Purpose To investigate the clinical utility of DAB2IP (DOC-2/DAB2 interactive protein)and β-catenin expression in bladder urothelial carcinoma (BUC).Methods The expression of DAB2IP and β-catenin was detected in 104 BUC cases and 40 peritumorial tissues using EnVision two-step immunohistochemical method,and the association with BUC clinicopathological parameters was analyzed.Results The expression of DAB2IP in BUC was significantly less than that of peritumorial normal tissues,and the expression of β-catenin in BUS was significantly higher than that of peritumorial normal tissues (P ＜ 0.05).DAB2IP expression and histologic grading,clinical pathologic staging and 5 years survival rate had statistical significance (P ＜ 0.05).No statistical significance with gender,age,tumor diameter and in patients with incipient/recurrence.β-catenin expression and age,histologic grading,clinical pathologic staging,tumor diameter and 5 years survival rate have statistical significance (P ＜ 0.05).No statistical significance correlated with gender and in patients with incipient/ recurrence,DAB2IP and β-catenin expression in BUC are negatively correlated (P ＜ 0.05).Conclusion In bladder urothelial carcinoma,down-regulation of DAB2IP and up-regulation of β-catenin,are in a negative correlation.Abnormal expression of DAB2IP and β-catenin is correlated with histologic grading,clinical pathologic staging and prognosis.

Objective:To observe the effects of berberine on procoagulant and fibrinolytic inhibitory factors produced by rat type Ⅱ alveolar epithelial cell (AECⅡ) induced by lipopolysaccharide (LPS).Methods:AECⅡ cells (RLE-6TN cells) were cultured in vitro, and the cells in logarithmic growth phase were collected. The cytotoxicity text of berberine was detected by cell counting kit-8 (CCK-8) to determine the drug concentration range according to inhibition concentration of half cells (IC 50). The RLE-6TN cells were divided into five groups, the cells in blank control group were cultured in DMEM; the cells in LPS group were stimulated with 5 mg/L LPS; and the cells in berberine pretreatment groups were pretreated with 20, 50 and 80 μmol/L berberine for 1 hour, and then were co-cultured with 5 mg/L LPS. The cells were collected after LPS induced for 24 hours. The protein and mRNA expression levels of tissue factor (TF), tissue factor pathway inhibitor (TFPI) and plasminogen activator inhibitor-1 (PAI-1) in the cells were detected by Western blotting and real-time fluorescence quantification reverse transcription-polymerase chain reaction (RT-qPCR). The levels of activated protein C (APC), precollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT) and antithrombin Ⅲ (ATⅢ) in the cell supernatant were measured by enzyme linked immunosorbent assay (ELISA). Results:According to the inhibition rate curve, the IC 50 of berberine on RLE-6TN cells was 81.16 μmol/L. Therefore, 20, 50 and 80 μmol/L were selected as the intervention concentration of berberine. Compared with the blank control group, the expression and secretion of procoagulant and fibrinolytic inhibitory factors were abnormal in RLE-6TN cells after LPS induced for 24 hours. The protein and mRNA expression levels of TF and PAI-1 in the LPS group were significantly increased, but the protein and mRNA expression levels of TFPI were significantly decreased. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly decreased, while the levels of PⅢP and TAT were significantly increased. After pretreatment with berberine, the abnormal expression and secretion of procoagulant and fibrinolytic inhibitory factors induced by LPS were corrected in a dose-dependent manner, especially in 80 μmol/L. Compared with the LPS group, the protein and mRNA expression levels of TF and PAI-1 in the berberine 80 μmol/L group were significantly decreased [TF protein (TF/GAPDH): 0.45±0.02 vs. 0.55±0.03, TF mRNA (2 -ΔΔCt): 0.39±0.08 vs. 1.48±0.11, PAI-1 protein (PAI-1/GAPDH): 0.37±0.02 vs. 0.64±0.04, PAI-1 mRNA (2 -ΔΔCt): 1.14±0.29 vs. 4.18±0.44, all P < 0.01] and those of TFPI were significantly increased [TFPI protein (TFPI/GAPDH): 0.53±0.02 vs. 0.45±0.02, TFPI mRNA (2 -ΔΔCt): 0.94±0.08 vs. 0.40±0.05, both P < 0.01]. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly increased [APC (μg/L): 1 358.5±26.0 vs. 994.2±23.1, ATⅢ (μg/L): 118.0±7.4 vs. 84.4±2.7, both P < 0.01], while those of PⅢP and TAT were significantly decreased [PⅢP (μg/L): 11.2±0.4 vs. 18.6±0.9, TAT (ng/L): 222.1±2.8 vs. 287.6±7.0, both P < 0.01]. Conclusions:Berberine could inhibit the LPS-induced expressions of procoagulant and fibrinolytic inhibitory factors in rat AECⅡ cells and promote the expressions of anticoagulant factors in a dose-dependent manner. Berberine may be a new therapeutic target for alveolar hypercoagulability and fibrinolysis inhibition in acute respiratory distress syndrome (ARDS).

Objective:To determine the effect of andrographolide (AD) on the expression of procoagulant and fibrinolytic inhibitory factors in rat type Ⅱ alveolar epithelial cells (AECⅡ) stimulated by lipopolysaccharide (LPS).Methods:The AECⅡ cells RLE-6TN in the logarithmic growth phase were divided into 5 groups: the normal control (NC) group, the LPS group, and the 6.25, 12.5, and 25 mg/L AD groups (AD 6.25 group, AD 12.5 group, AD 25 group). The NC group was cultured with RPMI 1640 conventional medium. In the LPS group, 5 mg/L LPS was added to the RPMI 1640 conventional medium for stimulation. Cells in the AD groups were treated with 6.25, 12.5, and 25 mg/L AD in advance for 1 hour and then given LPS to stimulate the culture. The cells and cell culture supernatant were collected 24 hours after LPS stimulation. The protein and mRNA expressions of tissue factor (TF), tissue factor pathway inhibitor (TFPI), and plasminogen activator inhibition-1 (PAI-1) in cells were detected by Western blotting and real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The levels of procollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT), antithrombin Ⅲ (AT-Ⅲ) and activated protein C (APC) in the cell supernatant were detected by enzyme linked immunosorbent assay (ELISA).Results:Compared with the NC group, the protein and mRNA expressions of TF and PAI-1 in the LPS group were significantly increased, and the protein and mRNA expressions of TFPI were significantly reduced. At the same time, the levels of PⅢP and TAT in the cell supernatant were significantly increased, the levels of AT-Ⅲ, APC were significantly reduced. Compared with the LPS group, the protein and mRNA expressions of TF and PAI-1 in AD 6.25 group, AD 12.5 group, AD 25 group were significantly reduced [TF/GAPDH: 0.86±0.08, 0.45±0.04, 0.44±0.04 vs. 1.32±0.10, TF mRNA (2 -ΔΔCt): 2.59±0.25, 2.27±0.05, 1.95±0.04 vs. 4.60±0.26, PAI-1/GAPDH: 2.11±0.07, 1.45±0.04, 0.86±0.09 vs. 2.56±0.09, PAI-1 mRNA (2 -ΔΔCt): 3.50±0.22, 2.23±0.29, 1.84±0.09 vs. 6.60±0.27, all P < 0.05], while the protein and mRNA expressions of TFPI were significantly increased [TFPI/GAPDH: 0.78±0.05, 0.81±0.03, 0.84±0.07 vs. 0.36±0.02, TFPI mRNA (2 -ΔΔCt): 0.46±0.09, 0.69±0.07, 0.91±0.08 vs. 0.44±0.06, all P < 0.05]. Also the levels of PⅢP and TAT in the cell supernatant were significantly reduced, and the levels of AT-Ⅲ and APC were significantly increased [PⅢP (μg/L): 13.59±0.23, 12.66±0.23, 10.59±0.30 vs. 15.82±0.29, TAT (ng/L): 211.57±6.41, 205.69±4.04, 200.56±9.85 vs. 288.67±9.84, AT-Ⅲ (μg/L): 102.95±3.86, 123.92±2.63, 128.67±1.67 vs. 92.93±3.36, APC (μg/L): 1 188.95±14.99, 1 366.12±39.93, 1 451.15±29.69 vs. 1 145.55±21.07, all P < 0.05]. With the increase of the dose of AD, the above-mentioned promotion and inhibition effects became more obvious. In the AD 25 group, TF, PAI-1 protein and mRNA expressions decreased, TFPI mRNA expression increased, PⅢP level in the supernatant decreased and AT-Ⅲ, APC levels increased compared with AD 6.25 group, the difference was statistically significant, and the decrease of PAI-1 protein expression and PⅢP level in the supernatant were also statistically significant compared with AD 12.5 group. Conclusions:Andrographolide in the dose range of 6.25-25 mg/L can dose-dependently inhibit the expression and secretion of procoagulant and fibrinolytic inhibitor-related factors in AECⅡ cells RLE-6TN stimulated by LPS, and promote the secretion of anticoagulant factors. 25 mg/L has the most obvious effect.

Objective To compare the influence of sevoflurane inhalation sedation and propofol intravenous sedation on duration of endotracheal intubation as well as the length of intensive care unit (ICU) stay and total length of hospital stay in postoperative critical patients.Methods Six databases including CNKI,Wanfang data,PubMed,Embase,Cochrane Library and Web of Science were searched for randomized controlled trials (RCTs) about the influence of sevoflurane inhalation sedation or propofol intravenous sedation on the sedation time,the duration of endotracheal intubation,the length of ICU stay,the total length of hospital stay and the adverse effects rate in postoperative critical patients from the time of database establishment to July 2018.At the same time,the reference materials of included literature were retrieved manually.All literatures were screened by three independent reviewers,and the data extraction and quality evaluation of the included studies were conducted.Meta-analysis was used for RCT that met the quality standards.Results A total of 7 RCT studies were enrolled involving 537 patients who were all transferred into ICU after surgery with trachea cannula.Among the patients,272 received sevoflurane sedation while the other 265 received propofol sedation.All the included studies were well designed and of high quality.The results of Meta-analysis showed that compared with propofol sedation,sevoflurane sedation could significantly shorten the duration of endotracheal intubation [standardized mean difference (SMD) =-0.60,95％ confidence interval (95％CI) =-0.88 to-0.31,P ＜ 0.000 1]and the total length of hospital stay (SMD =-0.36,95％CI =-0.61 to-0.12,P =0.003),and lower the cardiac troponin T (cTnT) within 12-24 hours after ICU admission (SMD =-0.61,95％CI =-0.85 to-0.36,P ＜ 0.000 01).There was no significant difference in the sedation time (SMD =-0.07,95％CI =-0.29 to 0.15,P =0.52),the length of ICU stay (SMD =-0.19,95％CI =-0.39 to 0.01,P =0.06),the incidence of nausea and vomiting [odds ratio (OR) =1.19,95％CI =0.61 to 2.32,P =0.61] or incidence of delirium (OR =0.80,95％CI =0.34 to 1.90,P =0.62) between sevoflurane group and propofol group.Conclusions Sevoflurane inhalation sedation may lead to shorter duration of endotracheal intubation and total length of hospital stay,and had better protection for myocardium as compared with propofol intravenous sedation.The above conclusions needed further study to confirm,due to the lack of literature enrolled in this Meta-analysis.

Objective:To explore the effect of small dose of low molecular weight heparin on the prognosis of elderly patients with severe pneumonia using systematic evaluation method.Methods:Databases including Wanfang data, VIP, CNKI, SinoMed, PubMed, Embase and Cochrane Library were searched for randomized controlled trial (RCT) studies about the comparison of conventional therapy and low molecular weight heparin on prognosis of elderly patients with severe pneumonia from the time of database establishment to August 2019. The patients in conventional treatment group were treated by improving ventilation, anti-infection, eliminating phlegm, relieving asthma and maintaining homeostasis while those in low molecular weight heparin group were subcutaneously injected with low molecular weight heparin of 4 000 U, once a day for 7 days. The patients' main outcomes included the oxygenation index (PaO 2/FiO 2) after 7 days of treatment, duration of mechanical ventilation, mortality in hospital, and secondary outcomes included acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score and coagulation function after 7 days of treatment, the length of intensive care unit (ICU) stay, and incidence of bleeding. Data extraction and quality evaluation were conducted. The Meta-analysis of included studies that met the quality standards was performed using RevMan 5.3 software. Funnel diagram analysis was used to analyze the parameters with no less than 10 studies enrolled. Results:A total of 14 RCT studies were enrolled involving 1 173 elderly patients with severe pneumonia, among whom 590 received low molecular weight heparin while the other 583 received conventional therapy. All the included studies were well designed and of high quality. The results of Meta-analysis showed that compared with conventional therapy, small dose of low molecular weight heparin significantly elevated PaO 2/FiO 2 after 7 days of treatment [mean difference ( MD) = 19.25, 95% confidence interval (95% CI) was 16.88 to 21.61, P < 0.000 01], shortened the duration of mechanical ventilation ( MD = -48.88, 95% CI was -67.42 to -30.33, P < 0.000 01), and decreased mortality in hospital [odds ratio ( OR) = 0.40, 95% CI was 0.22 to 0.73, P = 0.003] and APACHEⅡ score after 7 days of treatment ( MD = -3.38, 95% CI was -3.94 to -2.83, P < 0.000 01), and shortened the length of ICU stay ( MD = -4.51, 95% CI was -5.75 to -3.27, P < 0.000 01). There was no significant difference in the changes of coagulation parameters after 7 days of treatment or the incidence of bleeding between low molecular weight heparin group and conventional therapy group [7-day thrombin time (TT): MD = 0.57, 95% CI was -0.15 to 1.28, P = 0.12; 7-day prothrombin time (PT): MD = 0.32, 95% CI was -0.35 to 0.98, P = 0.35; 7-day fibrinogen (FIB): MD = -0.17, 95% CI was -0.45 to 0.10, P = 0.22; incidence of bleeding: OR = 0.86, 95% CI was 0.36 to 2.07, P = 0.74]. The funnel diagram showed that there was publication bias of included 10 studies about APACHEⅡ score after 7 days of treatment. Conclusion:Small dose of low molecular weight heparin can improve the prognosis of elderly patients with severe pneumonia and it has no obvious side-effect on coagulation function.

Objective To explore the impacts of clinical pulmonary infection score (CPIS) on duration and defined daily doses (DDDs) of antibiotics in patients with bacterial severe pneumonia in intensive care unit (ICU). Methods Patients with severe pneumonia, whose antibiotic usage was prescribed with the guide of CPIS, and admitted to ICU severe respiratory and infectious disease ward of Guizhou Medical University Affiliated Hospital from May 2017 to October 2017 were enrolled as CPIS group. Patients with the first CPIS score > 5 were given antimicrobial therapy, and the score was dynamically evaluated every 2-3 days. If the CPIS score < 5, the score was evaluated again after 2 days. If the score was still < 5, the antimicrobial drugs were discontinued. Patients admitted to the same ward from November 2016 to April 2017 were regarded as controls, of whom the antibiotic usage was completely conducted by the clinical experience of the chief physician. The duration and DDDs of antibiotics were compared between patients in two groups. At the same time, the usage of ventilator and prognostic indicators (the length of ICU stay, ICU mortality) were recorded. Kaplan-Meier survival curve was drawn, and the cumulative survival rates of 28 days, 90 days and 12 months were analyzed and compared between the two groups. Results In our department, 177 and 182 patients were admitted to ICU from November 2016 to April 2017 and from May 2017 to October 2017, respectively, of whom 101 and 65 patients with severe pneumonia were collected respectively during the two stages. There was no significant difference in gender composition, age, underlying diseases, vital signs, acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score, or peripheral blood routine at admission between the two groups, indicating that the baseline data of the two groups were equally comparable. During the treatment process, there was no significant difference in the duration of mechanical ventilation [hours: 126.0 (69.0, 228.8) vs. 120.0 (72.0, 192.0)], the length of ICU stay [days: 7.0 (5.0, 11.0) vs. 8.0 (5.0, 14.0)], or ICU mortality [18.8% (19/101) vs. 26.2% (17/65)] between the control group and CPIS group (all P >0.05). Kaplan-Meier survival curve analysis showed that there was no significant difference in the cumulative survival rate of 28 days (log-rank test: χ2 = 0.540, P = 0.462), 90 days (log-rank test: χ2 = 0.332, P = 0.564) or 12 months (log-rank test: χ2 = 0.833, P = 0.362). Patients from CPIS guided group, however, had a shorter duration of antibiotics usage (days: 7.54±4.81 vs. 9.88±4.96, P < 0.01), and had a lower DDDs of antibiotics (17.58±13.09 vs. 22.73±18.31, P < 0.05) as compared with those in the control group. Conclusion CPIS-guided therapeutic regimen shortens antibiotic duration and decreases antibiotic DDDs in patients with severe pneumonia in ICU, indicating the values of CPIS in guiding antibiotics usage in these patients.

OBJECTIVE: To investigate the differentially expressed microRNAs(miRNAs) in human embryonic lung fibroblast MRC-5 cells stimulated by transforming growth factor-β1(TGF-β1) using microarray chip, and screen for key genes and signaling pathways of fibroblast trans-differentiation. METHODS: The miRNA expression gene chip dataset GSE43992 on TGF-β1 stimulated MRC-5 cells were downloaded from high-throughput Gene Expression Omnibus(GEO) database of National Center for Biotechnology Information of the United States. The R language Limma package was used to screen the differentially expressed miRNAs. Corresponding target genes were predicted by miRWalk database performed by Gene Ontology(GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) signaling pathway enrichment analysis. The protein-protein interaction(PPI) network was constructed by the search tool for the Retrieval of Interacting Genes database. RESULTS: A total of five differentially expressed miRNAs were identified, including four up-regulated miRNAs and one down-regulated miRNA; and 42 corresponding differentially expressed target genes were predicted. GO analysis indicated that the target genes were significantly enriched in collagen catabolic process, extracellular matrix organization, membrane organization, collagen fibril organization, and cellular response to amino acid stimulus. The results of KEGG pathway analysis showed that the signaling pathways corresponding to miRNAs and target genes were mainly concentrated in 18 signaling pathways, that were mainly related to the age-ethnic signaling pathways and protein digestion and absorption miRNAs in tumors and diabetic complications. The core genes transfected into the myofibroblasts by the three fibroblasts screened by the PPI network were threonine kinase 1, estrogen receptor 1 and β-catenin. CONCLUSION: Five differentially expressed miRNAs, 42 target genes, 18 signaling pathways, and 3 core genes related to TGF-β1-induced MRC-5 cell trans-differentiation were screened. It can provide new reference for the treatment and research of many diseases including pneumoconiosis and pulmonary fibrosis.

Objective: To explore the therapeutic effect of Xuebijing on patients with acute paraquat poisoning (APP) by using systematic evaluation method. Methods: PubMed, Cochrane Library, Embase, Wanfang database, China National Knowledge Infrastructure (CNKI), VIP database (VIP) and China Biology Medicine (CBM) were searched using the computers to find the literatures published about the Xuebijing injection for the treatment of APP. Randomized controlled trials (RCT) were retrieved from the establishment of the database to August 2019. Patients in experimental group were treated with Xuebijing injection combined with conventional treatment, while the patients in control group were only given conventional treatment. The patients' outcome included the 14-day mortality, arterial oxygen saturation (SaO₂) and incidence of pulmonary fibrosis. In addition, the 6-month survival rate, alanine aminotransferase (ALT), serum creatinine (SCr), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), malondialdehyde (MDA) and superoxide dismutase (SOD) between the two groups were compared. The literature data were extracted by two researchers independently, and the quality of the literatures was evaluated according to the Cochrane 5.1 handbook. The Meta-analysis was performed by using RevMan 5.3 software. The results stability of Meta-analysis was tested by sensitivity analysis. The publication bias was analyzed through drawing of funnel diagram. Results: Twenty-seven RCT studies in total were enrolled, of which 26 were in Chinese and 1 was in English. A total of 1 429 patients were enrolled, among whom 726 were in experimental group and another 703 were in control group. Meta-analysis showed that compared with the control group, the 14-day mortality [relative risk (RR) = 0.62, 95% confidence interval (95%CI) was 0.54 to 0.72, P < 0.000 01] and incidence of pulmonary fibrosis (RR = 0.67, 95%CI was 0.53 to 0.85, P = 0.000 9) of patients in the experimental group were significantly lowered, while SaO₂ at 7 days and 14 days were significantly increased [7 days: mean difference (MD) = 16.86, 95%CI was 9.89 to 23.83, P < 0.000 01; 14 days: MD = 16.51, 95%CI was 10.22 to 22.80, P < 0.000 01]. Compared with the control group, the survival rate within 6 months (RR = 1.55, 95%CI was 1.41 to 1.71, P < 0.000 01) and SOD (MD = 13.88, 95%CI was 7.43 to 20.33, P < 0.000 1) of patients in the experimental group were significantly increased, ALT at 14 days (MD = -78.35, 95%CI was -127.35 to -29.34, P = 0.000 5), SCr at 7 days and 14 days (7 days: MD = -135.13, 95%CI was -219.09 to -51.17, P = 0.002; 14 days: MD = -206.05, 95%CI = -290.13 to -121.96, P < 0.000 01), CRP (MD = -11.55, 95%CI was -17.77 to -5.33, P = 0.000 3), TNF-α (MD = -9.27, 95%CI was -15.48 to -3.96, P = 0.000 9) and MDA (MD = -1.27, 95%CI was -1.57 to -0.96, P < 0.000 01) were significantly lowered. The overall effect value of the parameters with high heterogeneity was not significantly changed after furtherMeta-analysis excluding any one of the studies, suggesting that the result was relatively stable. Funnel chart analysis was used to analyze the parameters from more than 10 articles enrolled, and it showed that there was publication bias. Conclusion Xuebijing injection can reduce the mortality of patients with APP, which may because that it can improve liver and kidney function, reduce inflammation and oxidative stress damage, inhibit pulmonary fibrosis and increase oxygenation level.

In order to prepare antioxidant peptide through hydrolyzing low-value protein resources with bacterial extracellular proteases and to discover novel proteases, crude extracellular protease from Pseudoalteromonas sp. SHK1-2 was obtained through fermentation which was used to hydrolyze collagen extracted from Cirrhinus molitorella skin. Small peptide fraction was isolated from hydrolysate by ultrafiltration and Sephadex LH-20 size exclusion chromatography and showed 1, 1-diphenyl-2-picrylhydrazyl radical scavenging activity (35.6%±7%), oxygen radical absorbance capacity and inhibition of DNA oxidation damage. The molecule weight was 776.2 Da, and amino acid sequence was Thr-Ala-Gly-His-Pro- Gly-Thr-His through liquid chromatography mass spectrum. Our findings suggest that peptide obtained from low-value protein of fish waste by hydrolysis with bacterial protease has antioxidant activity.
Amino Acid Sequence ; Animals ; Antioxidants ; chemistry ; Chromatography, Gel ; Collagen ; chemistry ; Cyprinidae ; Dextrans ; Hydrolysis ; Oxidation-Reduction ; Peptide Hydrolases ; Peptides ; chemistry ; Skin ; chemistry