1.The difference of gene expression profile of JAK/STAT signaling pathway in 5-fluorouracil resistant cell line of human gastric adenocarcinoma
Lifen YU ; Yunlin WU ; Yongping ZHANG
Chinese Journal of Digestion 2001;0(01):-
Objective To investigate the difference of gene expression in two human gastric adenocarcinoma cell lines, 5-fluorouracil resistant cell line SGC7901/R and its parental cell line SGC7901, and to screen related genes of JAK/STAT signaling pathway by gene chip. Methods The mRNAs of two cell lines were extracted and purified. The two cDNA probes were made from these two mRNAs which were labelled by (biotin-)16-dUTP, hybridized with human JAK/STAT signaling pathway gene array and scanned for intensity, respectively. The acquired images were analyzed by software. Different expression genes were then screened out. The mRNA expressions of Stat3, NF-?B1 and bcl-x were analyzed by semi-quantitative RT-PCR, (respectively.) Results A parallel comparison between the gene profiles of JAK/STAT signalling pathway in two cell lines showed that a total of 70 genes were screened out whose expressive level was more than 2 folds in 5-flurouracil resistant cell line SGC7901/R and its parental cell line SGC7901. Among these genes, 40 were upregulated and the other 30 were down-regulated. The results of RT-PCR were consistent with those of microarray scanning. Conclusions The knowledge of gene expression profile of JAK/STAT signaling pathway, which was changed in 5-fluorouracil resistant cell line of human gastric adenocarcinoma, has proven to be useful for illustrating the multi-drug resistance mechanisms of gastric cancer.
2.Expression of Bcl-2 of peripheral T, B lymphocytes in patients with liver fibrosis
Ji LI ; Yu ZHENG ; Yongping CHEN
Chinese Journal of Infectious Diseases 1997;0(04):-
Objective To investigate Bcl-2 expression in the peripheral blood mononuclear cells and its clinical significance of liver fibrosis (LF). Methods Tested Bcl-2 protein levels in T, B cells of 47 patients (male 17, female 30) with LF, 35 patients (male 24, female 11) with chronic hepatitis B and no LF and 41 cases (male 29, female 12) normal controls by two color cytofluorography. Results Among them, LF patients, chronic hepatitis B without LF and normal controls, the proposition of T cells (including CD3~+, CD4~+ and CD8~+ subgroups) and CD19~+ B cells expressed Bcl-2 protein increased significantly in LF patients (P
4.Analysis of the TCR β-chain gene rearrangement and CDR3 repertoire in patients with breast cancer
Jianbo ZHANG ; Yongping SONG ; Qingkai YU ; Jun HU ; Tao DONG
Cancer Research and Clinic 2010;22(3):179-181
Objective To analyze the clonal gene rearrangement and complementarity determining region 3 (CDR3) repertoire of TCR β-chain in fine needle aspiration biopsy (FNAB) specimens of lymph node metastasis in patients with breast cancer. Methods The TCR CDR3 region genes of 24 TCR Vβ subfamilies were amplified by utilizing RT-PCR technology, and the CDR3 lengths of TCR β-chain were analyzed with gene scan technology for 2 cases with lymph node reactive hyperplasia and 3 patients with lymph node metastasis of breast cancer. The clonality of T cells presumed by spectra typing was further confirmed by CDR3 sequencing. Results TCR β-chain presented specific repertoire skewing in metastatic lymph node,and only 3-5 TCR Vβ subfamily of T cells were identified, respectively. Clonal expanded T cells, including oligoclonal, polyclonal patterns, in one or more Vβ subfamilies were found in all cases. The oligoclonal expanded T cells had different CDR3 amino acid sequences. Conclusion There are characteristic T cells cloning proliferation and selected usage of TCR Vβ subfamily T cells could be found in metastatic lymph node.The sequences of CDR3 in different TCR clone proliferation are mostly different.
5.Effects of mesalazine combined with bifico on the treatment of ulcerative colitis and on TLR4 and MyD88
Yongping ZHANG ; Yifeng ZHOU ; Pingfan WANG ; Huyun GAO ; Xican YU
Journal of Chinese Physician 2014;(4):498-500
Objective To investigate the clinical efficacy of mesalazine combined with bifico in the treatment of ulcerative co -litis (UC) and its effect on toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88).Methods A total of 86 cases of UC was randomly divided into the observation and control groups with 43 cases in each group .The observation group was given me-salazine combined bifico treatment , and the control group was given mesalazine , and eight weeks as a course of treatment .The clinical efficacy was observed .Serum TLR4 and MyD88 were tested.Adverse reaction in treatment was recorded .Results The total effective rate (95.35%) in observation group was significantly higher than that (81.40%) in control group( P <0.05).The serum TLR4 and MyD88 after treatment of two groups were significantly lower than that before treatment , respectively ( P <0.05 or P <0.01 ) , and the decline in TLR4 of observation group was superior to that of control group ( P <0.05 ) .The difference of adverse reaction rate in two groups was not significant ( P >0.05 ) .Conclusions Mesalazine combined with bifico in the treatment of UC is safe and effective . The effect may be related to regulation of peripheral blood TLR 4 and MyD88 content .
6.Methylprednisolone pulse therapy of hematological disorders in systemic lupus erythematosus
Jihong YANG ; Xuewen WANG ; Yaping YU ; Yongping ZHAI ; Haining LIU
Chinese Journal of Postgraduates of Medicine 2006;0(34):-
Objective To explore methylprednisolone and conventional dose prednisone treatment for hematological damage in systemic lupus erythematosus(SLE) in the near future response. Methods Hemocytopenia in 147 patients with SLE were treated by intravenous injecting methylprednisolone and conventional dose prednisone and therapy response were observed in the tenth day after treatment. Results The responses were obtained in methylprednisolone and in conventional dose prednisone increased percentage of Hb were 34.8% and 14.0%,of WBC were 76.7% and 63.0%,of Pt were 66.7% and 27.3% in two group respectively. In comparison of values of Hb,WBC,and Pt before treatment with those after treatment showed significant difference in two groups(P
7.CT Diagnosis and Differential Diagnosis of Wilms′ tumors of Children
Qiuyan WNAG ; Yu GAO ; Biao JIN ; Yongping ZHANG
Journal of Practical Radiology 2001;17(5):333-335
Objective To investigate the importance of CT diagnosis and differential diagnosis in Wilms′ tumors of children.Methods 45 cases of Wilms′ tumors confirmed by clinical and pathology were retrospectively analyzed,CT characteristics of Wilms′ tumor,renal carcinoma(2 cases),renal rhabdomyosarcoma(1 case)in children were discussed.Results Wilms' tumors appeared in solid masses in 34 cases (76%),cysticsolid masses in 9 cases(20%),cystic masses in 2 cases (4%),7 cases (15.6%)had thrombosis in renal vein or IVC.Renal carcinoma shared the same appearance with Wilms′ tumor in image.Rhabdoid tumor of kidney appeard as cystic-solid mass with lineal calcification and subcapsular fluid accumulation.Conclusion Wilms′ tumor is the most popular malignant renal tumors in children.It is originate from renal parenchyma,and likely to invade renal pelvis.Hemorrhhage,necrosis or cystic formation in tumor is common.They are difficult to be distinguished from renal cell carcinomas in radiology.Rhabdoid tumor of kidney is rare and has characteristic features on CT.
8.Inhibition effect of adiponectin on the levels of PC Ⅲ and HA in HSC-T6 cell
Fujun YU ; Peihong DONG ; Xufei FAN ; Yongping CHEN ; Ji LI
Journal of Chinese Physician 2011;13(2):150-153
Objective To observe the effects of adiponectin on mRNA and protein expressions of connective tissue growth factor (CTGF) in hepatic stellate cells (HSCs) and the levels of procollagen type Ⅲ (PC Ⅲ) and hyaluronic acid (HA).Methods Cultured rat HSCs were treated with different concentrations of adiponectin.CTGF mRNA and protein expressions were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western Blot.The levels of PCⅢ and HA were detected by enzymelinked immunosorbent assay (ELISA).Results Compared with the control group,the result of RT-PCR showed that the four groups had different degrees of inhibitory effect,of which group D exhibited the strongest inhibitory effect.The absorbance ratio was 1.54 ±0.18,1.21 ±0.14,0.96 ±0.10,and 0.79 ± 0.08,respectively (t =2.42,P <0.05;t =2.73,P <0.05;t =3.28,P <0.01;t =4.67,P <0.01).Western Blot also indicated that four groups had different degrees of inhibitory effect,of which D group exhibited the strongest inhibitory effect.The ratio of integral absorption was 1.54 ± 0.18,1.21 ±0.14,0.96±0.10,and 0.79 ±0.08,respectively (t =2.84,P <0.01;t =4.05,P <0.01;t =6.25,P <0.01;t =9.72,P <0.01).The levels of PCⅢ and HA secreted in culture media were also decreased.It was significantly decreased with the concentration of adiponectin increased.Conclusion Adiponectin can inhibit the levels of PC Ⅲ and HA,which may be achieved through reducing CTGF mRNA and protein expressions.
9.Effect of small interfering RNA targeting transforming growth factor β receptor Ⅰ gene on the collagen synthesis of hepatic stellate cells in vitro
Fujun YU ; Didong LOU ; Zhuo LIN ; Peihong DONG ; Yongping CHEN
Chinese Journal of Infectious Diseases 2010;28(1):5-9
Objective To observe the effect of small interfering RNA(siRNA)expression plasmids targeting transforming growth factor p receptor(TαR)Ⅰ gene on the collagen synthesis of hepatic stellate cells(HSCs).Methods Three siRNA expression plasmids were designed and constructed according to TBR Ⅰ sequence.Then the plasmids were transfected into HSC-T6 using 1ipofectamine2000 reagent. The mRNA and protein expressions of TβR Ⅰ were analyzed by reverse transcription polymerase chain reaction(RT-PCR)and Western blot technique, respectively. The cell proliferation was detected using methylthiazo-lyldiphenyl-tetrazolium bromide(MTT)methods. Concentrations of haluronic acid and type Ⅲ pro-collagen in the supernatants were determined by radioimmunoassay. The data were analyzed using least significant difference(LSD).Results Three recombinant plasmids expressing siRNAs were successfully constructed and confirmed by restriction enzyme assay. Compared with the blank control,all the three recombinant plasmids could inhibit the expressions of TβR Ⅰ mRNA,of which plasmid expressing siRNA2 exhibited the strongest inhibitory effect(psiRNA1 group:t=7.354,P<0.01;psiRNA2 group:t=9.214,P<0.01;psiRNA3 group:t=5.967,P<0.01).The expressions of TβR Ⅰ protein were also reduced by all the three recombinant plasmids,of which the plasmid expressing siRNA2 showed the strongest inhibitory effect(psiRNA1 group: t=6.324,P<0.01;psiRNA2 group:t=8.741,P<0.01;psiRNA3 group:t=4.128,P<0.01).The proliferation activity and collagen synthesis of HSCs also decreased in all three HSC groups treated with recombinant plasmids, of which, again, plasmid expressing siRNA2 exhibited the strongest inhibitory effect. However, no significant change was observed in HSCs transfected with non-related siRNA. Conclusion Recombinant plasmids targeting TβR I can inhibit collagen synthesis, which suggests a novel target for gene therapy of liver fibrosis.
10.Liver regeneration after transplantation of microencapsulated hepatecytes in rats with acute liver failure
Yongping CHEN ; Lanman XU ; Yu HUANG ; Xiaodong WANG ; Feng GAO
Chinese Journal of Clinical Infectious Diseases 2008;1(2):91-95
Objective To investigate liver regeneration after transplantation of microencapsulated hepatocytes in rats with acute liver failure (ALF). Methods ALF rat model was established by intraperitoneal injection of D-galactosamine (D-GalN). After 18 h, rats were randomized into control group ( Ⅰ ), free hepatoeyte transplantation group ( Ⅱ ) and the microencapsulated hepatecyte transplantation group (Ⅲ). Six rats for each group were randomly selected and sacrificed at 6, 12, 24, 36, 48, 72, 120, 168 and 240 h after ALF induced and blood samples from inferior vena cava were collected. Liver functions were tested in blood samples, and the expression of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry. Results Ten-day survival rates of 3 groups were 26.7% (4/15), 40.0% (6/15) and 73. 3% (11/15), respectively (x2 = 9. 349,P = 0. 009). Survival rate of group Ⅲ was significantly higher than that of group Ⅰ and Ⅱ. Levels of ALT and AST in each group increased significantly at 6 h after ALF induced, and peaked between 48 ~ 72 h. Levels of ALT and AST in group Ⅱ and Ⅲ declined from 36 h, which was more significant in group Ⅲ. Tbil levels in group Ⅰ gradually increased after ALF induced and peaked at 72 h. Tbil in group Ⅱ and Ⅲ declined from 48 h, which was more markedly in group Ⅲ. In normal rats, the expression of PCNA protein was almost negative, but it was strongly expressed in ALF rats and peaked at 48 h. The number of positive cells in group Ⅲ was higher than that in group Ⅰ and Ⅱ, and the differences were of statistical signifieance. Conclusion The transplantation of microencapsulated hepatocytes can promote the regeneration of liver, and it can improve the liver function and prognosis in rats with ALF.