1.Optimization of Reverse Osmosis Process of Oil-Bearing Water Bodies Ultrafiltrate from Compound Chuanxiong Capsules
Ping XU ; Liwei GUO ; Zhifeng HAN
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(11):-
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.
2.Effects of basic fibroblast growth factor modified skeletal muscle satellite cell transplantation on neovascularization following acute myocardial infarction
Zhifeng XU ; Jinglai LI ; Zhen HAN ; Gang FENG ; Mingming REN
Chinese Journal of Tissue Engineering Research 2017;21(9):1397-1401
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.
3.Cationic liposome-mediated enhanced green fluorescent protein plasmid transferred into skeletal muscle satellite cells
Zhifeng XU ; Jinglai LI ; Zhen HAN ; Gang FENG ; Mingming REN
Chinese Journal of Tissue Engineering Research 2013;(46):8056-8061
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.
4.Clinical and imaging efficacy observation of closed reduction and plaster splint fixation in the treatment of ge-rontal patients with distal radius fractures
Daojun LU ; Caifeng ZHANG ; Dong HAN ; Zhifeng HAN ; Xi HAN ; Yan CHEN ; Zhong PENG
Chinese Journal of Primary Medicine and Pharmacy 2014;(5):677-678
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.
5.A new isolation method for peripheral blood circulating solid tumor cells with EpCAM antibody linked nanobeads
Chuanli REN ; Chongxu HAN ; Daxin WANG ; Buhai WANG ; Xingxiang XU ; Jiaxin ZHANG ; Lin ZHOU ; Zhifeng WU
Chinese Journal of Laboratory Medicine 2011;34(3):218-223
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.
6.A multicenter study of correlation between peripheral lymphocyte counts and CD4+ T cell counts in HIV/AIDS patients
Jing XIE ; Zhifeng QIU ; Yang HAN ; Yanling LI ; Xiaojing SONG ; Taisheng LI
Chinese Journal of Internal Medicine 2015;54(2):118-121
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.
7.Analysis on species classification and chracteristics of drug resistance and virulence in CTX-M-producing Escherichia coli isolated from urine culture
Yang LI ; Han SHEN ; Zhifeng ZHANG ; Li CHENG ; Xuejing XU ; Mingzhe NING ; Wanqing ZHOU ; Xiaoli CAO
International Journal of Laboratory Medicine 2017;38(16):2207-2209
Objective To analyze the species classification and chracteristics of drug resistance and virulence in CTX-M producing Escherichia coli isolated from urine culture.Methods Escherichia coli cultured by urine were collected from our hospital during 2014,the ring disk diffusion test was implemented to determine the bacterial susceptibility,the EBLs determination test was used to analyze the bacterial EBLs producing situation;the enterobactoer duplicated gene spacer consensus sequency PCR(ERIC-PCR) was adopted to perform the genetic relation analysis;PCR was used to amplify the CTX-M encoding genes and multiple virulence genes iutA,ompT,fyuA,fdeC,fimH,traT,cvaC,pap,kpsMT,pAI,usp,aer,hlyA,cnf and chuA;the multiple PCR was used to analyze the species calssification of CTX-M-producing Escherichia coli;these strains of bacteria were classified as the CTX-M-producing group and non-CTX-M-producing group according to the results of CTX-M coding gene detection,the differences in the antibacterial drug resistance and virulence genes between the two gorups were performed the contrastive analysis.Results One hundred and sixty-two strains of E.coli by urine culture had no genetic correlation,among 126 EBLs positive strains,91 strains produced CT-M,in which 57 strains of CT-M producing Escherichia coli belonged to type D,and 116 strains belong to Type B2.The statistical analysis found that the drug resistance rate in the CTX-M-producing group was significantly higher than that in the non-CT-M producing group (except for imipenem),the prevalence of virulence genes including iutA,chuA and traT in the CT-M producing bacteria group was significantly higher than that in the non-CTX-M-producing group(P=0.001,0.006,0.000)Conclusion CTX-M-producing E.coli is main pathogenic bacterium of urinary infection in our hospital,its majority belong to type D with increased drug resistance,moreover has close correlation with virulence genes iutA,chuA and traA and is a pertential threat in clinical treatment of urinary infection.
8.Changes of natural killer cells and γδT lymphocytes in human immunodeficiency virus infected patients with different disease status
Jing XIE ; Yahling LI ; Zhifeng QIU ; Yang HAN ; Lingyan ZUO ; Zhengyin LIU ; Taisheng LI
Chinese Journal of Infectious Diseases 2008;26(9):533-536
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.
9.The reconstitutional profiles of peripheral blood natural killer cell,γδT lymphocyte and CD4+T cell in human immunodeficiency virus infected patients during one-year antiretroviral therapy
Jing XIE ; Zhifeng QIU ; Yang HAN ; Lingyan ZUO ; Yanling LI ; Zhengyin LIU ; Taisheng LI
Chinese Journal of Internal Medicine 2008;47(9):750-753
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.
10.A study of HIV-related lipodystrophy syndrome in 55 HIV-infected Chinese adult patients
Lu ZHANG ; Ming LI ; Meimei TAO ; Zhifeng QIU ; Jing XIE ; Yang HAN ; Wei YU ; Taisheng LI
Chinese Journal of Internal Medicine 2009;48(2):118-121
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.