1.SRF expression and its biological significance in gastric carcinoma cells
Jing ZUO ; Liang LIU ; Min ZHAO ; Lianfu ZUO
Chinese Journal of Clinical Oncology 2014;(3):166-169
Objective:This study aims to explore the relationship between serum response factor (SRF) expression level and gas-tric cancer progression by detecting SRF expression level in cancer cells. Methods:The SRF gene in SGC-7901 cells was silenced by RNA interference. Transfection efficiency was detected by fluorescence microscopy, cell proliferation by CCK 8 method, SRF gene and protein expression level by real-time polymerase chain reaction and Western blot, and cell cycle by flow cytometry. Results:Cell treat-ment with siRNA-SRF induced significant reduction in SRF mRNA levels. Western blot analysis showed that SRF protein decreased by 40.1%in the siRNA group compared with that in the control group (P<0.05). Compared with the blank, negative, and mock transfection control groups, cell proliferation of the siRNA-SRF group decreased. The inhibition ratio reached 64.24%, as measured by the CCK-8 assay (P<0.05). Treatment with siRNA could block SGC-7901 cell cycle at G0/G1 phase (P<0.05). Conclusion:SRF expression is close-ly associated with gastric carcinoma cell proliferation. SRF protein level detection can provide a certain reference value in evaluating malignant gastric carcinoma progression. SRF is possibly an important target for the prevention and control of gastric cancers.
2.Expression of ABCG2 in Adriamycin-resistant Human Esophageal Cancer Cells
Liang LIU ; Jing ZUO ; Li ZHAO ; Jing WANG ; Jianwen GUO ; Jianghui LIU ; Lianfu ZUO
Journal of China Medical University 2010;(2):101-104
Objective To explore the expression of ATP-binding cassette transporter C2(ABCG2) in adriamycin(ADM)-resistant human esophageal cancer cells.Methods The ADM-resistant human esophageal cancer cell(Eca109/ADM) was induced by gradually increasing the ADM concentration in the culture medium of human esophageal cancer cell line(Eca109) and long time screening culture.ABCG2 mRNA and protein of ADM-resistant cells was detected by RT-PCR,flow cytometry(FCM) and Western blot.Drug excretion of Eca109/ADM cells was examined by FCM.The drug resistance index to ADM was detected by MTT.Results The expression of ABCG2 in Eca109/ADM cells was higher than that in Eca109 cells.The drug excretion of Eca109/ADM cells was stronger than Eca109 cells.The Ecal09/ADM cells drug resistance index to ADM was 3.29.Conclusion The ADM-resistant cell line Eca109/ADM was established successfully as an ideal chemoresistant cell model.ABCG2 might be involved in the drug resistance of esophageal cancer cell.
3.Effects of limited fluid resuscitation on immune function and inflammatory mediators in patients with multiple bone fractures complicated with traumatic hemorrhagic shock
Hualin TANG ; Liang WANG ; Zhenxin LIU ; Rongyue ZUO
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2017;24(4):393-395,400
Objective To observe the effects of limited fluid resuscitation (LFR) on immune function and inflammatory mediators in patients with multiple bone fracturescomplicated with traumatic hemorrhagic shock. Methods One hundred and two patients with multiple bone fractures complicated with traumatic hemorrhagic shock admitted to the Affiliated Hospital of Medical College of Ningbo University from January 2014 to June 2016 were enrolled, and they were divided into a LFR group and a early sufficient routine fluid resuscitation (RFR) group by random number table, each group 51 cases. After admission, the patients in the two groups underwent bandaging and hemostasis at the traumatic sites and preoperative management was prepared. The patients in RFR group were treated with early sufficient RFR, while LFR group was treated with LFR. The changes of hematocrit (HCT), blood platelet count (PLT), prothrombin time (PT), blood lactic acid and immune cells, inflammatory mediators and micro RNA-155 (miR-155) were observed in patients of the two groups at 4 hours after resuscitation.Results Compared with RFR group, the resuscitation time (hours: 3.67±1.45 vs. 5.14±1.61), levels of PT (s: 11.43±2.21 vs. 15.73±2.52), serum lactic acid (mmol/L: 3.35±0.15 vs. 3.81±0.25), tumour necrosis factor-α [TNF-α (ng/L): 14.10±3.39 vs. 16.28±3.47], interleukin [IL-10 (ng/L): 31.43±10.51 vs. 40.09±13.23, IL-6 (ng/L): 490.10±55.13 vs. 610.30±63.15] and endothelin-1 [ET-1 (pg/L): 183.35±30.51 vs. 250.01±31.23] in LFR group were significantly decreased (allP < 0.01), while PLT (×109/L: 134.58±28.13 vs. 108.12±30.35), HCT (×10-2: 0.34±0.04 vs. 0.24±0.05), miR-155 (0.15±0.02 vs. 0.08±0.02) and CD4+CD25+ regulatory T cell [CD4+CD25+Treg (×10-2): 2.28±0.47 vs. 2.10±0.39] in LFR group were obviously increased (allP < 0.01).Conclusions Using LFR in the emergency treatment of patients with multiple bone fractures complicated with traumatic hemorrhagic shock can effectively shorten the resuscitation time, regulate the patients' coagulation function, reduce the unnecessary excessive liquid infusion, improve immune status and decrease the degree of inflammatory reaction.
4.Expressions of IL-10, IL-23 and CD86 in lesions of epidermodysplasia verruciformis
Si LIANG ; Yuehua LIU ; Yagang ZUO ; Kai FANG ; Baoxi WANG
Chinese Journal of Dermatology 2010;43(3):196-198
Objective To measure the expressions of IL-10, IL-23 and CD86 in lesions of epidermodysplasia verruciformis (EV), and to explore the relationship between cellular immune abnormality and EV pathogenesis. Methods Immunohistochemistry was used to measure the expressions of IL-10, IL-23 and CD86 in tissue samples from 10 patients with EV and 10 normal human controls. Results Three cytokines were observed in all the samples of EV, with the expression score ranging from 3 to 6 and expression intensity from moderate to high. However, of the control specimens, only 1 was positive for IL-10 with the expression score being 3, and expression intensity being moderate. Conclusion The pathogenesis of epidermodysplasia verruciformis may be correlated with the expression abnormality of some cytokines secreted by keratinocytes.
5.Relationship between OPN gene expression, esophageal cancer metastasis and the growth inhibition effect of artesunate on esophageal cancer cells
Liang LIU ; Li WANG ; Qi YANG ; Lianfu ZUO
Medical Journal of Chinese People's Liberation Army 2017;42(1):23-28
Objective To study the relationship between osteopontin (OPN) gene expression and the development and metastasis of esophageal cancer, and explore the effect of growth inhibitory of artesunate (Art) on regulating the expression of OPN in esophageal carcinoma cells.Methods The expressions of OPN gene and protein were detected in normal esophageal tissue (24 cases), esophageal dysplasia (21 cases) and esophageal squamous cell carcinoma (ESCC) tissue (45 cases) byin situ hybridization and immunohistochemistry, and the relationships between the OPN gene and protein expressions and the pathological features were analyzed with SAS software. After intervention with different concentrations of Art (0, 30, 60, 120mol/L) on ESCC Eca109 cells for 24h, the levels of OPN expression and cell cycle were detected by flow cytometry.Results The expression levels of OPN gene and protein were significantly higher in esophageal cancer tissues than in esophageal dysplasia and normal esophageal tissue (P<0.01), and in esophageal dysplasia than in normal esophageal tissue (P<0.05). The expression levels of OPN gene and protein in esophageal cancer tissues showed no relations with patients' gender, age and the degree of tumor differentiation (P>0.05), but showed a positive correlation with lymph node metastasis and the invasion depth of esophageal cancer (P<0.05). After treatment by Art for 24h, the proliferation index of Eca109 cells and the expression of OPN protein decreased significantly (P<0.01) in a concentration dependent manner.Conclusions The abnormal high expression of OPN gene may participate in the occurrence, invasion and metastasis of esophageal cancer, so can be used as an objective index of metastasis for esophageal cancer. Art inhibits the growth of esophageal carcinoma cells by down-regulating the expression of OPN protein.
7.Early adverse outcomes in young patients with ischemic stroke:an analysis of the related factors
Meng LIANG ; Meng ZUO ; Nana ZHAO ; Dezhi LIU ; Peng WANG ; Yan MA ; Xinfeng LIU
Chinese Journal of Cerebrovascular Diseases 2017;14(8):393-398
Objective To investigate the related factors of early adverse outcomes in young patients with ischemic stroke.Methods From January 2006 to June 2016,685 young patients (18-45 years old) with acute ischemic stroke admitted to the Department of Neurology,Nanjing General Hospital of Nanjing Military Command were enrolled retrospectively.They were diagnosed as the first onset with head CT or MRI.According to the modified Rankin scale (mRS) at 90 d,the patients were divided into a favorable outcome (mRS 0-2) group (n=554) and a poor outcome (mRS 3-6) group (n=131).The collection of clinical data were completed on the day of admission,including the risk factors for cerebrovascular disease (oral contraceptives,etc),the National Institutes of Health stroke scale (NIHSS) score on admission,the mean systolic blood pressure (>140 mmHg was analyzed) and laboratory examination.The stroke subtypes were classified with the trial of org 10172 in acute stroke treatment (TOAST) classification criteria.Univariate analysis was used to analyze the difference of clinical data between groups,and multivariate logistic regression analysis was used to analyze the risk factors for early poor outcomes.Results Compared with the favorable outcome group,the patients with the ratio of mean systolic pressure >140 mmHg in the first 3 d after hospitalization (37.4% [49/131] vs.21.7% [120/554],χ2=14.131),NIHSS score on admission (10.0 [7.0,14.0] vs.1.5 [0,3.0],Z=-15.300),white blood cell count (7.5 [6.0,9.0] ×109/L vs.6.8 [5.7,8.2] ×109/L,Z=-3.157),fasting glucose (4.9 [4.6,6.0] mmol/L vs.4.8 [4.4,5.3] mmol/L,Z=-2.726),higher fibrinogen level (2.8 [2.3,3.4] g/L vs.2.6 [2.3,3.2] g/L,Z=-2.018,blood uric acid level (291[220,346] mmol/L vs.315 [261,374] mmol/L,Z=-3.443),and plasma albumin level (43.1[40.0,45.9] g/L vs.44.8 [42.4,47.4] g/L,Z=-4.708) were decreased in the poor outcome group.There were significant differences between the two groups (all P<0.05).TOAST classification comparison:the proportion of the patients with cardioembolism in the poor outcome group was higher than that in the favorable outcome group.There was significant difference between the two groups (6.9% [9/131] vs.2.5% [14/554];χ2=4.893,P<0.05).There were no significant differences in the remaining clinical data between the two groups (all P>0.05).Multivariate logistic regression analysis showed that the higher NIHSS score on admission (OR,1.474,95%CI 1.378-1.576,P<0.01),the mean systolic pressure >140 mmHg at the first 3 d after admission (OR,2.134,95%CI 1.210-3.764,P=0.009) and the cardioembolism(OR,4.902,95%CI 1.073-22.222,P=0.040) were the risk factors for early poor outcome,and the elevated plasma albumin level (OR,0.902,95%CI 0.850-0.956,P=0.001) was a protective factor of early favorable outcome.Conclusion The higher NIHSS score at admission,the cardioembolism and the increased mean systolic blood pressure in the first 3 d after admission may result in early poor outcome in young patients with ischemic stroke,while the elevated plasma albumin level is beneficial to the early outcome.
8.Inhibitory effect of artesunate on human esophageal carcinoma cells and the possible mechanism
Liang LIU ; Jing WANG ; Jian-Wen GUO ; Jiang-Hui LIU ; Lian-Fu ZUO ;
Chinese Journal of Cancer Biotherapy 1995;0(03):-
Objective: To explore the inhibitory effect of artesunate(Art)on human esophageal carcinoma cells and to study the related mechanism.Methods: Nude mice were inoculated with Eca109 cells subcutaneously on the left upper limbs to establish esophageal carcinoma model.The model mice were divided into 5 groups: first group received 100 mg/kg Art,second group 200 mg/kg Art,third group 300 mg/kg Art,forth group 3 mg/kg cisplatin(DDP),and the fifth group received normal saline.Mass and volume changes of transplant tumors in different groups were observed.Flow cytometry was used to detect the cell cycle,apoptosis,and the expression of CDC25A protein,Smad3 protein and TGF-?protein in the transplanted tumors in mouse model.RT-PCR was used to detect the expression of CDC25A,Smad3 and TGF-?mRNA in the transplanted tumors.Results: Nude mouse model bearing human esophageal carcinoma was success- fully created.Compared with the control group,the volume and mass of transplant tumors in Art groups were significantly smaller(P
9.Morphological characteristics for the magnetic resonance imaging assessment of discoid lateral meniscal tears in children
Xiaoxin SUN ; Wei ZHOU ; Shuping ZUO ; Hao LIU ; Jingfeng SONG ; Chunyu LIANG
Chinese Journal of Tissue Engineering Research 2017;21(7):1104-1109
BACKGROUND: Currently, the morphologic characteristics of the torn discoid lateral meniscus remain unclear, andmorphological indexes used to evaluate the discoid lateral meniscus tears still need to be explored. OBJECTIVE: To discuss the specificity and sensitivity of discoid lateral meniscal tears in children using morphological indexes. METHODS: Seventy-three patients (seventy-six knees) with torn discoid lateral meniscus underwentarthroscopy and MRI data of knee examinations were retrospectively reviewed. All MR images were double-blinded, independently, retrospectively analyzed by attending radiologist (doctor A) and sports physician (doctor B). The following morphological signs of discoid lateral meniscus, such as a internal displaced free edge sign, a hypertrophic free edge sign and a hypertrophied-horn sign, were selected to evaluate the discoid lateral meniscus tears. The likelihood of the discoid lateral meniscus tears was analyzed on basis of arthroscopic findings constituted the gold standard. Sensitivity, specificity and accuracy of MRI diagnosis of the discoid lateral meniscus tears by two physicians were calculated, and the consistency of diagnosis results by two physicians was evaluated using the Kappa statistics.RESULTS AND CONCLUSION: (1) The sensitivity, specificity and accuracy in the diagnosis of discoid lateral meniscustears by internal displaced free edge sign for doctor A were 19%, 94% and 53%, respectively, and for doctor B were 21%, 97% and 55%. (2) The sensitivity, specificity and accuracy in the diagnosis of discoid lateral meniscus tears by hypertrophic free edge sign for doctor A were 52%, 100% and 74%, respectively, and for doctor B were 57%, 97% and 75%. (3) The sensitivity, specificity and accuracy in the diagnosis of discoid lateral meniscus tears by hypertrophied-horn sign for doctor A were 26%, 97% and 58%, respectively, and for doctor B were 24%, 100% and 58%. (4) The sensitivity, specificity and accuracy in the diagnosis of discoid lateral meniscus tears by binding morphological index for doctor A were 86%, 91% and 87%, respectively, and for doctor B were 88%, 94% and 91%. (5) The Kappa statistics for discoid lateral meniscus tears respectively demonstrated a good inter-observer agreement (K > 0.75, P < 0.001). (6) These findings suggest that single morphological index used to evaluate the discoid lateral meniscus tears shows a good specificity, but which also shows a low sensitivity, while a binding morphological index can significantly improve the diagnostic sensitivity. Morphological characteristics may be reliable indexes to evaluate the discoid lateral meniscal tears in children.
10.Influence of interleukin-1beta on microRNA-27b and matrix metalloproteinase-13 expression in rat chondrocytes
Jing SHI ; Tong LI ; Canhui ZUO ; Liang ZHI ; Rong WEI ; Weimin WANG ; Yingjie LIU
Chinese Journal of Tissue Engineering Research 2016;20(29):4277-4283
BACKGROUND:Matrix metal oproteinase-13 is most active in the degradation of col agen type II in the extracel ular matrix of cartilage. Interleukin-1 (IL-1) is thought to be the inducer of matrix metal oproteinases, and participates in the degradation and degeneration of articular cartilage. OBJECTIVE:To study the influence of IL-1βon microRNA-27b (miR-27b) and matrix metal oproteinase-13 expression of chondrocytes in rats. METHODS:Chondrocytes isolated from seven male Wistar rats were cultured and divided into IL-1βstimulation group and control group. No stimulus was given in the control group;10μg/L of serum free medium was used to culture rat chondrocytes in the IL-1βstimulation group. Cel growth was observed at 0, 24, and 48 hours under an inverted microscope. miR-27b and matrix metal oproteinase-13 expression in the cultured chondrocytes were detected. RESULTS AND CONCLUSION:The relative expression of matrix metal oproteinase-13 in rat chondrocytes was gradual y increased when induced by IL-1βat 0, 24, and 48 hours (P<0.05). Expression of miR-27b and miR-31 in rat chondrocytes at 24 and 48 hours induced by IL-1βgradual y decreased (P<0.05);conversely, expression of MiR-26a, miR-26b, miR-23, and miR-204 gradual y increased (P<0.05). After 48 hours of IL-1βinduction, expression of miR-27b was the lowest in rat chondrocytes (P<0.05). These findings suggest that IL-1βinhibits miR-27b expression, strengthens the expression of matrix metal oproteinase-13, and damages chondrocytes, contributing to both the onset and progression of osteoarthritis.