2.Comparative evaluation of ATB FUNGUS 3 procedure and CLSI M27-A2 broth microdilution method for antifungal susceptibility testing of pathogenic yeasts
Wei LIU ; Jianjun QIAO ; Yan MA ; Zhe WAN ; Ruoyu LI
Chinese Journal of Laboratory Medicine 2009;32(9):1034-1037
to common antifungal drugs.
3.Agrobacterium tumefaciens-mediated transformation of uracil auxotroph Aspergillus fumigatus is an efficient method for target gene knockout
Jianjun QIAO ; Wei LIU ; Yan MA ; Zhe WAN ; Ruoyu LI
Journal of Peking University(Health Sciences) 2004;0(03):-
Objective:To investigate the efficiency of Agrobacterium tumefaciensmediated transfor-mation of Aspergillus fumigatus by using pyrG as a recessive selectable marker.Methods: FAP1 and SHO1 genes target sequences,composed of a selectable marker pyrG and the flanking sequences of the FAP1 and the SHO1 genes,were cloned into a binary plasmid pDHt/sk,respectively.The produced plasmids were transformed into A.tumefaciens.The A.tumefaciens and uracil auxotroph A.fumigatus were cocultured in induction medium without uricil and uridine at 24 ℃ for 48 h.To inhibit growth of A.tumefaciens and to select transformants,the cultures were transferred to 37 ℃ and incubated for another 48 h.Results: In this study,A.tumefaciens-medidated transformation of A.fumigatus produced high homologous recombination rates,which was 44%(7 of 16) for FAP1 and 35%(7 of 20) for SHO1.Conclusion: Our study showed that A.tumefaciens-medidated transformation by using pyrG as a recessive selectable marker is an efficient tool for target gene deletion of A.fumigatus.
4.NGX6 gene combined with cisplatin in treatment of lung cancer in vitro and in vivo
Jing ZHANG ; Zhe WANG ; Xiaoyu SHI ; Wei MENG ; Feng MA ; Jin WANG ; Yongming ZHAO
Chinese Journal of Biochemical Pharmaceutics 2015;37(5):33-36
Objective To evaluate the effect of NGX6 combined with cisplatin on the inhibition rate of A549 cells and NCI-H1975 cells and antitumor effects in vivo.Methods The NGX6 was loaded in the LPD, and prepare Liposome protamine DNA complexes.A549 cells and NCI-H1975 cells were seperately divided into NGX6 group ( 30 μg/mL NGX6 concentration ) , cisplatin group, NGX6 +cisplatin group, PBS as negative control group.The effect of cytotoxicity of four group on A549 cells and NCI-H1975 cell were evaluated by MTT assay.the clone forming rate and the inhibition rate were determined by Cell colony count.lung transplantation tumor model were successfully established, then nude mice were divided into four groups as abeve, each group of 10, tumor size and survival period were determined tumor cell apoptosis were observed.Results The cell viability of A549 cells and NCI-H1975 cells of (NGX6 +cisplatin) were lower than that of NGX6 group, cisplatin group and saline group, respectively(P<0.01).The cloning efficiency of A549 cells and NCI-H1975 cells of ( NGX6 +cisplatin) were lower than that of NGX6 group, cisplatin group and saline group, respectively(P<0.01).The tumor inhibitory rate was in vivo for (NGX6 +cisplatin) was higher than other group(P<0.01).The median survival of nude mice in (NGX6 +cisplatin), NGX6, cisplatin and saline group were 43,31,29 and 15 days.Conclusion NGX6 combination with cisplatin can inhibit the cell proliferate of lung cancer cells and inhibit the tumor growth and the combination of NGX6 and cisplatin may be a potentially effective treatment for lung cancer.
5.Cerebral Palsy Registration in Jiamusi, Heilongjiang
Dongmei MA ; Zhimei JIANG ; Yuting MA ; Shuxia LI ; Zhihai LV ; Lanmin GUO ; Lei XU ; Zhe WU ; Ruixue SUN ; Wei SUN
Chinese Journal of Rehabilitation Theory and Practice 2013;19(11):1096-1098
Objective To explore the methods of cerebral palsy registration. Methods The operation of cerebral palsy registration in Jiamusi was introduced, including the development of the management system, and contents, steps and notices of registration. Results 51 cases with cerebral palsy in Jiamusi were registered from Januaray 1st, 2002 to December 31st, 2011. Conclusion Less children with cerebral palsy had registered, and received insufficient rehabilitation.
6.Protective function of melatonin to acute lung injury and its mechanisms in rats caused by oleic acid.
Zhi ZHANG ; Lei GAO ; Chun-Hua DING ; Wen-Zhe MA ; Wei-Wei GU ; Yun-Lei MA
Chinese Journal of Applied Physiology 2011;27(4):480-483
OBJECTIVETo observe the expression of P-selectin (Ps), intercellular adhesion molecule-1 (ICAM-1) and nuclear factor-kappa B (NF-kappaB) in lung tissues of acute lung injury (ALI) rat model induced by oleic acid (OA) and to explore the protective effects of melatonin (MT) in lung tissues in rats.
METHODSAll rats were randomly divided into four groups: control group, OA group, MT + OA group and SB203580 + OA group. Rat model of ALI was established by intravenous injection of oleic acid (OA). Lung coefficient was measured, lung tissues were imbedded by paraffin to observe morphological changes and the expression of Ps, ICAM-1 and NF-kappaB in lung tissues by means of immunohistochemistry staining.
RESULTSCompared with control group, the lung coefficient increased significantly in OA group (P < 0.05). Alveolar septum thickened significantly in OA group, there had many infiltrated inflammatory cells and collapsed alveoli of lung; positive expression of Ps, ICAM-1 and NF-kappaB were very obvious (P < 0.05); the administration of MT and SB203580 mitigated above changes significantly (P < 0.05).
CONCLUSIONMT possesses obviously protective effect on lung tissues during ALI, its protective mechanism might be related to the inhibition of the expression of Ps, ICAM-1 and NF-kappaB.
Acute Lung Injury ; chemically induced ; physiopathology ; prevention & control ; Animals ; Down-Regulation ; drug effects ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Melatonin ; pharmacology ; therapeutic use ; NF-kappa B ; metabolism ; Oleic Acid ; adverse effects ; P-Selectin ; metabolism ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley
7.Effect of amiloride on the invasion capacity of esophageal carcinoma EC9706 cell line and its possible mechanisms
Jiantao JIANG ; Bin ZHOU ; Shuqun ZHANG ; Shaomin LI ; Wei ZHANG ; Jin ZHANG ; Zhe QIAO ; Ranran KONG ; Yuefeng MA
Cancer Research and Clinic 2011;23(10):657-660
Objective To investigate the effect of amiloride on the invasion capacity of esophageal carcinoma EC9706 cell line in vitro and to elucidate its possible mechanism.Methods The invasion capacities of EC9706 cells pretreated with amiloride were measured by transwell chamber assay. The urokinase-type plasminogen activator (uPA) transcription were determined by RT-PCR.The protein expression of uPA were assessed by Western blot.Results After the EC9706 cells were pretreated with amiloride at different concentrations,the number of invaded cells was obviously less than those of control group with obvious dosage dependent pattern (96±7,78±6,57±6,33±4,15±3,F =43.46,P < 0.01).The transcription levels of uPA mRNA and the protein expression levels of uPA in EC9706 cells decreased significantly compared with the control (mRNA:0.623±0.065,0.526±0.054,0.389±0.041,0.312±0.038,0.247±0.025,F =6.71,P <0.01; protein:0.732±0.064,0.644±0.057,0.533±0.058,0.391±0.036,0.267±0.043,F =6.71,P <0.01).Conclusion Amiloride inhibits the invasion capacity of esophageal carcinoma EC9706 cells.The mechanism might be associated with down-regulation of the expression of uPA.
8.Expression of urokinase-type plasminogen activator (uPA) and vascular endothelial growth factor (VEGF) in esophageal cancer and their influence to tumor angiogenesis
Jiantao JIANG ; Bin ZHOU ; Shunqun ZHANG ; Shaomin LI ; Wei ZHANG ; Jin ZHANG ; Zhe QIAO ; Ranran KONG ; Yuefeng MA
Cancer Research and Clinic 2011;23(6):372-375
Objective To investigate the expression and influence to tumor angiogenesis of urokinase-type plasminogen activator (uPA) and vascular endothelial growth factor (VEGF) in esophageal carcinoma. Methods The expression of uPA and VEGF in the tissue of normal (18 cases) and esophageal carcinoma (68 cases) were evaluated by SP immunohistochemistry, CD34 was detected as marking tumor microvessel density (MVD). uPA and VEGF expression were assessed as to the pathologically biological features of esophageal cancer and to the influence to tumor angiogenesis. Results The positive rates of uPA were 27.8 % (5/18) and 70.6 % (48/68) in the tissue of normal and esophageal carcinoma, respectively, there was significant difference in two tissues (x2 =11.63, P <0.05). The positive rates of VEGF were 22.2 % (4/18)and 63.2 % (43/68) in the tissue of normal and esophageal carcinoma, respectively, there was significant difference in two eissues (x2 =9.78, P <0.05). The expressions of uPA and VEGF in esophageal carcinoma were uniformity (x2 =9.72, P <0.05). The mean of MVD was 42.38±11.62. The positive rates of uPA and VEGF were higher in the high MVD group than those in the low MVD group (x2 =6.13, P <0.05, x2 =10.12, P <0.05,respectively). uPA and VEGF expressions in malignant tumors weren' t associated with age, gender and pathological types (P >0.05), but associated with clinical stage, histologic grading and lymph node metastasis (P <0.05). Conclusion Rising expression levels of uPA and VEGF are common in esophageal carcinoma. Altered expression of uPA and VEGF may contribute to tumor angiogenesis of esophageal carcinoma, whose overexpression indicate worse prognosis.
9.Experimental study on rad submandibular gland cell and silk fibroin-chitosan in vivo
Wei-Wei HUANG ; Xue-Xin TAN ; Bo LI ; Zhe WANG ; Xin-Yu LI ; Li MA
Chinese Journal of Stomatology 2011;46(10):608-612
Objective To evaluate the feasibility of the tissue engineered submandibular gland constructed in vivo based on submandibular gland cells and silk fibroin-chitosan (SFCS).Methods Submandibular gland cells were obtained and purified.The second generation cells labeled by 5′-BrdU were seeded on SFCS(5 mm× 5 mm × 5 mm).Submandibular gland cells seeded on SFCS was implanted beneath the skin on the back.At 3,7,14 d post-implantation,implant sites were examined local and systemic responses.After paraffin embedding,serial sections 6 mm thick were cut and stained with either hematoxylin and eosin or brdu tissue stain for immunohistochemical studies and examined the responses of tissue.Scanning electron microscope was used to observe the growth behavior submandibular gland cells on SFCS scaffolds.Results General observation:at the 3,7,14 d after in vivo implantation,capsule formed in the surface of insert.Histological observation:in experimental group,submandibular gland cells proliferate on the SFCS scaffold fused to form unit 14 d after implantation.Brdu immunohistochemical observation:the results of labelled cells were positive by immunohistochemical method at each time point.Cytokeratin-8 (CK-8) immunohistochemical observation:the results of labelled cells were positive by immunohistochemical method at each time point.With time,the positive cells gradually increased.Scanning electron microscope:the shape of the SFCS scaffold was mesh.At earlier,submandibular gland cells presence in disorder at attach to the SFCS.At the 14 d submandibular gland cells proliferate on the SFCS scaffold and form functional unit.Conclusions Such constructed tissue engineered submandibular gland based on submandibular gland cells and SFCS is promising.
10.Effect of micronutrients on the immune status of human immunodeficiency virus-positive individuals.
Fei ZHAO ; Xue-Ling FENG ; Wei XU ; Yan-Min MA ; Zhe WANG ; Wen-Jie LI
Acta Academiae Medicinae Sinicae 2010;32(3):340-342
OBJECTIVETo evaluate the effect of micronutrients on the immune status of human immunodeficiency virus (HIV)-positive individuals.
METHODSTotally 102 HIV-positive individuals were randomly divided into supplementation group (received micronutrients supplement) and control group (received placebo). Physical examinations were performed at baseline and at the end of the trial. Immune status were determined in both two groups.
RESULTSAge, height, weight, and sex ratio were not significantly different between two groups (all P>0.05). Baseline CD3+, CD4+, and CD8+ T lymphocytes and IgA, IgG, IgM, C3 levels were not significantly different between two groups (all P>0.05), while the levels of CD3+, CD4+, and CD8+ T lymphocytes and IgA, IgG, IgM, C3 were significantly higher in supplementation group than in control group(all P0.05).
CONCLUSIONSupplementation of micronutrients can improve the immune status of HIV-positives individuals.
Adult ; Dietary Supplements ; HIV Infections ; drug therapy ; immunology ; Humans ; Micronutrients ; therapeutic use ; Middle Aged