Objective To investigate the effect and mechanism of action of epigallocatechin-3-gallate(EGCG)in the treatment of experimental metabolic dysfunction-associated steatohepatitis(MASH),and to provide a basis for clinical development and application.Methods A total of 32 experimental C57BL/6J mice were randomly divided into normal diet group(Con group with 8 mice)and model group with 24 mice.The mice in the model group were given a high-trans fatty acid high-carbohydrate(HFHC)diet for 24 weeks to establish a model of MASH,and at the end of week 24,the mice in the model group were further divided into HFHC group,EGCG treatment group(100 mg·kg-1·d-1),and obeticholic acid treatment group(10 mg·kg-1·d-1),with 8 mice in each group.After 6 weeks of treatment,samples were collected to observe the general conditions of mice;the content of triglycerides(TG)and hydroxyproline in liver tissue and the serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured;HE staining,oil red O staining,and picrosirius red staining were used to observe liver histopathological changes.In the in vitro experiment,L02 cells were induced with free fatty acid(FFA)to establish a model of lipid deposition,and the cells were divided into Con group,FFA group,and EGCG group.The content of TG in cells was measured,as well as the results of oil red O staining and the relative mRNA expression levels of TNF-α,CCL2,and CXCL10.The transcriptomics technique was used to identify differentially expressed genes between the Con group,the HFHC group,and the EGCG group and perform the GSEA analysis,and pathways with a P-adjust value of<0.05 that were associated with MASH were further classified into metabolism-related pathways and inflammation-related pathways.The specific signaling pathways in each category were ranked based on the degree of enrichment,and key genes in the top three pathways were verified by PCR in vivo.Key genes in the NOD-like receptor signaling pathway were verified by Western blotting.A one-way analysis of variance was used for comparison of normally distributed continuous data with homogeneity of variance between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the HFHC group,the EGCG group had significant reductions in the content of TG in liver tissue(P<0.05)and the serum levels of ALT and AST(P<0.05).Oil red O staining showed significant alleviation of hepatocyte fatty degeneration in the EGCG group,HE staining showed that EGCG effectively alleviated inflammation,and picrosirius red staining showed a significant reduction in the number of fibrous tissue after EGCG treatment.There was a significant reduction in the content of hydroxyproline in liver tissue after EGCG intervention(P<0.01).Cell experiments showed that compared with the FFA group,the EGCG group had a significant reduction in the content of TG,and oil red O staining showed the disappearance of lipid droplets in the EGCG group compared with the FFA group,with significant reductions in the relative mRNA expression levels of the inflammatory factors TNF-α,CCL2,and CXCL10(all P<0.01).The transcriptomics analysis identified 230 differentially expressed genes between the HFHC group and the EGCG group,among which there were 108 upregulated genes and 122 downregulated genes.EGCG significantly reduced the levels of the key proteins TLR4,NLRP3,and IL-1β in the NOD-like receptor signaling pathway in liver tissue(all P<0.05).Conclusion EGCG can significantly alleviate lipid deposition,inflammation,and fibrosis in the mouse model of MASH and improve lipid deposition and inflammatory injury in L02 cells,possibly by regulating the NOD-like receptor signaling pathway.

Objective To investigate the effect and mechanism of action of epigallocatechin-3-gallate(EGCG)in the treatment of experimental metabolic dysfunction-associated steatohepatitis(MASH),and to provide a basis for clinical development and application.Methods A total of 32 experimental C57BL/6J mice were randomly divided into normal diet group(Con group with 8 mice)and model group with 24 mice.The mice in the model group were given a high-trans fatty acid high-carbohydrate(HFHC)diet for 24 weeks to establish a model of MASH,and at the end of week 24,the mice in the model group were further divided into HFHC group,EGCG treatment group(100 mg·kg-1·d-1),and obeticholic acid treatment group(10 mg·kg-1·d-1),with 8 mice in each group.After 6 weeks of treatment,samples were collected to observe the general conditions of mice;the content of triglycerides(TG)and hydroxyproline in liver tissue and the serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured;HE staining,oil red O staining,and picrosirius red staining were used to observe liver histopathological changes.In the in vitro experiment,L02 cells were induced with free fatty acid(FFA)to establish a model of lipid deposition,and the cells were divided into Con group,FFA group,and EGCG group.The content of TG in cells was measured,as well as the results of oil red O staining and the relative mRNA expression levels of TNF-α,CCL2,and CXCL10.The transcriptomics technique was used to identify differentially expressed genes between the Con group,the HFHC group,and the EGCG group and perform the GSEA analysis,and pathways with a P-adjust value of<0.05 that were associated with MASH were further classified into metabolism-related pathways and inflammation-related pathways.The specific signaling pathways in each category were ranked based on the degree of enrichment,and key genes in the top three pathways were verified by PCR in vivo.Key genes in the NOD-like receptor signaling pathway were verified by Western blotting.A one-way analysis of variance was used for comparison of normally distributed continuous data with homogeneity of variance between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the HFHC group,the EGCG group had significant reductions in the content of TG in liver tissue(P<0.05)and the serum levels of ALT and AST(P<0.05).Oil red O staining showed significant alleviation of hepatocyte fatty degeneration in the EGCG group,HE staining showed that EGCG effectively alleviated inflammation,and picrosirius red staining showed a significant reduction in the number of fibrous tissue after EGCG treatment.There was a significant reduction in the content of hydroxyproline in liver tissue after EGCG intervention(P<0.01).Cell experiments showed that compared with the FFA group,the EGCG group had a significant reduction in the content of TG,and oil red O staining showed the disappearance of lipid droplets in the EGCG group compared with the FFA group,with significant reductions in the relative mRNA expression levels of the inflammatory factors TNF-α,CCL2,and CXCL10(all P<0.01).The transcriptomics analysis identified 230 differentially expressed genes between the HFHC group and the EGCG group,among which there were 108 upregulated genes and 122 downregulated genes.EGCG significantly reduced the levels of the key proteins TLR4,NLRP3,and IL-1β in the NOD-like receptor signaling pathway in liver tissue(all P<0.05).Conclusion EGCG can significantly alleviate lipid deposition,inflammation,and fibrosis in the mouse model of MASH and improve lipid deposition and inflammatory injury in L02 cells,possibly by regulating the NOD-like receptor signaling pathway.

Objective To investigate the effect of NOD-like receptor family pyrin domain containing 3(NLRP3)knockdown on a mouse model of nonalcoholic steatohepatitis(NASH)induced by high-fat high-carbohydrate(HFHC)diet.Methods A total of 44 mice were randomly divided into normal diet group(CON group)with 20 mice and HFHC group with 24 mice.At the end of week 14 of modeling,4 mice were randomly selected from the HFHC group for the pre-experiment of adeno-associated virus(AAV)by tail vein injection,and NLRP3 knockdown was verified after 4 weeks.After NLRP3 knockdown was verified at the end of week 18,the remaining 40 mice were given a single tail vein injection of AAV,and then they were divided into CON+NLRP3 knockdown negative control group(CON+NLRP3-NC group),CON+NLRP3 knockdown group(CON+NLRP3-KD group),HFHC+NLRP3-NC group,and HFHC+NLRP3-KD group,with 10 mice in each group.At the end of week 24,the activation of NLRP3 inflammasome was observed;related indicators were measured,including body weight,liver weight,liver index,and glucose metabolism(fasting blood glucose,fasting insulin,and Homeostasis Model Assessment of Insulin Resistance[HOMA-IR]index);the indicators of liver lipid content(liver triglyceride[TG]and oil red O staining),liver inflammation(serum alanine aminotransferase[ALT]activity,HE staining,and inflammation-related genes),and liver fibrosis(Sirius Red staining and fibrosis-related genes)were measured.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the CON+NLRP3-NC group based on the results of Western Blot,the HFHC+NLRP3-NC group had significant increases in the protein expression levels of NLRP3,pro-Caspase1,Caspase1,ASC,and IL-1β,while the HFHC+NLRP3-KD group had significant reductions in these levels(all P<0.05).The HFHC+NLRP3-NC group showed varying degrees of increase in body weight,liver weight,liver index,and glucose metabolism indicators,while the HFHC+NLRP3-KD group showed significant improvements in these indicators(all P<0.05).As for hepatic fat deposition,compared with the CON+NLRP3-NC group,the HFHC+NLRP3-NC group had a significant increase in liver TG,with a large number of red lipid droplets shown by oil red O staining,and the HFHC+NLRP3-KD group had significant reductions in liver TG and the number of lipid droplets in the liver(all P<0.01).In terms of liver inflammation,compared with the CON+NLRP3-NC group,the HFHC+NLRP3-NC group had significant increases in serum ALT,NAFLD activity score,and inflammation-related genes,while the HFHC+NLRP3-KD group had significant reductions in these indicators(all P<0.01).As for liver fibrosis,compared with the CON+NLRP3-NC group,the HFHC+NLRP3-NC group had significant increases in collagen fiber area and fibrosis-related genes,and the HFHC+NLRP3-KD group had significant reductions in fibrosis-related genes(all P<0.05)and a tendency of reduction in collagen fiber area(P>0.05).Conclusion NLRP3 knockdown can significantly improve hepatic fat deposition and inflammation in a mouse model of HFHC-induced NASH.

Objective:To investigate the effects of a perioperative whole course composite thermal insulation strategy on complications of cesarean section, maternal coagulation function and serum inflammatory indexes.Methods:A total of 250 pregnant women who were subjected to cesarean section in Zhoushan Hospital between June 2020 and August 2021 were included in this study. The 125 pregnant women who gave birth using a routine simple thermal insulation strategy from June to November 2020 were assigned to the routine simple thermal insulation group, and those who gave birth using a perioperative whole course composite thermal insulation strategy were assigned to whole course composite thermal insulation group. Two groups of pregnant women underwent cesarean section under subarachnoid block. Volume of intraoperative blood loss was recorded. The incidence of complications such as shivering and postoperative infection was calculated. Platelet count, prothrombin time, activated partial thromboplastin time, thrombin time measured before surgery and 48 hours after surgery were compared between the two groups. Peripheral blood white blood cell count, neutrophil count (N%), C-reactive protein, procalcitonin, interleukin-6 measured 48 hours after surgery were compared between the two groups.Results:Volume of intraoperative blood loss in the whole course composite thermal insulation group was significantly lower than that in the routine simple thermal insulation group [(393.84 ± 79.78) mL vs. (434.80 ± 123.49) mL, t = 3.11, P < 0.05). The incidence of shivering and postoperative infection in the whole course composite thermal insulation group was 10.4% (13/125) and 7.2% (9/125), respectively, which was significantly lower than that in the routine simple thermal insulation group [25.6% (32/125), 18.4% (23/125), χ 2 = 9.78, 7.02, both P < 0.05]. At 48 hours after surgery, prothrombin time, activated partial thromboplastin time, thrombin time in the whole course composite thermal insulation group were (10.28 ± 0.48) seconds, (26.97 ± 2.27) seconds, and (14.09 ± 1.36) seconds, respectively, which were significantly shorter than those in the routine simple thermal insulation group [(11.71 ± 0.27) seconds, (27.96 ± 2.25) seconds, (15.91 ± 1.09) seconds, t = 7.34, 3.43, 11.66, all P < 0.05]. At 48 hours after surgery, white blood cell count, neutrophil count, C-reactive protein, procalcitonin, and interleukin-6 in the whole course composite thermal insulation group were (10.38 ± 2.38) ×10 9/L,(0.79 ± 0.06), (52.79 ± 20.73) mg/L, (0.13±0.42) μg/L, and (55.73 ± 24.38) ng/L, respectively, which were significantly lower than those in the routine simple thermal insulation group [(12.24 ± 7.05) × 10 9/L, 0.81 ± 0.05, (65.38 ± 25.92) mg/L, (0.20 ± 0.97) μg/L, (76.22 ± 39.08) ng/L, t = 2.79, 2.92, 4.24, 8.12, 4.97, all P < 0.05]. Conclusion:Perioperative whole course composite thermal insulation strategy can improve the coagulation function of pregnant women who are subjected to cesarean section under subarachnoid block, reduce volume of intraoperative blood loss, and decrease incidence of shivering, inflammatory reaction, and postoperative infection.

Objective:To investigate the effects of perioperative whole course thermal insulation strategy on extubation, recovery time, coagulation index and the incidence of complications in patients undergoing thoracoscopic surgery.Methods:A total of 121 patients who underwent thoracoscopic surgery who received treatment in Zhoushan Hospital from October 2016 to February 2018 were included in this study. Among them, 59 patients who underwent thoracoscopic surgery from October 2016 to February 2017 were included in the simple thermal insulation group, and 62 patients who underwent thoracoscopic surgery from October 2017 to February 2018 were included in the whole process thermal insulation group. Before and 24 hours and 72 hours after surgery, platelet count, prothrombin time, activated partial thromboplastin time and thrombin time were compared between the two groups. Postoperative extubation time and recovery time were recorded. The incidence of shivering, restlessness and other complications was analyzed.Results:Postoperative extubation time and recovery time in the whole process thermal insulation group were significantly shorter than those in the simple thermal insulation group [(8.06 ± 4.60) min vs. (13.98 ± 7.22) min, (47.19 ± 12.97) min vs. (56.84 ± 17.49) min, t = 5.40, 3.47, both P < 0.05). At 24 and 72 hours after surgery, platelet count and activated partial prothrombin time in the whole process thermal insulation group were significantly shorter than those in the simple thermal insulation group [(12.55 ± 0.88) s vs. (13.11 ± 0.97) s, (27.44 ± 2.43) s vs. (29.03 ± 2.14) s, (11.42 ± 0.73) s vs. (11.87 ± 0.74) s, (27.44 ± 1.96) s vs. (28.80 ± 2.22) s, t = 3.32, 3.81, all P < 0.05). The incidence of postoperative shivering and restlessness in the whole process thermal insulation group was significantly lower than that in the simple insulation Group (7 cases vs. 27 cases, 5 cases vs. 22 cases, χ2 = 17.782, 14.894, 3.33, 3.57, all P < 0.05). Conclusion:Perioperative whole course thermal insulation strategy can shorten extubation and recovery time, inhibit perioperative coagulation dysfunction, and decrease the incidence of perioperative restlessness, shivering and other complications caused by hypothermia. The effect of perioperative whole course thermal insulation is superior to that of simple thermal insulation.

OBJECTIVETo provide a theoretical basis for surface modification of titanium implants, the effects of the stiffness of polyelectrolyte multilayer films on titanium surface on bacterium adhesion was explored.

METHODSVia layer-by-layer technique, catechol functionalized polyelectrolyte multilayer film (cPEM) was constructed on titanium surface by using catechol functionalized hyaluronic acid (cHA) and lipopolysaccharide-amine nanopolymersomes (NP). The stiffness of cPEM was controlled by adjusting the catechol substitution degree of cHA (5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%). Titanium samples covered with cPEM were selected as test group, and the cPEM was constructed with the lowest, medium and highest stiffness. The polished titanium was used as a control. The surface topography of titanium before and after film construction was observed by scanning electron microscopy (SEM). At 1 and 24 h after incubation, the adhesion and clonal formation of Streptococcus mutans (S. mutans) on different titanium surfaces were quantified, and their morphology and survival status were observed by SEM and laser scanning confocal microscope (LSCM).

RESULTSWhen the catechol grafting ratio was 5%, 30% and 70%, the lowest, medium and highest cPEM stiffness were obtained, and the cPEM stiffness were (10.69±4.54) GPa(cPEM-L), (20.99± 5.81) GPa (cPEM-M) and (32.57±6.93) GPa (cPEM-H) respectively, and the stiffness of polished titanium was (107.12±8.68) GPa (P<0.05). SEM observation showed that after cPEM coating, the titanium surface became smoother. After incubation for 1 and 24 h, the amount of adhesion and clonal formation of S. mutans on cPEM were higher than those on control titanium, and the difference was statistically significant (P<0.05). SEM images showed that for 1 h incubation, softer surfaces were beneficial for S. mutans adhering and agglomerating, while this difference nearly disappeared at 24 h. Observation under LSCM revealed that most of bacteria were alive on titanium disks at 1 h, and their amount decreased with the increase of stiffness. At 24 h, the living/dead bacterium ratios on cPEM-L and control titanium was higher than that on cPEM-M and cPEM-H, and cPEM-L surface was dominated by living bacteria, while stiffer cPEM-M and cPEM-H had more dead bacteria than living bacteria.

CONCLUSIONSIncreasing the stiffness of polyelectrolyte films on titanium limits the adhesion of S. mutans. As an independent factor, stiffness influences the bacterium adhesion.

Bacterial Adhesion ; Catechols ; Elasticity ; Hyaluronic Acid ; Lipopolysaccharides ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Nanoparticles ; Polymers ; chemistry ; Streptococcus mutans ; physiology ; Surface Properties ; Time Factors ; Titanium ; chemistry

We immobilized carbonic anhydrase (CA) onto the surface of membrane oxygenator of polymethyl pentene (PMP) to enhance the removal of carbon dioxide in blood by two steps. We first introduced hydroxyl groups onto PMP surface by water plasma treatment, and then coupled CA onto PMP surface by using cyanate bromide (CNBr) as a crosslinker. After plasma treatment, the contact angle with water and chemical composition of PMP surface were characterized by analysis system of surface contact angle and XPS. Using p-nitrophenyl acetate (p-NPA) as a substrate, the activity, concentration, storage stability and re-usability of immobilized CA on PMP hollow fibers were studied by ultraviolet spectrophotometer. The preliminary data showed that hydroxyl groups could be introduced on the surface of PMP by water plasma treatment, and CA with catalysis activity could be successfully introduced onto PMP surface in high immobilization efficiency. The activity of covalently immobilized CA increased with the increase of concentration of CNBr, and the maximum was 73% of the theoretical activity of CA spread on PMP surface in monolayer in studied range. Covalently immobilized CA showed higher reusability compared to physically adsorbed CA, and higher storage stability compared to free CA in solution at 37 degrees C. The method would be used potentially in the membrane oxygenator to improve the capacity of removal of carbon dioxide in blood in the future.
Carbonic Anhydrases ; chemistry ; metabolism ; Enzymes, Immobilized ; Membranes, Artificial ; Oxygenators, Membrane ; Polyenes ; chemistry

Objective To investigate the impact of symbiotic probiotic fermentation on anti-tumor effect of Agaricus blazei polysaccharide. Methods Probiotic fermentation and non-fermentation of Agaricus blazei polysaccharide samples were divided into large (0.8 g/kg), middle (0.4 g/kg), small (0.2 g/kg) dose groups, and negative control group for a total of 7 group. The anti-tumor effect on tumor-bearing mice was observed. Results The highest inhibition rate of probiotic fermentation Agaricus blazei polysaccharides group was 53.71%, while that of non-fermatation of Agaricus blazei polysaccharide group was 38.90%. Conclusion Inhibition rate of probiotic fermentation samples of Agaricus blazei polysaccharide was significantly higher than non-fermentation samples. Probiotic fermentation can significantly improve the anti-tumor effect of the Agaricus blazei.

3-year practice and questionnaire of the graduates majoring in ophthalmology indicate ophthalmologic typical case image implicates very much clinical information,can satisfy ophthalmologic clinical skills test,reflect examinee's clinical knowledge and technical skill level and guide intern to cultivate clinical thinking capability.

Copoly (benzyl glutamate-hydroxy ethyl glutamine) were prepared by partially aminolysis of poly benzyl glutamate with hydroxyl ethylamine at 60 degrees C for predeternined period. As aminolysis was proceeding, the content of hydroxy ethyl glutamine in copolymer increased and the yield of aminolysis decreased. When aminolysis time reached 16.5 hours, a copolymer with 0.357 mole fraction of hydroxy ethyl glutamine was obtained. After 16.5 hours a soluble yellow viscous product of reaction was obtained. The aminolysized specimens displayed more swelling degree in water, that is, more hydrophilits. The meassurement of solid-liquid contact angles showed that an increase in critical surface tention with content of hydroxy ethyl glutamine in copolymer was observed. Likewise a obvious increase in polar component and a slight decrease in dispersive of surface free energy, thus an increase in total surface free energy with content of hydroxy ethyl glutamine was found. The platelet adhesion and deformation test indicated that less platelets were adhered to surfaces of all aminolysized specimens than that of both poly benzyl glutamate and polydimethylsilicone. The least adhered platelets on surface of copolymer with 0.133 mole fraction of content of hydroxy ethyl glutamine were observed. The partiall aminolysis of poly benzyl glutamate films is an effective method to improve its hydrophility and antithrombogenicity.
Animals ; In Vitro Techniques ; Male ; Materials Testing ; Platelet Adhesiveness ; Polyglutamic Acid ; analogs & derivatives ; chemistry ; Rabbits ; Surface Properties