1.Effect of miR-5590-3p on the invasion and proliferation of gastric cancer cell line HS-746T by regulating TGFBR2 expression
Mingming DONG ; Geng HUANG ; Haiping LUO ; Xinming LI ; Youneng YUAN
Journal of Chinese Physician 2021;23(3):384-388
Objective:To explore the inhibition of microRNA (miRNA, miR)-5590-3p on the expression of transforming growth factor beta typeⅡreceptor (TGFBR2) gene and its effect on the invasion and proliferation of gastric cancer cell line HS-746T.Methods:The gastric cancer cell line was cultured in vitro. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to analyze the expression of miR-5590-3p in gastric cancer tissues and cell lines. The miR-NC and miR-5590-3p mimic were transfected into gastric cancer cell line HS-746T by lipofection, and named as miR-NC group and miR-5590-3p group, respectively. qRT-PCR was used to measure transfection effects. Transwell assay and cell counting kit-8 (CCK-8) assay were used to detect cell invasion and proliferation after transfection. The bioinformatics software predicts and the dual luciferase reporter gene system validates the target gene of miR-5590-3p. qRT-PCR and western blot were used to measure the expression of TGFBR2 and its downstream proteins in the transfected cells. Results:The expression of miR-5590-3p in gastric cancer tissues was significantly lower than that in adjacent tissues ( P<0.01). The expression of miR-5590-3p in gastric cancer cell lines was significantly lower than that in normal gastric mucosal epithelial cells ( P<0.05), and lowest in HS-746T cells ( P<0.01). After transfection, the expression of miR-5590-3p in miR-5590-3p group (11.76±0.21) was significantly higher than that in miR-NC group (1.06±0.21), with statistically significant difference ( P<0.01). The number of invasive cells in miR-NC group and miR-5590-3p group were (101.20±15.47) and (26.53±6.53), respectively, and the invasion ability of miR-5590-3p group was significantly decreased ( P<0.01). Compared with the miR-NC group, the cell proliferation ability of the miR-5590-3p group was significantly decreased ( P<0.05). Bioinformatics software showed that the target gene for miR-5590-3p is TGFBR2. The dual luciferase reporter gene system confirmed that miR-5590-3p can target the TGFBR2 gene ( P<0.01). Western blot results showed that compared with miR-NC group, the expression of TGFBR2 in HS-746T cells in miR-5590-3p group was significantly decreased ( P<0.01), the expression of ZEB-1 and ZEB-2, and the expression of CDK1 and cyclin B proteins were decreased as well. Conclusions:miR-5590-3p can inhibit the invasion and proliferation of gastric cancer cell HS-746T by targeting and regulating the expression of TGFBR2 gene.
2.Risk factors related to postoperative death in patients with rectosigmoid junction tumor perforation
Xinming LI ; Shiqiang SHEN ; Youneng YUAN ; Nian LING ; Mingming DONG
Chinese Journal of General Surgery 2017;32(4):297-300
Objective To investigate risk factors related to postoperative death of patients with rectosigmoid junction tumor perforation.Methods The clinical data of 76 cases with rectosigmoid junction tumor perforation confirmed by laparotomy from January 2000 to October 2015 were collected.Results Of the 76 cases,17 patients died postoperatively,the mortality rate was 22%,the single factor analysis showed that age(x2 =4.649,P =0.031),duration of abdominal pain(x2 =8.218,P =0.016),severe heart and lung diseases(x2 =11.996,P =0.007),circulatory and renal function(x2 =10.360,P =0.016),serum albumin(x2 =7.252,P =0.027),white blood cell count(x2 =7.633,P =0.022),Perforation diameter (x2 =9.770,P =0.008),Geroge grade of intraperitoneal contamination (x2 =10.086,P =0.006) were related to postoperative death (P < 0.05).Multivariate analysis showed that complicating severe heart and lung diseases,preexisted circulatory and renal dysfunction,white blood cell count < 4 × 109/L,size > 3 cm,intraperitoneal contamination larger than one quadrant were independent risk factors for postoperative death.Conclusion Risk factors related to postoperative death of rectosigmoid junction tumor perforation were preoperative important organ dysfunction and intraperitoneal infection.
3.Application of a self-assembly adjustable traction frame for patients with severe trauma and ;fracture of lower limb in ICU
Yinglan DONG ; Mingming WANG ; Xiaochun LIU ; Qiaoyun ZHANG ; Jianfang WEI
Chinese Journal of Practical Nursing 2016;32(32):2514-2517
Objective To introduce a self- assembled adjustable traction frame for the patients with severe trauma and lower limb fracture in ICU, compared with Brawns traction frame. Methods 96 patients with severe trauma and lower limb fracture diagnosed by clinical signs and X-ray who had to be transferred to ICU were randomly divided into the study group (46 cases) and the control group (50 cases) by sealing letters method.The study group were tracted by the self-assembled adjustable traction frame, and the control group were tracted by the Brawns traction frame. The fracture alignment, degree of pain, incidence of pressure ulcers, nursing limit time of changing the sheets of the two groups were observed. Results The results declared that 37 cases with 2 pain score points, 9 cases with 0-1 points in the study group and 19 cases with 2 pain score points, 3 cases with 0-1 points, 28 cases with Grade 3 in the control group (χ2=38.683, P<0.01). In the control group, pressure ulcer were occurred, 5 of them at Level one, 1 of them at Level two, 1 of them at Level three and the incidence rate was 14.00% (7/50). In the study group, pressure ulcer were occurred, 1 of them at Level two and the incidence rate was 2.17%(1/46). The pain level and incidence of pressure sore in the study group were decreased and the difference between the two groups was significant (χ2=8.197, P < 0.05). Conclusions The self-assembly adjustable traction rack can not only ensure the same effect as the Brawns traction frame but also reduce the deficiency caused by Brown traction frame. It is safer and more comfortable for the patients and can reduce nursing complications and improve the quality and efficiency of the nursing work.
4.Study on alexithymia and its affecting factors in continuous ambulatory peritoneal dialysis patients
Lei LIU ; Yiming ZHANG ; Mingming WEI ; Dong WANG ; Xinjian LI
Chinese Journal of Behavioral Medicine and Brain Science 2015;24(8):710-714
Objective To evaluate the alexithymia in the continuous ambulatory peritoneal dialysis (CAPD) patients in our peritoneal dialysis center,and determine the factors affecting alexithymia in CPAD patients.Methods The patients with end-stage renal disease treated by CAPD were divided into alexithymia group (total score≥61)and non-alexithymia group(total score≤51),according to the Toronto Alexithymia Scale(TAS-20).Risk factors for alexithymia were analyzed by Logistic regression statistical model.Results (1)The TAS-20 score was(55.8±9.2) in 86 cases of patients with CAPD.Among them,TAS-20 scores of 38 cases were ≥61 points with an average score (64.5 ±4.4),and the scores of 30 cases were ≤ 51 points with an average score (48.9±5.3),which had a significant difference(t=10.1,P<0.01).(2) There were significant differences in the pay-way and the residual urine volume between the two groups(P<0.01).There were significant correlations of alexithymia with the pay-way and the residual urine volume(P<0.01).(3) The level of serum albumin was significantly lower in alexithymia groups than that in non-alexithymia(P<0.01).The level of serum calcium,phosphorus and parathormone were significantly higher in alexithymia group than that in non-alexithymia group(P<0.05).There were significant negative correlations of alexithymia with the level of serum albumin(P<0.01) and positive correlations of alexithymia with the level of serum calcium,phosphorus and parathormone(P<0.05).(4) There were significant differences score in SF-36 total score,Physical Function,Bodily Pain,General Health,Vitality,Social Function,Role-Emotional and Mental Health (P< 0.01);There were significant negative correlations of alexithymia with SF-36,Physical Function,Bodily Pain,General Health,Vitality,Social Function,Role-Emotional and Mental Health (P< 0.05).(5) The Kt/v was 1.65±0.47 in alexithymia group and 2.13±0.69 in non-alexithymia group.There were significant differences in the two groups(t=2.5,P=0.017).There were significant negative correlations of alexithymia with Kt/v(r=-0.417,P=0.007)by Pearson correlation analysis.Logistic regression analysis showed that the level of serun albumin,SF-36 scores,residual urine volume,the pay-way and Kt/v were the risk factors for alexithymia.Conclusion The incidence rate of alexithymia is higher.The level of serum albumin,SF-36 scores,residual urine volume,the pay-way and Kt/v are the risk factors for alexithymia.
5.Replication Kinetics of Coxsackievirus A16 in Human Rhabdomyosarcoma Cells
Jun JIN ; Mingming HAN ; Lin XU ; Dong AN ; Wei KONG ; Chunlai JIANG
Virologica Sinica 2012;27(4):221-227
Coxsackievirns A16(CVA16),together with enterovirus type 71(EV71),is responsible for most cases of hand,foot and mouth disease(HFMD) worldwide.Recent findings suggest that the recombination between CVA16 and EV71,and the co-circulation of these two viruses may have contributed to the increase of HFMD cases in China over the past few years.It is therefore important to further understand the virology,epidemlology,virus-host interactions and host pathogeuesis of CVA16.In this study,we describe the viral kinetics of CVAI6 in human rhabdomyosarcoma(RD) cells by analyzing the cytopathic effect(CPE),viral RNA replication,viral protein expression,viral RNA package and viral particle secretion in RD cells.We show that CVA16 appears to first attach,uncoat and enter into the host cell after adsorption for 1 h.Later on,CVA16 undergoes rapid replication from 3 to 6 h at MOI 1 and until 9 h at MOI 0.1.At MOI 0.1,CVA16 initiates a secondary infection as the virions were secreted before 9 h p.i.CPE was observed after 12 h p.i.,and viral antigen was first detected at 6 h p.i.at MOI 1 and at 9 h p.i.at MOI 0.1.Thus,our study provides important information for further investigation of CVA16 in order to better understand and ultimately control infections with this virus.
6.A preliminary study on chemical bile duct embolization for chemical hepatectomy
Fuyu LI ; Sheng HE ; Ning LI ; Jingqiu CHENG ; Jiahong DONG ; Mingming ZHANG ; Lisheng JIANG ; Nansheng CHENG
Chinese Journal of General Surgery 1997;0(04):-
Objective To investigate the feasibility, effect and mechanism of chemical bile duct embolization for chemical hepatectomy. Methods Phenol or absolute ethanol plus cyanonacrylate were employed as embolization agents. Histology, Fas and TGF-? 1 measurement were used to evaluate the result. Results Phenol plus cyanonacrylate effectively destroyed and embolized intrahepatic biliary duct, leading to complete disappearance of hepatocytes in the periphery of embolization lobe and thereby achieving the effect of chemical hepatectomy. Expression of Fas and TGF-? 1 in phenol embolism group (88.90?38.10, 185.22?70.39) and ethanol embolism group (72.39?29.51, 163.56?51.75) were higher than those in biliary duct ligated group (26.31?12.07, 74.84?40.73) ( P
7.Effects of Qingfei Oral Liquid on the expressions of IL-10 and IL-17 in the lung tissue and those of Treg and Th17 in the spleen of RSV-infected mice
Wenge DONG ; Bin YUAN ; Lihua ZHOU ; Jianya XU ; Jiangquan LI ; Mingming WANG ; Fei KONG
Journal of Medical Postgraduates 2015;(12):1242-1245
Objective The action mechanisms of Qingfei Oral Liquid ( QFOL) in the treatment of respiratory syncytial virus ( RSV) infection need to be studied more deeply.The aim of this study is to examine the expressions of interleukin ( IL)-10 and IL-17 in the lung tissue and those of Treg and Th17 in the spleen tissue of RSV-infected mice before and after treated with QFOL, and to explore the action mechanisms of QFOL from the perspective of the Treg/Th17 cy-tokines balance. Methods Fifty BABL/c mice were equally ran-domized to five groups: blank control, RSV model, Ribavirin, low-dose QFOL, and high-dose QFOL.Models of RSV ( long strain) infec-tion were made in the latter four groups.At 48 hours after viral activa-tion, the mice of the control and RSV model groups were treated intragastrically with 0.9%normal saline and those in the Ribavirin and QFOL groups with Ribavirin at 0.0025 g/mL and QFOL at 1.33 g/mL and 4 g/mL, respectively, all for 72 hours.Then all the mice were killed and the lung tissue harvested from 5 animals in each group for pathological analysis, while the levels of IL-10 and IL-17 in the bronchoalveolar lavage fluid of the other 5 detected by ELISA.The expressions of the cytokines Treg and Th17′in the spleen from 4 mice in each group were determined by flow cytometry. Results Compared with the RSV models, pathologic changes were significantly re-duced in the mice of the QFOL, Ribavirin and control groups (P<0.01), the expression of IL-10 remarkably up-regulated in the low-dose QFOL, high-dose QFOL, Ribavirin, and control groups ([39.21 ±1.57] vs [43.54 ±1.03], [46.64 ±0.48], [47.83 ±0.87], and [50.44 ±1.04] ng/L, all P<0.01), while the level of IL-17 markedly down-regulated ([70.96 ±0.53] vs [55.92 ±0.83], [33.66 ±0.70], [21.92 ±1.38], and [9.42 ±0.59] pg/mL, all P<0.01).The expressions of Treg/Th17′were significantly in-creased in the low-dose QFOL, high-dose QFOL, Ribavirin, and control groups (2.89 ±0.52, 6.38 ±0.36, 3.95 ±0.26, and 3.54 ± 0.85) as compared with that in the RSV models (0.96 ±0.16) (all P<0.01).Both low-and high-dose QFOL groups showed statisti-cally significant differences from the Ribavirin group in the levels of Treg, Th17, and Treg/Th17 (P<0.05). Conclusion QFOL can regulate the balance of Treg/Th17, increase the expression of IL-10 and decrease that of IL-17 in the lung tissue of RSV-infected mice, which further proves the efficacy of QFOL in the treatment of RSV-induced pneumonia.
8.Effect of Acupuncture on Serum TNF-?and IL-6 Contents in Cerebral Hemorrhage Rats
Qiuxin CHEN ; Wei ZOU ; Mingming NIU ; Peng LIU ; Dong YAO ; Yu BAO ; Xiaowei SUN
Shanghai Journal of Acupuncture and Moxibustion 2015;(10):1006-1008
ObjectiveTo investigate the effect of acupuncture on motor function and serum expressions of TNF-aand IL-6 contents in cerebral hemorrhage rats.MethodSeventy-two male Wistar rats were randomized into sham operation, model and acupuncture groups. A rat model of cerebral hemorrhage was made by intracerebral injection of autoblood. Baihui-to-Qubin acupuncture was performed after successful model making. The TNF-aand IL-6 contents of rat serum were measured by enzyme linked immunosorbent assay at three time points: one, three and seven days.ResultBaihui-to-Qubin acupuncture markedly increased the neurological function score and reduced neurologic deficits in cerebral hemorrhage rats. At the same time point, serum TNF-aand IL-6 contents were significantly lower in the acupuncture group of rats thanin the model and acupuncture group; there were statistically significant differences (P<0.01).ConclusionBaihui-to-Qubin acupuncture can reduce neurologic deficits in cerebral hemorrhage rats. It may be related to decreases in serum TNF-aand IL-6 contents after acupuncture.
9.Clinicopathologic features and prednisone therapeutic effect of 50 cases of proliferative sclerosing IgA nephropathy
Mingming MA ; Baozhang GUAN ; Lihua LUO ; Bo HU ; Xiangnan DONG ; Lianghong YIN
Chinese Journal of Nephrology 2016;32(8):568-572
Objeetive To analyze the clinicopathologic features of proliferative sclerosing IgA nephropathy and the efficacy of prednisone therapy.Methods A retrospective analysis was conducted,enrolling 50 patients with biopsy-proven primary proliferative sclerosing IgA nephropathy who were admitted in the Hospital from January 2005 to June 2015-26 males and 24 females,mean age (36.8 ± 10.4) years.Clinicopathologic features and prednisone therapeutic effect were analyzed.Results The clinical manifestations of 50 cases were nephritis syndrome with varying degrees of renal insufficiency,including 32 cases (64.0%) with hypertension,15 cases (30.0%) with microscopic hematuria.Renal biopsy showed the incidence of glomerular global sclerosis was 17.0%-47.2%,tubular atrophy/ interstitial fibrosis outstanding (T0 50%,T1 32%,T2 18%).After prednisone treatment,compared with sustained remission group and relapse group,invalid patients had higher incidence of hypertension (P < 0.05),relatively lower Hb (P < 0.01) and serum albumin,more significant renal dysfunction (P < 0.01),more severe glomerular global sclerosis,segmental sclerosis,tubular atrophy/ interstitial fibrosis,while the lower interstitial inflammatory cell infiltration.During the follow-up,which lasted from 6 to 132 months (median 27.3 months),the effective rate of treatment was 74.0% after sufficient prednisone or half dose prednisone therapy.Repeated recurrence rate was 32.0%.At the end of the follow-up period,13(26.0%) patients entered the stage of uremia.Conclusions Application of glucocorticoids in the treatment of proliferative sclerosing IgA nephropathy can protect renal function and delay the progression of renal impairment.The efficacy of glucocorticoids therapy is significantly associated with the presence or absence of hypertension,the degree of renal function impairment,and the severity of the onset of renal pathology.
10.The effects of nourishing lung and kidney formulas on inflammatory response of alveolar epithelial cells stimulated by monocytes conditioned medium
Yanqin QIN ; Haoran DONG ; Jiansheng LI ; Yulong CHEN ; Xiaoning MAO ; Mingming WU
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2017;24(1):63-67
Objective To observe the effects of nourishing lung and kidney formulas on inflammatory response of alveolar epithelial cells stimulated by monocytes conditioned medium and study the anti-inflammatory mechanism of the formulas for the treatment of chronic obstructive pulmonary diseases (COPD).Methods The reproduction of inflammation models of A549 cells were stimulated by monocyte THP-1 cell strain conditioned medium.A549 cells were randomly divided into blank control group (20% blank rabbits serum),model group (20% blank rabbits serum+25.0% THP-1 cell conditioned medium),nuclear transcription factor activator protein-1 (AP-1) pathway inhibitor T-5224group (20% blank rabbits serum+25.0% THP-1 cells conditioned medium+100 μmol/L T-5224),nourishing lung and kidney group (20% rabbits serum with nourishing lung and kidney formulas+25.0% THP-1 cells conditioned medium).The contents of interleukins (IL-6,IL-8),tumor necrosis factor-α (TNF-αα),matrix metalloprotein-9 (MMP-9) in cell culture supernatant were detected with enzyme linked immunosorbent assay (ELISA),the supernatant content of malondialdehyde (MDA) was detected with thibabituric acid (TBA) method,the total activity of superoxide dismutase (T-SOD) was detected with hydroxylamine method,and the activity of AP-1 pathway was detected with electrophoretic mobility shift assay (EMSA) method.Results Compared with the blank control group,the A549 cell proliferation were significantly increased at 24 hours,48 hours stimulation by 25.0% cell conditioned medium (A value:24 hours was 0.41 ± 0.02 vs.0.37 ± 0.04,48 hours was 1.30 ± 0.09 vs.1.15 ± 0.19).Compared with the blank control group,the contents of IL-6,IL-8,TNF-αα,MMP-9,MDA,AP-1 expression were significantly increased in model group [IL-6 (ng/L):35.00±3.63 vs.23.15±1.72,IL-8 (ng/L):273.09± 164.36 vs.231.45±33.90,TNF-α(ng/L):51.61 ± 9.51 vs.28.87 ± 3.34,MMP-9 (ng/L):442.85 ± 78.86 vs.235.60 ± 14.62,MDA (μmol/L):6.90 ± 0.11 vs.6.01 ± 0.12,AP-1 expression (A value):2.260 ± 0.062 vs.1.000 ± 0.000],MDA/T-SOD ratio was increased (4.43 ± 0.05vs.3.96 ± 0.06).Compared with model group,the levels of IL-8 (ng/L:100.29 ± 17.03),TNF-α (ng/L:25.13 ± 0.46),AP-1 expression (A value:1.38 ± 0.02),and the MDA/T-SOD ratio (4.23 ± 0.23) in T-5224 group,and MMP-9 (ng/L:195.44±9.80),MDA (μmol/L:5.86±0.30),MDA/T-SOD ratio (3.56±0.41),AP-1 expression (A value:0.76 ± 0.01) in nourishing lung and kidney group were all reduced significantly (all P < 0.05).Conclusion Nourishing lung and kidney formulas can suppress the inflammatory response through regulating the alveolar epithelial cells AP-1 signaling pathways.