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 study the prevalence, clinical characteristics and risk factors of HIV-related lipodystrophy syndrome (HIV-LD) in our cohort of HIV-1 infected Chinese adults. Methods In a cross-sectional study, 55 HIV-infected patients were recruited from the HIV clinic of Peking Union Medical College Hospital; most of them were undergoing the first-class highly active antiretroviral therapy (HAART) of today in China. Lipoatrophy or lipohypertrophy was defined if there was concordance between the report of fat change and clinical examination of the participants. Whole body dual-energy X-ray absorptiometry (DEXA) scanning was performed. Results Prevalence of clinical body fat redistribution in the present study was 47.3%. Comparing with non-LD patients, HIV-LD patients had elder age and longer exposure to HAART(P<0.05). HAART exposure and stavudine(d4T) usage were two independent risk factors for HIV-LD. Conclusions HIV-related fat redistribution does exist in Chinese HIV population. Peripheral lipoatrophy occurs commonly in HIV-infected adults but is not associated with increased trunk fat. HAART exposure and especially d4T usage are independent risk factors for HIV-LD.

Objective To study the profile of peripheral natural killer cells(NK cells)and γδT lymphocytes in human immunodeficiency virus(HIV)infected patients with different disease status and to explore the pathogenesis of acquired immunodeficiency syndrome(AIDS).Methods Three hundred and eleven HIV/AIDS patients without antiviral treatment were enrolled in this study.The percentages and absolute numbers of CD4+T lymphocytes,NK cells,and γδT ceils were measured by flow cytometry.The patients were divided into 3 groups based to their CD4+T lymphocytes counts:low CD4+T lymphocytes group(L),patients with CD4+T lymphocytes <0.20×109L;middle CD4+T lymphocytes group(M),CD4+T lymphocytes counts between 0.20×109and 0.35×109L;high CD4+T lymphocytes group(H),patients with CD4+T lymphocytes counts >0.35×109L.Rank sum test of independent samples of two-group and multiple-group was performed using Mann-Whitney U test and Kruskal-Wallis test.Correlation analysis was done by Spearman and Pearson test. Results The median percentage and cell counts of NK cells(8.4%,103×106L) and γδT cells(3.4%,41×106L)in HIV/AIDS patients were all significantly lower than those of healthy individuals(Z=-5.029,Z=-7.723,Z=-2.437,Z=-6.063;all P<0.01).The median cell counts of CD4+T lymphocytes in L,M,H groups were 0.062×109L,0.276×109L and 0.482×109L,respectively.The median cell counts of NK cells in these 3 groups were 89×106L,97×106L and 146×106L,respectively.NK cell counts were not significantly different between L and M groups,whereas both of them were much lower than that of H group(Z=-3.392,P=-0.001,Z=-4.849,P<0.01,respectively).The median γδT cell counts of L,M and H group were 29×106L,43×106L and 59×106L,respectively.The differences between any 2 groups were not significant.Conclusion These data suggest that the decreasing levels of peripheral NK cells and γδT cells are different after HIV infection.

To investigate the different reconstitutional profiles for acquired(CD4+ T cell)and innate(NK cell,γδT lymphocyte)immunity after highly active antiretroviral therapy(HAART).Methods The CD4+ CD4+,CD3+ CD4- CD8-,CD3- CD16/CD56+,CD4+ CD45 RA+ CD62 L+ and CD4+ CD45 RA- subsets were measured by flow cytometry.The dynamic changes of these subsets after HAART initiation were assessed in 59 patients who were followed for 12 months in resular 3-month visits.Results At baseline the cell counts of CD4+ T cells including its na(I)ve and memory subsets,NK cell and γδT cells in HIV/AIDS patients were all significantly lower than those of healthy individuals.There was a decrease of 2.33 lg copies/ml in HIV-1 RNA from baseline noted 1 month after initiation of treatment which was sustained through 12 months.CD4+ T cell count showed a bi-phase increase during treatment.The first rapid increase was mainly memory CD4+T cells and this followed by the second slow but steady increase of na(I)ve CD4+ T cells.Increases in NK cell and γδT cell were noted at 3 months of HAART and this restoration were different quantitatively when compared with the oge in CD4+ T cells.Conclusion HAART could induce a different quantitative restorational patterns in peripheral CD4+ T cells,NK cells and γδT cells.

Objective To investigate the prevalence of primary HIV-1 genotype drug-resistance and viral subtype from 237 treatment-naive patients in China.nethods CD4+ T cell counts,plasma HIV-1 viral load and HIV-1 gene sequencing in total 237 treatment-naive patients enrolled from 20 provinces/regions were detected for the evaluation of primary HIV genotypic drug-resistance.Results The survey of 237 treatment-naive patients from muhicenter areas including Henan Province,Yunan Province,and Shanghai showed that 9 subtypes of HIV-1 strains were finally identified.Most of patients were infected before 2003.Only 3 cases had genotypic mutations associated resistance to antiretroviral drugs,with high resistance to nucleoside reverse transcriptase inhibitors(NRTIs),moderate resistance to protease inhibitors(PIs)and high resistance to non-nucleoside reverse transcriptase inhibitors(NNRTIs).respectively.The prevalence of primary genotypic drug resistance was 1.3%(3/237)in this study.Conclusions The rate of HIV-1 primary genotypic drug-resistance is still relatively low in treatment-naive HIV/AIDS patients while 9 subtypes of HIV-1 strain was diseovered.

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.