1.Effects of silencing STIL by RNA interference on biological behaviors of gastric cancer cell line
Ju WANG ; Yong ZHANG ; Xiaoping GAO
International Journal of Surgery 2015;42(10):684-688
Objective To investigate the expression pattern of STIL in gastric cancer and effect of silencing STIL on biological behaviors of SGC-7901 cell line.Methods The expression of STIL at mRNA and protein level were detected by qPCR and Western Blot,respectively.SiRNA against STIL was constructed and transfected into SGC-7901 cell line.The effect of Si-STIL on cell proliferation,cloning ability,cycle distribution and apoptosis was detected by MTT,colony-forming assay and flow cytometry,respectively.Results STIL was aberrantly expressed in gastric cancer compared with that in paratumor tissues.Downregulation of STIL level,inhibition of proliferation and cloning ability,increasing apoptosis and blocked cell cycle at S phase were observed after SGC-7901 cell line was treated with Si-STIL.Conclusion STIL was overexpressed in gastric cancer.STIL gene silencing effectively inhibited cell growth,transition of metaphase and promoted apoptosis of SGC-7901 cell line.
2.Role of syndecan-4 in inflammatory responses of rats with ventilator-induced lung injury
Yali GE ; Ju GAO ; Cunjin WANG
Chinese Journal of Anesthesiology 2017;37(6):666-669
Objective To evaluate the role of syndecan-4 (SDC-4) in inflammatory responses of rats with ventilator-induced lung injury (VILI).Methods Twenty-four healthy male wild type Sprague-Dawley rats and 24 male SDC-4 gene knockout Sprague-Dawley rats,aged 2-3 months,weighing 200-220 g,were assigned into 2 groups (n =12 each) using a random number table:control group (group C)and VILI group.The animals were anesthetized with pentobarbital sodium and tracheostomized.The rats kept spontaneous breathing in group C.The rats were mechanically ventilated for 4 h in group VILI.Blood samples were taken from the femoral artery at the end of mechanical ventilation for detection of arterial oxygen partial pressure (PaO2).The animals were sacrificed after blood sampling.The left lung was lavaged,and the broncho-alveolar lavage fluid (BALF) was collected for determination of the tumor necrosis factor-α (TNF-α),interleukin-1beta (IL-1β) and IL-18 concentrations (by enzyme-linked immunosorbent assay) and total protein concentrations (by BCA assay).The right lungs were removed for determination of the wet to dry weight ratio (W/D ratio) and expression of TNF-α,IL-1β and IL-18 mRNA in lung tissues (by real-time polymerase chain reaction) and for examination of the pathological changes (with a light microscope).The lung injury scores were recorded.Results Compared with group C of wild type rats,PaO2 was significantly decreased,W/D ratio and lung injury scores were increased,the concentrations of total protein,TNF-α,IL-1β and IL-18 in BALF were increased,and the expression of TNF-α,IL-1β and IL-18 mRNA in lung tissues was up-regulated in group VILI of wild type rats (P<0.05),and no significant change was found in the variables mentioned above in group C of gene knockout rats (P>0.05).Compared with group C of gene knockout rats,PaO2 was significantly decreased,W/D ratio and lung injury scores were increased,the concentrations of total protein,TNF-α,IL-1β and IL-18 in BALF were increased,and the expression of TNF-α,IL-1β and IL-18 mRNA in lung tissues was up-regulated in group VILI of gene knockout rats (P<0.05).Compared with group VILI of wild type rats,PaO2 was significantly decreased,W/D ratio and lung injury scores were increased,the concentrations of total protein,TNF-α,IL-1β and IL-18 in BALF were increased,and the expression of TNF-α,IL-1β and IL-18 mRNA in lung tissues was up-regulated in group VILI of gene knockout rats (P<0.05).Conclusion SDC-4 can inhibit inflammatory responses of rats with VILI and is involved in the endogenous protective mechanism.
3.Effect of selective gut decontamination in regulation of inflammatory reaction in patients with systemic ;inflammatory response syndrome
Youli WANG ; Shuqiang LU ; Ju GAO ; Zhichao WANG ; Aibing CAI
Chinese Critical Care Medicine 2015;(6):484-488
Objective To explore the effect of selective gut decontamination in regulation of inflammatory reaction compared with rhubarb and glycerine enema for catharsis in patients with systemic inflammatory response syndrome ( SIRS ), and to discuss its mechanisms. Methods A prospective randomized controlled trial was conducted. Fifty-seven patients with SIRS admitted to Department of General Surgery of Aviation General Hospital from June 2009 to June 2014 were enrolled. The patients were randomly divided into rhubarb decontaminate group, traditional decontaminate group and blank control group, with 19 cases in each group. Besides the treatment for primary disease, including anti-infection, operation, alleviate pain, nutritional support, and maintaining water and electrolyte balance, the patients in rhubarb decontaminate group received aqueous extract from rhubarb 15-20 g by gastric tube, enema, or peros, twice a day;and those in traditional decontaminate group received glycerine enema or glycerol enema, twice a day; while no gavage or enema was prescribed in blank control group. Peripheral blood was collected before and 72 hours after treatment. Enzyme linked immunosorbent assay ( ELISA ) was used to determine the concentration of lipopolysaccharide ( LPS ) and inflammatory mediators. Results Compared with blank control group and traditional decontaminate group, the levels of interleukins ( IL-1, IL-8 ), LPS, platelet activating factor ( PAF ), tumor necrosis factor-α( TNF-α), andγ-interferon ( IFN-γ) before treatment was similar to that of rhubarb decontaminate group [ IL-1 ( ng/L ): 53.154±5.783, 50.564±5.771, 51.082±6.403, F = 0.994, P = 0.377; IL-8 ( ng/L ): 70.492±6.146, 68.376±6.112, 68.673±8.384, F=0.514, P=0.601;LPS (μg/L ):11.630±2.449, 10.858±2.307, 10.463±2.145, F = 1.261, P = 0.291; PAF (μg/L ): 4.173±0.395, 4.051±0.362, 4.078±0.487, F = 0.446, P = 0.642; TNF-α( ng/L ):132.498±10.772, 129.735±12.881, 127.207±11.514, F=0.963, P=0.388;IFN-γ(μg/L ):45.645±4.558, 43.692±5.578, 43.767±5.028, F = 0.904, P = 0.411 ]. The above parameters after treatment were significantly lower than those before treatment in three groups. The effect on the LPS and pro-inflammatory factors of the rhubarb decontaminate group was more obvious than that of the blank control group and traditional decontaminate group [ LPS (μg/L ): 7.571±1.113 vs. 9.008±1.904, 8.874±1.808, F = 4.416, P = 0.017; IL-1 ( ng/L ): 45.309±3.563 vs. 48.731±4.466, 46.112±4.322, F = 3.557, P = 0.035; IL-8 ( ng/L ): 60.492±5.346 vs. 65.553±5.384, 63.437±5.462, F = 4.213, P = 0.020; PAF (μg/L ): 3.519±0.250 vs. 3.832±0.356, 3.766±0.309, F = 5.450, P = 0.007; TNF-α ( ng/L ): 114.988±8.772 vs. 123.230±10.433, 118.534±9.519, F = 3.525, P = 0.036; IFN-γ(μg/L ):38.683±3.190 vs. 41.831±4.122, 39.161±3.972, F=3.820, P=0.028 ]. Conclusion The usage of selective gut decontamination can inhibit the release of endotoxin and inflammatory mediators in patients with SIRS, and it will get a better effect using rhubarb, and the mechanism may be related to the protection of intestinal mucosal barrier function.
4.Study on relationship between polymorphism of TNF-?and IgA nephropathy of Han nationality Chinese in inner Mongolia Autonomous Region
Cheng WANG ; Caili WANG ; Lijun GAO ; Tianlin JU ; Wenbin QIN
Chinese Journal of Clinical Laboratory Science 2006;0(03):-
Objective To study the relationship of tumor necrosis factor beta (TNF-?) polymorphism with human IgA nephropathy of Chinese Han nationality in Inner Mongolia Autonomous Region. Methods The A→G single base mutation polymorphism in TNF-?1096 locus were analyzed among 80 normal controls and 79 IgA nephropathy patients by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results The genotype frequencies of TNF?* 1/1 and TNF?* 2/2 in IgA nephropathy patients were significantly higher than that in normal controls (x1/12=5.58,P
5.Mixed culture of Madin-Darby canine kidney cells, human epidermoid cancer cells and African green monkey kidney cells for detection of common respiratory viruses and enteroviruses
Yingyang GAO ; Chenyan JIANG ; Lufang JIANG ; Qianli WANG ; Liwen JU
Chinese Journal of Infectious Diseases 2011;29(6):321-324
Objective To establish a clinical test assay for detecting common respiratory viruses and enteroviruses (EV) by using mixed cultured Madin-Darby canine kidney cells (MDCK), human epidermoid cancer cells (Hep-2) and African green monkey kidney cells (Vero) to isolate common respiratory viruses and enteroviruses. Methods Throat swabs with influenza A and B viruses,adenovirus and EV71 were incubated with mixed cultured MDCK, Hep-2 and Vero in a single vial to observe the presence of cytopathic effects. Polymerase chain reaction (PCR), reverse transcription (RT)-PCR and monoclonal antibody-based immunofluorescene assay were also used for confirmatory test. Results The sensitive cell lines developed obvious cytopathic effects to the corresponding viruses, which were confirmed by the specific green particles observed by immunofluorescence assay and specific target PCR segments. Conclusions The shell-vial of mixed cells can simultaneously isolate different common respiratory viruses and EV. The isolated pathogens can be further confirmed by antigen test and PCR. This assay may improve the diagnosis of clinical viral diseases.
6.Effects of mechanical stretch preconditioning on pathological stretch-induced activation of NF-κB and STAT3 signaling pathways in human type Ⅱ alveolar epithelial cells
Xiangzhi FANG ; Tianfeng HUANG ; Yang ZHANG ; Cunjin WANG ; Ju GAO
Chinese Journal of Anesthesiology 2015;35(8):1003-1006
Objective To evaluate the effects of mechanical stretch preconditioning on pathological stretch-induced activation of nuclear factor kappa B (NF-κB) and signal transducer and activator of transcription 3 (STAT3) signaling pathways in human type Ⅱ alveolar epithelial cells.Methods Human type Ⅱ alveolar epithelial cell line A549 cells cultured in vitro were randomly divided into 3 groups (n =24 each) using a random number table: control group (group Ⅰ), pathological stretch group (group Ⅱ), and mechanical stretch preconditioning group (group Ⅲ).In group Ⅰ , A549 cells were cultured routinely without receiving pathological stretch.In group Ⅱ , A549 cells were exposed to 20% cyclic stretch at 0.3 Hz for 6 h.In group Ⅲ , A549 cells were exposed to 5% cyclic stretch at 0.3 Hz for 60 min, and then exposed to 20% cyclic stretch at 0.3 Hz for 6 h.After the end of the treatment, the cells were collected for determination of the cell viability (by methyl thiazolyl tetrazolium assay) and lactate dehydrogeuase (LDH)release (by colorimetric method).The concentrations of tumor necrosis factor-alpha (TNF-α),interleukin-8 (IL-8) and high mobility group box 1 (HMGB1) in the culture medium were detected using enzyme linked immunosorbent assay.The expression of total NF-κB, phosphorylated NF-κB, total STAT3 and phosphorylated STAT3 was detected using Western blot.The ratios of phosphorylated NF-κB to total NF-κB and phosphorylated STAT3 to total STAT3 were calculated to reflect the activation.Results Compared with group Ⅰ , the cell viability was significantly decreased, the amount of LDH released was increased, and the concentrations of TNF-α, IL-8 and HMGB1, and activation of NF-κB and STAT3 were increased in Ⅱ and Ⅲ groups.Compared with group Ⅱ , the cell viability was significantly increased, the amount of LDH released was decreased, and the concentrations of TNF-α, IL-8 and HMGB1, and activation of NF-κB and STAT3 were decreased in group Ⅲ.Conclusion The mechanism by which mechanical stretch preconditioning attenuates pathological stretch-induced damage to human type Ⅱ alveolar epithelial cells is related to inhibited activation of NF-κB and STAT3 signaling pathways.
7.Role of spinal RhoA/ROCK signaling pathway in development of lipopolysaccharide-induced inflammatory pain in rats
Cunjin WANG ; Shuangming KONG ; Ju GAO ; Yali GE ; Xiangzhi FANG
Chinese Journal of Anesthesiology 2015;35(8):969-971
Objective To evaluate the role of spinal RhoA/ROCK signaling pathway in the development of lipopolysaccharide (IP)-induced inflammatory pain(IP) in rats.Methods Fifty-two male Sprague-Dawley rats, weighing 180-220 g, were equally randomized into 4 groups using a random number table: normal saline group (group NS) , LPS group, RhoA inhibitor C3 exoenzyme group (group LC) , and ROCK inhibitor Y27632 group (group LY).Inflammatory pain was induced by injecting LPS 25 μl (300 ng) into the plantar surface of hindpaws in IP, LC and LY groups, while the equal volume of normal saline was injected instead in NS group.C3 exoenzyme 10 pg and Y27632 10 nmol were injected intrathecally at 30 min prior to LPS administration in LC and LY groups, respectively.Before LPS injection (T0) , and at 1, 3, 5, 12 and 24 h after LPS injection (T1-5) , the mechanical and thermal pain thresholds were measured.Five rats in each group were sacrificed after pain thresholds were measured at T3, and L4.5 segments of the spinal cord were removed for determination of tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) mRNA expression in spinal dorsal horns by real-time reverse transcriptase-polymerase chain reaction.Results Compared with group NS, the mechanical and thermal pain thresholds were significantly decreased at T2-5in IP, LC and LY groups, and TNF-α and IL-1β mRNA expression was up-regulated at T3 in IP group.Compared with group IP, the mechanical and thermal pain thresholds were significantly increased at T2-5, and TNF-α and IL-1β mRNA expression was down-regulated at T3 in LC and LY groups.Conclusion Spinal RhoA/ROCK signaling pathway is involved in the development of LPS-induced IP in rats.
8.Regulatory effect of TGF-beta 1 antibodies on extracellular matrix reconstruction in rats with lung injury fibrosis
Haiyan WANG ; Chuang HE ; Jianbin XIAO ; Ju GAO
Journal of Medical Postgraduates 2014;(4):368-372
Objective Transforming growth factor-β( TGF-β) can regulate the expressions of matrix metalloproteinase (MMP) and matrix metalloproteinase (TIMP) inhibitors, thus affecting the reconstruction of extracellular matrix (ECM) after lung in-jury.So far, little is known about the regulation of TGF-β1 and MMPs/TIMPs ratio in endotoxin-induced lung injury as well as about the exact mechanisms of ECM reconstruction disorders .This study was to investigate the effects of TGF-β1 antibodies on the expressions of MMP and TIMP inhibitors and lung injury fibrosis in rats with endotoxin-induced acute lung injury . Methods Fifty-six male SD rats were randomly divided into three groups, normal (n=16), LPS (n=20), and LPS+TGF-β1(L+T, n=20).The rats of the LPS and L+T groups received intraperitoneal and intra-tracheal injection of endotoxin to induce lung injury fibrosis , the latter intrave-nously injected with TGF-β1 antibodies previously .The normal rats received intra-tracheal and intra-abdominal injection of the same a-mount of saline.At 1, 5, 9 and 14 days after modeling, all the rats were killed and blood and lung tissue samples obtained to detect the wet/dry lung tissue ( W/D ) ratio, diffuse alveolar damage ( DAD) score, hydroxyproline activity ( Hpy) , and the expressions of TGF-β1 , MMP-2 and TIMP-1. Results The DAD score, Hpy ac-tivity, and expressions of TGF-β1 , MMP-2 and TIMP-1 were signifi-cantly higher in the LPS group than in the normal control ( P <
0.05), and so were the DAD score, Hpy activity, MMP-2, and TIMP-1 in the L+T group than in the controls (P<0.05).Com-pared with the LPS group, Hpy, TGF-β1 and TIMP-1 were markedly decreased in the L +T group (P<0.05).The expression of MMP-2 was gradually increased at 1, 5 and 9 days after modeling , reaching the peak at 9 days and then beginning to decline .The Hpy activity, TGF-β1 and TIMP-1 expressions kept rising after modeling . Conclusion The MMP/TIMP imbalance and ECM reconstruc-tion disorders mediated by the TGF-β1 signaling pathway may be an important cause of fibrosis in endotoxin-induced lung injury . TGF-β1 antibodies can exert a protective effect by alleviating lung injury fibrosis .
9.Laparoscopic surgery for the treatment of achalasia
Hongwei JIANG ; Ju WANG ; Haijun LI ; Jikui PENG ; Xiaoping GAO
Chinese Journal of Postgraduates of Medicine 2014;37(5):8-10
Objective To analyze the complications and short-term effects of laparoscopic surgery for achalasia.Methods The clinical data of 41 patients with achalasia who were treated by laparoscopic surgery were analyzed retrospectively.Results The 41 patients were no deaths.All patients underwent Heller cardiomyotomy and fundoplication,39 patients with Dor anterior fundoplication,2 patients had the presence of hiatal hernia with Toupet fundoplication.The mean surgical time was 142 min.Intraoperative complications occurred in 7 cases,including 6 cases of mild complications.The mean follow-up was 21 months.The clinical efficacy excellent in 27 cases,good in 7 cases,generally in 5 cases,poor in 2 cases.Conclusions Heller cardiomyotomy should be the treatment of choice in achalasia patients,because of its short and medium term outcomes,and its low morbidity.
10.Clinical study on procedure for prolapse and hemorrhoids (PPH) in treating hemorrhoids
Hongwei JIANG ; Ju WANG ; Haijun LI ; Jikui PENG ; Xiaoping GAO
Chinese Journal of Primary Medicine and Pharmacy 2014;21(11):1601-1603,后插1
Objective To observe the clinical effect of procedure for prolapse and hemorrhoids (PPH) in treating hemorrhoids.Methods 78 patients with symptomatic Ⅱ,Ⅲ and Ⅳ degree hemorrhoids undergoing PPH were selected in this study.The diagnosis,medical history,operative time,blood loss and perioperative complications were recorded.The World Health Organization Quality of Life Scale(WHOQOL-BREF) was observed preoperation and 6 hours,24 hours,1 month postoperation.Results The postoperative pain score measured by visual analogue scale (VAS) at postoperative six hours was (7.6 ± 2.1),which reduced to (1.3 ± 1.1) at 24 hours.There was significant improvement in the overall perception of QOL,health,and in physical and psychological domains (t =1.901,1.805,1.714,P =0.041,0.038,0.029,all P < 0.05).There was modest improvement in environmental domain,while no change was found in social domain (P > 0.05).Conclusion PPH surgery achieves good clinical results,improved quality of life of patients,with a low incidence of complications,patients with hemorrhoids should be recommended for this preferred surgical approach.