1.The separation of side population cells in multiple myeloma cell line PRIM8226 and their biological characteristics
Zhanping HUANG ; Ziyan WANG ; Jinxiang FU
Journal of Leukemia & Lymphoma 2014;23(8):460-464
Objective To detect and separate the side population cells(SP) from multiple myeloma (MM) cell lines PRIM8226,and to study their biological characteristics.Methods Fluorescence activated cell sorter (FACS) and Hoechst33342/PI dye were used to sort SP cells of PRIM8226.The multiplication capacity was tested by the growth curve and MTT test,SP cells proportion and the cell cycle were analyzed by flow cytometry,the colony-formtion ability was compared in terms of colony forming experiment,the expression of c-myc,KIF4,SOX2,OCT4 was tested by RT-PCR.The oncogenicity of the cells was analyzed by nude mouse transplantation tumor experiment in vivo.Results The ratio of SP cells in PRIM8226 was (1.78±0.89) %.More SP cells in the G0 / G1 period,(44.34±3.09) % vs (28.49±1.97) %,P < 0.05,and fewer cells in S phase than MP cells,(38.83±3.69) % vs (51.49±4.62) %,P < 0.05.There were no difference in the expression of CD38 and CD138 between SP cells and MP cells,respectively,(78.5±8.5) % vs (82.0±4.0) % and (72.3±15.7) % vs (84.3±11.9) %,P > 0.05.Colony formation assay showed that the colony forming efficiency of the SP cells was higher than the MP cells,single cell clone diameter,the number of clone forming,the clone formtion rate were significantly higher than MP cells,0.280±0.016 vs 0.118±0.019,1 722±127 vs 358±14,(86.1±3.46) % vs (17.9±1.88) %,P < 0.05.The mRNA expression levels of c-myc,KIF4,SOX2,OCT4 in SP cells were higher than those in MP cells,c-myc (29.90±3.73) % vs (16.84±2.35) %,KIF4 (29.97±2.89) % vs (19.06±1.23) %,SOX2 (40.00±4.58) % vs (16.62±2.09) %,OCT4 (32.96±0.92) % vs (23.27±0.92) %,all P < 0.05.Nude mouse transplantation tumor experiment in vivo showed the oncogenicity of the SP cells was more stronger than MP cells (5×103 vs 5×l05).Conclusion There are notably difference in the cell cycle,colony formation assay,the mRNA expression levels and oncogenicity,but no difference in the expression of CD38 and CD138,the proliferation ability between SP cells and MP cells.
2.Study and Exploration of Examination Forms of University Credit System
Cuijuan LIU ; Aimin SHEN ; Jinxiang HUANG
Chinese Journal of Medical Education Research 2003;0(04):-
Credit system is suitable for all-round development of students and teaching personality and it is an important component of the modern university system.The paper mainly studies the credit system as a learning management system and explores the form of the examination and assessment patterns.It also puts forward some specific measures and ideas about how to improve and perfect examination and assessment modes.
3.Matrix metalloproteinases-11 collabrotive matrix metalloproteinases-14 promote development of breast carcinoma
Hongyan HUANG ; Jinxiang TAN ; Guosheng REN
Chinese Journal of Immunology 2016;32(7):996-999
Objective:To investigate the expression pattern of MMP-11 and MMP-14 in breast carcinoma, and the effect of MMP-11 on breast carcinoma cell migration and invasion. Methods:MMP-11 and MMP-14 expression were examined in 161 invasive breast carcinoma tissue samples and 10 normal breast tissue samples. siRNA was used to knockout MMP-11 in breast carcinoma cell line MB-231 and Transwells were used to evaluate changes in migration ability and invasion ability. Results:Both MMP-11 and MMP-14 were highly expressed in breast carcinoma samples,122 and 149 samples out of 161,respectively. The expression of both proteins were correlated with lymph node metastasis and TNM staging. After knockout of MMP-11,the expression of both proteins decreased in MB-231 cell line and experiments show that the cell′s migration and invasion abilities were significantly weakened. Conclusion:MMP-11 and MMP-14 could promote invasion and metastasis of breast carcinoma. Knockout of MMP-11 results in the downregulated expression of MMP-14,and the inhibition of breast carcinoma cell′s migration and invasion. They could be potential prognostic markers and treatment targets for of breast carcinoma.
4.Association between polymorphisms of CYP3A5,MDR1,COX-2 and chemotherapy outcomes of advanced NSCLC
Jihong PAN ; Jinxiang HAN ; Jianmei WU ; Lijun SHENG ; Hainan HUANG
Journal of International Oncology 2008;35(5):395-399
Objective To investigate whether genotypes of CYP3A5,MDR1 and cyclooxygenase-2 are associated with the sensitivity of vinorelbine-platinum to NSCLC.Methods The genotypes of CYP3A5(*3),MDR1 (2677G>T at exon 21 and 3435C>T at exon 26 and their haplotypes),cyclooxygenase-2 (-1 195G>A) were determined by RFLP-PCR and chemotherapy responses were analyzed in 69 non-small-celllung cancer (NSCLC) Chinese Han patients.They received a combination chemotherapy of vinorelbine-cispla-tin.Chi-square test was used to investigate the potential association of genotype with chemotherapy response.OR and 95% C1 were calculated.Results The 3435 CC genotype was associated with a significantly betterchemotherapy response compared with the combined 3435 CT and 1Tr genotypes(P=0.033).The 2677 GG genotype was also associated with a significantly better chemotherapy response compared with the combined 2677 GT and IT genotype(P=0.012).Moreover.patients with the 2677 G-3435 C haplotype seemed to have a better response to chemotherapy compared with those with the other haplotypes(P=0.063).CYP3A5*3 was not likely to correlate with sensitivity of vinorelbine-platinum to NSCLC.Cyclooxygenase-2-1195G>A was likely to have better response to vinorelbine but not statistically significant(P=0.067).Conclusion Polymor-Dhisms of MDR1 3435 C>T and MDR1 2677 G>A/T can be used for predicting treatment response to vinorel-bine-cisplatin chemotherapy in NSCLC patients.
5.Changes of microRNA-206 in peripheral blood mononuclear cells from children with respiratory syncytial virus bronchiolitis
Hong ZHOU ; Qinglei XU ; Xiaobo MA ; Guoyu WEI ; Jinxiang HUANG
Chinese Journal of Applied Clinical Pediatrics 2015;30(22):1709-1712
Objective To explore the role of microRNA-206 (miR-206) in peripheral blood mononuclear cells (PBMCs) in infantile bronchiolitis caused by respiratory syncytial virus (RSV).Methods Thirty-five cases of infantile bronchiolitis and 25 cases of healthy controls were enrolled into the current study.PBMCs were isolated from the peripheral blood of both healthy subjects and those with infantile bronchiolitis in the acute and the convalescent stages.Total RNAs were extracted from PBMCs which were stimulated by phorbol-12-myristate-13-acetate (PMA) and Ionomycin, and then the RNA was transcribed reversely into cDNA.The expressions of miR-206 and Kruppel-like transcription factor 4 (KLF4) were detected by real-time quantitative polymerase chain reaction (qRT-PCR) method.Plasma interleukin-17 (IL-17) was determined by enzyme linked immunosorbent assay (ELISA).Results There was a significant difference in miR-206 levels of children with RSV bronchiolitis in the acute stage(0.055 ±0.018) and the convalescent stage(0.187 ±0.069) as well as the healthy controls(0.204 ± 0.075).Through pairwise comparison, the miR-206 levels in the children in the acute stage were significantly lower than those in the convalescent stage and healthy control group (P < 0.01), but no statistical significance was found between the convalescent stage group and healthy control group(P > 0.05).The levels of KLF4 mRNA of children in the acute stage,convalescent stage as well as the healthy subjects were 0.588 ± 0.161,0.086±0.024,0.075 ±0.019, respectively,which was significantly difference (P < 0.01).The levels of IL-17 were (58.26 ±25.88) ng/L, (9.87 ± 3.01) ng/L, (7.65 ± 2.16) ng/L, respectively (P < 0.01).Compared to the convalescent and the normal control group,both the KLF4 mRNA and IL-17 levels were markedly higher in the acute stage (P < 0.01), but there were no significant differences between children with RSV bronchiolitis in convalescent stage and in the healthy controls (P > 0.05).Furthermore, the result of this study showed a negative correlation between the expression of miR-206 and KLF4(r =-0.624 ,P <0.01)and IL-17 (r =-0.609 ,P <0.01) in children in the acute stage and a positive correlation between KLF4 mRNA and IL-17 in children in the acute stage (r =0.662, P < 0.01).Conclusion The levels of miR-206 may play a role in the onset of RSV associated post-bronchiolitis (PB) and the low expression of miR-206 in children infected with RSV may increase the susceptibility to PB.
6.Simultaneous Determination of Six Saponins Constituents in Xinling Pills by HPLC-ELSD
Jinxiang PENG ; Feng WU ; Huabin HUANG ; Yuancui XU ; Jin YANG
China Pharmacist 2017;20(6):1133-1135
Objective: To develop an HPLC-ELSD method for the simultaneous determination of six saponins constituents including notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc and ginsenoside Rd in Xinling pills.Methods: HPLC-ELSD was used, and the chromatographic separation was performed on an Agilent Eclipse XBD-C18 column (150 mm×4.6 mm,5 μm) with acetonitrile-water as the mobile phase with gradient elution at the flow rate of 1.0 ml·min-1, the column temperature was maintained at 20℃, the drift tube temperature was 60℃, and the gas pressure was 4.00 bar.Results: Notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Rb1, ginsenoside Rc and ginsenoside Rd was linear within the range of 0.30-6.00μg(r=0.999 5), 1.14-22.80μg (r=0.999 6), 0.17-3.40 μg (r=0.999 7), 0.81-16.20 μg (r=0.999 7), 0.08-1.60 μg (r=0.999 8) and 0.07-1.40 μg (r=0.999 8), respectively.The average recovery was 98.23%, 97.98%, 99.14%, 99.15%, 98.72% and 98.37%, and the RSDs were 1.56%, 1.31%, 1.16%, 1.07%, 0.73% and 0.92%(n=6), respectively.Conclusion: The method is convenient, accurate and reproducible in the quality control of saponins components in Xinling pills
7.Epidemiological Characters of Yunnan Sudden Death Syndrome and Its Research Progress
Jinxiang ZHENG ; Su ZHAO ; Wenli HUANG ; Lei HUANG ; Shuangbai TANG ; Chi BEN ; Jianding CHENG
Journal of Forensic Medicine 2014;(2):122-125
Yunnan sudden death syndrome (YSDS) is an abruptly fatal disease of unknown etiology, found mostly in central or northwestern mountain area (with altitude between 1 815 and 2 225 meters ) of Yunnan province from June to September every year. It occurs mostly in young female adults, with high incidences in Lisu, Yi and Miao ethnics and high familial aggregation. The clinical manifestation of YSDS is changeful and the pathological characteristic is lack of specificity. The pathogenesis may be at-tributed to several factors including poor hygiene and lower socioeconomic conditions, lack of Selenium or Chromium, infection of Coxsackie B virus, mushroom consumption and special geological conditions. This article reviews the epidemiologic features, clinical manifestations, pathological features, etiology and hypothesis in order to provide clues for the research of YSDS.
8.Impairment in immunoregulatory capability of mesenchymal stem cells from multiple myeloma patients and role of it in the pathogenesis of myeloma bone disease
Bingzong LI ; Wenzhuo ZHUANG ; Ping CHEN ; Hong ZHANG ; Haiwen HUANG ; Jinxiang FU
Chinese Journal of Microbiology and Immunology 2010;30(9):853-859
Objective To deplore the immunoregulatory function changes of mesenchymal stem cells(MSCs)from multiple myeloma(MM)patients and its effects on the pathogenesis of myeloma bone disease.Methods MSCs from MM patients and normal controls were isolated and the immunophenotype was detected.Real-time PCR was performed to detect the expressions of TGF-β1,TGF-β2,TGF-β3,IL-6,IL3,TNF-α,FasL and RANKL of MSCs.Transwell coculture systems were performed between MSCs and T cells.Lymphocyte proliferative assay was employed to detect the effect of MSCs on T cell proliferation.The effect of MSCs on T cell cycle and T cell activation markers CD25 and CD69 expression were analyzed by flow cytometry.Cleaved caspase 3 protein by western blot and hoechst 33258 staining were employed to detect the apotosis of T cells.Influence of T cells on the osteogenesis potential of MSCs were detected by Von kossa stain,real-time PCR and Western blot.Results MSCs from both MM patients and normal controls possessed similar morphology and immunophenotypes.MM derived MSCs exhibited increased expressions of TGF-β1,IL-6,IL-3,TNF-α and RANKL and decreased expression of TGF-β2,TGF-β3 and FasL.The inhibitory effect of MM derived MSCs on T cell proliferative ability was attenuated compared to control MSCs.MSCs from normal controls silence more T cells in Go/G1 phase than those from MM patients.The daupening effect of MM derived MSCs on activation-induced T apoptosis seemed to be enhanced.Expression of T cell activation markers were significantly inhibited by MSCs from normal controls.Both T cells cocultured with MM deprived MSCs and T cells directly from MM patients inhibited osteogenesis potential of MSCs from normal controls.Conclusion MSCs from MM patients showed impaired immunoregulatory capability on T cells.The activated T cells,in turn,inhibited the osteogenesis potential of MSCs.This may participate in the pathogenesis of myeloma bone disease.
9.In vitro short term cartilage tissue engineering with oriented cartilage extracellular matrix scaffolds and bone marrow mesenchymal stem cells
Jun YAO ; Shibi LU ; Jiang PEN ; Quanyi GUO ; Li ZHANG ; Jinxiang HUANG ; Aiyuan WANG ; Wenjing XU
Chinese Journal of Orthopaedics 2010;30(12):1217-1222
Objective To fabricate cartilage extracellular matrix (ECM) oriented scaffolds and investigate the attachment, proliferation, distribution and orientation of bone marrow mesenchymal stem cells (BMSCs) cultured within the scaffolds in vitro. Methods Cartilage slices were shattered in sterile phosphate-buffered saline (PBS) and the suspension were differentially centrifugated untill the micro- fiber of the cartilage extracellular matrix was disassociated from the residue cartilage fragments. At last the supernatant were centrifugated, the precipitation were collected and were made into 2%-3% suspension. Using unidirectional solidification as a freezing process and freeze-dried method, the cartilage extracellular matrix derived oriented scaffolds was fabricated. The scaffolds were then cross-linked by exposure to ultraviolet radiation and immersion in a carbodiimide solution. By light microscope and scan electron microscope (SEM) observation, histological staining, and biomechanical test, the traits of scaffolds were studied. After being labelled with PKH26 fluorescent dye, rabbit BMSCs were seeded onto the scaffolds. The attachment, proliferation and differentiation of the cells were analyzed using inverted fluorescent microscope. Results The histological staining showed that toluidine blue, safranin O, alcian blue and anti-collagen Ⅱ immunohistochemistry staining of the scaffolds were positive. A perpendicular pore-channel structures which has a diameter of 100 μm were verified by light microscope and SEM analysis. The cell-free scaffolds showed the compression moduli were (2.02±0.02) MPa in the mechanical testing. Inverted fluorescent microscope showed that most of the cells attached to the scaffold. Cells were found to be widely distributed within the scaffold, which acted as a columnar arrangement. The formation of a surface cells layer was found on the surface of the scaffolds which resembled natural cartilage. Coclusion The cartilage extracellular matrix derived oriented scaffolds have promising biological, structural, and mechanical properties.
10.Theoretical study of corresponding points for pain.
Lou BIDAN ; Ping GONG ; Wei ZHANG ; Jinxiang LI ; Jie HUANG ; Zhaoan YU
Chinese Acupuncture & Moxibustion 2015;35(1):77-79
The theoretical basis of "corresponding points" originates from opposing needling, contralateral needling and distal needling in acupuncture. It is to select points in corresponding area that is distant from diseased re gion to balance yin and yang and to activate meridians. Acupuncture at corresponding points, through reflex regu- lation of nervous system. could activate protective inhibition of cerebral cortex and cutoff of local malignant stimu lation, leading to quick elimination of pain. In clinic, it is mostly used for pain-related diseases.
Acupuncture Points
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Acupuncture Therapy
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Humans
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Medicine in Literature
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Pain Management