Objective To study the effect of circumferential pulmonary vein isolation(CPVI) guided by Carto-Merge and Lasso catheter for the treatment of atrial fibrillation(AF).Methods Fifty-six drug refractory patients(41 male) with paroxysmal AF were enrolled into this study with a mean age of(58.5?12.7) years.Ostia of pulmonary veins(PVs) were identified by Carto-Merge.Then circumferential pulmonary vein isolation was performed.The endpoint of ablation was abolishment or dissociation of the pulmonary vein potentials(PVPs).Results Mean distance between electroanatomic mapping points and MRI surfaces was(1.79?0.33) mm as a whole in this group.Abolishment or dissociation of PVPs was accomplished in 52 patients(92.9%) during the first procedure.After a mean(18.3?5.7) months of follow up,41 patients(73.2%) maintained sinus rhythm.Ten patients received a second ablation procedure,and 8 were free of atrial tachyarrhythmias after a mean(8.2?6.9) months of the second follow up.No PV stenosis was identified after ablation procedure.Conclusion Combination of Lasso catheters with Carto-Merge to guide the CPVI procedure can confirm the isolation of PVs and lower the incidence of PVs stenosis.

BACKGROUND:Renal ischemia-reperfusion injury has been shown to exhibit gender difference, but its precise mechanisms deserve further investigations. OBJECTIVE:To investigate the differential expression of microRNAs in the kidney between female and male mice in order to study the effects and mechanisms of microRNA in pathogenesis of ischemia-reperfusion injury between different genders. METHODS:Male and female mice received kidney ischemia for 45 minutes and reperfusion injury for 24 hours. Simultaneously, male and female sham surgery groups served as controls. The microRNA gene chip technology was used to detect the differences of microRNA expression in the kidney of male and female mice at 45 minutes after ischemia and 24 hours of reperfusion as wel as after sham surgery. The threshold of difference in expression among samples was double. RESULTS AND CONCLUSION:Five microRNAs were up-regulated between female and male ischemia-reperfusion injury groups. Twenty-nine microRNAs differential y expressed in the female ischemia-reperfusion group and female sham surgery group, including 25 up-regulated microRNAs and 4 down-regulated microRNAs. Thirty-eight microRNAs differential y expressed in male ischemia-reperfusion injury group and male sham surgery group, including 9 up-regulated microRNAs and 29 down-regulated microRNAs. 102 microRNAs differential y expressed in the female sham surgery group and male sham surgery group, including 22 up-regulated microRNAs and 80 down-regulated microRNAs. Results suggested that there was differential expression in microRNAs in the kidney before and after renal ischemia-reperfusion in male and female mice. These differential y expressed microRNAs may be lead to different sensitivity and tolerance to the ischemia-reperfusion injury in the kidney of male and female mice.

AIM: To construct the prokaryotic expression plasmid of His-tagged human high mobility group box 1 fusion protein (hHMGB1) and to express the fusion protein in E. coli for the affinity purification. METHODS: The cDNA coding region of HMGB1 was amplified by PCR from pGEX4T-HMGB1 and cloned into a modified pET14b vector following the routine procedure. After identification by enzyme digestion, PCR and sequencing, the plasmid was transformed into BL21 (DE_3) competent cells, and the His-HMGB1 fusion protein was induced for expression with isopropyl-beta-D-thiogalactopyranoside (IPTG), and further purified by Ni-NTA affinity chromatography. The protein was filtered for sterilization and used to stimulate human umbilical vein endothelial cells (HUVECs). 24 hours later, the cultured supernatant of HUVECs was collected for the detection of cytokines/chemokines with LiquiChip system. RESULTS: The His-HMGB1 fusion protein expression plasmid was identified by enzyme digestion and sequencing. The purified His-tagged fusion protein was analyzed by SDS-PAGE and Western blotting with specific anti-His antibody. It was found that the production of IL-8 from HUVECs was highly induced in a dose-dependent manner by HMGB1. CONCLUSION: The His-tagged HMGB1 fusion protein expression plasmid was successfully constructed, and purified. Recombinant HMGB1 protein has a high bioactivity on the induction of cytokines in HUVECs, which may significantly facilitate the future study of HMGB1 biological functions.

TLR4-MD-2 complex plays a key role in LPS recognition and its signal transduction. These steps are the vital elements of the host's defensive reaction. Studying the functional domain of TLR4 and MD-2 is very important to further understand the mechanism of LPS signal transduction. It was studied the interaction domain of TLR4 and MD-2 in living cells based on gene mutation, gene transfection and fluorescence resonance energy tramsfer(FRET) which is considered as one of the best methods used for intracellular protein-protein interaction study. CY-15P which was fused by CFP and YFP through 15 neutral amino acids was used as positive control, while co-expressed CFP and YFP proteins were used as negative control. The results showed that the ability of TLR4 binding to MD-2 decreased dramatically after the deletion of Glu~(24) ～ Met~(41) in N terminal of TLR4. Aggregation of TLR4 to LPS stimulation was observed, however, TLR4 without the Glu~(24)～ Met~(41) mutation did not aggregate. All these results indicated that TLR4 Glu~(24)～ Met~(41) might be the interaction domain of TLR4 binding to MD-2 and participate in the aggregation effect of TLR4 upon LPS stimulation.

Objective To investigate the effects of interleukin-34 (IL-34) on prostaglandin E2 (PGE2)/cyclo-oxygenase-2 (COX-2) expression on fibroblast-like synoviocytes (FLS) in patients with rheumatoid arthritis (RA). Methods FLS was isolated from 6 RA patients and stimulated with IL-34 (50 ng/ml), IL-34 receptor antagonist (25 ng/ml) and IL-34 (50 ng/ml), inhibitors of signaling pathway (10 μmol/L) and IL-34 (50 ng/ml) in vitro respectively. The expression of COX-2 mRNA was detected by reverse transcription polymerase chain reac-tion (RT-PCR). The level of PGE2 in the supernatant of RA FLS culture was measured by Enzyme linked immunosorbent assay (ELISA). Statistical analysis between groups were performed by t test. Results Com-pared to unstimulated FLS, COX-2 and PGE2 expression was increased dramatically on IL-34-stimulated FLS, most evidently in 48 hours [(139±24) pg/ml vs (201±8) pg/ml, t=-6.177, P<0.01]; Moreover, the level of PGE2 was decreased when anti-IL-34 antibody was added to the IL-34-stimulated RA FLS at 24 hours, 48 hours, 72 hours [(250 ±58) pg/ml vs (100 ±28) pg/ml, t=5.742, P<0.01; (375 ±24) pg/ml vs (97 ±23) pg/ml, t=20.564, P<0.001; (357 ±21) pg/ml vs (94 ±18) pg/ml, t=22.353, P<0.01]; In the presence of SB203580 and IKK-16, PGE2 level produced by IL-34-stimulated FLS was obviously decreased [(279 ±37) pg/ml vs (63 ±17) pg/ml, t=12.806, P<0.01;(279±37) pg/ml vs (77±16) pg/ml, t=6.177, P<0.01]. Conclusion Binding of IL-34 with its receptor may promote the secretion of PGE2 via NF-κB and P38 MAPK signaling pathway in RA FLS, suggesting that it might be involved in the pathogenesis of RA.

Objective To investigate and find a multiple treatment to reduce the mortality of sepsis patients induced by severe surgical abdominal infection.Methods While treating to severe surgical abdominal infection,inflammatory mediator bacteria,extoxin and endotoxin,immunity,dysfunction of microcirculation,nutrition and metabolism and the function of organs should be paid more attention on and considered as a whole.We also carried out 14 concrete treating measurement.Combined high dosage of anisodaminum and dexamethason were used in sbort-term.Bring forward nourishment support according to different stage of MODS and applying it in clinic could significantly reduce the companion syndrome.Oral administration of“JIE-DU-GU-BEN-TANG”which developed by our division could regulate the imbalance of immunity and inflammatory mediator.Results There were 46 patients died in 368 patients,mortality was 12.50%. Conclusion It was difficult to treat sepsis patients induced by severe abdominal infection and our new multiple treatment could significantly reduce the mortality of severe sepsis.

Objective To preliminarily investigate the levels of interleukin (IL)-1 family and IL-34 in serum of patients with ankylosing spondylitis (AS) and their roles.Methods Serum IL-1 family levels were detected from 6 AS patients and 4 healthy controls by using protein-chip technique.Enzyme-linked immunosorbent assay (ELISA) method was used to detect the levels of serum IL-34 from 65 AS patients and 85 healthy controls and the relationships of serum IL-34 levels and clinical or laboratory features were analyzed.T test and Spearman correlation were used for statistical analysis.Results IL-1Ra [(3302±1352) pg/ml vs (10778±2764) pg/ml]and IL-36Ra [(1363±194) pg/ml vs (3875±996) pg/ml] levels were significantly down-regulated in AS patients compared with that of healthy controls (t=5.363 and 4.289 respectively,both P＜0.05).The levels of IL-1α,IL-18,IL-36α and IL-37 were increased more remarkable in AS patients than in healthy controls (t=-2.532,-5.400,-5.023 and-5.783 respectively,both P＜0.05).Moreover,serum IL-34 levels were elevated more significantly in AS patients than in healthy controls [(169±153) pg/ml vs (54±31) pg/ml,t=6.722,P＜0.01] and were positively correlated with the levels of CRP and ESR.Serum IL-34 levels were markedly up-regulated in human leukocyte antigen (HLA)-B27 positive patients than in HLA-B27 negative patients(P＜0.05).Conclusion Part of IL-1 family and IL-34 may be involved in inflammatory or immunological process of AS.

Objective To study the impacts of alcohol dependence on the anticonvulsant effect of diazepam. Meth?ods Kunming mice (n=36) were divided into 3 groups (n=12 in each group), Alcohol Dependence Group(A group), Diaze?pam Group(D group)and Normal Saline Group(N group). A group received an intraperitoneal injection with a 0.2 mL dose of 0.8%alcohol in NS (normal saline) , while both D and N group received an injection with a 0.2 mL dose of NS without alco?hol , twice a day. Mice’s autonomic activities were monitored every day. After 7 days, the electroconvulsive experiment was performed. Both A and D group were given a weight-based dose of 0.05 mL/10 g of 0.05%diazepam via intraperitoneal injec? tion, while N group was given a 0.05 mL/10 g dose of NS. Before administration and after 15, 30, 60 min of administration, the convulsion threshold of each group was measured. Results The count of autonomic activity of mice in A group was less than that of mice in D and N group during the 2nd day to 6th day(P<0.05). On the 1st and 7th day, the difference of the count of autonomic activity of mice between A group and the other two groups was not statistically significant(P>0.05). The convulsion threshold of mice in A group was higher than that of mice in D and N group before administration(P<0.05). Af?ter administration, the convulsion threshold of mice in N group didn’t show statistically significant difference from that of mice before administration(P>0.05). After 15 min of administration, the convulsion threshold of mice in D group was high?er than that of mice in A and N group(P<0.05), while the convulsion threshold of mice in A group was higher than that of mice in N group(P<0.05). After 30 min and 60 min of administration, both the convulsion thresholds of mice in A and D group were higher than that of mice in N group(P<0.05). However, at this point, the difference of the convulsion thresholds of mice between A and D group was not statistically significant(P>0.05). Conclusion Alcohol dependence has anticon?vulsant effect. Alcohol dependence weakens the anticonvulsant effect of diazepam.

Background and Objectives: The immunomodulatory potential of mesenchymal stem cells (MSCs) can be regulated by a variety of molecules, especially cytokines. The inflammatory cytokine, TNF-like ligand 1A (TL1A), has been reported as an inflammation stimulator in-multiple autoimmune diseases. Here, we studied the effects of TL1A/TNF-receptor 2 (TNFR2) pathway on the therapeutic potency of bone marrow-derived MSCs (BMSCs). Methods: and Results: BMSCs, fibroblast-like synoviocytes (FLSs), and H9 and jurkat human T lymphocytes were used in this study. BMSCs paracrine activities, differentiation, proliferation, and migration were investigated after stimulation with TL1A, and intervened with anti-TNFR2. Additionally, the effects of TL1A on BMSCs therapeutic potency were evaluated by treating RA-FLSs, and H9 and jurkat T cells with TL1A-stimulated BMSCs conditioned medium (CM). Indian hedgehog (IHH) involvement was determined by gene silencing and treatment by recombinant IHH (rIHH). TL1A induced BMSCs stemness-related genes, COX-2, IL-6, IDO, TGF-β and HGF through TNFR2. Also, TL1A corrected biased differentiation and increased proliferation, and migration through TNFR2. Meanwhile, CM of TL1A-stimulated BMSCs decreased the inflammatory markers of RA-FLSs and T cells. Moreover, TL1A-stimulated BMSCs experienced IHH up-regulation coupled with NF-κB and STAT3 signaling up-regulation, while p53 and oxidative stress were down-regulated. Furthermore, treatment of BMSCs by rIHH increased their anti-inflammatory effects.More importantly, knockdown of IHH decreased the ability of TL1A-stimulated BMSCs to alleviating the inflammation in RA-FLSs and T cells. Conclusions This study reports the effects of TL1A/TNFR2 pathway on the biological behaviors and therapeutic potency of BMSCs through IHH. These findings could introduce novel procedures to increase the stemness of MSCs in cellular therapy.

Objective:To investigate the clinical value of peripheral blood circulating tumor cells (CTC) in the diagnosis and treatment of prostate cancer.Methods:Sixty-four patients with prostate cancer who received treatment in Xinjiang Production and Construction Corps Hospital, China between June 2018 and May 2020 were included in the cancer group. An additional 35 patients with benign prostatic lesions who concurrently received treatment in the same hospital were included in the benign disease group. Twenty male patients with non-prostate disease were included in the control group. Cell enrichment, separation, staining and identification together with Gleason score and pathological stage were subjected to one-way analysis of variance.Results:The percentage of patients with CTC count ≥ 3 in the cancer, benign disease and control groups was 73.43% (47/64), 17.14% (6/35) and 10.00% (2/20), respectively. The level of prostate-specific antigen in patients with CTC was significantly higher than that in patients without CTC ( t = 2.89, P < 0.05). There was significant difference in CTC count between different Gleason score groups ( F = 3.25, P < 0.05) and between different pathological stage groups ( F = 3.42, P < 0.05). Conclusion:Peripheral blood CTC measurement can be used as an auxiliary method for the differentiation of benign and malignant prostate diseases. CTC count in patients with prostate cancer is correlated with prostate-specific antigen level, Gleason score, and pathological stage. Therefore, peripheral blood CTC measurement plays an auxiliary role in predicting prognosis in patients with CTC. This study is innovative and scientific.