Background::Erosive esophagitis (EE) is a gastroesophageal reflux disease characterized by mucosal breaks in the esophagus. Proton pump inhibitors are widely used as maintenance therapy for EE, but many patients still relapse. In this trial, we evaluated the noninferiority of vonoprazan vs. lansoprazole as maintenance therapy in patients with healed EE. Methods::We performed a double-blind, double-dummy, multicenter, phase 3 clinical trial among non-Japanese Asian adults with endoscopically confirmed healed EE from April 2015 to February 2019. Patients from China, South Korea, and Malaysia were randomized to vonoprazan 10 mg or 20 mg once daily or lansoprazole 15 mg once daily for 24 weeks. The primary endpoint was endoscopically confirmed EE recurrence rate over 24 weeks with a noninferiority margin of 10% using a two-sided 95% confidence interval (CI). Treatment-emergent adverse events (TEAEs) were recorded.Results::Among 703 patients, EE recurrence was observed in 24/181 (13.3%) and 21/171 (12.3%) patients receiving vonoprazan 10 mg or 20 mg, respectively, and 47/184 (25.5%) patients receiving lansoprazole (differences: -12.3% [95% CI, -20.3% to-4.3%] and -13.3% [95% CI, -21.3% to -5.3%], respectively), meeting the primary endpoint of noninferiority to lansoprazole in preventing EE recurrence at 24 weeks. Evidence of superiority (upper bound of 95% CI <0%) was also observed. At 12 weeks, endoscopically confirmed EE recurrence was observed in 5/18, 2/20, and 7/20 of patients receiving vonoprazan 10 mg, vonoprazan 20 mg, and lansoprazole, respectively. TEAEs were experienced by 66.8% (157/235), 69.0% (156/226), and 65.3% (158/242) of patients receiving vonoprazan 10 mg, vonoprazan 20 mg, and lansoprazole, respectively. The most common TEAE was upper respiratory tract infection in 12.8% (30/235) and 12.8% (29/226) patients in vonoprazan 10 mg and 20 mg groups, respectively and 8.7% (21/242) patients in lansoprazole group.Conclusion::Vonoprazan maintenance therapy was well-tolerated and noninferior to lansoprazole for preventing EE recurrence in Asian patients with healed EE.Trial Registration::https://clinicaltrials.gov; NCT02388737.

OBJECTIVE: To compare a new thermosensitive recombinant human amelogenin (rhAm) carrier and traditional propylene glycol alginate (PGA) carrier for their characteristics, antibacterial activity, and biocompatibility with human periodontal membrane fibroblasts. METHODS: PGA-rhAm was prepared by mixing 3.3% PGA and rhAm, and CS-βGP-rhAm was prepared by mixing 2% chitosan (CS) with rhAm and then with 60% β-sodium glycerophosphate solution (βGP) as the crosslinking agent. The biophysical properties of the prepared carriers were characterized, and their antibacterial activity was assessed by observing Staphylococcus aureus growth. The biocompatibility of the carriers was evaluated in human periodontal membrane fibroblasts (hPDLFs) using CCK8 assay and scratch test, and mRNA and protein expressions of osteogenic genes of the cells incubated with the carriers were detected using RT-qPCR and Western blotting; osteogenic differentiation of the cells was detected using alkaline phosphatase staining. RESULTS: PGA-rhAm had a viscosity value of 3.262±0.055 Pa.s. CS-βGP-rhAm had a solidification capacity of 6 min at 37 ℃ with a pH value close to that of the oral cavity and a swelling rate of about 90%. CS-β GP-rhAm maintained sustained release of rhAm for over 2 weeks with a self-degradation time over 3 weeks. CS-βGPrhAm more effectively inhibited the growth of S. aureus than rhAm-loaded PGA. While PGA did not obviously affect the proliferation of hPDLFs, both CS-βGP and CS-βGP-rhAm significantly promoted the cell proliferation(P < 0.001). Scratch test showed that after rhAm loading, both CS-βGP and PGA promoted cell migration (P < 0.01). CS-βGP-rhAm significantly enhanced the mRNA expressions of RUNX2 and OCN mRNA level and the protein expressions of Ki67, RUNX2, collagen I, and β-catenin (P < 0.05); PGA-rhAm only enhanced RUNX2 (P < 0.05) and OCN (P < 0.01) mRNA expressions without significant effects on the protein expressions. Alkaline phosphatase staining results showed that CS-βGP, but not PGA, promoted osteogenic differentiation of hPDLFs. CONCLUSION CS-βGP carrier is capable of sustained release of rhAm, inhibiting the growth of S. aureus, and improving the biological activity of hPDLFs without affecting the bioactivity of rhAm after drug loading.
Alginates ; Alkaline Phosphatase ; Amelogenin ; Anti-Bacterial Agents/pharmacology* ; Cell Differentiation ; Cells, Cultured ; Chitosan/pharmacology* ; Collagen ; Core Binding Factor Alpha 1 Subunit ; Delayed-Action Preparations ; Glycerophosphates ; Humans ; Ki-67 Antigen ; Osteogenesis ; Periodontal Ligament ; RNA, Messenger ; Staphylococcus aureus ; beta Catenin

OBJECTIVE: High-fat diet (HFD) and inflammation are two key contributors to nonalcoholic fatty liver disease (NAFLD). Shenling Baizhu powder (SLBZP), a classical herbal compound, has been successfully used to alleviate NAFLD. However, its specific mechanisms are not fully understood. In this study, we assessed the anti-NAFLD effect of SLBZP in vivo. METHODS: Rats were fed an HFD with or without SLBZP or with probiotics. At the end of week 16, an echo magnetic resonance imaging (EchoMRI) body composition analyser was used to quantitatively analyse body composition; a micro-computed tomography (micro-CT) imaging system was used to evaluate whole body and liver fat; and the Moor full-field laser perfusion imager 2 was used to assess liver microcirculation, after which, all rats were sacrificed. Then, biochemical indicators in the blood and the ultrastructure of rat livers were evaluated. Protein expression related to the liver Toll-like receptor 4 (TLR4)/Nod-like receptor family pyrin domain-containing 3 (NLRP3) signalling pathway was assessed using Western blot analysis. Further, high-throughput screening of 29 related inflammatory factors in liver tissue was performed using a cytokine array. RESULTS: SLBZP supplementation reduced body weight, serum free fatty acid, and insulin resistance index (P < 0.05). It also ameliorated liver microcirculation and ultrastructural abnormalities. EchoMRI and micro-CT quantitative analyses showed that treatment with SLBZP reduced fat mass and visceral fat (P < 0.05 and P < 0.01, respectively). In addition, SLBZP decreased the expression of lipopolysaccharide (LPS)-activated TLR4/NLRP3 signalling pathway-related proteins and altered the expression levels of some inflammatory cytokines in liver tissues. CONCLUSION SLBZP can inhibit NLRP3 inflammasome activation and interleukin-1β release by suppressing LPS-induced TLR4 expression in rats with HFD-induced NAFLD. Thus, SLBZP may be beneficial for the prevention and treatment of inflammatory damage and associated diseases.
Animals ; Liver ; NLR Family, Pyrin Domain-Containing 3 Protein ; Non-alcoholic Fatty Liver Disease/drug therapy* ; Powders ; Rats ; Toll-Like Receptor 4 ; X-Ray Microtomography

Objective: To investigate the inhibitory effect on Burkholderia pseudomallei (B. pseudomallei) strain HNBP001 of a bacillomycin D-like cyclic lipopeptide compound named bacillomycin DC isolated from Bacillus amyloliquefaciens HAB-2. Methods: The antibacterial effect of bacillomycin DC on B. pseudomallei was determined using the disk diffusion method. The minimum inhibitory concentrations were evaluated by microdilution assay. In addition, transmission electron microscopy was performed and quantitative real-time polymerase chain reaction assay was carried out to determine the expression of MexB, OprD2, and qnrS genes. Results: Bacillomycin DC produced an inhibition zone against B. pseudomallei with minimum inhibitory concentration values of 12.5 μg/mL 24 h after treatment and 50 μg/mL at 48 and 72 h. Transmission electron microscopy showed that bacillomycin DC resulted in roughening cell surface and cell membrane damage. Quantitative real-time polymerase chain reaction analysis showed low expression of MexB, OprD2 and qnrS genes. Conclusions: Bacillomycin DC inhibits the growth of B. pseudomallei and can be a new candidate for antimicrobial agents of B. pseudomallei. Rajaofera Mamy 1 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan Kang Xun 2 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan Jin Peng-Fei 3 Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests (Hainan University), Ministry of Education, Haikou 570228, Hainan Chen Xin 4 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan Li Chen-Chu 5 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan Yin Li 6 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan Liu Lin 7 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan Sun Qing-Hui 8 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan Zhang Nan 9 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan Chen Chui-Zhe 10 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan He Na 11 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan Xia Qian-Feng 12 Key Laboratory of Tropical Translational Medicine of Ministry of Education and School of Tropical Medicine and Laboratory Medicine, Hainan Medical University, Haikou, Hainan Miao Wei-Guo 13 Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests (Hainan University), Ministry of Education, Haikou 570228, Hainan Kung CT, Lee CH, Li CJ, Lu HI, Ko SF, Liu JW. 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Secondary metabolites from Bacillus amyloliquefaciens isolated from soil can kill Burkholderia pseudomallei. Amb Express 2017; 7(1):16. Kang X, Fu Z, Rajaofera MJN, Li C, Zhang N, Liu L, et al. Whole-genome sequence of Burkholderia pseudomallei strain HNBP001, isolated from a melioidosis patient in Hainan, China. Microbiol Resour Announc 2019; 8(36): e00471-19. Liu L, Sun QH, Pei H, Chen CZ, Xiu H, Zhang N, et al. Multilocus sequence typing of Burkholderia pseudomallei collected in Hainan, China. Chin J Zoono 2019; 35(06): 514-517+524. Gay K, Robicsek A, Strahilevitz J, Park CH, Jacoby G, Barrett TJ, et al. Plasmid-mediated quinolone resistance in non-Typhi serotypes of Salmonella enterica. Clini Infect Dis 2006; 43(3): 297-304. Fu QY, Chen CY, Wu J, Wu Q, Qin X, Qian SY, et al. Establishment and evaluation of real-time PCR for rapid and quantitative detection of Burkholderia pseudomallei. J Third Mil Med Univ 2015; 17: 1734-1738. Serra C, Bouharkat B, Tir Touil-Meddah A, Guénin S, Mullié C. MexXY multidrug efflux system is more frequently overexpressed in ciprofloxacin resistant french clinical isolates compared to hospital environment ones. Front Microbiol 2019; 10: 366. Cai S, Chen Y, Song D, Kong J, Wu Y, Lu H. Study on the resistance mechanism via outer membrane protein OprD2 and metal ß-lactamase expression in the cell wall of Pseudomonas aeruginosa. Exp Ther Med 2016; 12(5): 2869-2872. Kamjumphol W, Chareonsudjai P, Chareonsudjai S. Antibacterial activity of chitosan against Burkholderia pseudomallei. Microbiologyopen 2018; 7(1). Doi: 10.1002/mbo3.534 Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-AAC) method. Methods 2001; 25(4): 402-408. Baindara P, Mandal SM, Chawla N, Singh PK, Pinnaka AK, Korpole S. Characterization of two antimicrobial peptides produced by a halotolerant Bacillus subtilis strain SK.DU.4 isolated from a rhizosphere soil sample. AMB Express 2013; 3(1): 2. Chalhoub H, Sáenz Y, Nichols WW, Tulkens PM, Van Bambeke F. Loss of activity of ceftazidime-avibactam due to Mex-AB-OprM efflux and overproduction of AmpC cephalosporinase in Pseudomonas aeruginosa, isolated from patients suffering from cystic fibrosis. Int J Antimicrob Agents 2018; 52(5): 697-701. Verchère A, Picard M, Broutin I. Functional investigation of the MexA-MexB-OprM efflux pump of Pseudomonas aeruginosa. Biophysic J 2013; 104(2): 286a. Van Duin D, Lok JJ, Earley M, Cober E, Richter SS, Perez F. Colistin versus ceftazidime-avibactam in the treatment of infections due to carbapenem-resistant Enterobacteriaceae. Clin Infect Dis 2018; 66(2): 163-171. Schweizer HP. Mechanisms of antibiotic resistance in Burkholderia pseudomallei: Implications for treatment of melioidosis. 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Objective: To analyze the comprehensive intervention effect of post-traumatic stress disorder (PTSD) in patients with acute hand injury, and its correlation with hand function recovery. Methods: A total of 120 patients with hand trauma admitted to Wenzhou Hospital of Traditional Chinese Medicine from April 2016 to April 2018 were selected.The patients were randomly divided into observation group (60 cases) and control group (60 cases) according to the digital table.The observation group received routine treatment and comprehensive intervention.The control group was given routine treatment.The PDST scale (CAPS), depression self-rating scale (SDS) and Hamilton anxiety scale (HAMA) rating scale used by clinicians were used to assess the psychological status, depression and anxiety of patients with PTSD, and to evaluate the upper limb function of patients. Results: After intervention, the scores of CAPS, SDS and HAMA in the control group were (24.51±8.43)points, (50.61±7.59)points and (10.63±2.11)points, respectively, which were significantly higher than those in the observation group[(16.53±7.62)points, (40.26±4.18)points, (8.24±1.86)points](t=5.61, 5.59, 4.73, all P<0.05). The CAPS, SDS and HAMA scores of the two groups were significantly improved compared with before intervention (t=4.11, 5.04, 3.98, 5.18, 3.86, 4.72, all P<0.05). After intervention, the upper limb function scale (DASH) scores of the observation group and the control group were (89.27±6.08)points and (82.11±7.92)points, respectively.The DASH scores of the two groups were significantly higher than those before intervention (t=6.14, 5.28, all P<0.05). The DASH score of the observation group was superior to the control group (t=3.96, P<0.05). The DASH score of patients with hand injury was negatively correlated with CAPS score and SDS score (r=-0.362, -0.425, P=0.012, 0.008). Conclusion Comprehensive intervention for PTSD in patients with acute hand injury can significantly improve the degree of depression and anxiety, and improve the upper limb function of patients.

Adult ; Aged ; Alprostadil ; therapeutic use ; Female ; Hemorheology ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; blood ; drug therapy ; physiopathology ; Respiratory Function Tests

OBJECTIVETo construct a stable nm23-H1-knock-down cell model with K562 cell line and study its differentiation toward megakaryocyte.

METHODSEukaryotic expression vector pSilencer 4.1-CMV-sinm23 expressing siRNA targeting nm23-H1 was transfected into K562 cells with lipofectamine2000. Cells with stably nm23-H1 silence were screened out by G418. Real-time quantitative PCR, immunocytochemistry, western blot were used to confirm the nm23-H1-knock-down K562 model. Cell differentiation capacity was detected by NBT reduction assay. Surface antigen Gp IIb-IIIa (CD41) of knock-down cells treated with phorbol 12-myristate 13-acetate was analyzed by flow cytometry. Western blot was used to detect the ERK1/2 signal pathway after the stimulation of phorbol 12-myristate 13-acetate.

RESULTSEndogenous nm23-H1 was silenced by pSilencer 4.1-CMV-sinm23 and the silence efficiency was up to 75% and 70% in mRNA and protein levels respectively compared with the mock cells. Under phorbol 12-myristate 13-acetate treatment, the knock-down cells displayed a significantly increased differentiation ability toward megakaryocyte compared with control. The NBT reduction values were (0.31 +/- 0.07) and (0.23 +/- 0.05) respectively. Further results revealed that nm23-H1 gene regulating the megakaryocytic differentiation was due in part to the increased ERK1/2 phosphorylation.

CONCLUSIONSA stable nm23-H1-knock-down K562 cell model is successfully constructed. nm23-H1 involves in regulating the megakaryocytic differentiation of K562 cell line.

Cell Differentiation ; genetics ; Gene Knockdown Techniques ; Humans ; K562 Cells ; Megakaryocytes ; cytology ; NM23 Nucleoside Diphosphate Kinases ; genetics ; RNA Interference

To purify recombinant human nucleoside diphosphate kinase A (rhNDPK-A) efficiently in pilot scale, cells of rhNDPK-A producing E. coli were homogenized by high pressure under 4 degrees C, 950 Pa. The insoluble debris was removed by microfiltration and the soluble portion was concentrated by ultrafiltration. The resulted crude sample was loaded on DEAE-sepharose Fast Flow. The target fraction was collected and then load on Cibacron Blue 3GA Sepharose CL-4B. Eluted with buffer containing ATP from the AC column, rhNDPK-A was polished with ultrafiltration. The results showed that after homogenized 2 rounds, 1500g cells of E. coli brought crude sample containing 47.6g NDPK-A. Treated with microfiltration and ultrafiltration, 27.3g of NDPK-A were recovered from this bacteria homogenate. After 2-step purification with column chromatography and then polished with ultrafiltration, 17.2 g rhNDPK-A were collected with purity of 96.3%. The recovery of the whole purification process was 36.2%, and the productivity of rhNDPK-A was 1.15 g per 100 g wet cells. Comparing the recovery of each purification step, it was found that the recovery of polish is higher than that of affinity chromatography, which is higher than that of ion exchange chromatography. The limit step was the process of sample pretreatment among the 4 purification steps. Combine with the fermentation results reported before, it was deduced that the productivity of rhNDPK-A was 510 mg/L. In conclusion, an easily controlled purification condition with high yield provides material for the translation researches of NDPK; In addition, it was suggested the crucial step determine the recovery of non-secretive recombinant proteins might be the process of sample pretreatment, not be the process of column chromatography.
Chromatography, Affinity ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Humans ; NM23 Nucleoside Diphosphate Kinases ; genetics ; metabolism ; Pilot Projects ; Recombinant Proteins ; isolation & purification ; metabolism ; Ultrafiltration

To investigate the relationship between nm23-H1 gene and human chronic myeloblastic leukemia we designed siRNAs which target nm23-H1 gene. According to the principles of designing siRNA, we selected three siRNAs and transfected them into K562 cells by lipofectamine2000. The expression levels of nm23-H1 mRNA were detected by reverse transcriptase polymerase chain reaction after transfection for 24 hours. The expression levels of nm23-H1 protein were assayed by immunocytochemical method after transfection for 48 hours. And after transfection for 24, 48 and 72 hours, cell proliferation was determined by MTT method. Among the three siRNAs, siNM526 can effectively inhibit the expression of nm23-H1 on mRNA and protein levels. The growth of K562 cells was suppressed after transfection of siNM526. These results suggest that low expression level of nm23-H1 in K562 cells inhibited cell proliferation, namely reduced malignant degree of them. Therefore nm23-H1 gene might be a potential target of leukemia treatment.
Cell Proliferation ; Humans ; K562 Cells ; Leukemia, Erythroblastic, Acute ; genetics ; pathology ; NM23 Nucleoside Diphosphate Kinases ; biosynthesis ; genetics ; RNA Interference ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Alkaloids ; pharmacology ; Animals ; Antiviral Agents ; pharmacology ; Benzylisoquinolines ; Cercopithecus aethiops ; Dose-Response Relationship, Drug ; SARS Virus ; drug effects ; Vero Cells