Objective To observe the expression of zinc finger protein A20(A20), NF-κB and related inflammatory factors before and after lipopolysaccharide (LPS) stimulates degeneration of rabbit intervertebral disc nucleus pulposus cells.Methods The normal and degenerative nucleus pulposus cells were isolated and cultured, then divided into normal group,degenerative group,LPS stimulation group and NF-κB inhibition group.HE staining observe the morphological changes of nucleus pulposus and annulus fibrosus,immunohistochemistry was used to detect the expression of A20,NF-κB/p65 and COL-Ⅱ.Real-time PCR was employed to analyze the expression of A20,IL-1β,TNF-α,NF-κB and COL-Ⅱ,Western blot was used to observe the A20 protein,p65 and COL-Ⅱexpression in the four groups, and TNF-α, IL-1β in cell supernatant was determined by ELISA.Results The number of nucleuspulposus cells significantly decreased, aggregation occured in the degenerative group.COL-Ⅱ was obvious lower and A20, p65 significantly higher than that in normal group by immunohistochemical staining.Compared with the normal group,A20,TNF-α,IL-1β,p65 expression was significantly increased and COL-Ⅱ decreased in the mRNA and protein levels in degenerative group.Above indexes changed more significant in LPS stimulation group than in degenerative group.The expression of A20, TNF-α, IL-1β, p65 in the NF-κB inhibitor group was lower than that in the LPS group, and the expression of type Ⅱ collagen increased(P<0.05).Conclusions Intervertebral disc inflammatory response is closely related to the development of intervertebral disc degeneration, A20 may play an important role.

Objective To assess the feasibility and accuracy of transesophageal echocardiography (TEE) in screening patients,intraoperative guidance of occluder releasing and efficacy assessment in the patients of percutaneous left atrial appendage (LAA) occlusion with Watchman.Methods Ultrasonic instrument Philips iE33(proble model X7-3t) were used to measure the maximum diameter and the depth of LAA ostium in enrolled patients by TEE,and intraoperative guided puncture of interatial septum,positioning Watchman transmission system and instruction closure release under TEE,immediate evaluate the postoperative therapeutic effect and record the complications by TEE.At 45 days after implantation,patients should repeat TEE to assess the efficacy.Results Eighteen patients underwent device implantation.Seventeen patients implant successfully,except one patient can't implant device beacuse of the LAA morphologe was not suitable.There was one patient showing thrombus formation on the surface of device,one patient of LAA and the device axial angulation was 90 °.At 45 days after implantaion,16 patients were fullowed up,there were no major stroke at all.Conclusions TEE has important application value in screening of preoperative patients,intraoperative guidance of occluder releasing and efficacy assessment in the patients of percutaneous left atrial appendage occlusion with Watchman.

Objective:To explore the application value of transesophageal echocardiography (TEE)in secondum atri-al septal defect (ASD)intervention and minimally invasive occlusion surgery in adult patients.Methods:A total of 134 cases adult patients (>15 years)with pure secondum ASD (including 91 cases undergoing interventional occlu-sion and 43 cases undergoing minimally invasive occlusion surgery),who were screened via transthoracic echocardio-graphy (TTE)and TEE from Jan 2012 to Dec 2012,were selected.Intervention group and minimally invasive sur-gery group received TTE and TEE monitoring respectively during operation.Preoperative TTE and TEE parameters and operation results were compared and analyzed.Results:The parameters of ASD diameter measured by TEE were significantly higher than those measured by TTE in two groups (P <0.01 all).Maximum defect diameter of ASD measured by TEE was significantly higher than that of measured by TTE [(19.8±5.2)mm vs.(18.7±4.9)mm], P <0.01. The correlation between maximum defect diameter measured by TTE and occluder size (intervention group r =0.926,minimally invasive surgery group r =0.215)was all lower than that of TEE (intervention group r=0.965,minimally invasive surgery group r =0.627),P <0.01 both.Conclusion:TEE is better than TTE in as-sessment of ASD size.It should be regarded as a routine preoperative screening method for adult ASD patients.It has important value in real-time monitoring,guidance and immediate evaluation of therapeutic effects during mini-mally invasive ASD occlusion surgery.

Objective:To investigate the effects of growth hormone combined with triptoreline acetate on growth and sex hormone in girls with central precocious puberty.Methods:Sixty-two girls with central precocious puberty who received treatment in Xin'an International Hospital from January 2017 to December 2019 were included in this study. They were randomly assigned to receive treatment with either triptorelin acetate (control group, n = 31) or triptorelin acetate plus growth hormone (observation group, n = 31) for 12 successive months. Before and after treatment, bone age difference/chronological age difference (△BA/△CA), body height, body weight, uterine and ovarian volume and sex hormone level were compared between the control and observation groups. Results:△BA/△CA in the observation group was significantly lower than that in the control group [(0.64 ± 0.17) vs. (0.95 ± 0.13), t = 8.065, P < 0.05). Body height and weight in the observation group were (127.32 ± 1.08) cm and (33.42 ± 2.37) kg, respectively, which were significantly higher than those in the control group [(126.34 ± 0.87) cm and (31.01 ± 2.15) kg, t = 3.934 and 4.193, both P < 0.05]. Uterine and ovarian volume in the observation group were (1.68 ± 0.29) cm 3 and (1.26 ± 0.18) cm 3, respectively, which were significantly lower than those in the control group [(2.41 ± 0.46) cm 3 and (1.83 ± 0.26) cm 3, t = 7.474 and 10.036, both P < 0.05). After treatment, there were no significant differences in serum estradiol and luteinizing hormone levels between the two groups (both P > 0.05). Conclusion:Growth hormone combined with triptoreline acetate has a good effect on central precocious puberty in girls because it can improve the growth and development of girls and reduce serum levels of estradiol and luteinizing hormone.

Objective To find the relationship between nuclear factor kappB (NF-κB) activation and cell apoptosis.Methods Degenerative human nucleus pulposus cells were cultured in vitro.Useing CCK-8 to observe the proliferative effect of LPS on the nucleus pulposus cells in vitro, in the concentration of 100, 200, 500 and 1 000 μg/mL and choose the most apropriate concentration.The experiment was divided into blank control group, LPS(500 μg/mL)groups, and NF-κB inhibitor(BAY11-7082)plus LPS(500 μg/mL)group,Annexin V-FITC/PI flow cytometry and Hoechest33258 staining was used to analyze apoptosis.The expression of cleaved caspase-3,PARP,P65,P-P65 proteins were detected by Western blot respectively.Results When LPS was 500 μg/mL, the cell vitality was obviously declined.Compared with the blank control group, cell apoptosis rate of the LPS group is increasing (P<0.05), and the expression of P-P65,cleaved caspase-3, cleaved PARP were alsohigher (P< 0.05).Compared with the LPS group, cell apoptosis rate of the NF-κB inhibitor plus LPS group is significantly lower (P<0.05) and the expression of P-P65,cleaved caspase-3,cleaved PARP were also lower (P< 0.05).Conclusions NF-κB signaling pathway may be associated with the nucleus pulposus cell apoptosis in disc degeneration.

Objective:To investigate the effect of montelukast sodium combined with glucocorticoid on acute attack of children with bronchial asthma and its influence on cytokines and immune function.Methods:A total of 122 children with acute asthma attack admitted to Xin'an International Hospital from September 2018 to September 2019 were divided into observation group(61 cases) and control group(61 cases) according to the random digial table mrethod.The control group was treated with glucocorticoid, and the observation group was treated with montelukast sodium on the basis of the control group.The course of treatment in both two groups was 7 days.The therapeutic effect, lung function, cytokines and immune function of the two groups were compared before and after treatment.Results:The total effective rate of the observation group(91.80%) was higher than 73.77% of the control group (χ 2=6.960, P<0.05). The peak expiratory flow rate(PEFR)[(2.93±0.21)L/s], forced expiratory volume in the first second(FEV 1%)[(82.54±5.37)%] and forced expiratory volume/forced vital capacity in the first second(FEV 1/FVC)[(90.32±4.36)%] in the observation group were higher than those in the control group[(2.48±0.16)L/s, (75.42±3.28)% and (83.98±3.42)%] ( t=13.313, 8.837, 8.936, all P<0.05). The levels of VEGF[(129.83±17.64)ng/L], TGF-β1[(83.21±16.79)ng/L] and IL-6[(24.32±4.19)ng/L] in the observation group were lower than those in the control group [(210.37±25.43)ng/L, (176.48±23.12)ng/L and (48.39±5.47)ng/L] ( t=20.325, 25.494, 27.284, all P<0.05). The CD 3+ [(75.72±3.46)%], CD 4+ [(42.56±3.18)%] and CD 4+ /CD 8+ (1.97±0.19) in the observation group were higher than those in the control group [(66.81±4.80)%, (37.87±2.63)% and (1.62±0.16)] ( t=11.761, 8.877, 11.005, all P<0.05). Conclusion:Montelukast sodium combined with glucocorticoid has obvious therapeutic effect on children with acute attack of asthma.It can reduce inflammatory reaction and enhance immune function, which is worthy of clinical reference.

OBJECTIVESTo examine the anti-oncogenic effects of promyelocytic leukemia (PML) on bladder cancer and to explore its molecular mechanisms of growth suppression.

METHODSWild-type PML was transfected into bladder cancer cells (5637 cell) and expressed in a replication-deficient adenovirus-mediated gene delivery system and introduced into human bladder cancer cells (5637 cell) in vitro and in vivo. The effect and mechanisms of the PML gene in cell growth, clonogenicity, and tumorigenicity of bladder cancer cells were studied using in vitro and in vivo growth assays, soft agar colony-forming assay, cell cycle analysis, apoptosis assay and in vivo tumorigenicity assay.

RESULTSOverexpression of PML in 5637 cells significantly reduced their growth rate and clonogenicity on soft agar. PML suppressed bladder cancer cell growth by inducing G1 cell cycle arrest and apoptosis. Adenovirus-mediated PML (Ad-PML) significantly suppressed the tumorigenicity and growth of bladder cancer cells. Intratumoral injection of Ad-PML into tumors induced by 5637 cells dramatically suppressed their growth.

CONCLUSIONSThe results indicated that overexpression of PML protein may promote efficient growth inhibition of human bladder cancer cells by inducing G1 cell cycle arrest and apoptosis, and adenovirus-mediated PML (Ad-PML) expression efficiently suppresses human bladder cancer growth.

Adenoviridae ; Animals ; Apoptosis ; physiology ; Cell Division ; physiology ; Cells, Cultured ; Humans ; Male ; Mice ; Mice, Nude ; Neoplasm Proteins ; analysis ; Nuclear Proteins ; Promyelocytic Leukemia Protein ; Transcription Factors ; analysis ; Transfection ; Tumor Cells, Cultured ; Tumor Suppressor Proteins ; Urinary Bladder Neoplasms ; pathology

Objective To investigate the mechanism of miRNA-19a impacting on biological characteristics of colon cancer Lovo cell.Methods miRNA-19a mimics was transfected into human colon cancer cell line LoVo cells by liposome mediated transfection.The expression of miRNA-19a gene after transfection was detected.Cell proliferation was detected by CCK8 Kit.The effect of apoptosis was detected by TUNEL method.In vitro invasiveness of cells was detected by cell invasion chamber method.Hsp90/Akt signaling pathways changes were detected by RT-PCR and Western blot.Results The expression of miRNA-19a were increased.The ability of proliferation were promoted in vitro.The cell apoptosis was reduced.The cell invasion was increased.The expression of Hsp90 and Akt gene and protein were increased.Conclusion miRNA-19a could enhance Lovo cell invasion ability by the method of raising the expression of Hsp90 and Akt signaling pathways,accelerating LoVo cell proliferation,reducing cell apoptosis.

Objective: To investigate the action and antioxidant effects of CB2 agonist AM-1241 on rat hepatic stellate cell line (HSC-T6). Methods: HSC-T6 was randomly divided into four groups: control group, oxidative stress group, AM-1241 intervention group and AM-1241+AM-630 antagonist group. Survival rate of HSC-T6 was detected by thiazolyl blue assay under 24 h interventions with 0, 20, 50, 80 μmol/L AM-1241 and 0, 10, 20, 30, 40 μmol/L AM-630, respectively. Besides control group, the remaining groups were well cultured in low-glucose DMEM containing 100 mU/L glucose oxidase (GO) for 12 h to prepare the oxidative stress model. Then, AM-1241 intervention group was treated with 50 μmol/L low-glucose DMEM medium. After incubation for 12 h, the AM-1241+AM-630 antagonist group was treated with CB2 antagonist AM-630 (20 μmol/L) for 2 h, and cultured with 50 μmol/L AM-1241 in complete low-glucose medium for 12 h. The optimal drug concentration was selected according to the cell viability considered by the experiment results. Type III collagen (C III) content in the HSC-T6 supernatant was detected by enzyme-linked immunosorbent assay. Glutathione (GSH) content in HSC-T6 was detected by spectrophotometry. CB2 and heme oxygenase-1(HO-1) in each group of HSC-T6 were detected by western blotting. Results: HSC-T6 proliferation was inhibited in each group of AM-1241 in a concentration-dependent manner (P < 0.05). The inhibition was highest at 80μmol/L, and the cell survival rate was (41.61% ± 3.13%) (P < 0.05). AM-630 concentration group had no significant inhibitory effect on the proliferation of HSC-T6 (P > 0.05). HSC-T6 expressed CB2 receptor in each group. The expression level of CB2 in the AM-1241 intervention group was higher compare with control group (P < 0.05).The expression of Col III were significantly higher in oxidative stress group (P < 0.05) than in control group, and the expression of Col III of AM-1241 intervention group was significantly lower than that in oxidative stress group (P < 0.05). Col III level in AM-1241+AM-630 antagonistic group was significantly higher than that in AM-1241 intervention group (P < 0.05). There was no significant difference between AM-1241+AM-630 antagonistic group and oxidative stress group (P > 0.05). The content of GSH and HO-1 in oxidative stress group was higher (P < 0.05) than control group. The content of GSH and HO-1 in the AM-1241 intervention group was higher compared with oxidative stress group, while content of AM-1241 + AM-630 antagonist group was lower compared to AM-1241 intervention group (P < 0.05), and the differences were not statistically significant for oxidative stress group. Conclusion CB2 agonist AM-1241 can inhibit the proliferation and activation of HSC-T6 and its mechanism may activate the nuclear translocation of Nrf2 binding to HSC-T6, initiating the up-regulation of antioxidant enzymes HO-1 and GSH protein expression, and thus increase the antioxidant effect of HSC-T6.