Researchers commonly use cyclization recombination enzyme/locus of X-over P1 (Cre/loxP) technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and Leydig cells within the testis. However, the shortcomings of these genetic tools include high costs, lengthy experimental periods, and limited accessibility for researchers. Therefore, exploring alternative gene silencing techniques is of great practical value. In this study, we employed adeno-associated virus (AAV) as a vector for gene silencing in Sertoli and Leydig cells. Our findings demonstrated that AAV serotypes 1, 8, and 9 exhibited high infection efficiency in both types of testis cells. Importantly, we discovered that all three AAV serotypes exhibited exquisite specificity in targeting Sertoli cells via tubular injection while demonstrating remarkable selectivity in targeting Leydig cells via interstitial injection. We achieved cell-specific knockouts of the steroidogenic acute regulatory ( Star ) and luteinizing hormone/human chorionic gonadotropin receptor (Lhcgr) genes in Leydig cells, but not in Sertoli cells, using AAV9-single guide RNA (sgRNA)-mediated gene editing in Rosa26-LSL-Cas9 mice. Knockdown of androgen receptor ( Ar ) gene expression in Sertoli cells of wild-type mice was achieved via tubular injection of AAV9-short hairpin RNA (shRNA)-mediated targeting. Our findings offer technical approaches for investigating gene function in Sertoli and Leydig cells through AAV9-mediated gene silencing.
Animals ; Male ; Leydig Cells/metabolism* ; Mice ; Dependovirus/genetics* ; Sertoli Cells/metabolism* ; Gene Silencing ; Genetic Vectors ; Testis/cytology*

The aim of this study was to identify the Salmonella strains isolated from an outbreak of foodborne illness in a seafood buffet restaurant and analyze their pathogenic characteristics.Epidemiological data,fecal samples from patients and chefs,and food/environmental samples from the restaurant were analyzed.Research methods included bacterial culture,serotyping,quadruple fluo-rescence PCR identification,whole-genome sequencing and antibiotic susceptibility testing.The results showed that 4 S.Java strains(serotype 1,4,12∶b∶1,2;ST42)were isolated from two outbreak cases and two sporadic cases.All isolates exhibited similar genomic features,harboring 9 virulence islands and 98 virulence genes.Antimicrobial resistance profiling revealed streptomycin monoresis-tance,mediated by aac(6′)-Iy and aac(6′)-Iaa genes.In conclusion,this event was the first reported outbreak of foodborne illness caused by S.Java in China,indicating that S.Java may be prevalent in the surveyed district.The catering industry should optimize food handling and processing procedures and enhance the surveillance of high risk pathogens.Meanwhile,further studies should ad-dress differential diagnosis and pathogenic mechanism differences between S.Java and S.paratyphi B,which will facilitate evidence-based monitoring in China.

This article comprehensively discusses the concept of workplace ostracism, measurement tools, theoretical basis, the current situation of workplace ostracism among nurses, and research on related variables. It proposes targeted strategies to address workplace ostracism among nurses, aiming to reduce the occurrence of workplace ostracism incidents among nurses. This is of great significance for constructing a harmonious work environment and promoting individual mental health.

This article comprehensively discusses the concept of workplace ostracism, measurement tools, theoretical basis, the current situation of workplace ostracism among nurses, and research on related variables. It proposes targeted strategies to address workplace ostracism among nurses, aiming to reduce the occurrence of workplace ostracism incidents among nurses. This is of great significance for constructing a harmonious work environment and promoting individual mental health.

Objective To systematically review the safety of disialoganglioside(GD2)targeted therapies in the treatment of pediatric neuroblastoma(NB).Methods PubMed,Embase,Cochrane Library,CNKI,WanFang Data,SinoMed databases and ClinicalTrials.gov clinical trial registration system were electronically searched to collect clinical studies of GD2-targeted therapies for pediatric NB from inception to November 30,2024.Two reviewers independently screened the literature,extracted data and assessed the risk of bias of the included studies.Meta-analysis was performed by using R 4.3.1 software.Results A total of 21 studies involving 2,438 patients were included.Meta-analysis results showed that the overall incidence(95％CI)of serious adverse drug events(SADEs)in GD2-targeted therapies for pediatric NB was 51.67％(35.43％to 67.92％).The serious fever and alanine aminotransferase(ALT)increase were common with the incidence(95％CI)of 7.80％(0.80％to 18.85％)and 5.10％(2.02％to 10.55％)respectively.The incidences(95％CI)of grade 3 to 5 adverse drug reactions(ADRs):fever 18.81％(8.00％to 29.61％),pain 30.32％(16.21％to 44.44％),hypokalemia 18.96％(6.44％to 31.49％),ALT increased 10.41％(2.96％to 17.86％),hypotension 8.48％(3.78％to 13.18％),anemia 16.62％(5.97％to 27.26％),platelet count decreased 16.46％(5.55％to 27.37％),lymphocyte count decreased 10.42％(0.97％to 26.09％),neutrophil count decreased 13.37％(3.12％to 23.62％).Among the intervention subgroups,the incidences of grade 3 to 5 ADRs were relatively low in dinutuximab β group and GD2-chimeric antigen receptor T cells(GD2 CART)group.Conclusions The incidences of SADEs and grade 3 to 5 ADRs are high in pediatric NB patients receiving GD2-targeted therapy,which should be paid attention to.The safety profile of GD2 targeted therapies under development for pediatric NB are even better than those of GD2 targeted drugs already on the market,especially GD2 CART,which shows good safety potential.

Objective To systematically review the safety of disialoganglioside(GD2)targeted therapies in the treatment of pediatric neuroblastoma(NB).Methods PubMed,Embase,Cochrane Library,CNKI,WanFang Data,SinoMed databases and ClinicalTrials.gov clinical trial registration system were electronically searched to collect clinical studies of GD2-targeted therapies for pediatric NB from inception to November 30,2024.Two reviewers independently screened the literature,extracted data and assessed the risk of bias of the included studies.Meta-analysis was performed by using R 4.3.1 software.Results A total of 21 studies involving 2,438 patients were included.Meta-analysis results showed that the overall incidence(95％CI)of serious adverse drug events(SADEs)in GD2-targeted therapies for pediatric NB was 51.67％(35.43％to 67.92％).The serious fever and alanine aminotransferase(ALT)increase were common with the incidence(95％CI)of 7.80％(0.80％to 18.85％)and 5.10％(2.02％to 10.55％)respectively.The incidences(95％CI)of grade 3 to 5 adverse drug reactions(ADRs):fever 18.81％(8.00％to 29.61％),pain 30.32％(16.21％to 44.44％),hypokalemia 18.96％(6.44％to 31.49％),ALT increased 10.41％(2.96％to 17.86％),hypotension 8.48％(3.78％to 13.18％),anemia 16.62％(5.97％to 27.26％),platelet count decreased 16.46％(5.55％to 27.37％),lymphocyte count decreased 10.42％(0.97％to 26.09％),neutrophil count decreased 13.37％(3.12％to 23.62％).Among the intervention subgroups,the incidences of grade 3 to 5 ADRs were relatively low in dinutuximab β group and GD2-chimeric antigen receptor T cells(GD2 CART)group.Conclusions The incidences of SADEs and grade 3 to 5 ADRs are high in pediatric NB patients receiving GD2-targeted therapy,which should be paid attention to.The safety profile of GD2 targeted therapies under development for pediatric NB are even better than those of GD2 targeted drugs already on the market,especially GD2 CART,which shows good safety potential.

The aim of this study was to identify the Salmonella strains isolated from an outbreak of foodborne illness in a seafood buffet restaurant and analyze their pathogenic characteristics.Epidemiological data,fecal samples from patients and chefs,and food/environmental samples from the restaurant were analyzed.Research methods included bacterial culture,serotyping,quadruple fluo-rescence PCR identification,whole-genome sequencing and antibiotic susceptibility testing.The results showed that 4 S.Java strains(serotype 1,4,12∶b∶1,2;ST42)were isolated from two outbreak cases and two sporadic cases.All isolates exhibited similar genomic features,harboring 9 virulence islands and 98 virulence genes.Antimicrobial resistance profiling revealed streptomycin monoresis-tance,mediated by aac(6′)-Iy and aac(6′)-Iaa genes.In conclusion,this event was the first reported outbreak of foodborne illness caused by S.Java in China,indicating that S.Java may be prevalent in the surveyed district.The catering industry should optimize food handling and processing procedures and enhance the surveillance of high risk pathogens.Meanwhile,further studies should ad-dress differential diagnosis and pathogenic mechanism differences between S.Java and S.paratyphi B,which will facilitate evidence-based monitoring in China.

Objective To investigate the clinical characteristics and prognosis of pneumococcal meningitis(PM),and drug sensitivity of Streptococcus pneumoniae(SP)isolates in Chinese children.Methods A retrospective analysis was conducted on clinical information,laboratory data,and microbiological data of 160 hospitalized children under 15 years old with PM from January 2019 to December 2020 in 33 tertiary hospitals across the country.Results Among the 160 children with PM,there were 103 males and 57 females.The age ranged from 15 days to 15 years,with 109 cases(68.1% )aged 3 months to under 3 years.SP strains were isolated from 95 cases(59.4% )in cerebrospinal fluid cultures and from 57 cases(35.6% )in blood cultures.The positive rates of SP detection by cerebrospinal fluid metagenomic next-generation sequencing and cerebrospinal fluid SP antigen testing were 40% (35/87)and 27% (21/78),respectively.Fifty-five cases(34.4% )had one or more risk factors for purulent meningitis,113 cases(70.6% )had one or more extra-cranial infectious foci,and 18 cases(11.3% )had underlying diseases.The most common clinical symptoms were fever(147 cases,91.9% ),followed by lethargy(98 cases,61.3% )and vomiting(61 cases,38.1% ).Sixty-nine cases(43.1% )experienced intracranial complications during hospitalization,with subdural effusion and/or empyema being the most common complication[43 cases(26.9% )],followed by hydrocephalus in 24 cases(15.0% ),brain abscess in 23 cases(14.4% ),and cerebral hemorrhage in 8 cases(5.0% ).Subdural effusion and/or empyema and hydrocephalus mainly occurred in children under 1 year old,with rates of 91% (39/43)and 83% (20/24),respectively.SP strains exhibited complete sensitivity to vancomycin(100% ,75/75),linezolid(100% ,56/56),and meropenem(100% ,6/6).High sensitivity rates were also observed for levofloxacin(81% ,22/27),moxifloxacin(82% ,14/17),rifampicin(96% ,25/26),and chloramphenicol(91% ,21/23).However,low sensitivity rates were found for penicillin(16% ,11/68)and clindamycin(6% ,1/17),and SP strains were completely resistant to erythromycin(100% ,31/31).The rates of discharge with cure and improvement were 22.5% (36/160)and 66.2% (106/160),respectively,while 18 cases(11.3% )had adverse outcomes.Conclusions Pediatric PM is more common in children aged 3 months to under 3 years.Intracranial complications are more frequently observed in children under 1 year old.Fever is the most common clinical manifestation of PM,and subdural effusion/emphysema and hydrocephalus are the most frequent complications.Non-culture detection methods for cerebrospinal fluid can improve pathogen detection rates.Adverse outcomes can be noted in more than 10% of PM cases.SP strains are high sensitivity to vancomycin,linezolid,meropenem,levofloxacin,moxifloxacin,rifampicin,and chloramphenicol.[Chinese Journal of Contemporary Pediatrics,2024,26(2):131-138]

Lung cancer is the top cause of cancer deaths globally.Advances in immune checkpoint inhibitors(ICIs)have transformed cancer treatment,but their use in lung cancer has led to more side effects.This study examined if past pulmonary tuberculosis(TB)affects ICIs'effectiveness and safety in lung cancer treatment.We reviewed lung cancer patients treated with ICIs at Beijing Chest Hospital from January 2019 to August 2022.We compared outcomes and side effects between patients with and without prior TB.Of 116 patients(40 with TB history,76 without),prior TB didn't reduce treatment effectiveness but did increase severe side effects.Notably,older patients(≥65 years)faced a higher risk of severe side effects.Detailed cases of two patients with severe side effects underscored TB as a risk factor in lung cancer patients receiving ICIs,stressing the need for careful monitoring and personalized care.

Lipopolysaccharides (LPS) are major outer membrane components of Gram-negative bacteria. Unlike most Gram-negative bacteria, Acinetobacter baumannii can still survive and acquire polymyxin resistance after complete loss of LPS. Previous studies of LPS-deficient Acinetobacter baumannii mostly focused on LPS-deficient strains induced by polymyxin, whose background was complex and unstable. To investigate LPS loss-mediated polymyxin resistant-Acinetobacter baumannii, this study constructed a stable and clear background LPS-deficient strain by knocking out lpxC gene in Acinetobacter baumannii ATCC 19606 with CIRSPR/Cas9, and then studied the phenotypic changes of the lpxC-deficient strain including morphology, growth rate, antibiotic susceptibility, virulence, membrane permeability, and membrane potential. Animal experiments were approved by the Animal Care and Welfare Committee Institute of Medicinal Biotechnology, CAMS and PUMC (approval number: IMB-20240119D₉). The results indicated that the lpxC gene of Acinetobacter baumannii ATCC 19606 was successfully knocked out. After losing the lpxC, the strain underwent the morphological change from rod-shaped to spherical. Furthermore, it leads to reduced growth rate, enhanced membrane permeability, decreased membrane potential, lower virulence, and increased antibiotic susceptibility to β-lactams, quinolones, aminoglycosides, macrolides, glycopeptides. The lpxC deletion results in significant changes in membrane homeostasis and adaptability of Acinetobacter baumannii. Understanding the phenotypic changes of colistin-resistant Acinetobacter baumannii mediated by LPS loss is useful for exploring the resistance mechanism of Acinetobacter baumannii and developing new therapeutic strategies.