significance.Intergroups contrast,showing significant difference (P<0.05).Conclusion For the patients who request to deal with primary disease,splanchnicectomy is manipulated easy,short of complication,significantly relief of pain,and deserve to be spreaded.

Objective To compare the effect of different cryoprotectants and different concentrations on controlled?rate freezing of human ovarian tissues. Methods Ovarian tissues were sampled from 15 patients undergoing benign ovarian tumor surgery. Cortical slices were frozen by controlled?rate freezing using three cryoprotectants,propanediol,ethanediol,and dimethylsulphoxide,and the concentration of each cryoprotectant was 1.5 mol/L or 2.0 mol/L. Cortical slices obtained from each patient were processed with each cryopreservation procedure simultaneously. Morphology of fol?licles was studied by light and electron microscopy and the normal rate was compared with that of the fresh tissues from the patient. Results There were no significant differences in distribution of follicles of different developmental stages between each group(P<0.05). Light microscopy showed 1.5 mol/L EG and 2.0 mol/L EG groups had the best freezing effect,and the difference in the morphologically normal rate of follicles was not statisti?cally significant compared to fresh controls(P>0.05). However,the difference was statistically significant for 1.5 mol/L PROH,2.0 mol/L PROH, 1.5 mol/L DMSO and 2.0 mol/L DMSO groups(P<0.05). Electron microscopy showed the oocyte quality declined after controlled?rate freezing pro?cedure. However,the statistical analysis was not conducted due to little data. The viability of granulosa cells was significantly declined after all the freezing procedures compared to that of the fresh control tissues(P<0.01). The number of morphologically normal granulosa cells was slightly higher in the tissues which had been cryopreserved with 2.0 mol/L PROH and 1.5 mol/L EG,but no significant differences were found between any of two frozen groups(P>0.05). Conclusion Controlled?rate freezing using 1.5 mol/L EG as the cryoprotectant can better save oocytes and granulosa cells. It is a preferable freezing procedure for ovarian tissues.

Objective:To investigate the effects of ovarian cancer ascites-derived exosomes on the stem cell properties and invasion ability of ovarian cancer stem-like cell (OCS-LC).Methods:(1) A2780 cells were induced into OCS-LC in serum-free medium, and authenticating their stem-like properties by sphere-forming test, differentiation test and CD 133 marker detection. (2) Exosomes from ascites and A2780 cell were extracted by ultracentrifugation, then authenticating them. The morphology of exosomes was observed by the transmission electron microscope, exosome particle size was measured by nanoparticle tracking analysis (NTA). The expressions of heat shock protein 70 (HSP-70), CD 63 and CD 9 were detected by western blot. (3) The exosomes from ovarian cancer ascites and tumor cell supernate were co-cultured with OCS-LC. The groups were divided into control group, ascites-derived exosomes (ADE) group (ADE+OCS-LC group), and cells-derived exosomes (CDE) group (CDE+OCS-LC group). The sphere-forming ability was evaluated by sphere-forming cycle, maximum sphere diameter and sphere-forming rate of each group; the expression of CD 133 was detected by immunofluorescence staining under microscope; quantitative real-time (qRT)-PCR was used to detected the expression levels of octamer-4 (Oct-4), Nanog mRNA of the signature genes in the stem cells of each group; the metastasis ability of each group was measured by transwell assay. Results:(1) Identification of OCS-LC: sphere-forming experiment showed that the suspension of OCSC single cells was grown in serum-free medium in secondary sphere-forming. Differentiation function experiment showed that OCS-LC were differentiated into adherent A2780 cells by changing the growth mode in serum containing medium. Flow cytometry showed that the proportion of CD 133 positive ( CD133+) cells in OCS-LC group was (18.9±0.9)%, significantly higher than that of control group (0.6±0.5)% ( t=38.570, P<0.01). (2) Under transmission electron microscope, clear lipid bilayer structure was observed in ADE and CDE, and one side presented a concave hemispheric or cup like structure. NTA showed that the diameter of exosomes mainly ranged from 30 to 100 nm, with an average diameter of 67.2 nm. Western blot analysis showed that the specific protein molecules HSP-70, CD 63 and CD 9 were positive. (3) Three groups′ OCS-LC could continue to grow into spheres, and the group of ADE+OCS-LC showed two growth modes, most of the cells continued to grow into spheres, while a small part of cells grew in adherent differentiation. The sphere-forming rate of OCS-LC in the control group, ADE+OCS-LC group, and CDE+OCS-LC group were (1.05±0.20)%, (4.15±0.10)%, and (10.45±0.25)%, the sphere-forming cycle of OCS-LC in the three groups were (15.3±1.5), (10.3±0.6), and (6.7±0.6) days, and the maximum diameters of OCS-LC were (100.3±3.2), (145.2±5.1) and (170.0±2.1) μm, respectively. And the differences were statistically significant (all P<0.05). After co-culture of exosomes with OCS-LC, the sphere-forming ability of cells in the group of CDE+OCS-LC was prior to ADE+OCS-LC group (all P<0.05). Immunofluorescence staining showed that the number of CD 133 green fluorescent chromophore cells in OCS-LC groups [(46.2±2.1)%, (58.4±2.2)%] was significantly higher than that in the control group [(26.6±1.5)%] after the addition of exosomes in co-culture, the positive rate of CD 133 was higher than that in the control group( F=187.588, P<0.05). The qRT-PCR results showed that the expression levels of Oct-4 mRNA in ADE+OCS-LC and CDE+OCS-LC groups were 3.46±0.24, 4.03±0.31, compared with that in control group (1.04±0.12), the differences were statistically significant ( F=134.932, P<0.05). The mRNA expression levels of Nanog were 1.57±0.32, 2.66±0.15, which were significantly higher than that in the control group (1.00±0.07), and the differences were statistically significant ( F=49.329, P<0.05). And the expression of both in CDE+OCS-LC group increased more significantly than ADE+OCS-LC group (all P<0.05). The number of invasive cells in the three groups of OCS-LC were: control group 30±5, ADE+OCS-LC group 102±4, CDE+OCS-LC group 210±7, and there were statistically significant differences among three groups ( F=820.800, P<0.05). Compared with the control group, the number of invaded cells in the co-culture group were significantly increased ( P<0.05), and the CDE+OCS-LC group had the higher cell invasion ability then the ADE+OCS-LC group ( t=23.202, P<0.05). Conclusions:Exosomes derived from ovarian cancer ascites could enhance and maintain the stemness of OSC-LC, and promote the invasion of tumor cells. Moreover, CDE is superior to ADE.

AIM To investigate the effects of the novel antihypertensive drug iptakalim hydrochloride on potassium currents in artery smooth muscle cells. METHODS The effects of iptakalim hydrochloride on potassium currents in smooth muscle cells derived from rat pulmonary arteries were observed by using patch clamp technique(whole cell recording) after application of the drug in the bath. RESULTS The potassium current-voltage curves ( I-U curves) of smooth muscle cells derived from normotensive rat pulmonary arteries were up-ward shifted by iptakalim hydrochloride(0.1,1,10 and 100 ?mol?L -1 ). Within 5 minutes after application of the drug, the current amplitude could increase to[(118.6?15.9)%, P

AIM:To explore the depression-like behavior and cystathionine β-synthase (CBS)/hydrogen sulfide (H2S) levels of amygdala in posttraumatic stress disorder (PTSD) rats and to study the effect of exogenous H2S on PTSD rats.METHODS:Single prolonged stress paradigm was adopted to prepare PTSD animal model.Forced swimming test and sucrose preference test were used to evaluate the depression-like behavior.The content of CBS/H2S in amygdala tissue was measured by Western blot and methylene blue method.In vivo extracellular single unit recordings was used to examine the frequency of spontaneous discharges of amygdala neurons.RESULTS:The immobility time in forced swimming test of PTSD group increased and sucrose preference in sucrose preference test of PTSD group decreased compare with normal group (P<0.01).CBS/H2S level in amygdala tissue of PTSD group decreased compared with normal group (P<0.01).The immobility time of the rats in forced swimming test of NaHS+PTSD group decreased and the sucrose prefe-rence in sucrose preference test of NaHS+PTSD group increased compare with PTSD group (P<0.01).L-cysteine increased the frequency of spontaneous discharges of amygdala neurons (P<0.01).CONCLUSION:Depression-like behavior is aggravated in PTSD model rats owing to the inhibition of CBS/H2S content in amygdala tissue.The mechanism of behavior-improving effect of H2S on PTSD model rats is possibly related to excitating amygdala neurons and increasing the frequency of spontaneous discharges.

Objective: To exmine the status and perspectives of e-health among patients in Shanghai community health center. Methods: In November 2014, a semi-open and in-depth qualitative interview was conducted among 40 patients from two community health service centers in Shanghai. According to the results of the interview, a self-made questionnaire was designed and a cross-sectional survey was carried out from March 2015 to May 2015. Eight districts or counties of Shanghai were randomly selected, and one community health service center was randomly selected from each district or county. Total 480 patients who visited the selected center participated in the survey. The main contents of the interviews and questionnaires included: the current status of communication using information and communication technologies (ICT) in patient daily life; the status of the communication with medical personnel by using ICT; the attitudes and perspectives of patients in applying ICT for medical service; and views on e-health development. Results: Among the participants, 25.0%(10/40) of the interviewees and 16.7%(62/371)of the quesitionnaire surveyees had experience of using e-health. Among the questionaired patients, telephone-call was the most common method for communication with doctors (90.3%, 335/371), followed by short message (16.1%, 60/371) and wechat (12.9%, 48/371). Illness consulting is the most common perpurse for communication(64.5%, 40/62), followed by medicine advice(40.3%, 25/62) and sudden physical help(24.2%, 15/62). Among the participants, 56.3%(209/371)of whom thought that e-health should be promoted in the community, 69.4% (145/209) of whom thought that it could save time and effort; among the patients not using or considering unnecessary to use e-health, 87.6% (142/162) of whom thought they lived closer to the hospital and easy to access. Conclusion At present, less patients have the experience of using e-health, but more than half of them believe that it is necessary to use e-health and it should be promoted in the community healthcare.

The chromatographic fingerprint was established for evaluating and controlling the quality of germplasm resource of Ophiopogon japonicus in Sichuan Basin by reversed-phase high performance liquid chromatography and gas chromatography mass spectrometry. The results showed that each sample the characterized by the peak area of 28 HPLC peaks and 13 GC peaks in each program and these peaks were employed for hierarchical cluster analysis. Furthermore, the discrimination of the sample from different regions was achieved by hierarchical cluster analysis via recognizing the 16 (41 datamatrix. This was the first report of hierarchical cluster analysis of the wild germplasm resource of Ophiopogon japonicus according to their chemical fingerprints. Thus, the results proved it is a simple, rapid and accurate method suitable for the quality control of the traditional Chinese medicines.
Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Gas Chromatography-Mass Spectrometry ; methods ; Medicine, Chinese Traditional ; Ophiopogon ; chemistry

OBJECTIVETo compare and appraise the quality of germplasm resource of Ophiopogon japonicus in Sichuan basin.

METHODAccording to the main contents and yield traits, 24 wild germplasm resources of O. japonicus from different areas of Sichuan basin were comprehensively compared by the SPSS 17.0 software with principal component analysis and cluster analysis.

RESULTThe six samples of Ziyang, Jianyang, Leshan, Yibin, Chongqing, Mianyang, their comprehensive evaluation value of quality were higher than the others, and the sample of Ziyang had the best quality, the sample of Dazhou had the least quality, the results of the cluster analysis to raw data were also shown a similar results as principal component analysis.

CONCLUSIONThe wild resources of O. japonicus in Sichuan basin is rich, there are much differences among their quality; the method, through principal component analysis to study the comprehensive evaluation of the O. japonicus quality, is reliability and the results of cluster analysis is also support the conclusions, it could be able to provide a reference to select high O. japonicus quality resources.

Objective To examine the contribution of CD4+ CD25+ regulatory T cells to liver transplant tolerance. Methods After injection of anti-CD25 monoclonal antibody (mAb, PC61), mouse orthotopic liver transplantation was performed and survivals were determined. The paraffin-embedded sections of hepatic allografts were cut and stained with hematoxylin and eosin (HE). Furthermore, the effect of CD4+ CD25+ regulatory T cells on proliferative response of CD4+ T cells and cytotoxicity of CD8+ T cells was examined by depleting these regulatory T cells. Results Depletion of these cells in the recipients but not in the donors before liver transplantation caused rejection. Histological analyses of hepatic allografts with PC61 treatment showed extensive leukocyte infiltration and tissue destruction, whereas those in the control group showed minimal changes. Moreover, elimination of CD4+CD25+ T cells resulted in the enhancement of both proliferative response of CD4+ T cells and cytotoxicity of CD8+ T cells against donor-type alloantigen. Conclusions These results suggest that CD4+CD25+ regulatory T cells were important for tolerance induction to hepatic allografts.