1.Effect of costimulatory molecule B7-H3 on the biological characteristics of esophageal cancer Eca-109 cell line
Nana CAO ; Ling WANG ; Baoen SHAN
China Oncology 2014;(8):561-567
Background and purpose:Esophageal cancer is a serious disease threatening human health, and it is very difficult to understand the development mechanism and find the therapeutic methods for esophageal cancer. In recent years, B7-H3, as a new member of B7 immunoregulatory superfamily, overexpressed in multiple tumor types, is considered to be a new tumor marker and potential therapeutic target. This study aimed to detect the expression of B7-H3 in esophageal cancer cell lines TE-1, TE-13, Eca-109 and exploring the effect of B7-H3 siRNA on cell proliferation, migration and invasionin vitro in human esophageal cancer Eca-109 cell line. Methods:The expression of B7-H3 in esophageal cancer cell lines TE-1, TE-13 and Eca-109 were detected by reverse transcription polymerase chain reaction (RT-PCR). B7-H3 siRNA and control siRNA were transfectedin vitro into human esophageal cancer Eca-109 cells using LipofectamineTM 2000. The expressions of B7-H3 mRNA and protein in Eca-109 cells were analyzed by RT-PCR and Western blot. The proliferation, migration and invasion abilities of Eca-109 cells were measured by MTT assay, wound scrape assay and transwell invasion assayin vitro,respectively.Results:All tested cultured esophageal cancer cell lines constitutively expressed B7-H3 mRNA under normal conditions (TE-1 0.382±0.008, TE-13 0.399±0.008, Eca-109 0.428±0.012). After transfection, the expression of B7-H3 mRNA levels decreased in B7-H3 siRNA transfected group, compared with control siRNA transfected group (0.128 5±0.000 2vs 0.532 4±0.000 7,P<0.01) and untransfected group (0.128 5±0.000 2vs 0.540 3±0.001 3,P<0.01), while its protein expression levels were also signiifcantly lower than the control transfection group (0.421 4±0.004 8vs 0.500 6±0.012 9,P<0.05) and untransfected group (0.421 4±0.004 8vs 0.492 1±0.014 8, P<0.05). Compared with control transfected and untransfected cells, Eca-109 cell migration and invasion abilities decreased significantly (P<0.05) by siRNA interference, but no significant difference was observed between their proliferative capacity (P>0.05).Conclusion:All tested esophageal cancer cell lines constitutively express B7-H3 mRNA. B7-H3 siRNA interference inhibits Eca-109 cell migration and invasion abilities. B7-H3 may have a critical role in regulating Eca-109 cell progression.
2.Dietary fiber and stroke
Wanying SHAN ; Ling WANG ; Gelin XU
International Journal of Cerebrovascular Diseases 2017;25(1):84-90
Epidemiological studies indicated that dietary tiber was inversely associated with stroke risk.Epidemiological studies have confirmed that increased dietary fiber intake can reduce the risk of stroke.Animal experiments and clinical studies have shown that dietary fiber intake may reduce the incidence of stroke by reducing vascular risk factors,such as hypertension,diabetes,dyslipidemia,and obesity.The direct mechanism of increased dietary fiber intake reducing stroke risk remains to be further studied.Clinicians should pay attention to the important role of dietary fiber,guide the patients with stroke and individuals at risk of stroke to increase consumption of fiber-rich fruits and vegetables.
3.Effects of mild moxibustion on imbalance of Th1/Th2 in rabbits with atherosclerosis and its regulating rules.
Shan WU ; Ling-Ling WANG ; Hai-Hong CAI ; Wen JIANG
Chinese Acupuncture & Moxibustion 2014;34(2):163-167
OBJECTIVETo observe effects of mild moxibustion and lovastatin on immunologic function in rabbits with chronic hyperlipidaemia and atherosclerosis (AS) to initially explain regulating rules of mild moxibustion on immunologic function.
METHODSAmong thirty-two Japanese male big-ear rabbits, 8 rabbits were randomly selec ted as a blank group, the rest 24 rabbits were fed with method of endothelial injury and high-fat diet to establish AS model. The blank group was raised with normal diet and free water. After ten weeks of model establishment, the rest 24 rabbits were randomly divided into a model group, a moxibustion group and a medicine group, eight rabbits in each one. Moxibustion was applied at "Shenque" (CV 8) and "Zusanli" (ST 36) for 10 min per acupoint per day in the moxibustion group, while intragastric administration of 3.6 mg/kg lovastatin capsule was applied in the medicine group. After treatment, serum was acquired. Spectrophotometry method was adapted to measure cholesterol (TC) and high-density lipoprotein (HDL-C) and evaluated atherosclerosis index (AI), while enzyme-linked immunosorbent assay method was used to measure interferon-gamma (IFN-gamma) and interleukin-4 (IL-4).
RESULTS(1) The serum TC and HDL-C in the model group were significantly higher than those in the blank group, moxibustion group and medicine group (all P < 0.01). The mean value of AI was 1.683 +/- 0.486 in the moxibustion group, which was obviously lower than 20.301 +/- 4.022 in the model group (P < 0.01). (2) The ratio of Th1/Th2 was 0.569 +/- 0.143 in the moxibustion group and 0.445 +/- 0.079 in the medicine group, which were significantly lower than 0.917 +/- 0.255 in the model group (both P < 0.01), but there was no statistically significant difference between the moxibustion group and the medicine group (P > 0.05).
CONCLUSIONThe moxibustion for AS could reduce atherosclerosis index, influence drift and bias of helper T cell and regulate balance between humoral immunity and cellular immunity. As a result, status of relative balance of immunity is acquired, which could slow down the development of atherosclerosis and process of thrombus burst.
Animals ; Atherosclerosis ; immunology ; therapy ; Humans ; Hyperlipidemias ; immunology ; therapy ; Interferon-gamma ; immunology ; Interleukin-4 ; immunology ; Male ; Moxibustion ; Rabbits ; Th1 Cells ; immunology ; Th2 Cells ; immunology
4.Serum proteomic spectra of esophagial carcinoma patients and a corresponding esophagial carcinoma diagnostic model
Lihua LIU ; Baoen SHAN ; Shijie WANG ; Jun MENG ; Ling WANG
Chinese Journal of Cancer Biotherapy 1995;0(03):-
Objective:To examine the serum proteomic spectra of human esophagial carcinoma by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS),so as to set up a diagnostic model of esophagial carcinoma and to investigate its clinical value. Methods:Thirty-two esophagial carcinoma patients and 28 healthy controls were obtained from Fourth Affiliated Hospital of Hebei Medical University during May to September of 2008. Serum protein was extracted by weak cation exchange (WCX) protein chip system,and proteomic spectra was examined by MALDI-TOF MS. The obtained data were analyzed by ZUCI-protein chip data analyze system (ZUCI-PCDAS) and an esophagial carcinoma diagnostic model was established by genetic arithmetic (GA) combined support vector machine (SVM). The above 60 samples were randomly divided into training set and blinding test set,with training set including 21 esophagial carcinoma patients and 19 healthy controls and blinding test set including 11 esophagial carcinoma patients and 9 healthy controls,so as to examine the specificity and sensitivity of this diagnostic model. Results:Serum proteomic spectra of esophagial carcinoma patients and healthy controls were obtained by MALDI-TOF MS,and m/z (mass to charge) peaks of 44 differential proteins were obtained after analyzed by ZUCI-PCDAS software package (P
5.Effects of celecoxib on tumor growth and cell apoptosis in human triple-negative breast cancer xenografts in nude mice
Ling WANG ; Baoen SHAN ; Lihong LIU ; Jie LI ; Bin WANG
Cancer Research and Clinic 2011;23(11):739-742
Objective To evaluate the effects of celecoxib on tumor growth and cell apoptosis in human triple-negative breast cancer (TNBC) xenografts in nude mice.Methods Human TNBC MDA-MB-231 cells were inoculated subcutaneously into BALB/c nude mice.The mice (n=32) were then randomly divided into 4 groups,the control group and the celecoxib group (receiving 25,50,100 mg·kg-1·d-1 respectively).At the end of the study,tumor tissues were collected and tumor volume was measured.Cell apoptosis was determined by flow cytometry (FCM) analysis.NF-κB p65 and pS0 protein levels were measured by immunohistochemistry.Caspase-3 and survivin protein levels were detected by western blotting.Results celecoxib at dose of 25,50 and 100 mg·kg-1·d-1 inhibited the tumor growth significantly,compared with the control group.FCM results showed that apoptotic rates were (13.58±3.16) % and (21.91±4.75) % in moderate and high dose of celecoxib-treated group,compared with (3.15±1.73) % in control group (t =6.736,P < 0.05;t =12.151,P < 0.05).p65 expressions were 79.3 %,46.7 % and 23.9 % in low,moderate and high dose of celecoxib-treated group,compared with 89.7 % in control group (x2 =3.312,P < 0.05; x2 =10.785,P < 0.05;x2 =15.900,P < 0.05).Besides,western blotting analysis demonstrated that celecoxib significantly downregulated survivin expression,while upregulated the active form of caspase-3 expression.Conclusion Celecoxib could suppress TNBC tumor growth and induce cell apoptosis,which might be partially associated with inactivation of p65 and downregulation of survivin.
6.Antithymocyte globulin for graft-versus-host disease prophylaxis in hematopoietic stem cell transplantation from unrelated donors: a retrospective report
Hanbo DOU ; Jiehui SHAN ; Ling WANG ; Wei TANG ; Jiong HU
Journal of Leukemia & Lymphoma 2012;21(12):716-719,726
Objective To assess the impact of antithymocyte globulin (ATG) on the incidence of graft-vs-host disease (GVHD) in hematopoietic stem cell transplantation from unrelated donors.Methods A total of 92 patients with hematological malignancies including leukemia,myelodysplastic syndrome (MDS) and lymphoma who underwent hematopoietic stem cell transplantation from unrelated donors from January 1999 to December 2011 were included in this retrospective analysis.Patients were classified into ATG group (n =66)and non-ATG group (n =26) according to the GVHD prophylaxis regimen.The incidence of acute GVHD (aGVHD) and chronic GVHD (cGVHD),risk factors of aGVHD and cGVHD and impact of ATG on the overall survival (OS),treatment related mortality (TRM) and relapse rate were analyzed.Results Grade Ⅱ-Ⅳ aGVHD (26.7 % vs 44.0 %,P=0.12) or grade Ⅲ-Ⅳ aGVHD (13.3 % vs 8.0 %,P =0.74) were not significantly different between ATG and non-ATG group.However,the incidence of cGVHD in the ATG group was significantly lower (34.0 % vs 72.2 %,P =0.005) than non-ATG group.The incidence of extensive cGVHD was also significantly reduced (10.0 % vs 44.4 %,P =0.005) compared to non-ATG group.In multivariate analysis,the use of ATG prophylaxis significantly decreased the cGVHD (RR =0.22,95 %CI 0.081-0.599,P =0.003) while one allele mismatch of human leukocyte antigen (HLA) was associated with increased risk of cGVHD (RR =3.25,95 % CI 1.39-7.61,P =0.007).As to the extensive cGVHD,the use of ATG was the only independent factor (RR =0.05,95 % CI 0.009-0.240,P < 0.001).With a median follow-up of 12 months (1-84 months),ATG prophylaxis had no impact on OS rate (60.4 % vs 43.1%,P =0.41),TRM rate (19.8 % vs 34.3 %,P =0.43) and relapse rate (40.6 % vs 33.6 %,P=0.54).Conclusion In hematopoietic stem cell transplantation from unrelated donors,ATG prophylaxis total dose of 6 mg/kg may significantly decrease the incidence of cGVHD and extensive cGVHD without increase of TRMand relapse rate and impairment of OS.
7.Outcome of autologous peripheral blood stem cell transplantation for T cell lymphoma
Jiehui SHAN ; Ling WANG ; Wei TANG ; Zhixiang SHEN ; Jiong HU
Journal of Leukemia & Lymphoma 2012;21(3):137-140
Objective A retrospective analysis of patients with T-cell lymphoma (TCL) received autologous peripheral blood stem cell transplantation (APBSCT) was performed to evaluate the outcome of APBSCT.Methods A total of 22 patients who underwent APBSCT from September 2006 to December 2011 in Ruijin hospital were enrolled in the study,including 6 cases of lymphoblastic lymphoma and 16 of peripheral T-cell lymphoma (8 anaplastic large cell lymphoma, 4 PTCL-u, 1 subcutaneous panniculitis-like T-cell lymphoma, 2 nasal type extranodal NK/T and 1 primary cutaneous T-cell lymphoma). All patients were diagnosed based on the WHO Classification of Tumors of Hematopoietic and Lymphoid Tissues. Conditioning regimens were high-dose chemotherapies alone which include 13 cases with BEAM, 4 with ICE and 5 with CBV. The outcomes of the treatment were evaluated according to the revised International Working Group criteria.Results With a median follow-up of 13.1(1-60) months,the predicted 2-year overall survival (OS) and progression-free survival (PFS) after transplantation were (67.6±11.0) % and (71.1±11.1) %,respectively.A total of 6 patients experienced disease progression and 5 patients eventually died of disease. When all these patients based on the remission status before APBSCT (CR1 vs non-CR1) and chemosensitivity (sensitive vs refractory) were further classified, the PFS rates and OS rates were 100 % and 91.7 % respectively in CR1 or chemosensitive patients which were significantly higher than patients not in CR1 (42.6 % ) or with chemoresistant disease (19.0 % ). Conclusion Remission status and chemosensitivity at the time of transplantation significantly affect the outcome of APBSCT for TCL patients, thus it can be recommend to perform APBSCT for patients either in CR1 or early stage when the disease remain sensitive to chemotherapy.
8.Ultrastructural analysis of Barrett's esophagus under magnifying chromoendoscopy
Shan YAN ; Hong LING ; Weiping YANG ; Yongcheng XU ; Hong WANG
Chinese Journal of Primary Medicine and Pharmacy 2011;18(6):736-738
Objective To study Barrett's esophageal consistency with its histological findings,which were found with magnifying chromoendo6copy,and to advance the endoscopic diagnosis of Barrett's esophagus. Methods The patients with BE were diagnosed by endoscopy and histology, and their histological findings and endoscopic appearances of a total of 67 patients with BE were observed. Results Endoscopic appearances type of BE showed island (28 cases) ,tongue(7 cases) ,and circum ference(32 cases). Three types of BE under magnifying chromoendoscopy,and in magnifying endoscope group,type Ⅲ which were found with magnifying endoscope were verified intestinalisation epithelium pat ho-type, compared with type I and type Ⅱ,the discrepancy of them had statistical significance(P<0.01). Conclusion Magnifying chromoendoscopy had a relatively better consistency in the diagnosis of Barrett's esophagus with histological findings.
10.Expression of S100B during heart failure in rats
Zhenni JIANG ; Ling CHENG ; Jiang SHAN ; Jianan WANG
Chinese Journal of Emergency Medicine 2008;17(5):475-478
Objective To evaluate the value of S100B gene on cardiovascular remodeling in rats with abdominal aorta coarctation.Method Sprague-Dawley rats(sanitary degree,male),weighring(220~240)g,were used in the study.They were provided by College of Medicine,Zhejiang University.The abdominal aortas of rats were isolated and constricted so as to establish models of heart failure;the abdominal aortas of another ten rats were isolated but not ligated(sham-operation group).After one week,the 20 rats were alive and randomly divided into 2 groups:operation group(n=10)and Carvidilol-operation group(Car group,n=10).Car Group was treatedwith 2 mg·kg-1per day Carvedilol and operation group was administered with the same doses of normal saline.After 4 weeks,a catheter was inserted through the right jngular artery into the left ventricle to record hemodynamic.Then all rats were euthanized,and the heart tissues were rapidly excised,rinsed with PBS.The left ventricles were then cut into two parts at the equator:the upper sections were fixation,the lower sections were stoted at-80℃,at 100 mg per tube.LV transverse section(4μm thick)was subjected to S100B immunehistoc- hemistry.RNA was iselated from tissues by Trlzol to determine levels of S100BmRNA and β-actinmRNA by RT- PCR.The data was expressed as(x±s);One-way analysis of variance was used for comparison among groups, and Student-Newman-Keulsa test for comparison between two groups.Statistical significance was set at P<0.05. Results In Operation Group,there was a decrease in maximal rate of systolic and diastolic of left ventricular pressure(±dp/dtmax)[(1543.6±277.9)mmHg/s vs.(2640.4±481.3)mmHg/s and(-1352.5±202.3) mmHg/s vs.(-1873.2±412.3)mmHg/s];and an increase in left ventricular end diastolic pressure(LVEDP) [(24.8±5.2)mmHg vs.(2.1±0.7)mmHg]compared with sham-operation group(P<0.01,P<0.01 and P<0.05).And in Car group,the level of LVEDP was just in the midst of the Operation Group and sham-opera- tion group,and had statistical significance(P<0.01);while±dp/dtmac[(2372.3±92.6)mmHg/s and (-1786.4±62.6)mmHg/s]were much higher than those in operation group(P<0.01).There were no any S100B positive cells and expression of S100B mRNA in sham-operation group;while there were much more S100B positive cells in operation group than those in Car group(P<0.01).The expression of S100B mRNA in operation group was more pronounced than that in Car group.Changes in expression of S100BmRNA were positively correlat- ed with changes in LVEDP(r=0.847,P<0.01);while changes in expression of S100BmRNA were negatively correlated with changes in±dp/dt max(r=-0.853 and-0.689,beth P<0.01).Conclusions There was hish expression of S100B in myocytes from rats with experimental heart failure and negative correlation between the expression of S100B and heart function.It indicated that S100B could play a negative role in heart failure.