Objective To analyze drug resistance and clustering of environmental and clinical isolates of Acinetobacter baumannii (A. baumannii) in ICU of a medical institution in Shanghai. Methods The isolates of A. baumannii from ICU environments and clinic were used to analyze the contamination and distribution in 2021-2024. Antimicrobial susceptibility testing was carried out with microbroth dilution method. Whole genome sequencing was performed out of strains for MLST typing and SNP clustering. Results The detection rate of contamination in ICU environment was 7.67%, and the most serious contamination was found in pillows, bedding, hospital gowns and other items that patients directly contacted. Clinical isolates were predominantly from sputum specimens. The environmental and clinical isolates had a high level of resistance to third generation cephalosporins, third generation quinolones and carbapenems (more than 85%). Environmental isolates had a low level of resistance to polymyxin B, but none of the clinical isolates were resistant. MLST typing showed that ST2 was the dominant clone (66.67%), and SNP clustering found that isolates from different sources but with the same ST type were clustered together. Conclusion ST2 is the dominant clone of A. baumannii isolates in this medical institution, and there is cross-contamination between different samples. Monitoring of drug resistance and disinfection should be further strengthened to prevent the emergence and spread of pan-resistant or even fully resistant strains.

OBJECTIVE To investigate the effect and mechanism of Jingangteng capsules in the treatment of non-alcoholic fatty liver disease （NAFLD）. METHODS Thirty-two SD rats were randomly divided into normal group and modeling group. The modeling group was fed a high-fat diet to establish a NAFLD model. The successfully modeled rats were then randomly divided into model group， atorvastatin group[positive control， 2 mg/（kg·d）]， and Jingangteng capsules low- and high-dose groups [0.63 and 2.52 mg/（kg·d）]， with 6 rats in each group. The pathological changes of the liver were observed by hematoxylin-eosin staining and oil red O staining. Enzyme-linked immunosorbent assay was performed to determine the serum levels of triglycerides （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， alanine transaminase （ALT）， aspartate transaminase （AST）， tumour necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， IL-6， IL-18. 16S rDNA amplicon sequencing and metabolomics techniques were applied to explore the effects of Jingangteng capsules on gut microbiota and metabolisms in NAFLD rats. Based on the E-mail：591146765@qq.com metabolomics results， Western blot analysis was performed to detect proteins related to the nuclear factor kappa-B （NF-κB）/NOD-like receptor family protein 3 （NLRP3） signaling pathway in the livers of NAFLD rats. RESULTS The experimental results showed that Jingangteng capsules could significantly reduce the serum levels of TG， TC， LDL-C， AST， ALT， TNF-α， IL-1β， IL-6， IL-18， while increased the level of HDL-C， and alleviated the hepatic cellular steatosis and inflammatory infiltration in NAFLD rats. They could regulate the gut microbiota disorders in NAFLD rats， significantly increased the relative abundance of Romboutsia and Oscillospira， and significantly decreased the relative abundance of Blautia （P＜0.05）. They also regulated metabolic disorders primarily by affecting secondary bile acid biosynthesis， fatty acid degradation， O-antigen nucleotide sugar biosynthesis， etc. Results of Western blot assay showed that they significantly reduced the phosphorylation levels of NF-κB p65 and NF-κB inhibitor α， and the protein expression levels of NLRP3， caspase-1 and ASC （P＜0.05 or P＜0.01）. CONCLUSIONS Jingangteng capsules could improve inflammation， lipid accumulation and liver injury in NAFLD rats， regulate the disorders of gut microbiota and metabolisms， and inhibit NF-κB/NLRP3 signaling pathway. Their therapeutic effects against NAFLD are mediated through the inhibition of the NF-κB/NLRP3 signaling pathway.

Aim To investigate the effect of p-AMPK activity on autophagy in different subtypes of MDA-MB-231(triple-negative breast cancer cells)and MCF-7(estrogen receptor-positive cells)and its regulatory mechanism.Methods MDA-MB-231 cells were trea-ted with EBSS,Baf-A1,and EBSS+Baf-A1 for four hours,and MCF-7 cells for eight hours.The effects of autophagy on cell proliferation and apoptosis were ob-served,mitochondrial morphology was examined,and the expression of autophagy markers LC3B,P62,LAMP1,TOM20,AMPK,p-AMPK,ULK1,and Bec-lin1/VPS34 proteins was detected.The autophagy pathway was validated by inhibiting AMPK activity.Results Breast cancer cells underwent autophagy af-ter starvation induction(EBSS),with inconsistent au-tophagy processes observed in different subtypes of breast cancer cells.Autophagy inhibited cell prolifera-tion.In MDA-MB-231 cells,autophagy led to an in-crease in p-AMPK levels and a decrease in ULK1 lev-els,initiating autophagy through p-AMPK activation of ULK1.In MCF-7 cells,both p-AMPK and ULK1 levels decreased after autophagy,suggesting that autophagy might not be mediated by p-AMPK activation.Conclu-sions MDA-MB-231 cells primarily initiate autophagy by directly activating ULK1 by p-AMPK,independent of the MTOR pathway.In MCF-7 cells autophagy might be triggered by inhibiting MTOR through AMPK activity or directly activating MTOR through other up-stream factors.Regulating p-AMPK activity based on the autophagy pathways in different cell subtypes could enable more precise targeting and treatment of different types of breast cancer.

Objective:To investigate the effects of smoking cessation on lung function improvement in patients with chronic obstructive pulmonary disease (COPD) and the related mechanisms.Methods:A case-control study was conducted involving 184 patients with COPD admitted to the Department of Respiratory and Critical Care Medicine at Lishui Central Hospital from January 2022 to May 2023. Based on smoking behavior following 6-month smoking cessation intervention, the patients were categorized into three groups: 56 patients who continued to smoke (control group), 63 patients who completely quit smoking (observation group), and the remaining 65 patients who were partial quitters. Forced vital capacity (FVC), forced expiratory volume in 1 second (FEV 1), and peak expiratory flow rate were monitored before and after the intervention in both the control and observation groups. Additionally, serum levels of interleukin-6 (IL-6), interleukin-1 beta (IL-1β), tumor necrosis factor alpha, nuclear factor kappa B (NF-κB), malondialdehyde, superoxide dismutase, and glutathione peroxidase were measured. Western blotting was performed to detect the protein levels of phosphoinositide 3-kinase (PI3K), serine/threonine kinase (Akt), Forkhead box protein O1 (FoxO1), IL-6, IL-1β, NF-κB, nuclear factor erythroid 2-related factor 2, and heme oxygenase-1 in peripheral blood cells. FoxO1 chromatin immunoprecipitation sequencing was performed to analyze FoxO1-bound chromatin. Results:After smoking cessation intervention, the FEV 1, FVC, FEV 1/FVC ratio, and peak expiratory flow rate in the observation group were (1.29 ± 0.32) L, (1.96 ± 0.36) L, (71.81 ± 8.57)%, and (2.58 ± 0.72) L/s, respectively, which were significantly higher than those in the control group [(1.10 ± 0.37) L, (1.72 ± 0.34) L, (63.17 ± 8.82)%, (2.20 ± 0.71) L/s, t = -3.00, -3.73, -5.42, -2.89, all P < 0.01]. The serum levels of IL-6, NF-κB, IL-1β, and tumor necrosis factor alpha in the observation group were (21.67 ± 3.25) ng/L, (19.58 ± 4.02) ng/L, (24.30 ± 4.03) ng/L, and (270.14 ± 32.49) ng/L, respectively, which were significantly lower than those in the control group [(39.18 ± 4.34) ng/L, (35.48 ± 4.17) ng/L, (34.42 ± 4.05) ng/L, (445.04 ± 39.12) ng/L, t = 25.08, 21.16, 13.64, 26.63, all P < 0.001]. Additionally, the serum malondialdehyde level in the observation group was significantly lower than that in the control group ( t = 29.08, P < 0.001). In contrast, the levels of superoxide dismutase and glutathione peroxidase in the observation group were significantly higher than those in the control group ( t = -9.21, -9.59, both P < 0.001). The phosphorylation levels of PI3K, Akt, and FoxO1 in peripheral blood cells were significantly lower in the observation group compared with the control group ( t = 6.64, 9.35, 7.12, all P < 0.001). FoxO1 bound to genes involved in inflammation and oxidative stress signaling pathways. The levels of nuclear factor erythroid 2-related factor 2 and heme oxygenase-1 in the observation group were significantly higher than those in the control group ( t = -4.97, -10.49, both P < 0.05). Conclusions:Smoking cessation intervention can inhibit PI3K/Akt phosphorylation in patients with COPD, and then activate FoxO1, exert anti-inflammatory and antioxidant effects, and inhibit the deterioration of lung function in patients with COPD who smoke.

Creutzfeldt-Jakob disease(CJD)is a rapidly progressive and fatal neurodegenerative disorder caused by prions,with certain infectivity and iatrogenic transmission risks.With the rapid progress and application of new dia-gnostic biomarkers and detection methods,as well as the construction and improvement of surveillance and reporting systems,the detection of CJD in patients domestically and internationally has shown an increasing trend year by year.Due to its long incubation period and heterogeneity of early symptoms,early identification and diagnosis of the disease is difficult,increasing the risk of transmission within medical institutions.Currently,there is a lack of con-sensus on the infection prevention and control of CJD.In order to timely identify and diagnose CJD as well as effec-tively block its transmission in medical institutions,this consensus summarizes 15 clinical concerns and formulates 24 specific recommendations based on the latest domestic and international research findings and clinical evidence,as well as combines with clinical practice,aiming to standardize healthcare-associated infection prevention and control measures for CJD and reduce its transmission risk in medical institutions.

Objective To investigate the factors that influence ocular surface biology and visual acuity in individuals with diabetic dry eye(DDE)and analyze how these factors contribute to changes in visual quality.Methods Based on the disease duration,fasting blood glucose(FBG),and glycated hemoglobin(HbA1c)levels of patients with type 2 diabe-tes mellitus(T2DM),the DDE patients were divided into different groups.Logistic regression analysis was used to identify influencing factors related to ocular surface biology and visual quality in each group of DDE patients.Tear film stability was evaluated based on the tear film rupture time(BUT),Schirmer I test(SIt),and ocular surface disease index(OSDI).Lip-iview? Surface interferometers were used to measure tear film lipid layer thickness(LLT),meibomian gland loss rate(MGP),meibomian gland opening number(MGYLS),and meibomian gland secretion score(MGYSS).Wavefront aber-rometry was used to measure corneal wavefront aberration values at 4 mm and 6 mm pupil diameters.Ocular response ana-lyzer(ORA)was adopted to analyze corneal hysteresis(CH)and corneal resistance factor(CRF).Moreover,ELISA ex-periment to evaluate the trend of changes in inflammatory factors in tears.Results Logistic regression analysis revealed that T2DM duration,smoking history,FBG,HbA1c,total cholesterol(TC),triglycerides(TG),OSDI score,LLT,BUT,SIt,MGP,MGYLS,MGYSS,total higher-order aberrations,spherical aberration,coma aberration,trefoil aberration,tumor necrosis factor-α,interleukin-6,matrix metalloproteinase-9,receptor for advanced glycation end products,and insu-lin were all influencing factors for the risk of DDE(all P＜0.05).As the T2DM course prolonged and FBG or HbA1 c levels rose,tear film-related indicators(LLT,BUT,and SIt)and meibomian gland-related indicators(MGYLS and MGYSS)inpa-tients gradually decreased,while OSDI scores and MGP gradually increased(all P＜0.05).As the T2DM course prolonged and FBG or HbA1c levels rose,the total higher-order aberrations,spherical aberration,coma aberration,and trefoil aber-ration in DDE patients under 4 mm and 6 mm pupil diameters gradually increased;Meanwhile,best corrected visual acuity,corneal hysteresis,and corneal resistance factor gradually decreased;The contents of tumor necrosis factor-α,interleukin-6,matrix metalloproteinase-9,receptor for advanced glycation end products,and insulin in tears all gradually increased,while mucin-5AC gradually decreased(all P＜0.05).Conclusion With the prolongation of T2DM duration and the in-crease of FBG or HbA1c,the ocular surface inflammatory response in DDE patients gradually worsens,corneal biological function decreases,and visual quality deteriorates.Timely systemic and local interventions are of great significance for im-proving dry eye symptoms and visual quality in DDE patients.

Creutzfeldt-Jakob disease(CJD)is a rapidly progressive and fatal neurodegenerative disorder caused by prions,with certain infectivity and iatrogenic transmission risks.With the rapid progress and application of new dia-gnostic biomarkers and detection methods,as well as the construction and improvement of surveillance and reporting systems,the detection of CJD in patients domestically and internationally has shown an increasing trend year by year.Due to its long incubation period and heterogeneity of early symptoms,early identification and diagnosis of the disease is difficult,increasing the risk of transmission within medical institutions.Currently,there is a lack of con-sensus on the infection prevention and control of CJD.In order to timely identify and diagnose CJD as well as effec-tively block its transmission in medical institutions,this consensus summarizes 15 clinical concerns and formulates 24 specific recommendations based on the latest domestic and international research findings and clinical evidence,as well as combines with clinical practice,aiming to standardize healthcare-associated infection prevention and control measures for CJD and reduce its transmission risk in medical institutions.

The infectious clone plasmid of genotype Ⅰ Japanese encephalitis virus GX strain was suc-cessfully obtained by reverse transcription-polymerase chain reaction,where the cDNA of GX strain is divided into three fragments for amplification and the three fragments were sequentially cloned into pBR322 vector.After the infectious clone plasmid pGX was sequenced correctly,the pGX and pCAGGS-T7 eukaryotic plasmid were co-transfected into BHK-21 cells for virus rescue.The experimentalresults indicated that the Japanese encephalitis virus could be successfully res-cuedfrom BHK-21 cells.The plaque experiment and mouse experiment indicated that the rescued virus had similar replication ability and pathogenicity with wild type virus.It was confirmed that the infectious clone of genotype Ⅰ Japanese encephalitis virus GX strain was successfully construc-ted in this study.

Aim To investigate the neuroprotective effect of Takeda G protein-coupled receptor-5(TGR5)activated by INT-777 on hypoxic-ischemic encephalop-athy(HIE)in neonatal rats.Methods Seven-day-old SD rats were randomly divided into the sham opera-tion group(Sham,S),model group(HIE,G),INT-777 low-dose(L),medium-dose(M),and high-dose(H)groups.The modified Rice-Vanucci method was used to construct the HIE model and Intranasal admin-istration 1 h after modeling.Short-term neurobehavioral tests were performed 48 h after modeling to evaluate the neurological function of neonatal rats,TTC staining was used to determine the volume of cerebral infarction,dry and wet specific gravity was used to determine the brain water content,ferrous ion kit was used to deter-mine the brain ferrous ion content,HE staining was used to observe the pathological damage of brain tis-sue,Nissl staining was used to observe the loss of Nissl substance,Transmission electron microscopy(TEM)was used to observe the mitochondrial morphological changes of cortical neurons,and Western blot was em-ployed to detect the expression of ferroptosis-related proteins TFR1 and GPX4.Results Compared with group S,group G had increased short-term neurobehav-ioral test consumption time,higher scores,increased cerebral infarct volume,brain water content,and brain ferrous iron content,significant brain tissue damage on the affected side,severe loss of Nissl substance,smaller neuronal mitochondria,decreased mitochondrial cris-tae,and increased expression of TFR1 and reduced ex-pression of GPX4.Compared with group G,the INT-777 administration group had a shorter consumption time for short-term neurobehavioral tests,lower scores,the cerebral infarction volume,brain water content,and brain ferrous ion content decreased,the brain tissue damage on the affected side was reduced,and there was insignificant loss of Nissl substance,larger neuronal mi-tochondrial volume,increased mitochondrial cristae,re-duced expression of TFR1,and increased expression of GPX4.Conclusions INT-777 agonist TGR5 has a protective effect against hypoxic-ischemic encephalopa-thy in neonatal rats,and its mechanism of action may be related to the inhibition of neuronal ferroptosis.

BACKGROUND:The development of calcium phosphate bone cement is limited due to its poor mechanical properties and weak osteogenic ability.Silicate bioactive glass is highly favored due to its excellent biological activity and osteogenic ability.Simultaneously,fiber structures can enhance the mechanical strength of materials.OBJECTIVE:To investigate the mechanical properties,biocompatibility,and osteogenic effect of silicate bioactive glass fiber composite calcium phosphate bone cement.METHODS:Different mass percentages(0％,10％,and 20％)of silicate bioactive glass fiber were added to the solid phase of calcium phosphate bone cement,mixed with the liquid phase and cured for 48 hours to obtain silicate bioactive glass fiber composite calcium phosphate bone cement.The mechanical properties,setting time,and ion precipitation of the cement were characterized.The three groups of bone cement extracts were co-cultured with MC3T3-E1 cells.The cell compatibility of the materials was evaluated by CCK-8 assay,live/dead staining,and phalloidin staining.After osteogenic induction,the osteogenic induction ability of the materials was evaluated by alkaline phosphatase staining,alizarin red staining,RUNX2 immunofluorescence staining,and RT-PCR.RESULTS AND CONCLUSION:(1)With the increase of silicate bioactive glass fiber content,the compressive strength and flexural strength of bone cement increased,and the setting time was prolonged.When bone cement was immersed in simulated body fluid,the precipitation of silicon ions,calcium ions,and phosphorus ions could be detected.Moreover,with the increase of silicate bioactive glass fiber content,the mass concentration of silicon ions and phosphorus ions released by bone cement increased,and the mass concentration of calcium ions decreased.(2)Live/dead staining and phalloidin staining results exhibited that silicate bioactive glass fiber composite calcium phosphate bone cement had no toxic effect on MC3T3-E1 cells.CCK-8 assay results showed that silicate bioactive glass fiber composite calcium phosphate bone cement could promote the proliferation of MC3T3-E1 cells.(3)With the increase of silicate bioactive glass fiber content in bone cement,the alkaline phosphatase activity and extracellular calcium deposition of MC3T3-E1 cells increased,the expression of RUNX2 protein increased,and the expression of alkaline phosphatase,osteocalcin,osteopontin,and RUNX2 mRNA expression increased.(4)The results indicate that silicate bioactive glass fibers can enhance the mechanical properties and osteogenic induction ability of calcium phosphate bone cement,among which 20％silicate bioactive glass fibers have a more obvious effect.