Objective To investigate the effect of liquid temperature, pressure, concentration, pH parameters on the membrane process and rejection in the process of using reverse osmosis membrane to concentrate oil-bearing water bodies ultrafiltrate of Compound Chuanxiong Capsules, and optimize the process. Methods With oil-bearing water bodies ultrafiltrate of Compound Chuanxiong Capsules volatile oil as the research object, the changes in flux under different operating parameters was measured. Results For this system, suitable operation parameters was as follows: pressure was 1.6 MPa, temperature was 35～ 40 ℃, pH was 9～10. Conclusion Using reverse osmosis membrane to concentrate oil-bearing water bodies ultrafiltrate of Compound Chuanxiong Capsules volatile oil, better flux and rejection can be obtained at the appropriate operating conditions.

BACKGROUND:Skeletal muscle satel ite cells are totipotential stem cells with multi-directional differentiation potential, locate in skeletal muscle interstitium, have a certain tolerance to ischemia and hypoxia, and are important cells in stem cellengineering.
OBJECTIVE:To establish a thrifty, convenient culture procedure and create a simple, efficient method to transfect skeletal muscle satel ite cells, and investigate genetic expression after the transfection for cellular cardiomyoplasty.
METHODS:Skeletal muscle satel ite cells were isolated from rabbit thigh and cultured. Their growth curves were determined by CKK-8 method. Grouped by different proportions of the plasmid and liposome, skeletal muscle satel ite cells were transfered by the enhanced green fluorescent protein plasmid based on liposome. After transfection, the efficiency and character of target genetic expression was determined.
RESULTS AND CONCLUSION:Satel ite cells were isolated, cultured and transfected successful y. In suitable ratio of plasmid and liposomes, the transfection efficiency reached up to above 35%. The target protein was expressed within 12 hours after transfection, reached maximum in 48-72 hours and decreased gradual y after one week. The expression stil could be observed two weeks latter. The enhanced green fluorescent protein plasmid conducted by cationic liposome could be transfered into skeletal muscle satel ite cells efficiently. The transfection efficiency was correlated closely to the ratio of plasmid and lipofectamine. The change of target gene expression depended on time.

BACKGROUND: Previous studies have found that skeletal muscle satellite cell transplantation can induce angiogenesisin myocardial infarction area, reduce infarct size and improve cardiac function. But the overall effect is not satisfactory.OBJECTIVE: To observe the survival of basic fibroblast growth factor (bFGF) gene modified skeletal muscle satellitecells in acute myocardial infarction and to observe the expression of bFGF gene and the effect of cell transplantation onangiogenesis in myocardial infarction area.METHODS: Eighteen New Zealand white rabbits were divided into three groups by random: skeletal muscle satellite cellgroup (control group), bFGF gene enhanced skeletal muscle satellite cell group (experimental group) and blank controlgroup. The left anterior descending branch of the coronary artery of the rabbits was ligated so as to establish an animalmodel of acute myocardial infarction in the former two groups. After labeled by DAPI before transplantation, the skeletalmuscle satellite cells, bFGF gene modified skeletal muscle satellite cells and the equivalent amount of DMEM/F12 wereinjected into the local infarct myocardium correspondingly. Samples were taken 4 weeks after transplantation. Then, thesurvival of skeletal muscle satellite cells and the expression of bFGF gene were observed under light microscope andfluorescence microscope, and the neovascularization in the myocardial infarction area was examined byimmunohistochemical staining.RESULTS AND CONCLUSION: No DAPI-labeled cells were visible in the blank control group, but in the other twogroups, a large amount of DAPI-labeled skeletal muscle satellite cells were seen in the infarction area. Enhanced greenfluorescent protein was highly expressed in the experimental group. Microvessel density in the infarction area washighest in the experimental group followed by the control and blank control groups (P < 0.05). These findings indicatethat bFGF gene modified skeletal muscle satellite cells can survive and promote neovascularization in the acutemyocardial infarction area.

Objective To discuss the clinical efficacy of closed reduction and plaster splint fixation ( CRPSF) in the treatment of gerontal patients with distal radius fractures ( DRF) .Methods 76 elderly patients with DRF who treated by CRPSF were selected .According to AO classification of fractures ,the patients were divided into three groups,the A group had 27 cases;B group had 26 cases,C group had 23 cases.The treatment effect was evalua-ted by analyzing the follow-up data,the corresponding imaging measurement parameters and clinical scores .Results All patients had 12-month clinical and imaging follow-up.In the last follow-up, the arm-shoulder-hand dysfunction score and palmar angle ,ulnar deviation angle of A and B group were significantly better than those of C group , the differences were statistically significant(all P<0.05),the difference between A and B group was not statistically sig-nificant (P>0.05).In the last follow-up,the satisfaction score of C group was slightly lower than that of the A or B group,but had no statistically significant difference (P>0.05).Conclusion CRPSF in the treatment of gerontal pa-tients with DRF has good function and imaging effects ,and the improvement level has a certain relationship with the degree of fracture,but has no significant impact on the patients'satisfaction.

Objective To investigate the short-term acute rejection incidence of the recipients under the steroid-free immunosuppressive therapy after liver transplantation. Methods This retrospective study included 186 patients who were divided into two groups by random number table.The patients in no steroid group (the study group, n =94) received tacrolimus (Tac) with mycophemolate mofetil (MMF) or cyclosporine with MMF,and those in the steroid group (the control group,n =92) received the aforementioned immunosuppressive therapy combined with steroids.The acute rejection incidence was analyzed during six months post-transplantation.Results There was no significant difference in the gender,age,indication for transplantation,Child-Pugh score,MELD score,operating time,bleeding and transfusion volume during the operation,warm ischemia time and cold ischemia time between the two groups (P＞0.05).Liver biopsy was done on 9 cases of each group.The acute rejection incidence had no significant difference between the study group and the control group (5/94 vs 4/92,5.3％ vs 4.4％,P＞0.05).Conclusion The steroid-free immunosuppressive therapy after liver transplantation did not increase the short term acute rejection incidence.

Objective To establish an isolation method for solid GTC in peripheral blood using EpCAM antibody-linked nanobeads and evaluate the sensitivity of the method and its application significance. Methods Five, ten, twenty, fifty and one hundred MCF7 (breast cancer), KYSE70 (esophageal cancer), BxPC-3 (pancreatic cancer) and 9811P (stomach cancer) cells were added into 7. 5 ml erythrocyte lysed peripheral blood obtained from healthy volunteers respectively. EpCAM antibodylinked nanobeads were used to enrich cancer cells. The recovery rates of the in vitro added cancer cells were evaluated by fluorescence microscopy. Then, the untreated thirty cases of esophageal cancer (six cases at stage Ⅰ and Ⅱ, twenty-four cases at stage Ⅲ and Ⅳ), thirty-five cases of breast cancer (fifteen cases at stage Ⅰ and Ⅱ , twenty cases at stage Ⅲ and Ⅳ), thirty cases of pancreatic cancer (five cases at stage Ⅰ and Ⅱ , twenty-five cases at stage Ⅲ and Ⅳ), thirty-three gastric cancer (thirteen cases for stage Ⅰ and Ⅱ ,twenty cases at stage Ⅲ and Ⅳ) were enrolled to enrich the peripheral blood CTC. Thirty healthy volunteers and thirty gastritis patients served as two groups of control. Meanwhile the enriched CTC was identified by IF and HE staining. FISH was used to analyze the copy number of chromosome 8 and chromosome 20 in two hundred esophageal cancer, breast cancer, pancreatic caner and gastric cancer CTC. Results After DAPI staining and mixing with 7.5 ml peripheral blood from healthy donors, the average cell recovery rates of KYSE70, MCF7, BxPC-3 and 9811P cells evaluated under fluorescence microscope were 87%, 87%, 86% and 88% (within group), and the recovery rates of 5 gradient dilution levels were 88%, 85%, 87%, 88% and 87% (intergroup). With a high sensitivity, this method was able to isolate one cancer cell in 107 white blood cells of peripheral blood. The positive rates of more than 2 CTC in the peripheral blood detected by this method were 50% (15/30) of esophageal cancer, 63% (22/35) of breast cancer, 70% (21/30) of pancreatic cancer and 61% (20/33) gastric cancer patients respectively,but no CTC was detected in the peripheral blood of healthy volunteers and gastritis patients (P = 0. 000).The aneusomy of chromosome 8 and chromosome 20 were found in 80% esophageal cancer, 75% breast cancer, 65% pancreatic cancer and 59% gastric cancer. Conclusions The CTC isolation technique with EpCAM antibody-linked nanobeads is sensitive and accurate. The aneusomy of chromosome 8 and 20 is frequent in CTC from esophageal cancer, breast cancer, pancreatic cancer and gastric cancer.

Objective To evaluate the accuracy of lymphocyte count as a surrogate for CD4+T cell count in treatment-na(i)ve HIV-infected adults.Methods A total of 2 013 HIV-infected patients were screened at 23 sites in China.CD4+ T cell counts were measured by flow cytometry.Correlation between CD4+ T cell count and peripheral lymphocyte count were analyzed by spearman coefficient.AUCRoc were used to evaluate the performance of lymphocyte count as a surrogate for CD4+ T cell count.Results The lymphocyte count and CD4+T cell count of these 2 013 patients were (1 600±670) × 106/L and (244 ± 148) × 106/L respectively.CDa+ T cell count were positively correlated with lymphocyte count(r =0.482,P ＜0.000 1).AUCROC of lymphocyte count as a surrogate for CD4+ T cell counts of ＜ 100 × 106/L,＜200 × 106/L and ＜ 350 × 106/L were 0.790 (95％ CI 0.761-0.818,P ＜ 0.000 1),0.733 (95％ CI 0.710-0.755,P ＜ 0.000 1) and 0.732 (95％ CI 0.706-0.758,P ＜ 0.000 1) respectively.Conclusion Lymphocyte count could be considerad as a potential surrogate marker for CD4+ T cell count in HIV/AIDS patients not having access to T cell subset test by flowcytometry.

Objective To evaluate the diagnostic and therapeutic effect of endoscopy on pharyngeal papillomas. Methods Data of patients with pharyngeal papillomas diagnosed and treated by endoscopy be?tween March 2009 and December 2014 were analyzed retrospectively, including dissection, treatment and follow?up results. Results The rate of endoscopic diagnosis of pharyngeal papillomas was 0?9%( 65/6 927) . Endoscopic biopsy forceps resection was performed successfully in 54 patients. Other patients ( n=11) were treated by endoscopic snare resection. There was hemorrhage of different degrees after resec?tion. Argon hemostasis was used in 6 patients for errhysis after resection. Supportive treatment was not given and no severe hemorrhage or death was seen. A total of 41 patients were followed up and endoscopic examina?tion was performed 2 months later. Pharyngeal papillomas were found again at the same site in two patients and endoscopic biopsy forceps resection was performed. Pharyngeal papillomas were not found in these two patients in the next endoscopic examination. Conclusion Endoscopy is a safe and effective diagnosis and treatment method for pharyngeal papillomas.

Objective To analyze the susceptibilities of Escherichia coli isolates collected from blood and the prevalence of ESBLs encoding genes.Methods A total of 121 Escherichia coli isolates collected from blood during 2012 were analyzed for antimicrobial susceptibilities by software of WHONET 5.6,the production of ESBLs was confirmed by confirmatory pheno-typic testing,PCR and DNA sequence were further implemented to analyze the ESBLs-encoding genes.Results 121 E.coli isolates displayed high resistance towards broad spectrum penicillin and 2nd or 3rd generation cephalosporins,levofloxacin and cotrimoxazole,with the resistance rates being more than 40%,susceptibilities to imipenem,piperacillin/tazobactam,ami-kacin were observed,with the resistance rates to be less than 12%,86(88.7%)out of 121 isolates were found to produce ESBLs.Among them,59.5% (72),38.8% (47)and 4.1% (5)were confirmed to carry blaCTX-M,blaTEM and blaSHV genes.Additionally,2(1.7%)isolates carried all the genes detected,30(24.8%)isolates carried both of blaCTX and bla-TEM,1(0.8%)isolate carried both of blaSHV andblaTEM.Conclusion Most of the E.coli isolates from the blood culture in Nanjing Gulou Hospital produce ESBLs,and displayed resistance towards most of the penicillins,cephalosporins and sin-gle amide antimicrobial agents should be chosen according to susceptibility results.

Objective To analyze the correlation between CD4+ T cell count and HLA-DR or CD38 expression on peripheral CD8+ T cells in treatment-naive Chinese HIV/AIDS patients.Methods A total of 471 treatment-naive HIV-infected patients and 53 healthy volunteers were enrolled.HLA-DR or CD38 expression on peripheral CD8+ T cells, CD4+ T cell count, naive CD4+ T cell subsets and CD28 expression on T cells were measured by flow cytometry.Plasma HIV RNA load was quantified by real-time PCR (COBAS TaqMan RT-PCR).Mann-Whitney U test and Spearman's rank correlation test were used for statistical analysis.Results CD38 expression intensity on CD8+ T cells was negatively correlated with CD4+ T cell count (r =-0.53, P ＜ 0.001) and naive CD4+ T cell count (r =-0.48, P ＜ 0.001).CD38 expression on CD8+ T cells also displayed significantly negative correlation with CD28 expression on CD8+ T cells (r =-0.46, P ＜ 0.001).HLA-DR expression on CD8+ T cells did not show significant correlation with CD4+ T cell count.Only weak positive correlation was found between CD38 intensity on CD8+ T cells and plasma HIV RNA load.Conclusions Compared with HLA-DR, CD38 expression on CD8+ T cells shows more significant negative correlation with CD4+ T cell count in treatment-naive patients.CD38 and HLA-DR may play different roles on the persistent immune activation in HIV infection.