1.Three-dimensional conformal radiotherapy for loco-regionally recurrent esophageal cancer after initial radiotherapy
Wenbin SHEN ; Shuchai ZHU ; Jun WAN ; Juan LI ; Jingwei SU ; Yuxiang WANG ; Ren LI
Chinese Journal of Radiation Oncology 2010;19(2):111-114
Objective To evaluate the feasibility, therapeutic effects and normal tissue complications of three-dimensional conformal radiotherapy (3DCRT) for loco-regionally recurrent esophageal cancer after initial radiotherapy. Methods Between March 2001 and May 2007, 42 patients with loco-reigonal recurrent esophageal cancer after initial radiotherapy were treated with 3DCRT, including 27 male and 15 female with a median age of 67.5 years. Radiotherapy was delivered at 1.8 -2.0 Gy per fraction, 5 fractions per week, with a median total dose of 54 Gy. Treatment outcomes and normal tissue complications were assessed with WHO and RTOG/EORTC criteria. Results By December 31,2008, the follow-up rate was 100%. Twenty patients had follow-up time of 1 year and the remaining 22 had 2 years. The clinical symptom relief rate was 60%, and the response rate was 90.5% with a complete remission rate of 17% and partial remission rate of 74%. The overall 1- and 2-year survival (OS) rates were 60% and 24%. Grade 2 and grade 3 acute radiation esophagitis developed in 31% and 5% of the patients, respectively. Grade 2 and grade 3 acute radiation pneumonitis developed in 19% and 2% , respectively. Grade 2 and grade 3 acute radiation hematology toxicities developed in 5% and 2%, respectively. Conclusions For patients with loco-regional recurrences of esophageal cancer after initial radiotherapy, 3DCRT is feasible, with a good clinical symptom relief rate and immediate tumor response. However,the complication rate was high and the clinical indications should be strictly controlled.
2.Effect of Chinese herbal therapy on T-lymphocytes of IgA nephropathy patients: a clinical observation.
Xiao-Juan CHEN ; Da-Jun YU ; Ren-Huan YU ; Qing-Min SU ; Yong-Gang XU ; Yan HE ; Qiao-Qiao LIU
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(7):786-789
OBJECTIVETo observe the effect of Chinese herbal therapy on T-lymphocyte subsets in patients with IgA nephropathy (IgAN).
METHODSTotally 36 inpatients and outpatients at Department of Nephropathy, Xiyuan Hospital, China Academy of Chinese Medical Sciences, from June 2011 to June 2013 were recruited in the treatment group, while 20 volunteers were recruited as the healthy control group. Patients in the IgAN group only took Chinese herbal decoctions by syndrome typing for 3 months (except those accompanied with hypertension additionally took antihypertensive agents such as angiotensin-converting enzyme inhibitor and/or dihydropyridines calcium antagonist). No intervention was performed in the healthy control group. The values of Th1, Th2, and CD4+ CD25+ Treg, and red blood cell number in urine were detected using flow cytometry before and after treatment. 24 h urine protein was detected using inmmunoturbidimetry.
RESULTSCompared with the healthy control group, the CD4+ CD25+ Treg level obviously decreased in the IgAN group, showing statistical difference (P < 0.01). In the IgAN group, Th1, 24 h urine protein, and urine red blood cell counts were obviously lower after treatment, showing statistical difference when compared with before treatment (all P < 0.05).
CONCLUSIONChinese herbal therapy could reduce urine erythrocyte number and 24 h urine protein of IgAN patients, and down-regulating Th1 expression might be its mechanism.
Adult ; Case-Control Studies ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerulonephritis, IGA ; drug therapy ; Humans ; Male ; Middle Aged ; Phytotherapy ; T-Lymphocyte Subsets ; drug effects ; Young Adult
3.Effect on retardation of G2/M phase in esophageal carcinoma cells transfected with CHK1 and CHK2 shRNA after irradiation.
Yu-xiang WANG ; Shu-chai ZHU ; Wei FENG ; Juan LI ; Jing-wei SU ; Ren LI
Chinese Journal of Oncology 2006;28(8):572-577
OBJECTIVETo observe the effect of RNA interference on CHK1 and CHK2 expression and change of G2/M phase arrest in esophageal carcinoma cells after irradiation.
METHODSFour sequences short hairhip RNA (shRNA) of each CHK1 and CHK2 genes were constructed and connected with vector of pENTR/U6 plasmid, respectively, and then transfected into Eca109 cells with lipofectamine 2000 reagent. Protein and mRNA expression of CHK1 and CHK2 genes were detected with Western blotting and RT-PCR, respectively. Cell cycling was measured by flow cytometry after 5 Gy irradiation. Cell survival rate after 5 Gy irradiation was evaluated by clonegenetic assay.
RESULTSFour shRNA vector each of CHK1 and CHK2 genes were successfully constructed and transfected into Ecal09 cells, respectively. Protein expression of CHK1 and CHK2 were obviously decreased. Their mRNA expressions were also decreased after transfected with shRNA of CHK1 and CHK2. Arrest of G2/M stage in Eca109 cells were obviously decreased only in cells transfected with CHK1 shRNA but not with CHK2 shRNA at 12 h after 5 Gy irradiation. In first progeny Eca109 cells transfected with CHK1 and CHK2 shRNA, expression of CHK1 and CHK2 protein was also decreased. The level of phosphorylated CHK2-T68 expression was decreased at 1 h after 5 Gy irradiation, and at 72 h only transfected with CHK2 shRNA but not with CHK1 shRNA. Phosphorylation level of CHK1-S345 was not increased after transfected with CHK1 or CHK2 shRNA, but arrest of G2/M stage still remained at 12 h after 5 Gy irradiation and at 72 h accordingly. The cell survival rate was decreased in Eca109 cells transfected with CHK1 or CHK2 shRNA after 5 Gy irradiation.
CONCLUSIONAfter transfected with shRNA of CHK1 or CHK2, their expressions of mRNA and protein in Ecal09 cells are markedly inhibited and this inhibition effect can be observed in their first progeny cells and at least hold for 3 days. Arrest of G2/M phase can be reduced after irradiation when teansfected with shRNA of CHK1 and the radiosensitivity of Ec109 cells can be increased.
Blotting, Western ; Cell Division ; genetics ; physiology ; radiation effects ; Cell Line, Tumor ; Cell Survival ; genetics ; physiology ; radiation effects ; Checkpoint Kinase 1 ; Checkpoint Kinase 2 ; Esophageal Neoplasms ; genetics ; pathology ; physiopathology ; G2 Phase ; genetics ; physiology ; radiation effects ; Gamma Rays ; Genetic Vectors ; Humans ; Protein Kinases ; genetics ; metabolism ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA Interference ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection
4.The clinical characteristics and molecular pathogenesis of a variant Glanzmann's thrombasthenia-like pedigree.
Su Juan LYU ; Wei Ru REN ; Huan Ling ZHU ; Ting LIU
Chinese Journal of Hematology 2018;39(10):807-811
Objective: To review the clinical characteristics of a pedigree with inherited hemorrhagic disease to explore its molecular pathogenesis. Methods: The clinical data of the pedigree with inherited hemorrhagic disease were collected. After extracting DNA, next generation sequencing was utilized to detect the potential gene mutation. The changes of RASGRP2 transcript of this proband and his parents were detected using RT-PCR to compare with normal control. Results: The phenotype of the proband in this pedigree with inherited platelet dysfunction and bleeding disorder was similar to variant Glanzmann's thrombasthenia, the maximum aggregations of platelet in response to the physiological agonists including ADP, epinephrine and arachidonic acid were significantly lower, leading to severe spontaneous mucosal bleeding. Integrin αIIbβ3 gene mutation was not detected, but another gene mutation RASGRP2 IVS3-1 stood out. The mutation was homozygous in the proband and heterozygosis in both of his parents. Two transcript types were detected in the proband, without transcripts coding functional RASGRP2 protein, however, his parents had functional transcripts and abnormal transcripts, with the normal transcripts in the majority. Conclusions: The RASGRP2 IVS3-1 gene mutation was responsible for the inherited hemorrhagic disease. The RASGRP2 IVS3-1 gene mutation led to abnormal alternative splicing, without formation of functional RASGRP2 protein. The RASGRP2 protein is at the nexus of calcium-dependent platelet activation and hemostasis after damage of blood vessels. Spontaneous mucosal bleeding was a result of the lack of the functional RASGRP2 protein. This was the first report of RASGRP2 gene mutation resulting in bleeding disorder in China, and also the first report of the mutation type of RASGRP2 IVS3-1.
Blood Platelets
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Guanine Nucleotide Exchange Factors
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Humans
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Pedigree
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Platelet Glycoprotein GPIIb-IIIa Complex
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Thrombasthenia
5.The role of Fas mutation on non-alcoholic steatohepatitis in mice.
Shan-shan SU ; Fang HAN ; Rong-qi WANG ; Wei-guang REN ; Wen-juan WU ; Ling-bo KONG ; Su-xian ZHAO ; Yue-min NAN
Chinese Journal of Hepatology 2011;19(9):653-657
OBJECTIVEOur previous study indicated that the death receptor Fas played a key role on hepatocyte apoptosis in nutritional steatohepatitis in mice. This study aimed to explore whether Fas mutation accelerated hepatic steatosis and inflammatory infiltration in methionine-choline deficient (MCD) diet feeding mice.
METHODSMice homozygous for the lymphoproliferation spontaneous mutation (C57BL/6J-Faslpr) and wild type C57BL/6J mice were fed with MCD diet for three weeks to induce non-alcoholic steatohepatitis (NASH). Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG) and total cholesterol (TC) levels were detected by an Olympus AU5400 automatic chemical analyzer. The role of Fas gene mutation on NASH was assessed by comparing the severity of hepatic steatosis and inflammation in the liver sections, the mRNA and protein expressions of hepatic inflammatory and fibrogenesis related factors, proliferating cell nuclear antigen (PCNA) and transforming growth factor beta 1 (TGFb1).
RESULTSThe serum ALT levels of the wild type and Faslpr mice fed with MCD were significant higher than that of the control mice (126.33+/-10.50 U/L vs (25.00+/-10.14) U/L, (160.33+/-48.29) U/L vs (18.33+/-9.08) U/L, with the LSD-t value 12.02, 5.08 respectively, the P value<0.001, 0.007 respectively. The serum ALT levels showed no significant difference between the Faslpr and wild type mice fed with MCD, with the LSD-t value 1.19, the P value 0.229. The serum AST, TG and TC levels showed neithere significant difference among the four groups. MCD diet induced hepatic steatosis and inflammatory infiltration in both of the wild type and Faslpr mice. Especially, severer hepatic injury was observed in Faslpr mice as compared with wild type mice. The mRNA expression levels of cell proliferation factor PCNA and fibrogenesis growth factor TGF b1 in wild type mice fed with MCD were significantly higher than that of the control mice (2.84+/-0.73, 2.77+/-0.54 vs 1.31+/-0.18, 0.89+/-0.18), with the LSD-t value 4.99, 8.08 respectively, the P value 0.001, <0.001 respectively. The mRNA expression levels of PCNA and TGFb1 in Faslpr mice fed with MCD were significantly higher than that of the Faslpr control mice and the wild type mice fed with MCD (5.57+/-1.13, 5.73+/-0.89 vs 1.04+/-0.16, 0.85+/-0.11 and 2.84+/-0.73, 2.77+/-0.54), with the LSD-t value 10.15, 13.19 and 5.33, 6.91 respectively, the P value<0.001. The protein expressions levels of PCNA and TGFb1 were concordant with the mRNA.
CONCLUSIONSFaslpr promoted hepatic steatosis and inflammatory infiltration in mice fed with MCD diet, which might associated with excessive release of cell proliferative, inflammatory and fibrogenesis factors.
Animals ; Fatty Liver ; chemically induced ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Mutation ; Non-alcoholic Fatty Liver Disease ; Proliferating Cell Nuclear Antigen ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; fas Receptor ; genetics
6.Analysis of HBV rtS213T mutation during interferon therapy.
Wei-Hong REN ; Su-Ling ZHAO ; Zhi-Juan ZHAO ; Yan-Qing LI ; Hui-Qing TAO ; Hui XIONG
Chinese Journal of Hepatology 2010;18(3):227-228
Adolescent
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Adult
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Aged
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Antiviral Agents
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therapeutic use
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DNA, Viral
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Female
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Hepatitis B virus
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genetics
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Hepatitis B, Chronic
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drug therapy
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virology
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Humans
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Interferons
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therapeutic use
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Male
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Middle Aged
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Mutation
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Young Adult
7.Expression of astrocyte elevated gene-1 protein and its clinical significance in laryngeal squamous cell carcinoma.
Yong LIU ; Guo LI ; Zhong-wu SU ; Yuan-zheng QIU ; Xin ZHANG ; Chang-yun YU ; Xiao-juan ZHOU ; Shu-ling REN ; Dong-hai HUANG ; Yong-quan TIAN
Chinese Journal of Pathology 2013;42(2):111-115
OBJECTIVETo assess the protein expression of astrocyte elevated gene-1 (AEG-1) in tissue specimens of laryngeal squamous cell carcinoma (LSCC), and to correlate its expression with clinicopathological parameters and prognosis in patients with LSCC.
METHODSRT-PCR was used to assay the expression of AEG-1 mRNA in 13 pairs of LSCC tissues and their corresponding noncarcinoma epithelia. Immunohistochemistry was performed on paraffin-embedded tissue specimens to investigate the protein expression of AEG-1 in 88 cases of LSCC specimens and 15 cases of adjacent epithelial samples.
RESULTSThe expression of AEG-1 mRNA was significantly increased in LSCC tissues compared to adjacent noncarcinoma epithelial tissues (0.81 ± 0.17 vs. 0.23 ± 0.10;t = 10.337, P < 0.001). Meantime, the positive rate of AEG-1 protein in 88 cases of LSCC was 87.5% (77/88). However, 15 cases of adjacent noncarcinoma epithelial merely demonstrated negative or mild expression of AEG-1 protein. AEG-1 overexpression was closely correlated with T stage (χ(2) = 6.289, P = 0.018), clinical stage (χ(2) = 11.049, P < 0.01), metastasis (χ(2) = 20.859, P < 0.01) and recurrence(χ(2) = 13.459, P < 0.01). The overall survival rates of patients with AEG-1 overexpression and low expression were 35.9% and 86.4%, respectively (χ(2) = 23.409, P < 0.01). Multivariate Cox regression analysis revealed that AEG-1 expression was an independent prognostic factor (P = 0.016).
CONCLUSIONAEG-1 protein may play a critical role in the initiation and progression of LSCC, implicating its predictive value in prognosis.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; surgery ; Cell Adhesion Molecules ; genetics ; metabolism ; Female ; Follow-Up Studies ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Proportional Hazards Models ; RNA, Messenger ; metabolism ; Survival Rate
8.Effect of miR-24 on chemotherapy sensitivity in lung cancer A549 cells
Shu-Juan SU ; Tao FAN ; Yi-Chang SUN ; Jin-Shan REN
Chinese Journal of Pathophysiology 2018;34(6):1042-1048
AIM:To study the effect of microRNA (miR)-24 on chemotherapy sensitivity and its possible mechanisms in human lung adenocarcinoma A549 cells. METHODS:The expression of miR-24 in the A549 cells and A549/DDP cells was determined by real-time PCR. Transfection of miR-24 inhibitor was used to down-regulate the miR-24 level in the A549/DDP cells. The viability and apoptosis rate were measured by CCK-8 assay and flow cytometry, respec-tively. The protein levels of Bcl-2, Bax, cleaved caspase-3, cleaved caspase-9, cytochrome C (Cyt C), phosphorylated extracellular signal regulated kinase (p-ERK) and P53 were detected by Western blot. Luciferase reporter assay was used to predict and identify the target genes of miR-24. RESULTS:The expression of miR-24 was significantly higher in the A549/DDP cells than that in the A549 cells (P<0.05). miR-24 inhibitor induced cell apoptosis and increased the sensi-tivity of the A549/DDP cells to cisplatin. Furthermore, miR-24 inhibitor down-regulated the ratio of Bcl-2/Bax, while up-regulated the protein levels of P53, p-ERK, cleaved caspase-9, cleaved caspase-3 and Cyt C. Incubation with U0126, a specific ERK inhibitor, partly reversed the viability of miR-24 inhibitor transfected A549/DDP cells. Bioinformatics analy-sis demonstrated that p53 was a potential target gene of miR-24. Co-teansfection of miR-24 inhibitor and P53 siRNA in A549/DDP cells partially reversed the effect of miR-24 inhibitor on cell viabiltiy. CONCLUSION:Down-regulation of miR-24 increases the sensitivity of A549/DDP cells to cisplatin. The mechanism may be related to directly targeting p53 gene and over-activation of ERK/P53 signaling pathway, thus promoting apoptosis via mitochondrial apoptosis pathway.
9.Effects of KIF23 Gene Silencing on Proliferation,Migration and Invasion of Human Hepatocellular Carcinoma HepG2 cells
Su-Juan LIU ; Qu LIN ; Ming-Jun BAI ; Chu-Ren ZHOU ; Jun-Wei CHEN ; Chun WU ; Ming-Sheng HUANG
Journal of Sun Yat-sen University(Medical Sciences) 2018;39(1):34-40
[Objective]To investigate the effect of KIF23 gene expression on the proliferation,migration and invasion of human hepatocellular carcinoma HepG2 cells in vitro,and to explore the possible mechanism.[Methods]The KIF23 siRNA was transfected into HepG2 cells by lipofectamine 3000.The expression of KIF23 mRNA and protein in HepG2 cells was de-tected by qRT-PCR and Western blot.The effect of silencing KIF23 on the proliferation of HepG2 cells was studied by CCK-8 assay and plate clone formation assay.The tumor cell abilities of migration and invasion after transfection were measured by scratch assay and Transwell assay.The expression of protein kinase B(PKB/Akt)and phosphorylated Akt(p-Akt)protein in HepG2 cells transfected with KIF23-siRNA2 was detected by Western blot.[Results]KIF23-siRNA could effectively si-lence the expression of KIF23 mRNA and protein in HepG2 cells(P<0.01).The results of CCK-8 assay,plate clone forma-tion assay,scratch assay and Transwell assay demonstrated that the cell proliferation,migration and invasion ability of the KIF23-siRNA2 interference group were significantly inhibited,compared to the negative control group and the blank control group(P<0.05).The expression level of total Akt protein in HepG2 cells was not changed,but the expression level of phos-phorylated Akt protein was down-regulated(P<0.05).[Conclusions]KIF23 may promote the proliferation,migration and in-vasion of human hepatocellular carcinoma cells by activating Akt signal transduction pathway.KIF23 is expected to be a new target for gene therapy of hepatocellular carcinoma.
10.Genetically modified myeloma cell vaccine inducing antitumor immune response in vivo.
Su-Ping REN ; Li-Sheng WANG ; Qiang GUO ; Hua WANG ; Xiang-Xu JIA ; Juan XU ; Heng-Xiang WANG ; Chu-Tse WU
Journal of Experimental Hematology 2006;14(1):54-60
This study was aimed to evaluate the in vivo antitumor effect of genetically modified myeloma cell vaccine on human myeloma xenografts implanted into nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mice. Human immune system was established in NOD/SCID mice by intraperitoneal injection of human peripheral blood lymphocytes (PBLs). After being inoculated subcutaneously with irradiated myeloma cell line sko-007, adenovirally transferred with GFP or p53, granulocyte-macrophage colony-stimulating factor (GM-CSF) and B7-1 genes, huPBL-NOD/SCID mice were challenged by subcutaneous injection of non-transferred sko-007 cells. The results indicated that Ad-p53/GM-CSF/B7-1-infected sko-007 cell vaccination significantly reduced local tumor growth compared with controls. Histopathological and immunohistochemical analysis showed that tumor tissues increasingly displayed diffuse necrosis, mainly caused by apoptosis, accompanied with significant fibroplasias and blood vessel hyperplasia, and human T cells infiltrated into the tumor tissues. It is concluded that transgenic p53, GM-CSF and B7-1 expression produces an immune response against myeloma cells and may be of therapeutic value for multiple myeloma in human being.
Adenoviridae
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genetics
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Animals
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B7-1 Antigen
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genetics
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immunology
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Cancer Vaccines
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immunology
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Genes, p53
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immunology
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Genetic Vectors
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Granulocyte-Macrophage Colony-Stimulating Factor
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genetics
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immunology
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Immunotherapy
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Mice
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Mice, Inbred NOD
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Mice, SCID
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Multiple Myeloma
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immunology
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Neoplasm Transplantation