ObjectiveTo analyze the genomic characteristics of Streptococcus pyogenes (GAS) isolated from children with respiratory tract infections in a tertiary hospital in Jinshan District of Shanghai during 2013‒2024, to compare the changes in trend for genomic characteristics before and after 2000, and to provide scientific data for the prevention and control of GAS infections. MethodsGAS strains isolated from children with respiratory tract infections in this hospital were collected from 2013 to 2024. Antimicrobial susceptibility of the isolated strains to 12 antibiotics, including penicillin, cefotaxime, cefepime, linezolid, vancomycin, meropenem, chloramphenicol, ofloxacin, levofloxacin, erythromycin, clindamycin, and tetracycline, was determined using broth microdilution plate method. Besides, whole genome sequencing (WGS) was used to analyze multilocus sequence type (MLST), emm typing, carriage of superantigen genes, mobile genetic element (MGE), carriage of virulence gene, and genomic phylogenetic tree of the isolated strains. ResultsA total of 50 GAS strains were collected and identified from children with respiratory tract infections aged 4‒14 years old, and the resistance rates of those isolates to erythromycin, clindamycin, and tetracycline were 100.00%, 100.00%, and 86.00%, respectively. There were two emm types in the GAS isolates; the emm12 type accounted for 76.00% (38/50), corresponding to ST36 type, and the emm1 type accounted for 24.00% (12/50), corresponding to ST28, ST1274, and new-1 types. There was a statistically significant difference in the constitution of the MLST before and after 2020 (P=0.015). All the isolates carried the superantigen genes speC, speG, ssa, and smeZ. The predominant emm12 isolates belonged to the Clade Ⅱ, carrying the mobile elements ICE-emm12 (harboring erythromycin-resistance gene ermB and tetracycline-resistance gene tetM) and ΦHKU.vir (carrying virulence genes speC and ssa). The emm1 isolates carried the mobile elements ICE-HKU488 (harboring erythromycin-resistance gene ermB and tetracycline-resistance gene tetM) and ΦHKU488.vir (carrying virulence genes speC and ssa), and had close phylogenetical relationships with isolates from Hong Kong, China. No M1_UK new clone strains were found. The ST1274 isolates of emm1 were newly discovered in 2020‒2024, and belonged to a separate phylogenetic clade. ConclusionGAS strains isolated from children with respiratory tract infections in a tertiary hospital in Jinshan District of Shanghai exhibit a high resistance to erythromycin, clindamycin, and tetracycline. It is recommended that the clinical treatments change to use other antimicrobial drugs, such as penicillin, third-generation cephalosporins, and fluoroquinolones. During 2020‒2024, a new ST1274 clone strain is discovered in emm1 GAS isolates, without M1_UK new clone strains being found. It is essential to continuously concern locally prevalent GAS strains and perform early identification of MLST types to promptly monitor the internal changes of the bacterial population and potential prevalence of new clones.

Although cisplatin is a widely used chemotherapeutic agent, it is severely toxic and causes irreversible hearing loss, restricting its application in clinical settings. This study aimed to determine the molecular mechanism underlying cisplatin-induced ototoxicity. Here, we established in vitro and in vivo ototoxicity models of cisplatin-induced hair cell loss, and our results showed that reducing STING levels decreased inflammatory factor expression and hair cell death. In addition, we found that cisplatin-induced mitochondrial dysfunction was accompanied by cytosolic DNA, which may act as a critical linker between the cyclic GMP-AMP synthesis-stimulator of interferon genes (cGAS-STING) pathway and the pathogenesis of cisplatin-induced hearing loss. H-151, a specific inhibitor of STING, reduced hair cell damage and ameliorated the hearing loss caused by cisplatin in vivo. This study underscores the role of cGAS-STING in cisplatin ototoxicity and presents H-151 as a promising therapeutic for hearing loss.
Cisplatin/toxicity* ; Animals ; Nucleotidyltransferases/antagonists & inhibitors* ; Membrane Proteins/antagonists & inhibitors* ; Signal Transduction/drug effects* ; Mice ; Hair Cells, Auditory, Inner/pathology* ; Antineoplastic Agents/toxicity* ; Mice, Inbred C57BL ; Hearing Loss/metabolism* ; Male ; Ototoxicity/metabolism*

Alzheimer's disease (AD) is the most prevalent neurodegenerative disorder worldwide, causing dementia and affecting millions of individuals. One prominent characteristic in the brains of AD patients is glucose hypometabolism. In the context of galactose metabolism, intracellular glucose levels are heightened. Galactose mutarotase (GALM) plays a crucial role in maintaining normal galactose metabolism by catalyzing the conversion of β-D-galactose into α-D-galactose (α-D-G). The latter is then converted into glucose-6-phosphate, improving glucose metabolism levels. However, the involvement of GALM in AD progression is still unclear. In the present study, we found that the expression of GALM was significantly increased in AD patients and model mice. Genetic knockdown of GALM using adeno-associated virus did not change the expression of amyloid precursor protein (APP) and APP-cleaving enzymes including a disintegrin and metalloprotease 10 (ADAM10), β-site APP-cleaving enzyme 1 (BACE1), and presenilin-1 (PS1). Interestingly, genetic overexpression of GALM reduced APP and Aβ deposition by increasing the maturation of ADAM10, although it did not alter the expression of BACE1 and PS1. Further electrophysiological and behavioral experiments showed that GALM overexpression significantly ameliorated the deficits in hippocampal CA1 long-term potentiation (LTP) and spatial learning and memory in AD model mice. Importantly, direct α-D-G (20 mg/kg, i.p.) also inhibited Aβ deposition by increasing the maturation of ADAM10, thereby improving hippocampal CA1 LTP and spatial learning and memory in AD model mice. Taken together, our results indicate that GALM shifts APP processing towards α-cleavage, preventing Aβ generation by increasing the level of mature ADAM10. These findings indicate that GALM may be a potential therapeutic target for AD, and α-D-G has the potential to be used as a dietary supplement for the prevention and treatment of AD.
Animals ; ADAM10 Protein/metabolism* ; Alzheimer Disease/pathology* ; Amyloid Precursor Protein Secretases/metabolism* ; Disease Models, Animal ; Humans ; Mice ; Amyloid beta-Peptides/metabolism* ; Male ; Mice, Transgenic ; Membrane Proteins/metabolism* ; Cognitive Dysfunction/pathology* ; Mice, Inbred C57BL ; Amyloid beta-Protein Precursor/metabolism* ; Female ; Hippocampus/metabolism* ; Long-Term Potentiation/physiology*

Objective To investigate the mechanism and protective effect of Humanin(HN)on rotenone(Rot)-induced toxic damage for dopamine neurons.Methods The Rot-poisened PC12 cell model was constructed,and the control group,the Rot poisening group,the HN pretreated Rot poisening group,and the HN treatment group were set up.ELISA was used to detect the content of HN inside and outside of Rot-infected cells,CCK-8 assay was used to detect cell viability,and ATP detection kit was used to detect the intracellular ATP content.Dichloro-dihydro-fluorescein diacetate(DCFH-DA)assay was used to detect the level of reactive oxygen species(ROS)in cells.Western blotting was performed to detect the expression level of mitochondrial autophagy regulatory proteins Pink1,Parkin,p62,LC3,mitochondrial biogenesis regulatory protein PGC1α,division/fusion regulatory proteins OPA1,MFN2,DRP1,p-DRP1 and antioxidant stress regulatory proteins Keap1 and Nrf2.HBAD-mcherry-EGFP-LC3 adenovirus transfected cells was used to observed the number of autophagosomes and autophagolysosomes.Results The results showed that the intracellular concentration of HN in PC12 in the Rot poisening group was significantly higher than that in the control group(P＜0.05);Compared with the control group,the Rot poisening group had significantly decreased activity of PC12 cells,decreased ATP content and increased production of ROS.After the poisen of Rot in PC12 cells,the expression of Pink1 and p-Parkin,the ratio of LC3Ⅱ/LC3Ⅰ and the expression of p-DRP1 in mitochondrial fusion protein was increased,while the expression of p62,the expression of mitochondrial biogenesis protein PGC1 α,mitochondrial fusion proteins MFN2 and OPA1,and antioxidant stress proteins Keap1 and Nrf2 were decreased(all P＜0.05).The number of autophagosomes and autophagolysosomes in PC12 cells in the Rot poisening group was higher than that in the control group(P＜0.05),and HN pretreatment(20 μmol/L)could significantly improve the changes mentioned above caused by Rot poisening(P＜0.05).Conclusion HN ameliorates Rot-induced toxic damage for dopamine neurons by inhibiting mitophagy and mitochondrial division and promoting mitochondrial biogenesis and fusion,and anti-oxidative stress.

The long-term efficacy and complications of implantable diaphragm pacer (IDP) in a child with cervical spinal cord injury (CSCI) in the Department of Spinal and Neural Functional Reconstruction, China Rehabilitation Research Center in September 2022 were retrospective analyzed.A male child had quadriplegia without an obvious cause at the age of 12 years, and he was then lived completely with the assistance of mechanical ventilation.At the age of 14 years, he could wean off the ventilator in unilateral diaphragmatic pacing mode.However, mechanical ventilation was re-given for months after 5 years due to pneumonia, and then the IDP was re-given with the self-felt decreased pacing effect.After hospitalization, the patient was examined with mild diaphragmatic atrophy, secondary flat chest, and mild scoliosis.After optimization of the transdiaphragmatic pacing threshold and rehabilitation, his respiratory function improved.IDP can be used in CSCI for long time, while flat chest and scoliosis that limited the expansion of the lungs should be considered.At the meantime, the increased abdominal spasm affected the abdominal compliance, leading to the decrease in the efficiency of the diaphragm.

Tuberous sclerosis complex(TSC)is a rare genetic disease that can lead to benign dysplasia in multiple organs such as the skin, brain, eyes, oral cavity, heart, lungs, kidneys, liver, and bones. Its main symptoms include epilepsy, intellectual disabilities, skin depigmentation, and facial angiofibromas, whilst incidence is approximately 1 in 10 000 to 1 in 6000 newborns. This case presents a middle-aged woman who initially manifested with epilepsy and nodular depigmentation. Later, she developed a lower abdominal mass, elevated creatinine, and severe anemia. Based on clinical features and whole exome sequencing, the primary diagnosis was confirmed as TSC. Laboratory and imaging examinations revealed that the lower abdominal mass originated from the uterus. CT-guided biopsy pathology and surgical pathology suggested a combination of leiomyoma and abscess. With the involvement of multiple organs and various complications beyond the main diagnosis, the diagnostic and therapeutic process for this patient highlights the importance of rigorous clinical thinking and multidisciplinary collaboration in the diagnosis and treatment of rare and challenging diseases.

Objective To analyze the changing prevalence and resistance profiles of multiple drug-resistant(MDR)bacteria in Minhang Hospital of Fudan University before and after the onset of COVID-19 pandemic.Methods The resistance profiles of 6 bacterial species were compared before(2017-2019)and after(2020-2022)the onset of COVID-19 pandemic.WHONET 5.6 software was used to statistically analyze the data of antimicrobial susceptibility testing.Results After the onset of COVID-19(2020-2022),the prevalence of carbapenem-resistant Klebsiella pneumoniae(CRKP)was significantly higher than that in the pre-pandemic period(2017-2019)(40.1％vs 27.9％,P＜0.01),while the prevalence of carbapenem-resistant Escherichia coli(CREC)decreased significantly(1.9％vs 3.1％,P＜0.05).The prevalences of carbapenem-resistant Acinetobacter baumannii(CRAB),carbapenem-resistant Pseudomonas aeruginosa(CRPA),methicillin-resistant Staphylococcus aureus(MRSA),and vancomycin-resistant Enterococcus(VRE)did not show significant difference before and after the onset of COVID-19 pandemic.MDR isolates were mainly isolated from respiratory tract samples either before or after COVID-19 pandemic(78.4％vs 78.5％).The prevalence of MDR in the intensive care unit(ICU)was significantly higher during the period from 2020 to 2022 compared to the pre-pandemic period(53.1％vs 35.5％,P＜0.01).The resistance rate of MRSA to methoprim-sulfamethoxazole decreased from 15.7％during 2017-2019 to 3.5％during 2020-2022(P＜0.01).Compared to the pre-pandemic period,the E.faecalis strains showed lower resistance rates to penicillin G,ampicillin,and levofloxacin during 2020-2022.The resistance rate of E.faecium to high-level gentamicin decreased significantly from 50.1％during 2017-2019 to 39.2％during 2020-2022(P＜0.01).The resistance rate of E.coli to imipenem decreased from 2.7％to 1.2％(P＜0.01),while A.baumannii and P.aeruginosa strains showed stable resistance rates to carbapenems(P＞0.05).Conclusions After the onset of COVID-19 pandemic,the prevalence of CREC decreased significantly.The prevalences of CRAB,CRPA,MRSA,and VRE also showed a decreasing trend.However,the prevalence and resistance rates of CRKP significantly increased.It is necessary to strengthen hospital infection control measures to curb the spread of MDR bacteria.

Objective To observe the effects of exercise combined with probiotic intervention on glycolipid metabolism,oxidative stress,and skeletal muscle satellite cell expression in type 2 diabetic rats.Methods From 60 8-week-old SPF-grade SD male rats,10 rats were randomly selected as the normal control group(NC),and the rest were injected intraperitoneally with streptozotocin,establishing a rat model of type 2 diabetes mellitus.The diabetic rats were randomly divided into model(TN),exercise+diabetes(YTN),probiotic+diabetes(GTN),and exercise+probiotics+diabetes(YGTN)groups.The exercise groups were subjected to 6 weeks of incremental load aerobic treadmill exercise(speed of 15 m/min for 30 min in week 1,15 m/min for 60 min in week 2,and 19.3 m/min for 60 min in weeks 3～6,with continuous training until week 6,5 days/week).The GTN and YGTN groups were administered 10.0 mL/(kg·d)of Lactibiane Iki probiotic solution(concentration of 107 CFU/mL)one hour before training.Glycolipid metabolism indexes,oxidative stress indexes,and expression of skeletal muscle satellite cell proteins were measured in rats.Results(1)The levels of fasting plasma glucose(FBG),fasting serum insulin(FINS),hemoglobin Alc(HBA1C)and(homeostasis model assessment-insulin resistance index(HOMA-1R)were higher in the TN,YTN,GTN,and YGTN groups than in the NC group(P＜0.01).The levels of FBG in the YGTN group were lower than those in the TN,YTN,GTN,and YGTN groups(P＜0.05).Serum FINS levels in YTN,GTN and YGTN groups were all decreased compared with those in the TN group(P＜0.01).Serum HBA1C levels were lower in the YGTN group than the TN,YTN,and GTN groups(P＜0.01).The HOMA-IR indices of rats in the YGTN group decreased more significantly compared with those in the TN,YTN,and GTN groups(P＜0.01).The result showed that FBG,FINS,HBA1C,and HOMA-IR decreased more significantly in rats in the aerobic exercise combined with probiotic intervention group than the other intervention groups.(2)Serum total cholesterol(T-CHO),triglyceride(TG),low density lipoprotein cholesterol(LDL)and free fatty acid(FFA)levels in the TN group were higher than those in the NC group(P＜0.05),and those of the YGTN group were lower than those of the TN group(P＜0.05)and basically restored to the level of the NC group.Serum high density lipoprotein cholesterol(HDL)levels were lower in the TN group than the NC group(P＜0.05),Serum HDL levels of rats in TN group were lower than those of the NC group(P＜0.05),and those in the YGTN group were higher than those of TN group(P＜0.05),which were basically restored to the level of NC group.The result showed that aerobic exercise combined with probiotic intervention reduced serum T-CHO,TG,LDL,FFA levels and increased HDL levels more significantly than other interventions in diabetic rats.(3)Serum malondialdehyde(MDA)and 8-iso-prostaglandin F2α(8-isoPGF2α)levels of rats in the TN group were higher than those in the NC group(P＜0.05 or P＜0.01),and those of rats in the YGTN group were lower than those in the TN group(P＜0.05 or P＜0.01).Serum catalase(CAT),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and total antioxidant capacity(T-AOC)in the TN group were lower than those in the NC group(P＜0.01),and those of rats in the YGTN group were higher than those of the NC group(P＜0.01)and were basically restored to the levels of NC group.(4)The protein expression levels of paired box gene 7(Pax7),myogenic determination gene(MyoD),myogenin(MyoG)and myogenic factor 5(Myf5)in the skeletal muscle of rats in the TN group were lower than those in the NC group(P＜0.05 or P＜0.01),and these levels were higher in the YGTN group than the TN group(P＜0.05 or P＜0.01);basically,the levels were restored to those of the NC group.In the TN group,myostatin(MSTN)protein expression was higher in the TN group than the NC group(P＜0.05),and that in the YGTN group was lower than that in the TN group(P＜0.05)and basically restored to the level of the NC group.Conclusions The result showed that probiotics and aerobic exercise had a synergistic effect in improving blood glucose and insulin levels,attenuating insulin resistance,improving lipids and oxidative radicals,and preventing the phenomenon of disturbed glucose metabolism,thus achieving dynamic equilibrium regulation in type 2 diabetic rats.Aerobic exercise combined with probiotics also promoted myoblast differentiation,which played a role in preventing T2DM-induced loss of muscle mass and strength as well as muscle tissue complications.

Objective To explore the improving effect and mechanism of saikosaponin A on insomnia rats based on cAMP/PKA/CREB pathway.Methods Seventy-five SD rats were randomly divided into blank group,model group,low-dose saikosaponin A group(0.625 mg·kg-1),high-dose saikosaponin A group(2.500 mg·kg-1)and Estazolam group(0.1 mg·kg-1),with 15 rats in each group.Insomnia rat model was established by intraperitoneal injection of Phenylalanine(PCPA,0.1 mg·kg-1).The general condition and circadian rhythm of rats were observed;the sleep latency and sleep duration of rats were measured by pentobarbital sodium righting experiment.The sleep phase of rats was observed,and the duration of slow wave sleep phase 1(SWS1),slow wave sleep phase 2(SWS2),rapid eye movement sleep phase(REMS)and total sleep time(TST)were recorded.The mRNA expression levels of hypothalamic circadian genes Clock,Bmal1 and clock-controlled genes Rev-erbα and Rorα were determined by qRT-PCR.The expression level of NeuN in hippocampus was determined by immunofluorescence.The level of cAMP in brain tissue was determined by ELISA.The expression levels of Clock,Bmal1,Rev-erbα,Rorα and cAMP/PKA/CREB pathway-related proteins in brain tissue were determined by Western Blot.Results Compared with the blank group,the rats in the model group had disordered circadian rhythms,extreme excitement,irritability,and reduced sleep;the sleep latency was significantly prolonged(P＜0.05),and the sleep duration and SWS1,SWS2,REMS and TST were significantly shortened(P＜0.05).The arrangement of neurons was disordered,and the IOD value of NeuN positive neurons was significantly decreased(P＜0.05).The mRNA and protein expression levels of Clock,Bmal1,Rev-erbα and Rorα in brain tissue were significantly decreased(P＜0.05).The expression levels of cAMP,p-PKA/PKA and p-CREB/CREB in brain tissue were significantly decreased(P＜0.05).Compared with the model group,the aggressiveness of the rats in the administration group was significantly weakened,the circadian rhythm was rhythmic,the activity was reduced,and the sleep was increased.The sleep latency was significantly shortened(P＜0.05),and the sleep duration and SWS1,SWS2,REMS and TST were significantly prolonged(P＜0.05).The disorder of neuronal arrangement was restored,and the IOD value of NeuN positive neurons was significantly increased(P＜0.05).The mRNA and protein expression levels of Clock,Bmal1,Rev-erbα and Rorα in brain tissue were significantly increased(P＜0.05).The protein expression levels of cAMP,p-PKA/PKA and p-CREB/CREB in brain tissue were significantly increased(P＜0.05).Conclusion Saikosaponin A may improve the circadian rhythm of insomnia rats by activating cAMP/PKA/CREB pathway.