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.

ObjectiveTo investigate the expression level of lysophosphatidyllecithin acyltransferase 4 （LPCAT4） in pancreatic cancer and its effect on the proliferation of pancreatic cancer cells. MethodsIn this study， the differentially expressed genes of patients with KRAS mutant and wild-type pancreatic cancer were analyzed by online database LinkedOmics. The LPCAT4 expression in pancreatic cancer tissues was analyzed online by the University of Alabama at Birmingham Cancer Data Analysis （UALCAN）， Sangerbox and gene expression profile interaction analysis 2 （GEPIA2）. Kaplan-Meier Plotter database was used to explore the correlation between LPCAT4 and the prognosis of patients with pancreatic cancer. The expression of LPCAT4 in human pancreatic cancer cells were detected by quantitative real-time PCR and Western blot analysis. LPCAT4 was knocked down in the high-expressing SW1990 cell line and overexpressed in the low-expressing MIA PaCa-2 cell line. The effects of LPCAT4 expression on cell proliferation were assessed using CCK-8 and EdU assays. STRING and GEPIA2 databases were used to obtain LPCAT4 binding and coexpressed genes in tumors， which were then analyzed by GO and KEGG. ResultsAnalysis of the LinkedOmics online database revealed a significant upregulation of LPCAT4 in patients with KRAS mutant pancreatic cancer compared to patients with KRAS wild-type pancreatic cancer. The online analysis of GEPIA2， UALCAN and Sangerbox 3.0 showed that the expression of LPCAT4 was higher in pancreatic cancer than in normal tissues. Analysis of the Kaplan-Meier Plotter database revealed that high LPCAT4 expression was associated with poorer prognosis in pancreatic cancer patients.Western blot and qPCR results showed that expression of LPCAT4 in pancreatic cancer cell lines was significantly higher than in normal pancreatic ductal epithelial cells. Knockdown of LPCAT4 in SW1990 cells inhibited proliferation， while overexpression in MIA PaCa-2 cells promoted proliferation. Enrichment analysis indicated that LPCAT4 was closely related to sulfur metabolism. ConclusionsLPCAT4 is highly expressed in pancreatic cancer and is associated with poor prognosis of patients. It plays a significant regulatory role in the proliferation of pancreatic cancer cells， with its expression level closely correlated with cell proliferation capacity. These findings reveal the critical role of LPCAT4 in the malignant progression of pancreatic cancer and provide important evidence for its potential as a therapeutic target.

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.

Objective To investigate the anti-tumor mechanism of natural compound toosendanin(TSN)combined with olaparib in triple-negative breast cancer(TNBC).Methods Human TNBC cell line MDA-MB-231 was cultured in vitro.Effects of TSN combined with olaparib on autophagy levels and cell viability in MDA-MB-231 cells were evaluated using 0.5,1.0,and 5.0 μmol/L olaparib alone or in combination.Surgical specimens from four TNBC patients who had residual tumors after neoadjuvant chemotherapy were selected to establish patient-derived organoid(PDO)models.The drug sensitivity of TSN combined with olaparib in TNBC patients was detected.Whether TSN combined with olaparib can exert autophagy inhibitory effects and tumor-killing effects in organoid model was verified.Results Olaparib induced autophagy in MDA-MB-231 cell line,and the combination of TSN and olaparib inhibited the proliferation of MDA-MB-231 cells(P＜0.01).In the TNBC PDOs model,the therapeutic effect of olaparib combined with TSN can significantly reduce the proliferation ability of tumor cells compared with olaparib alone.Conclusion The tumor-killing effect of TSN combined with olaparib is superior to that of olaparib alone,and the mechanism may be related to autophagy inhibition.

Objective To observe the differences in dynamic stability and kinematic/kinetic characteristics of the lower limbs between patients with knee osteoarthritis(KOA)and healthy individuals at the begining stage of practicing Tai Chi Yunshou movement.Methods Thirty Tai Chi beginners,including 15 patients with KOA and 15 healthy controls,were recruited to practice Tai Chi Yunshou movement for two hours under the guidance of a Tai Chi expert.A motion capture system and a three-dimensional force platform were used to collect and calculate dynamic stability parameter as well as kinematic and kinetic parameters of the left lower limb during the Yunshou movement.Results Compared with healthy controls,patients with KOA demonstrated a smaller center of mass(COM)-center of pressure(COP)inclination angle,reduced mean and peak ankle dorsiflexion angle,reduced peak hip adduction angle and increased peak knee flexion moment during the Yunshou movement(P<0.05).Conclusions Patients with KOA use adaptive postural strategies to maintain the lateral stability in Tai Chi Yunshou exercise,but a comprehensive training programme should be recommended to reduce the joint loading during flexion of the lower limbs at the benginning stages of this exercise.

Allergic diseases,also known as hypersensitivity diseases,are a systemic disorder affecting nearly 40%of the global population.Although they vary in affected sites and clinical manifestations,they share many common features.For example,allergic diseases are the same type Ⅰ hypersensitivity reaction;the disease mechanism is based on the inability of the positive deficiency to resist the evil,and the allergens induce the onset of the disease;the course of the disease is similar to the patient's physique;the onset of the disease is closely related to the taiyin meridian of the hand and the foot;and the disease locations in traditional Chinese medicine can be categorized as the"orifices",so that the different diseases can be treated with the same treatment.Chinese medicine believes that spleen deficiency is the initiating factor of immune dysfunction,and lung deficiency is the key to the occurrence of allergic diseases,and regulating the lung and spleen is the key point of treatment for allergic diseases.The method of"cultivating the earth and generating gold"is good at adjusting the spleen and stomach and restoring the function of the lungs,which is the key to"same treatment for different diseases"of allergic diseases.Therefore,the purpose of this article is to explore the commonalities of allergic diseases and the role of the method of"cultivating the earth and generating gold"in the treatment of allergic diseases under the perspective of"same treatment for different diseases".

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.