1.Distribution of free Ca~(2+) and characterization of Ca~(2+) channel in the cultured early-stage human embryonic retina
Wenwen LIU ; Ping XU ; Qiangsu GUO
Chinese Journal of Ocular Fundus Diseases 1996;0(01):-
Objective To investigate the distribution of free Ca 2+ and characterization of Ca 2+ channel in the cultured 11~15 week human embryonic retina. Methods The cells from 11~15 week embryonic retina were dissociated enzymatically and cultured on coated cover slides. Three weeks later, Fluo3, a Ca 2+ indicator, was added in Ca 2+ containing or no Ca 2+ Hepes buffer and incubated with retinal cells for 30 min, meanwhile with or without verapamil, perdipine, or dexamethasone adding. Ca 2+ staining and Ca 2+ transit before and after 10 ?mol/L or 50 ?mol/L KCl stimulation was observed and recorded using Zeiss LM510 confocal laser scanning microscopy. Results In response to K + exposure, a rapid rise of free Ca 2+ in cytoplasma and nuclei of neuron and glial cell was observed, but was repressed with the treatment of verapamil, perdipine, or dexamethasone. The obvious Ca 2+ influx from cytoplasma into nuclei was observed even in the absence of external Ca 2+ . Conclusion L type Ca 2+ channel was expressed and functionally matured in the cultured early stage human embryonic retina.
2.Role of P38 in endoplasmic reticulum stress-induced chemoresistance to cisplatin in gastric cancer cells
Ruo FENG ; Liping WANG ; Yurong CHAI ; Wenwen GUO ; Wenlong ZHAI
Chinese Journal of Clinical Oncology 2015;(13):637-641
Objective:To investigate the mechanism of endoplasmic reticulum (ER) stress-induced chemoresistance to cisplatin in gastric cancer cells. Methods:ER stress models were established in both BGC823 and SGC7901 gastric cancer cells. The expression of GRP78, an ER stress marker, was examined by Western blot analysis. Moreover, whether ER stress can decrease the sensitivity of gastric cancer cells to cisplatin and activate P38 was explored by flow cytometry and Western blot analysis, respectively. Whether ER stress-induced chemoresistance to cisplatin can be abrogated by blocking P38 activity in gastric cancer was also elucidated using flow cytometry. Results:GRP78 protein expression markedly increased after treating BGC823 and SGC7901 gastric cancer cells with tunica-mycin (TM) or thapsigargin (TG) for 8, 16, and 24 h (P<0.05), compared with that in the group treated for 0 h. The apoptotic rates of TM-(or TG)-, cisplatin-, and TM (or TG) plus cisplatin-treated groups significantly increased (P<0.05) in both BGC823 and SGC7901 gastric cancer cells compared with the rate in the control group. The apoptotic rate of TM (or TG) plus cisplatin-treated group signifi-cantly decreased (P<0.05) in both BGC823 and SGC7901 gastric cancer cells compared with that of the cisplatin-treated group. Com-pared with the group treated for 0 h, phospho-P38 expression markedly increased after treating BGC823 and SGC7901 gastric cancer cells with TM (or TG) for 8, 16, and 24 h (P<0.05). No difference in P38 protein expression was observed between each group in both BGC823 and SGC7901 gastric cancer cells (P>0.05). Both P38 inhibitors, either SB203580 or PD169316, can inhibit the activation of P38. The inhibition of P38 activity can overcome ER stress-induced chemoresistance to cisplatin in gastric cancer cells (P<0.05). Con-clusion:ER stress can trigger the chemoresistance to cisplatin by activating P38 in gastric cancer cells.
3.Analysis of complicating ascites of mini-percutaneous nephrolithotripsy
Yu LIU ; Zhenjia GUO ; Zengjun ZHU ; Wenwen SUN
Chinese Journal of Postgraduates of Medicine 2014;37(5):17-19
Objective To explore the clinical causes and preventive measures of complicating ascites of mini-percutaneous nephrolithotripsy (MPCNL).Methods Retrospective analysis of 285 patients with MPCNL for upper urinary tract calculus,which were divided into ascites group and no-ascites group.Results All the procedures were successful.Ascites group of 21 cases,no-ascites group of 264 cases.Univariate analysis showed that the diameter and number of calculus,perfusion pressure,perfusion time,pressure volume of irrigation fluid,preoperative upper urinary tract infection,history of treatment associated with complicating ascites (P< 0.05),with age,gender,body mass index no correlation (P> 0.05).Logistic regression analysis showed that perfusion pressure,perfusion time,pressure volume of irrigation fluid was independent risk factors after MPCNL concurrent ascites (P < 0.05).Conclusions MPCNL concurrent ascites are closely related to the large perfusion volume,the long operative perfusion time,the high perfusion pressure of irrigation fluid.On the premise of keeping the operative visual field clear,as far as possible to reduce the perfusion pressure,control irrigation fluid-flow rate,reduce the large peffusion volume.These could decrease the coincidence of the ascites.
4.Research progress on tumor-vessel associated extravascular tumor metastasis
Wenwen GUO ; Haiming WEI ; Xianglan MO ; Xiaogang ZHONG ; Xiaofeng DONG
Chinese Journal of Digestive Surgery 2017;16(3):313-315
The occurrence of malignant tunors is increasing annually around the world.In every year,8.2 million patients died of malignant tumors in which about 90% patients died of malignant tumors associated multiple organs metastases.Inhibiting tumor metastasis is the key event to prolong the survival time of patients.With the further research,tumor-vessel associated extravascular tumor metastasis is playing an increasingly important role in the clinical application.This paper summarizes the new developments of tumor-vessel associated extravascular tumor metastasis to provide possible ideas for the therapy of malignant tumors.
5.Effect of Everolimus on Radiosensitivity of Human Non_small Cell Lung Cancer Cell Line A549
Yu CHEN ; Qian CHU ; Juan GUO ; Yu HUANG ; Wenwen LI ; Yijun TIAN ; Shu XIA ; Shiying YU
Herald of Medicine 2014;(12):1541-1544
Objective To exPlore the effect of mammalian target of raPamycin ( mTOR ) inhibitor eVerolimus on radiosensitiVity of human non_small cell lung cancer cell line in vitro by using eVerolimus to inhibit mTOR signaling Pathway of A549. Methods Human non_small cell lung cancer cell line A549 was subjected to radiation alone or in combination with eVerolimus treatment. The 50%inhibition concentration ( IC50 ) of eVerolimus in A549 cells was detected by methylthiazol tetrazolium ( MTT) assay in vitro. EVerolimus at the 20%inhibition concentration ( IC20 ) was used to Pretreat A549 cells for 24 h. Cells were then irradiated by X_ray with 2,4,6,8 Gy. The cell surViVal fraction was comPuted by clone formation. Cell surViVal curVe was fitted by multitarget one_hit model, and mean lethal dose ( D0 ), dose quasithreshold ( Dq ), surViVal fraction at 2 Gy ( SF2 ), and sensitization enhancement ratio (SER) were calculated. The exPression ofγ_H2AX was determined by Western blotting and then the relatiVe gray Values were analyzed. Results EVerolimus significantly imProVed the sensitiVity of A549 cells to radiation. The D0 , Dq and SF2 of eVerolimus+irradiation grouP were significantly lower than those of irradiation grouP. The SER was 1. 36. The residual amount of γ_H2AX Protein in the eVerolimus + irradiation grouP was significantly higher than that of the irradiation grouP. Conclusion EVerolimus inhibiting mTOR signaling Pathway can increase the radiosensitiVity of A549 cells.
6.Investigation on the therapeutic effects and the mechanism of mesenchymal stem cells on acute immune liver injury in mice
Kui DONG ; Xuqian ZHANG ; Haiying GUO ; Weiqiang WANG ; Wenwen LI ; Bangmao WANG ; Wentian LIU
Chinese Journal of Digestion 2015;35(2):104-109
Objective To explore the mechanisms of mesenchymal stem cells (MSC) in the treatment of concanavalin A (ConA)-induced acute immune liver injury in mice.Methods MSC were isolated and cultured from bone of the four limbs of three-week-old C57BL/6 mice.The specific surface markers were identified and osteogenic,adipogenic differentiation ability were tested.A total of 15 six to seven-week old C57BL/6 mice were divided into control group,MSC treatment group and phosphate buffer saline (PBS) treatment group,five mice in each group.The mice of MSC treatment group was injected through tail firstly with ConA and then MSC,PBS treatment group was injected through tail firstly with ConA and then PBS,control group was injected through tail with PBS twice.The mice were sacrificed in 14 to 16 hours after injection.The level of alanine aminotransferase (ALT),aspartate transaminase (AST) in peripheral blood were detected and the pathological change in liver tissue was scored by Knodell score system.Activation rate of splenic CD4+ T cells and the proportion changes of T hepler cell (Th)1,Th2,Th17 and regulatory T cells (Treg) were detected by flow cytometry and the ratio of Th17/Treg was calculated.The levels of tumor necrosis factor α (TNF-α),interferon (IFN)-γ and interleukin (IL)-4 in peripheral blood were detected by enzyme linked immunosorbent assay (ELISA).Independent-sample t test was used for comparison between groups of measurement data.Results ALT,AST and Knodell score of MSC treatment group was (174.2± 46.9) U/L,(185.6± 71.6) U/L and 3.4±1.3,respectively,which were better than those of PBS treatment group ((647.0± 118.0) U/L,(749.0± 104.0) U/L and 5.2 ±0.8,respectively),and the differences were statically significant (t =8.33,9.98 and 2.55,all P<0.05).The activation rate of splenic CD4+ T cell of PBS treatment group was (26.10±2.17) %,the proportion of Th1 and Th2 in CD4+ T cell was (5.81±0.79) % and (5.98± 1.22)%,the ratio of Th17/Treg was 0.29±0.03,the levels of TNF-α,IFN-γ and IL4 in peripheral blood were (1 281.95±88.61) U/L,(1 838.66±196.91) U/L and (1 192.36±163.94) U/L,which were higher than those of control group ((13.74±1.59)%,(1.35±0.17)%,(2.13±0.17)%,0.15± 0.05,(21.71±2.50) U/L,(11.84±1.28) U/L and (24.46±3.96) U/L),and the differences were statistically significant (t=10.26,12.37,7.02,5.30,31.79,15.93 and 20.75,all P<0.01).There was no statistically significant difference in the activation rate of splenic CD4+T cell between MSC treatment group and PBS treatment group ((26.20±3.09)% vs (26.10±2.17)%,P>0.05).However,in MSC treatment group,the proportion of Th1 and Th2 in CD4+ T cell ((1.83±0.52) % and (2.75±1.06%)),the ratio of Th17/Treg (0.18±0.02) and the levels of TNF-α,IFN-γ and IL-4 in peripheral blood ((760.71± 73.19) U/L,(742.49±76.46) U/L and (825.76±101.74) U/L) significantly decreased compared with those of PBS treatment group,and the differences were statistically significant (t=9.45,4.48,6.41,10.14,5.56 and 10.22,all P<0.01).There was no significant difference in the ratio of Th17/Treg between MSC treatment group and control group (P>0.05).Conclusions The therapeutic effects of MSC on ConA induced acute immune liver injury were through influence splenic CD4+ T cell subsets by decreasing the proportion of Th1 and Th2 and then declining the levels of secreted cytokines such as TNF,IFN-γ and IL-4 in peripheral blood,increasing the proportion of Treg and decreasing the proportion of Th17 and keeping the balance of Th17/Treg.
7.Recent Progress of Colorimetric Assays Based on Gold Nanoparticles for Biomolecules
Wenwen CHEN ; Yongming GUO ; Wenshu ZHENG ; Yunlei XIANYU ; Zhuo WANG ; Xingyu JIANG
Chinese Journal of Analytical Chemistry 2014;(3):307-314
Biochemical analysis assays based on colorimetric methods using gold nanoparticles have many advantages including high sensitivity, good selectivity, naked-eyes readout and complex instruments free. These methods have good prospects in applications. The biomolecule assay is highly relative with human health. This review mainly focuses on colorimetric assays applying gold nanoparticles for biomolecules detection.
8.Relationship between the Cx37 polymorphism of connexin gene and essential hypertension in Kazak and Han population in Xinjiang
Bihan LU ; Wenwen ZHANG ; Wenyan SHI ; Ketao MA ; Li LI ; Shuxia GUO ; Junqiang SI
Tianjin Medical Journal 2015;(11):1326-1329,1330
Objective To investigate the association of the Cx37 polymorphism of connexin gene with essential hyper?tension (EHT) in Xinjiang Han and Kazak population. Methods In Xinjiang region, 500 EHT patients (EHT group) were in?cluded in this study including Kazak 250 cases and Han 250 cases. Five hundred healthy volunteers (NT) were used as NT group including Kazak 250 cases and Han 250 cases. The values of age, body mass index (BMI), systolic blood pressure (SBP) and diastolic blood pressure (DBP), and other general clinical features were compared between two groups. The poly?morphism of Cx37 gene rs1630310, rs697372 and rs705193 SNP were compared between EHT and NT groups in the two eth?nic groups. Hardy-Weinberg equilibrium was used to detect the representation, and differences of genotype frequencies and gene frequency were calculated in two groups of Kazak and Han groups. Results There were significant differences in BMI, SBP, DBP, apolipoprotein ratios and homocysteine between EHT group and NT group in Kazak and Han groups (P<0.05). There were no significant differences in genotype frequencies and gene frequencies of rs705193 between EHT and NT groups (P>0.05). The differences of Kazak rs1630310 genotype and gene frequency were statistically significant (P<0.01). The frequency of Kazak rs697372 locus genotype was not significantly different (P>0.05), but the difference of gene frequen?cy was statistically significant (P<0.05). There were no significant differences in rs1630310 and rs697372 locus genotype and gene frequency in two groups of Han group. Conclusion Cx37 gene polymorphism is associated with the occurrence of EHT in Xinjiang Kazak population, which may be related with the rs1630310 and rs697372 polymorphism.
9.PET/CT imaging and detection of levels of IL-6 and expression rates of ApoE in Alzheimer's disease patients with different body-mass indexes and their clinical significances
Guoxiu LU ; Guoxu ZHANG ; Shanhu HAO ; Jia GUO ; Wenwen ZHANG ; Jinghua ZHANG ; Zhiguo WANG
Journal of Jilin University(Medicine Edition) 2017;43(3):611-616
Objective:To detect the interleukin-6 (IL-6) levels and expressions of applipoprotein E (ApoE) in the Alzheimer's disease (AD) patients with different body-mass indexes (BMI),and to explore the diagnotic value of 11Pittsburgh comound-B (11C-PIB) combined with 18F-fluorodeoxyglucose (18F-FDG) PET/CT imaging in AD.Methods:A total of 58 AD patients were divided into four groups according to their BMI:low BMI group(BMI <18.5 kg·m-2,n=18),normal BMI group(18.5 kg·m-2≤BMI <24.9 kg·m-2,n=13),high BMI group(24.9 kg·m-2≤BMI <29.9 kg·m-2,n=12)and obese group(BMI ≥29.9 kg·m-2,n=15).All the patients underwent PET/CT imaging (11C-PIB and 18F-FDG).The sensitivity,specificity and accuracy rate,the expression rates of ApoE (ε2,ε3,and ε4) and the levels of serum IL-6 were detected.The relationship between BMI and the expression rates of ApoE and the serum levels of IL-6 were analyzed by Spearman analysis.Results:The sensitivity,specificity,and accuracy rate of the patients in low BMI group diagnosed by 11C-PIB and 18F-FDG PET/CT were 87.5%,80.0%,and 84.6%,which were higher than those diagnosed by 11C-PIB (55.6%,50.0%,and 53.8%) or 18F-FDG (42.9%,50.0%,and 46.2%)alone (P<0.05).The serum levels of IL-6 and BMI of the AD patients had a negative correlation(r=-0.407,P=0.002).The expression rate of ApoE ε4 allelic gene(60.3%) of the AD patients was higher than those of ε2(18.9%) and ε3 allelic genes(20.7%),but there was no correlation between the BMI and the expression rates of different ApoE allelic genes of the AD patients(r=-0.028,P=0.833).Conclusion:11C-PIB and 18F-FDG PET/CT has a high diagnotic value in the AD patients.11C-PIB and 18F-FDG combinated with serum IL-6 level and BMI could diagnose and evaluate AD more exactly.
10.Pathogenic role of Bcl-6/Blimp-1/IL-21 in primary Sj?gren′s syndrome
Chunqiao DUAN ; Wenwen YANG ; Lingyan LEI ; Huifang GUO ; Mingfeng ZHANG ; Lixia GAO ; Chao SUN
Chinese Journal of Immunology 2015;(3):384-388,392
Objective:To analyse the role of Bcl-6/Blimp-1/IL-21 in pathogenesis of primary Sj?gren′s syndrome(PSS),the relationship between Bcl-6/Blimp-1/IL-21 expression and labial gland biopsy grading.Methods:Immunohistochemical(IHC) method was used to detect the expression of Bcl-6/Blimp-1 in salivary gland between 30 cases pations with PSS in disease group and 11 cases patients with mucous cyst,lower lip trauma in control group,and the serum IL-21 levels between disease group of 40 cases patients with PSS and 30 cases healthy volunteers were measured by enzyme-linked immunosorbent assay( ELISA) ,relations among 15 patients with IL-21 expression levels with labial gland biopsy grading.Results:The expression levels of Bcl-6/Blimp-1/IL-21 in disease group were higher than control group;in disease group,with pathological grades increased,the expression levels of Bcl-6/Blimp-1/IL-21 were also raised.Conclusion:①Bcl-6/Blimp-1/IL-21 may participate in the pathogenesis of PSS;Bcl-6/Blimp-1/IL-21 is associated with infiltration lymphocytes of salivary gland.