1.Therapeutic effect of neuropeptide PACAP27 on Parkinson's disease in mice
Gang WANG ; Yu-Yan TAN ; Xiao-Kang SUN ; Ru-Jing REN ; Hai-Yan ZHOU ; Sheng-Di CHEN ;
Chinese Journal of Neurology 2005;0(12):-
Objective To investigate the effects of different doses of pituitary adenylate cyclase- activating polypeptide(PACAP)on the functional and morphological outcome in a mice model of Parkinson' s disease(PD)rendered by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP).Methods Male mice were treated with PACAP 0.02, 0.20 or 2.00 ?g by iv bolus for 7 days after MPTP was administered, and were compared with the saline-treated mice.The immunohistochemistry and Western blot were used to detect the alterations of PD biomarker including tyrosine hydroxylase(TH), dopamine transporter(DAT)and vesicular monoamine transporter2(VAMT2).In addition, monoamine neurotransmitters in the striatum of mice were measured by the high performance liquid chromatography (HPLC).Results TH immunohistochemistry indicated that the number of TH-positive neurons in the substantia nigra was increased in all PACAP-treated mice(PACAP(0.02 ?g/d)group was 93.33?4.87, F=85.85,P
2.Application of 293 cells in in vitro detection of carcinogens
Rui ZHAO ; Lei ZHANG ; Ru-Gang ZHONG ; Ling ZHOU ; Yi ZENG
Chinese Journal of Experimental and Clinical Virology 2009;23(1):47-49
Objective To investigate the application of 293 cells to detect suspected carcinogens and provide experimental evidence by using in vitro cell transformation assay and tumorigenicity study. Methods The transformation systems of cells cultured in vitro have been adopted to clarify the tumor promotive activity of Micrecystin LR (MC-LR). The malignant transformation of 293 cells induced by MC-LR is tested by several methods including clone forming in soft agarose, serum requirement assay and tumor forming in mice to defme the promotive activity of 293 cells. Results 293 cell acted like tumor cells after induced by MC-LR: serum dependence decreased, anchorage independence growth in soft agarose and formed cell clones, malignant tumors appeared in SCID mice. Conclusion 293 cells were easy to culture and sensitive to environmental carcinogens so that can be used in detection of suspicions carcinogens.
3.Observation on therapeutic effect of acupuncture at "cervical three points" combined with cake-separated moxibustion on cervical hyperosteogeny.
Min-yong LIU ; Rong-rong NIE ; Ru-gang ZHOU ; Wen-zhou LIAO
Chinese Acupuncture & Moxibustion 2008;28(12):877-879
OBJECTIVETo compare therapeutic effects of acupuncture at "cervical three points" combined with cake-separated moxibustion and routine acupuncture at Jiaji (EX-B 2) on cervical hyperosteogeny.
METHODSSingle blind, randomized and controlled methods were used and 91 cases were divided into an observation group of 46 cases and a control group of 45 cases. The observation group was treated by acupuncture at "cervical three points" combined with cake-separated moxibustion, i.e. acupuncture at Chonggu (EX), Dazhui (GV 14), Taodao (GV 13) and moxibustion at pain points with cake made by Rhizoma Atractylodis Macrocephalae and Radix Paeoniae Alba; the control group was treated with acupuncture at Jiaji (EX-B 2). Changes of sourness and pain of the neck, the shoulder and back, pressure pain and cervical activity scores were used for assessment of therapeutic effects.
RESULTSThe markedly cured rate of 76.1% in the observation group was better than 42.2% in the control group (P<0.05). After treatment, the scores of all the items were significantly improved in the two groups (all P<0.05), and the observation group in sourness and pain of the neck, the shoulder and back, and cervical activity were better than the control group (all P<0.05).
CONCLUSIONThe therapeutic effect of acupuncture at "cervical three points" combined with cake-separated moxibustion on cervical hyperosteogeny is superior to routine acupuncture at Jiaji (EX-B 2).
Acupuncture Points ; Acupuncture Therapy ; Adult ; Combined Modality Therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Spinal Osteophytosis ; therapy
4.Experimental research of promoting revascularization of tracheal transplantation by gene therapy.
Qi-xu ZHANG ; Gang ZHOU ; Chun-mei WANG ; Ai-ru ZHOU
Chinese Journal of Surgery 2004;42(10):622-626
OBJECTIVETo investigate methods of promoting revascularization of tracheal transplantation to increase the length of graft.
METHODSTransfer recombinant plasmid pcDNA3.1/myc-His(-)C-bFGF and pCD(2)-VEGF(121) into rabbit cervical muscle by direct injection of plasmid following electric pulses in vivo. Use histochemistry and immunohistochemistry analysis of muscles injected to show the transferred gene expression and the biological effect. Based on the former experiment, conduct gene therapy to the rabbit tracheal autotransplantation wrapped by cervical combined muscles by injecting plasmid DNA directly, combined with gene sutures following electric pulses. Observe and analyze the effect on trachea viability.
RESULTSThe recombinant plasmid, pcDNA3.1/myc-His(-)C-bFGF and pCD(2)-VEGF(121) was transferred into muscles flap in vivo successfully. The active protein bFGF and VEGF(121) were expressed at high levels. Blood vessels increased significantly in the muscles, and blood circulation was improved by local angiogenesis. Ten rings tracheal autograft wrapped by transgenic muscles integrating with gene structure revascularized completely, and the rabbit survived for a long period of time. There was significant difference between gene therapy group and control group (P < 0.01). There was no significant difference between bFGF gene therapy group and VEGF(121) gene therapy group. Almost rabbits in the control group died of graft necrosis.
CONCLUSIONTracheal grafts revascularization can be established early by the cervical combined muscles flap wrapping associated with single gene therapy. The length of the tracheal can be increased simultaneously.
Animals ; Female ; Fibroblast Growth Factor 2 ; genetics ; therapeutic use ; Genetic Therapy ; Male ; Neck Muscles ; Rabbits ; Surgical Flaps ; Trachea ; blood supply ; transplantation ; Transfection ; Transplantation, Autologous ; Vascular Endothelial Growth Factor A ; genetics ; therapeutic use
5.Comparative Study on Different Sample Pre-enrichment Methods for Flow Cytometry Cell Sorting
Hong-Yan YANG ; Li-Fang YAN ; Ru-Gang WANG ; Xin ZHOU
Journal of Modern Laboratory Medicine 2018;33(1):128-130
Objective To investigate the effect of different pre-enrichment methods on flow cytometry cell sorting of monocyte.Methods Peripheral blood of 15 volunteers were pre-enriched by three methods:erythrocyte lysis,hydroxyethyl starch sedimentation,density gradient centrifugation using Ficoll.Cell viability was detected by Trypan blue staining.The amount and the proportion of monocytes were analyzed by flow cytometry.Results The cell viability of the samples prepared by the 3 methods was above 95 %.There was no significant difference between the absolute count and grouping of monocytes in erythrocytes lysis and without pre-enrichment samples.The cell subgroup was not obvious by hydroxyethyl starch sedimentation.The number of monocytes obtained by Ficoll density gradient centrifugation was significantly lower than that without pre-enrichment samples.Conclusion Erythrocyte lysis was a high efficient method for monocytes flow cytometry cell sor ting.
6.Diagnostic value of the ratio of ligamentum flavum thickness to oblique diameter of lumbar canal in patients with ligamentum flavum hypertrophy
Hao-Zhi YIN ; Wan-Jun ZHOU ; Xi-He SUN ; Yong-Gang LIU ; Xiao-Qin FU ; Ru-Fen WANG ; Fu-Min LIANG ; Ming-Biao LIU ;
Chinese Journal of Radiology 2001;0(08):-
Objective To evaluate the diagnostic advantage of the ratio of ligamentum flavum(LF) thickness to oblique canal diameter(TODR)measured on CT images in patients with lumbar canal stenosis. Methods Seventy-one patients underwent CT and MRI examinations respectively,and they were divided into two groups,the positive group and negative group,according to the presence or absence of dural sac notch caused by the LF on bilateral parasagittal MR images.Meanwhile,50 volunteers without any symptom in the lumbar region or legs were examined by CT.TODRs were measured at the L3—S1 levels of the inferior margin of the intervertebral disc on transverse CT images,respectively.The results were further analyzed with the positive findings on MR images,clinical symptoms and physical examination,so as to find out the statistical correlation between them.Results LF thickness was(3.01?0.72)mm and TODR was 0.19?0.04 in the negative group,(3.94?0.84)mm and 0.28+0.06 in the positive group,and(3.16? 0.85)mm and 0.19?0.04 in the control group.There was significant difference between positive group and negative group or control group for LF thickness(P0.24, the sensitivity,specificity and positive predictive value were 74.8%,89.6% and 73.6% respectively. Positive correlation existed between LF thickness or TODR and clinical symptom(r=0.72,0.86,P
7.Diagnostic value of serum angiopoietin-2 level in pancreatic cancer.
Ru-Gen WAN ; De-Bao CHEN ; Yong-Gang LOU ; Mao-Feng WANG ; Qiao-Hong ZHANG ; Dan-Xia JIN ; Sun-Hong ZHOU
Chinese Journal of Oncology 2011;33(1):47-49
OBJECTIVETo evaluate the diagnostic value of serum angiopoietin-2 (Ang-2) level in pancreatic cancer patients.
METHODSSerum Ang-2 level was measured by enzyme-linked immunosorbent assays (ELISA) in samples from 116 patients with pancreatic cancer, 50 patients with chronic pancreatitis, and 50 normal control subjects.
RESULTSThe serum Ang-2 level in patients with pancreatic cancer [(1539.0 ± 449.3) ng/L] was significantly higher than those in patients with pancreatitis [(1044.6 ± 246.1) ng/L, P < 0.01] and normal control subjects [(1075.6 ± 228.2) ng/L, P < 0.01]. There was no significant difference of serum Ang-2 levels between patients with pancreatitis and normal controls. Pancreatic caner patients with lymph node metastasis had a significantly higher serum Ang-2 level [(1890.1 ± 354.9) ng/L] than those without metastasis [(1212.1 ± 224.2) ng/L, P < 0.01]. The area under ROC curve of serum Ang-2 level for diagnosis of pancreatic cancer was 0.819.
CONCLUSIONThe serum Ang-2 level can be a useful indicator for diagnosis of pancreatic cancer.
Adult ; Aged ; Angiopoietin-2 ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Pancreatic Neoplasms ; blood ; diagnosis ; pathology ; Pancreatitis ; blood ; ROC Curve ; Retrospective Studies
8.Effect of superparamagnetic iron oxide labeling on neural stem cell survival and proliferation.
Guang-Hui DAI ; Jun-Gang XIU ; Zhen-Jun ZHOU ; Zhong-Can CHEN ; Ru-Xiang XU ; Xiao-Dan JIANG ; Mou-Xuan DU
Journal of Southern Medical University 2007;27(1):49-55
OBJECTIVETo study the effect of superparamgnetic iron oxides (ferumoxides) on the survival and proliferation of neural stem cells (NSCs) and determine the optimal ferumoxides concentration for labeling.
METHODSBone marrow stromal cells (BMSCs) were obtained from rat femoral marrow and cultured in vitro to induce their differentiation into NSCs. Ferumoxides labeling of the NSCs was performed with different final concentrations of ferumoxides, and the labeling efficiency and viability of the labeled NSCs were evaluated by Prussian blue staining, MTT assay, flow cytometry and transmission electron microscope.
RESULTSThe NSCs could be effectively labeled with ferumoxides with a labeling efficiency of around 90%. Prussian blue staining showed numerous fine granules with blue staining in the cytoplasm of the labeled NSCs, and the intensity of the blue staining was in positive correlation with the ferumoxide concentration for labeling. Transmission electron microscopy of the labeled NSCs revealed the presence of numerous vesicles spreading in the cytoplasm and filled with electron-dense magnetic iron particles. The ferumoxides vesicles increased with the labeling concentration of ferumoxides, and at the final concentration exceeding 25 microg/ml, ferumoxides vesicles in the NSCs gave rise to conglomeration which hampered observation of the cellular ultrastructure by transmission electron microscope. The results of flow cytometry and MTT assay demonstrated that the cell viability, proliferation, differentiation and apoptosis of the labeled cells were affected by ferumoxides at the concentration above 25 microg/ml, but such effects could be minimal at lower concentrations.
CONCLUSIONFerumoxides might be feasible for in vitro labeling of the NSCs with the optimal concentration of 25 microg/ml.
Animals ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Dextrans ; Ferrosoferric Oxide ; Iron ; pharmacology ; Magnetite Nanoparticles ; Male ; Microscopy, Electron, Transmission ; Neurons ; cytology ; ultrastructure ; Oxides ; pharmacology ; Rats ; Stem Cells ; cytology ; ultrastructure
9.Characterization of a novel transplantable orthotopic nude mouse model with xenografted human bladder transitional cell tumor (BIU-87).
Chong LI ; Ru-Ping YAN ; Guo-Hong YUAN ; You-Gang FENG ; Jian-Song WANG ; Xiang GAO ; Yan-Shan HUANG ; Jin-Bao ZHOU ; Shu-Sheng XIE
Chinese Journal of Oncology 2006;28(10):733-736
OBJECTIVEA mouse model of orthotopic bladder cancer simulating its human counterpart is of great importance in preclinical evaluation of new treatment modalities such as immunotxin therapy. The aim of the present study is to establish a novel nude mouse model with xenografted human bladder cancer.
METHODSSingle cell suspension of an established human bladder transitional cell carcinoma (TCC) cell line BIU-87 was instilled into nude mouse bladders which were pretreated with mild acid washing. The tumor growth in mouse bladder was assessed weekly by magnetic resonance imaging (MRI). At intervals following implantation and MRI tumor detection, the animals were sacrificed for necropsy, histological examination and immunocytochemical studies.
RESULTSThe overall tumor establishment was 92.9% (52/56 mice) at 7 - 36 days, while in the subgroup of animals sacrificed at 12 - 13 days, 40 out of 42 animals (95.2%) developed TCC, the majority of which was superficial. The tumor stages were assessed by gross and histopathology. Histological examination confirmed the presence of grade II - III TCC. Immunocytochemistry confirmed that the tumor model maintained the biological and immunological features of BIU-87 cells. The changes seen on MRI images well correlated with the extent of tumor invasion identified by histology. Carcinoma in situ could be detected histologically at 7 - 9 days post-inoculation and progressed into papillary or invasive tumors thereafter.
CONCLUSIONThe orthotopic BIU-87 TCC model in nude mice is highly reproducible and is ideal for preclinical studies on experimental intravesical therapies.
Animals ; Antibodies, Monoclonal ; analysis ; Carcinoma, Transitional Cell ; immunology ; pathology ; Cell Line, Tumor ; Female ; Humans ; Immunohistochemistry ; Magnetic Resonance Imaging ; Mice ; Mice, Nude ; Neoplasm Staging ; Neoplasm Transplantation ; Neoplasms, Experimental ; pathology ; Transplantation, Heterologous ; Urinary Bladder Neoplasms ; immunology ; pathology
10.Observation of the changes in ventral prostatic microcirculation in castrated rats.
Ru-Zhu LAN ; Zhang-Qun YE ; Rong-Jing DENG ; Shao-Gang WANG ; Chun-Lian CHEN ; Sheng ZHOU
National Journal of Andrology 2009;15(8):708-712
OBJECTIVECastrated rats exhibit significant shrinkage of the ventral prostate and apoptosis of prostatic cells, which can be attributed to the reduced blood supply to the prostate. But what causes the blood decrease in the prostate remains unknown. This study aims to explore the molecular mechanism of the changes in the microcirculation of the ventral prostate of rats following castration.
METHODSWe randomized 24 male adult rats into 6 groups of equal number, and collected their ventral prostates at 0, 1/2, 1, 2, 3 and 7 d, respectively, after castration. Then we observed the changes of the microvessels under the transmission electron microscope, detected the apoptosis of endothelial cells by TUNEL, and determined the expressions of VEGF, endostatin, angiostatin and angiopoietin-2 by Western blot.
RESULTSThe castrated rats showed dramatic changes in the microvessels of the ventral prostate, obvious apoptosis of the endothelial cells, down-regulated expression of VEGF, and up-regulated expressions of endostatin and angiostatin, while angiopoietin-2 remained unchanged.
CONCLUSIONThe decreased level of VEGF and increased levels of endostatin and angiostatin might underlie the mechanism of the changes in the microcirculation of the ventral prostate of rats following castration.
Angiopoietin-2 ; metabolism ; Angiostatins ; metabolism ; Animals ; Endostatins ; metabolism ; In Situ Nick-End Labeling ; Male ; Microcirculation ; Orchiectomy ; Prostate ; blood supply ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; metabolism