Objective To explore the effect of curcumin(CUR)on oxidative damage of keratinocytes induced by ultraviolet B(UVB)irradiation through Toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)/nucleotide-binding oligomerization domain-containing protein 3(NLRP3)signaling pathway.Methods Human keratinocytes of HaCaT were cultured normally in vitro,and the keratinocyte oxidative damage model was established by the irradiation of 57 mJ/cm2 UVB.The cells with normal culture were as the control group,the cells treated after modeling were as the UVB group,the cells treated with 5 μmol/L CUR after modeling were as the CUR group,the cells treated with 100 μg/L TLR4 inhibitor of TAK-242 after modeling were as the TAK-242 group,and the cells treated with 5 μmol/L CUR and 100 nmol/L TLR4 activator of lipopolysaccharide(LPS)were as the CUR+LPS group.qRT-PCR was applied to detect the relative expression levels of TLR4,NF-κB,and NLRP3 mRNAs of cells in each group.CCK-8 was applied to detect the cell proliferation in each group.The relative content of reactive oxygen species(ROS),the viabilities of superoxide dismutase(SOD)and catalase(CAT),and the concentrations of glutathione(MDA)and glutathione(GSH)of cells in each group were detected by fluorescence assay according to the kit instruction.ELISA kit was used to detect the expression of inflammatory factors of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)of cells in each group.Flow cytometry was applied to detect the cell apoptosis in each group.Western blot was applied to detect the expression of proliferation related protein of proliferating cell nuclear antigen(PCNA),apoptosis related proteins[B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)],and TLR4/NF-κB/NLRP3 signaling pathway related proteins(TLR4,NF-κB and NLRP3)of cells in each group.Results Compared with the Control group,the cell survival rate,the expression levels of PCNA and Bcl-2 proteins,the viabilities of SOD and CAT,and the GSH concentration in the UVB group decreased,while the apoptosis rate,the level of Bax protein,the relative content of ROS,the concentration of MDA,the levels of TNF-α and IL-1β,and the mRNA and protein levels of TLR4,NF-κB and NLRP3 increased(P<0.05).Compared with the UVB group,the cell survival rate,the expression levels of PCNA and Bcl-2 proteins,the viabilities of SOD and CAT,and the GSH concentration in the TAK-242 group and CUR group increased,while the apoptosis rate,the level of Bax protein,the relative content of ROS,the concentration of MDA,the levels of TNF-α and IL-1β,and the mRNA and protein levels of TLR4,NF-κB and NLRP3 decreased(P<0.05).Compared with the CUR group,the cell survival rate,the expression levels of PCNA and Bcl-2 proteins,the viabilities of SOD and CAT,and the GSH concentration in the CUR+LPS group decreased,while the apoptosis rate,the level of Bax protein,the relative content of ROS,the concentration of MDA,the levels of TNF-α and IL-1β,and the mRNA and protein levels of TLR4,NF-κB and NLRP3 increased(P<0.05).Conclusion CUR can increase the antioxidant stress level of keratinocytes,alleviate inflammatory response,promote cell proliferation,and improve cell damage caused by UVB irradiation,which may be related to the inhibition of TLR4/NF-κB/NLRP3 signaling pathway.

Objective:To analyze changes of donor corneal endothelial cell density (ECD) and morphology from eye bank before and after keratolasty and the influencing factors.Methods:An observational case series study was performed.A total of 118 donor corneas, retrieved by the Shandong Province Eye Bank between July 2020 and June 2021 for penetrating keratoplasty (PKP) and endothelial keratoplasty (EK) were included.Among them, 99 corneas (83.90%) were used for PKP, and 19(16.10%) for EK.The basic information of donors and the results of corneal quality tests were analyzed and compared with ECD measured by endothelial microscopy one month after keratolasty.Morphological changes in endothelial cells before and after surgery were observed, and factors influencing corneal ECD and morphology were analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Shandong Eye Hospital (No.SDSYKYY20170319).Written informed consent was obtained from each subject.Results:Among the 99 donor corneas for PKP, there were statistically significant differences in preoperative donor corneal ECD and 1-month postoperative ECD of implant among different age groups ( F=18.136, 5.936; both P<0.01), which were lower in the 31-60-year-old group and the >60-year-old group than in the 0-30-year-old group and higher in the 31-60-year-old group than in the >60-year-old group, with statistically significant differences (all P<0.01).There were statistically significant differences in the preoperative donor corneal ECD among different donor cause of death groups ( F=4.524, P<0.01), which was higher in the traumatic accident group compared to the cardiovascular and cerebrovascular disease group, chronic organ failure group and malignant tumor group (all P<0.01).The preoperative donor ECD in the death-tissue retrieval time ≤6 hours group was (2 577.66±284.63)cells/mm 2, which was higher than (2 372.46±399.75)cells/mm 2 in the death-tissue retrieval time >6 hours group, with a statistically significant difference ( t=2.289, P<0.05).There were statistically significant differences in 1-month postoperative ECD among the preservation-surgery time ≤3 days, 3-6 days, and >6 days groups ( F=6.201, P<0.01), with higher ECD in preservation-surgery time ≤3 days groups than in 3-6 days and >6 days groups (both P<0.01).The preoperative donor corneal ECD applied to EK was significantly higher than that applied to PKP ( t=-2.660, P<0.01).ECD at 1 month after surgery applied to PKP was significantly higher than that applied to EK ( t=4.286, P<0.01).The ECD reduction rate was 7.14% (0.01%, 17.69%) and 31.07% (22.11%, 45.86%) in PKP group and EK group, respectively, with a statistically significant difference ( Z=4.969, P<0.01).The ECD was lower in the group with dark area than in the non-dark area group before PKP, with a statistically significant difference ( t=6.789, P=0.011).There was no significant difference in ECD at 1 month after keratoplasty between the two groups ( t=0.005, P=0.945).Multivariate logistic regression model results showed that preservation-surgery time >6 days and the cause of donor death being malignant tumor were risk factors for the appearance of dark areas in donor corneal endothelium ( OR=9.038, P=0.030; OR=6.577, P=0.018). Conclusions:The older the donor, the lower the ECD.Prolonged preservation-surgery time (>6 days) is the main factor contributing to the decline in ECD after keratolasty.Compared to PKP, there is a higher endothelial cell loss after EK.The tissue preservation-surgery time >6 days and the cause of donor death being malignant tumor are the main risk factors affecting the appearance of dark areas in the donor corneal endothelium.But the presence of physiological dark areas does not significantly influence the ECD after surgery.

Objective To explore the effect of curcumin(CUR)on oxidative damage of keratinocytes induced by ultraviolet B(UVB)irradiation through Toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)/nucleotide-binding oligomerization domain-containing protein 3(NLRP3)signaling pathway.Methods Human keratinocytes of HaCaT were cultured normally in vitro,and the keratinocyte oxidative damage model was established by the irradiation of 57 mJ/cm2 UVB.The cells with normal culture were as the control group,the cells treated after modeling were as the UVB group,the cells treated with 5 μmol/L CUR after modeling were as the CUR group,the cells treated with 100 μg/L TLR4 inhibitor of TAK-242 after modeling were as the TAK-242 group,and the cells treated with 5 μmol/L CUR and 100 nmol/L TLR4 activator of lipopolysaccharide(LPS)were as the CUR+LPS group.qRT-PCR was applied to detect the relative expression levels of TLR4,NF-κB,and NLRP3 mRNAs of cells in each group.CCK-8 was applied to detect the cell proliferation in each group.The relative content of reactive oxygen species(ROS),the viabilities of superoxide dismutase(SOD)and catalase(CAT),and the concentrations of glutathione(MDA)and glutathione(GSH)of cells in each group were detected by fluorescence assay according to the kit instruction.ELISA kit was used to detect the expression of inflammatory factors of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)of cells in each group.Flow cytometry was applied to detect the cell apoptosis in each group.Western blot was applied to detect the expression of proliferation related protein of proliferating cell nuclear antigen(PCNA),apoptosis related proteins[B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)],and TLR4/NF-κB/NLRP3 signaling pathway related proteins(TLR4,NF-κB and NLRP3)of cells in each group.Results Compared with the Control group,the cell survival rate,the expression levels of PCNA and Bcl-2 proteins,the viabilities of SOD and CAT,and the GSH concentration in the UVB group decreased,while the apoptosis rate,the level of Bax protein,the relative content of ROS,the concentration of MDA,the levels of TNF-α and IL-1β,and the mRNA and protein levels of TLR4,NF-κB and NLRP3 increased(P<0.05).Compared with the UVB group,the cell survival rate,the expression levels of PCNA and Bcl-2 proteins,the viabilities of SOD and CAT,and the GSH concentration in the TAK-242 group and CUR group increased,while the apoptosis rate,the level of Bax protein,the relative content of ROS,the concentration of MDA,the levels of TNF-α and IL-1β,and the mRNA and protein levels of TLR4,NF-κB and NLRP3 decreased(P<0.05).Compared with the CUR group,the cell survival rate,the expression levels of PCNA and Bcl-2 proteins,the viabilities of SOD and CAT,and the GSH concentration in the CUR+LPS group decreased,while the apoptosis rate,the level of Bax protein,the relative content of ROS,the concentration of MDA,the levels of TNF-α and IL-1β,and the mRNA and protein levels of TLR4,NF-κB and NLRP3 increased(P<0.05).Conclusion CUR can increase the antioxidant stress level of keratinocytes,alleviate inflammatory response,promote cell proliferation,and improve cell damage caused by UVB irradiation,which may be related to the inhibition of TLR4/NF-κB/NLRP3 signaling pathway.

Objective To explore the disinfection effect of periodic hypochlorous acid infusion on water supply pipelines.Methods Water specimens from 6 water outlets of central sterile supply department were collected at baseline for microbial culture.After introducing a slightly acidic hypochlorous acid water generator,water speci-mens were collected every 10 days.Continuous disinfection with hypochlorous acid was carried out on the 1st-3rd day,and discontinued on the 4th-10th day.Water specimens of 6 water outlets were collected on the 1st,4th,7th,and 10th day of each cycle,in total 3 cycles.Microbial culture and metagenomic next-generation sequencing(mNGS)analysis were conducted simultaneously on all water specimens from the outlets of the endoscope cleaning tank.Results Before disinfection of the pipeline,a total of 18 bottles of water specimens were collected.Bacterial count of water specimens from 10 bottles(55.56％)was between 102 and 104 CFU/mL,mainly Sphingomonas ad-hesins and Bacillus cereus.After disinfection,only 2 out of 72 water specimens(2.78％)exceeded the bacterial lim-it.The mNGS results showed that three most dominant bacterial genera in the pre-disinfection water specimens were Sphingomonas,Methylobacterium,and Ralstonia,with stringent mapped reads number(SMRN)ranging from 10 000 to 100 000;while the most dominant bacterial genera in post-disinfection water specimens were Lacto-baci-llus,Ralstonia,Acinetobacter,and Bradyrhizobium.PCoA analysis showed that there was a statistically sig-nificant difference in β-diversity of water specimens before and after disinfection(P＜0.05).Conclusion Periodic hypochlorous acid infusion disinfection can effectively remove bacterial biofilms in water pipelines,but there is a risk of bacterial biofilm recovery during disinfection suspension.In the future,monitoring and management of medical water should be strengthened.

Excessive fat accumulation,viral infections and sustained inflammatory responses caused by non-alcoholic and alcoholic factors can contribute to liver inflammation,fibrosis and carcinogenesis,promoting the development of chronic liver disease.Gaining an in-depth understanding of the etiologic factors and underlying mechanisms that lead to chronic liver disease can help identify potential therapeutic targets for targeted therapy.Lactate,as an important substance in hepatic metabolism,has been found to be involved in the process of chronic liver disease through various pathways,and this review will provide a useful reference for the prevention and treatment of chronic liver disease.

Objective:To analyze changes of donor corneal endothelial cell density (ECD) and morphology from eye bank before and after keratolasty and the influencing factors.Methods:An observational case series study was performed.A total of 118 donor corneas, retrieved by the Shandong Province Eye Bank between July 2020 and June 2021 for penetrating keratoplasty (PKP) and endothelial keratoplasty (EK) were included.Among them, 99 corneas (83.90%) were used for PKP, and 19(16.10%) for EK.The basic information of donors and the results of corneal quality tests were analyzed and compared with ECD measured by endothelial microscopy one month after keratolasty.Morphological changes in endothelial cells before and after surgery were observed, and factors influencing corneal ECD and morphology were analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Shandong Eye Hospital (No.SDSYKYY20170319).Written informed consent was obtained from each subject.Results:Among the 99 donor corneas for PKP, there were statistically significant differences in preoperative donor corneal ECD and 1-month postoperative ECD of implant among different age groups ( F=18.136, 5.936; both P<0.01), which were lower in the 31-60-year-old group and the >60-year-old group than in the 0-30-year-old group and higher in the 31-60-year-old group than in the >60-year-old group, with statistically significant differences (all P<0.01).There were statistically significant differences in the preoperative donor corneal ECD among different donor cause of death groups ( F=4.524, P<0.01), which was higher in the traumatic accident group compared to the cardiovascular and cerebrovascular disease group, chronic organ failure group and malignant tumor group (all P<0.01).The preoperative donor ECD in the death-tissue retrieval time ≤6 hours group was (2 577.66±284.63)cells/mm 2, which was higher than (2 372.46±399.75)cells/mm 2 in the death-tissue retrieval time >6 hours group, with a statistically significant difference ( t=2.289, P<0.05).There were statistically significant differences in 1-month postoperative ECD among the preservation-surgery time ≤3 days, 3-6 days, and >6 days groups ( F=6.201, P<0.01), with higher ECD in preservation-surgery time ≤3 days groups than in 3-6 days and >6 days groups (both P<0.01).The preoperative donor corneal ECD applied to EK was significantly higher than that applied to PKP ( t=-2.660, P<0.01).ECD at 1 month after surgery applied to PKP was significantly higher than that applied to EK ( t=4.286, P<0.01).The ECD reduction rate was 7.14% (0.01%, 17.69%) and 31.07% (22.11%, 45.86%) in PKP group and EK group, respectively, with a statistically significant difference ( Z=4.969, P<0.01).The ECD was lower in the group with dark area than in the non-dark area group before PKP, with a statistically significant difference ( t=6.789, P=0.011).There was no significant difference in ECD at 1 month after keratoplasty between the two groups ( t=0.005, P=0.945).Multivariate logistic regression model results showed that preservation-surgery time >6 days and the cause of donor death being malignant tumor were risk factors for the appearance of dark areas in donor corneal endothelium ( OR=9.038, P=0.030; OR=6.577, P=0.018). Conclusions:The older the donor, the lower the ECD.Prolonged preservation-surgery time (>6 days) is the main factor contributing to the decline in ECD after keratolasty.Compared to PKP, there is a higher endothelial cell loss after EK.The tissue preservation-surgery time >6 days and the cause of donor death being malignant tumor are the main risk factors affecting the appearance of dark areas in the donor corneal endothelium.But the presence of physiological dark areas does not significantly influence the ECD after surgery.

Objective To explore the disinfection effect of periodic hypochlorous acid infusion on water supply pipelines.Methods Water specimens from 6 water outlets of central sterile supply department were collected at baseline for microbial culture.After introducing a slightly acidic hypochlorous acid water generator,water speci-mens were collected every 10 days.Continuous disinfection with hypochlorous acid was carried out on the 1st-3rd day,and discontinued on the 4th-10th day.Water specimens of 6 water outlets were collected on the 1st,4th,7th,and 10th day of each cycle,in total 3 cycles.Microbial culture and metagenomic next-generation sequencing(mNGS)analysis were conducted simultaneously on all water specimens from the outlets of the endoscope cleaning tank.Results Before disinfection of the pipeline,a total of 18 bottles of water specimens were collected.Bacterial count of water specimens from 10 bottles(55.56％)was between 102 and 104 CFU/mL,mainly Sphingomonas ad-hesins and Bacillus cereus.After disinfection,only 2 out of 72 water specimens(2.78％)exceeded the bacterial lim-it.The mNGS results showed that three most dominant bacterial genera in the pre-disinfection water specimens were Sphingomonas,Methylobacterium,and Ralstonia,with stringent mapped reads number(SMRN)ranging from 10 000 to 100 000;while the most dominant bacterial genera in post-disinfection water specimens were Lacto-baci-llus,Ralstonia,Acinetobacter,and Bradyrhizobium.PCoA analysis showed that there was a statistically sig-nificant difference in β-diversity of water specimens before and after disinfection(P＜0.05).Conclusion Periodic hypochlorous acid infusion disinfection can effectively remove bacterial biofilms in water pipelines,but there is a risk of bacterial biofilm recovery during disinfection suspension.In the future,monitoring and management of medical water should be strengthened.

AIM To investigate the improvement effect of Qinhuo Formula on obese mice induced by high-fat diet.METHODS Obese mice model was induced by feeding with high-fat diet.The mice successfully established were randomly divided into model group,orlistat group(15.6 mg/kg)and low and high dose groups of Qinhuo Formula(13.5 and 22.5 g/kg),and normal mice were taken as control group,with 10 mice in each group,and the drugs were continuously intervened for 8 weeks.After the drug administration,the body weight,abdominal circumference,fasting blood glucose and glucose tolerance were measured,and area under the curve(AUC)of oral glucose tolerance test(OGTT),Lee's index and fat index were calculated.The serum levels of TC,TG,LDL-C and HDL-C were detected by blood biochemical analyzer.Serum levels of TNF-α,IL-1 β and IL-6 were detected by ELISA.HE staining was used to observe the white fat of epididymis and the pathological changes of liver tissue.RT-qPCR and Western blot were used to detect the mRNA and protein expressions of TLR4,MyD88 and NF-κB in liver tissue.RESULTS Compared with the model group,the body weight,abdominal circumference and Lee's index of mice in each dose group of Qinhuo Formula decreased(P＜0.01);OGTT-AUC decreased(P＜0.01);visceral fat index decreased(P＜0.05,P＜0.01);the levels of serum TG,TC,IL-6 and IL-1 β decreased(P＜0.01);the diameter of epididymal adipocytes decreased,arranged tightly and regularly,the lipid droplet cavities in liver tissue decreased,and the morphology of hepatocytes recovered;the mRNA and protein expressions of TLR4,MyD88 and NF-κB decreased(P＜0.05,P＜0.01).CONCLUSION Qinhuo Formula can effectively improve glucose-lipid metabolism and reduce inflammation in obese mice,and its mechanism may be related to the inhibition of TLR4/MyD88/NF-κB signaling pathway.

Objective To study the protective effects and mechanism of sacubitril(Sac)/valsartan(Val)on cardiomyocytes of rabbits with heart failure induced by doxorubicin(DOX).Methods Thirty New Zealand rabbits were selected to establish DOX-induced heart failure rabbit model.Twenty-five rabbits with successful modeling were randomly assigned to model group(DOX group,n=9),DOX+Val group(n=8),and DOX+Sac/Val group(n=8);and another 8 New Zealand rabbits were selected as blank group.The DOX+Val group was gavaged with 4.65 mg/kg Val suspension each time,the DOX+Sac/Val group was gavaged with 9.3 mg/kg Sac/Val suspension each time,and the blank group and DOX group were gavaged with equal volume of distilled water each time.Each group was gavaged twice a day for 8 weeks.After 8 weeks of administration,echocardiography was used to measure left ventricular end-diastolic diameter(LVDD),left ventricular end-systolic diameter(LVSD),left ventricular ejection fraction(LVEF),and left ventricular fractional shortening(LVFS).The heart mass index(HMI)and left ventricular mass index(LVMI)were calculated.The pathological morphology and myocardial fibrosis of myocardial tissue were observed by hematoxylin-eosin(H-E)and Masson staining.The ultrastructure of cardiomyocytes was observed by transmission electron microscope.Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining was used to observe cardiomyocytes apoptosis and apoptosis rate was calculated.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of N-terminal pro-brain natriuretic peptide(NT-proBNP),high-sensitivity cardiac troponin I(Hs-cTNI),angiotensin Ⅱ(Ang Ⅱ),aldosterone(ALD),atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),cyclic guanosine monophosphate(cGMP),and protein kinase G(PKG)in serum.Quantitative polymerase chain reaction(qPCR)was used to detect the expression of natriuretic peptide receptor A(NPR-A),cGMP-specific phosphodiesterase 5A(PDE5A[cGMP]),PKG,B-cell lymphoma 2(Bcl-2),Bcl-2 associated X protein(Bax),and cysteine aspartate protease 3(caspase 3)mRNA in myocardial tissue.Western blotting was used to detect the expression of phosphorylated cAMP response element-binding protein(p-CREB),phosphorylated Bcl-2 related death promoting factor(p-Bad),Bcl-2,Bax,and caspase 3 proteins in myocardial tissue.Results Compared with the blank group,the LVDD and LVSD in the DOX group were increased(both P<0.01),the LVEF and LVFS were decreased(both P<0.01)and the HMI and LVMI were increased(both P<0.01);the apoptosis and apoptosis rate of cardiomyocytes were increased(P<0.01);the levels of NT-proBNP,Hs-cTNI,Ang Ⅱ,ALD,ANP,BNP,cGMP and PKG and the expression of NPR-A,PDE5A(cGMP),PKG,p-CREB,Bax and caspase 3 were all increased(all P<0.01),while the expression of Bcl-2 was decreased(P<0.01),and the expression of p-Bad had no significant difference(P>0.05).Compared with the DOX group,the LVDD and LVSD of the DOX+Sac/Val group and DOX+Val group were decreased(all P<0.01),the LVEF and LVFS were increased(all P<0.01)and the HMI and the LVMI were decreased(all P<0.01);the apoptosis and apoptosis rate of cardiomyocytes were decreased(all P<0.01);the levels of NT-proBNP,Hs-cTNI,Ang Ⅱ,ALD,ANP,BNP,cGMP and PKG and the expression of NPR-A,PDE5A(cGMP),PKG,Bax and caspase 3 were all decreased(all P<0.01),while the expression of Bcl-2 was increased(P<0.01);and the expression of p-CREB and p-Bad was increased in the DOX+Sac/Val group(both P<0.01),but there was no significant difference in the DOX+Val group(both P>0.05).Compared with the DOX+Val group,the DOX+Sac/Val group showed a decrease in all indicators except for LVEF,LVFS,NPR-A,ANP,BNP,cGMP,PDE5A(cGMP),PKG,p-CREB,p-Bad,and Bcl-2,which were all elevated(all P<0.05).Myocardial pathology and transmission electron microscopy showed that Sac/Val effectively protected cardiomyocytes,reduced cardiomyocytes apoptosis and myocardial fibrosis,and these effects were significantly better than those of Val.Conclusion Sac/Val can effectively reduce cardiomyocytes apoptosis,improve cardiac function and reduce myocardial fibrosis in rabbits with heart failure,and these effects are superior to Val.Its mechanism may be related to activating the NPR-A/cGMP/PKG signaling pathway and inhibiting renin-angiotensin-aldosterone system.