A recombinant adenovirus(rAd)for expression of Mycobacterium tuberculosis(M.tb)c-di-AMP phosphodiesterase CnpB was constructed,and its induced humoral immune response was detected.The codon-optimized gene of M.tb CnpB was cloned into the adenoviral plasmid pcADV.The recombinant plasmid pcADV-CnpB was transfected into HEK293T cells,and expression was detected with Western blot.The recombinant plasmid pcADV-CnpB and the backbone plasmid were co-transfected into HEK293T cells to obtain the recombinant adenovirus rAd-CnpB.rAd-CnpB was amplified in HEK293T cells,and the target protein expression of rAd-CnpB was detected with Western blot and immunofluorescence.Mice were immunized with rAd-CnpB intranasally,and their sera and bronchoalveolar lavage fluid(BALF)were collected.ELISA was used to detect levels of antigen-specific antibodies.Restriction enzyme digestion and sequencing indicated that the recombinant plasmid pcADV-CnpB was successfully constructed and led to protein expression in eukaryotic cells.rAd-CnpB was packaged and produced in HEK293T cells.After amplification and purification,rAd-CnpB with a titer of 5.53×1010 PFU/mL was obtained.rAd-CnpB led to CnpB expression in HEK293T cells.Intranasal immunization with rAd-CnpB increased levels of IgG and secretory IgA in BALF and led to high levels of IgG in sera.rAd-CnpB,the recombinant adenovirus for expression of c-di-AMP phosphodiesterase CnpB was successfully constructed,and was found to induce antigen-specific humoral and mucosal immune responses through mucosal immunization.Thus,rAd-CnpB may be used in further research on new TB vaccine strategies.

BACKGROUND:Osteoporosis is a high-risk factor for dental implant treatment.The preparation of tissue engineering scaffolds with sustained-release Alendronate and its application in oral implant surgery for osteoporosis patients is a current hot topic in oral bone tissue engineering research.OBJECTIVE:To prepare alendronate/chitosan/polyvinyl alcohol composite hydrogel film and characterize its sustained release properties.METHODS:(1)Chitosan/polyvinyl alcohol composite hydrogel films with varying mass ratios(mass ratios of chitosan and polyvinyl alcohol were 3:7,5:5,and 7:3,respectively),chitosan and polyvinyl alcohol hydrogel films were prepared using a physical crosslinking method.By characterizing the morphology,water contact angle,mechanical properties,swelling rate,and cell compatibility of the hydrogel film,a suitable hydrogel film was screened as a carrier of alendronate.(2)Chitosan/polyvinyl alcohol composite hydrogel films containing 0,0.4,1.2,and 2.0 mg/L alendronate were prepared,and rat bone marrow mesenchymal stem cells were co-cultured with the four groups of hydrogel films.The cytocompatibility of the hydrogel films was detected by CCK-8 assay and CD44 immunofluorescence staining.The drug-loaded chitosan/polyvinyl alcohol composite hydrogel films were immersed in PBS.The drug release performance of the composite hydrogel films was detected by ultraviolet-visible spectroscopy.RESULTS AND CONCLUSION:(1)The characterization results showed that with the increase of the mass of polyvinyl alcohol in the hydrogel film,the structural density of the hydrogel film increased,the porosity decreased,the water contact angle(all within 90°),the elongation at break and the compressive strength increased,and the equilibrium swelling rate decreased.It had no effect on the proliferation of rat bone marrow mesenchymal stem cells.Chitosan hydrogel film could promote the adhesion of rat bone marrow mesenchymal stem cells.The 3:7 and 5:5 ratio composite hydrogel films did not affect the adhesion of rat bone marrow mesenchymal stem cells.Polyvinyl alcohol hydrogel film and 7:3 ratio composite hydrogel film inhibited the adhesion of rat bone marrow mesenchymal stem cells.Based on the above results,the 5:5 composite hydrogel film was selected as the carrier of alendronate.(2)CCK-8 assay results showed that the composite hydrogel film containing 0.4 and 1.2 mg/L alendronate had no cytotoxicity.CD44 immunofluorescence staining results showed that the composite hydrogel film containing 0.4 mg/L alendronate did not affect the adhesion of rat bone marrow mesenchymal stem cells,while the composite hydrogel film containing 1.2 and 2.0 mg/L alendronate inhibited the adhesion of rat bone marrow mesenchymal stem cells.Chitosan/polyvinyl alcohol composite hydrogel film containing 0.4,1.2,and 2.0 mg/L alendronate released alendronate in the first 6 hours,and then released alendronate evenly and stably,with a release time of more than 96 hours.(3)The results showed that the chitosan/polyvinyl alcohol composite hydrogel film with a ratio of 5:5 had good physical and chemical properties and cytocompatibility,and could be used as a sustained-release carrier of alendronate.

Objective To investigate the distribution and antimicrobial resistance profiles of common pathogens isolated from cerebrospinal fluid(CSF)in CHINET program from 2015 to 2021.Methods The bacterial strains isolated from CSF were identified in accordance with clinical microbiology practice standards.Antimicrobial susceptibility test was conducted using Kirby-Bauer method and automated systems per the unified CHINET protocol.Results A total of 14 014 bacterial strains were isolated from CSF samples from 2015 to 2021,including the strains isolated from inpatients(95.3％)and from outpatient and emergency care patients(4.7％).Overall,19.6％of the isolates were from children and 80.4％were from adults.Gram-positive and Gram-negative bacteria accounted for 68.0％and 32.0％,respectively.Coagulase negative Staphylococcus accounted for 73.0％of the total Gram-positive bacterial isolates.The prevalence of MRSA was 38.2％in children and 45.6％in adults.The prevalence of MRCNS was 67.6％in adults and 69.5％in children.A small number of vancomycin-resistant Enterococcus faecium(2.2％)and linezolid-resistant Enterococcus faecalis(3.1％)were isolated from adult patients.The resistance rates of Escherichia coli and Klebsiella pneumoniae to ceftriaxone were 52.2％and 76.4％in children,70.5％and 63.5％in adults.The prevalence of carbapenem-resistant E.coli and K.pneumoniae(CRKP)was 1.3％and 47.7％in children,6.4％and 47.9％in adults.The prevalence of carbapenem-resistant Acinetobacter baumannii(CRAB)and Pseudomonas aeruginosa(CRPA)was 74.0％and 37.1％in children,81.7％and 39.9％in adults.Conclusions The data derived from antimicrobial resistance surveillance are crucial for clinicians to make evidence-based decisions regarding antibiotic therapy.Attention should be paid to the Gram-negative bacteria,especially CRKP and CRAB in central nervous system(CNS)infections.Ongoing antimicrobial resistance surveillance is helpful for optimizing antibiotic use in CNS infections.

Objective To investigate the antimicrobial resistance of clinical isolates from elderly patients(≥65 years)in major medical institutions across China.Methods Bacterial strains were isolated from elderly patients in 52 hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program during the period from 2015 to 2021.Antimicrobial susceptibility test was carried out by disk diffusion method and automated systems according to the same CHINET protocol.The data were interpreted in accordance with the breakpoints recommended by the Clinical and Laboratory Standards Institute(CLSI)in 2021.Results A total of 514 715 nonduplicate clinical isolates were collected from elderly patients in 52 hospitals from January 1,2015 to December 31,2021.The number of isolates accounted for 34.3％of the total number of clinical isolates from all patients.Overall,21.8％of the 514 715 strains were gram-positive bacteria,and 78.2％were gram-negative bacteria.Majority(90.9％)of the strains were isolated from inpatients.About 42.9％of the strains were isolated from respiratory specimens,and 22.9％were isolated from urine.More than half(60.7％)of the strains were isolated from male patients,and 39.3％isolated from females.About 51.1％of the strains were isolated from patients aged 65-＜75 years.The prevalence of methicillin-resistant strains(MRSA)was 38.8％in 32 190 strains of Staphylococcus aureus.No vancomycin-or linezolid-resistant strains were found.The resistance rate of E.faecalis to most antibiotics was significantly lower than that of Enterococcus faecium,but a few vancomycin-resistant strains(0.2％,1.5％)and linezolid-resistant strains(3.4％,0.3％)were found in E.faecalis and E.faecium.The prevalence of penicillin-susceptible S.pneumoniae(PSSP),penicillin-intermediate S.pneumoniae(PISP),and penicillin-resistant S.pneumoniae(PRSP)was 94.3％,4.0％,and 1.7％in nonmeningitis S.pneumoniae isolates.The resistance rates of Klebsiella spp.(Klebsiella pneumoniae 93.2％)to imipenem and meropenem were 20.9％and 22.3％,respectively.Other Enterobacterales species were highly sensitive to carbapenem antibiotics.Only 1.7％-7.8％of other Enterobacterales strains were resistant to carbapenems.The resistance rates of Acinetobacter spp.(Acinetobacter baumannii 90.6％)to imipenem and meropenem were 68.4％and 70.6％respectively,while 28.5％and 24.3％of P.aeruginosa strains were resistant to imipenem and meropenem,respectively.Conclusions The number of clinical isolates from elderly patients is increasing year by year,especially in the 65-＜75 age group.Respiratory tract isolates were more prevalent in male elderly patients,and urinary tract isolates were more prevalent in female elderly patients.Klebsiella isolates were increasingly resistant to multiple antimicrobial agents,especially carbapenems.Antimicrobial resistance surveillance is helpful for accurate empirical antimicrobial therapy in elderly patients.

Objective To investigate the distribution and antimicrobial resistance profiles of common pathogens isolated from cerebrospinal fluid(CSF)in CHINET program from 2015 to 2021.Methods The bacterial strains isolated from CSF were identified in accordance with clinical microbiology practice standards.Antimicrobial susceptibility test was conducted using Kirby-Bauer method and automated systems per the unified CHINET protocol.Results A total of 14 014 bacterial strains were isolated from CSF samples from 2015 to 2021,including the strains isolated from inpatients(95.3％)and from outpatient and emergency care patients(4.7％).Overall,19.6％of the isolates were from children and 80.4％were from adults.Gram-positive and Gram-negative bacteria accounted for 68.0％and 32.0％,respectively.Coagulase negative Staphylococcus accounted for 73.0％of the total Gram-positive bacterial isolates.The prevalence of MRSA was 38.2％in children and 45.6％in adults.The prevalence of MRCNS was 67.6％in adults and 69.5％in children.A small number of vancomycin-resistant Enterococcus faecium(2.2％)and linezolid-resistant Enterococcus faecalis(3.1％)were isolated from adult patients.The resistance rates of Escherichia coli and Klebsiella pneumoniae to ceftriaxone were 52.2％and 76.4％in children,70.5％and 63.5％in adults.The prevalence of carbapenem-resistant E.coli and K.pneumoniae(CRKP)was 1.3％and 47.7％in children,6.4％and 47.9％in adults.The prevalence of carbapenem-resistant Acinetobacter baumannii(CRAB)and Pseudomonas aeruginosa(CRPA)was 74.0％and 37.1％in children,81.7％and 39.9％in adults.Conclusions The data derived from antimicrobial resistance surveillance are crucial for clinicians to make evidence-based decisions regarding antibiotic therapy.Attention should be paid to the Gram-negative bacteria,especially CRKP and CRAB in central nervous system(CNS)infections.Ongoing antimicrobial resistance surveillance is helpful for optimizing antibiotic use in CNS infections.

Objective To investigate the antimicrobial resistance of clinical isolates from elderly patients(≥65 years)in major medical institutions across China.Methods Bacterial strains were isolated from elderly patients in 52 hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program during the period from 2015 to 2021.Antimicrobial susceptibility test was carried out by disk diffusion method and automated systems according to the same CHINET protocol.The data were interpreted in accordance with the breakpoints recommended by the Clinical and Laboratory Standards Institute(CLSI)in 2021.Results A total of 514 715 nonduplicate clinical isolates were collected from elderly patients in 52 hospitals from January 1,2015 to December 31,2021.The number of isolates accounted for 34.3％of the total number of clinical isolates from all patients.Overall,21.8％of the 514 715 strains were gram-positive bacteria,and 78.2％were gram-negative bacteria.Majority(90.9％)of the strains were isolated from inpatients.About 42.9％of the strains were isolated from respiratory specimens,and 22.9％were isolated from urine.More than half(60.7％)of the strains were isolated from male patients,and 39.3％isolated from females.About 51.1％of the strains were isolated from patients aged 65-＜75 years.The prevalence of methicillin-resistant strains(MRSA)was 38.8％in 32 190 strains of Staphylococcus aureus.No vancomycin-or linezolid-resistant strains were found.The resistance rate of E.faecalis to most antibiotics was significantly lower than that of Enterococcus faecium,but a few vancomycin-resistant strains(0.2％,1.5％)and linezolid-resistant strains(3.4％,0.3％)were found in E.faecalis and E.faecium.The prevalence of penicillin-susceptible S.pneumoniae(PSSP),penicillin-intermediate S.pneumoniae(PISP),and penicillin-resistant S.pneumoniae(PRSP)was 94.3％,4.0％,and 1.7％in nonmeningitis S.pneumoniae isolates.The resistance rates of Klebsiella spp.(Klebsiella pneumoniae 93.2％)to imipenem and meropenem were 20.9％and 22.3％,respectively.Other Enterobacterales species were highly sensitive to carbapenem antibiotics.Only 1.7％-7.8％of other Enterobacterales strains were resistant to carbapenems.The resistance rates of Acinetobacter spp.(Acinetobacter baumannii 90.6％)to imipenem and meropenem were 68.4％and 70.6％respectively,while 28.5％and 24.3％of P.aeruginosa strains were resistant to imipenem and meropenem,respectively.Conclusions The number of clinical isolates from elderly patients is increasing year by year,especially in the 65-＜75 age group.Respiratory tract isolates were more prevalent in male elderly patients,and urinary tract isolates were more prevalent in female elderly patients.Klebsiella isolates were increasingly resistant to multiple antimicrobial agents,especially carbapenems.Antimicrobial resistance surveillance is helpful for accurate empirical antimicrobial therapy in elderly patients.

BACKGROUND:Osteoporosis is a high-risk factor for dental implant treatment.The preparation of tissue engineering scaffolds with sustained-release Alendronate and its application in oral implant surgery for osteoporosis patients is a current hot topic in oral bone tissue engineering research.OBJECTIVE:To prepare alendronate/chitosan/polyvinyl alcohol composite hydrogel film and characterize its sustained release properties.METHODS:(1)Chitosan/polyvinyl alcohol composite hydrogel films with varying mass ratios(mass ratios of chitosan and polyvinyl alcohol were 3:7,5:5,and 7:3,respectively),chitosan and polyvinyl alcohol hydrogel films were prepared using a physical crosslinking method.By characterizing the morphology,water contact angle,mechanical properties,swelling rate,and cell compatibility of the hydrogel film,a suitable hydrogel film was screened as a carrier of alendronate.(2)Chitosan/polyvinyl alcohol composite hydrogel films containing 0,0.4,1.2,and 2.0 mg/L alendronate were prepared,and rat bone marrow mesenchymal stem cells were co-cultured with the four groups of hydrogel films.The cytocompatibility of the hydrogel films was detected by CCK-8 assay and CD44 immunofluorescence staining.The drug-loaded chitosan/polyvinyl alcohol composite hydrogel films were immersed in PBS.The drug release performance of the composite hydrogel films was detected by ultraviolet-visible spectroscopy.RESULTS AND CONCLUSION:(1)The characterization results showed that with the increase of the mass of polyvinyl alcohol in the hydrogel film,the structural density of the hydrogel film increased,the porosity decreased,the water contact angle(all within 90°),the elongation at break and the compressive strength increased,and the equilibrium swelling rate decreased.It had no effect on the proliferation of rat bone marrow mesenchymal stem cells.Chitosan hydrogel film could promote the adhesion of rat bone marrow mesenchymal stem cells.The 3:7 and 5:5 ratio composite hydrogel films did not affect the adhesion of rat bone marrow mesenchymal stem cells.Polyvinyl alcohol hydrogel film and 7:3 ratio composite hydrogel film inhibited the adhesion of rat bone marrow mesenchymal stem cells.Based on the above results,the 5:5 composite hydrogel film was selected as the carrier of alendronate.(2)CCK-8 assay results showed that the composite hydrogel film containing 0.4 and 1.2 mg/L alendronate had no cytotoxicity.CD44 immunofluorescence staining results showed that the composite hydrogel film containing 0.4 mg/L alendronate did not affect the adhesion of rat bone marrow mesenchymal stem cells,while the composite hydrogel film containing 1.2 and 2.0 mg/L alendronate inhibited the adhesion of rat bone marrow mesenchymal stem cells.Chitosan/polyvinyl alcohol composite hydrogel film containing 0.4,1.2,and 2.0 mg/L alendronate released alendronate in the first 6 hours,and then released alendronate evenly and stably,with a release time of more than 96 hours.(3)The results showed that the chitosan/polyvinyl alcohol composite hydrogel film with a ratio of 5:5 had good physical and chemical properties and cytocompatibility,and could be used as a sustained-release carrier of alendronate.

Depression is a prevalent mental and emotional disor-der that often results in significant emotional disturbances,cog-nitive dysfunction,and memory impairments.It is characterized by a high incidence rate,a substantial disability burden,and limited therapeutic efficacy.Currently,the long-term use of medications for the treatment of depression can result in a range of adverse reactions,highlighting the urgent need to explore no-vel approaches that can effectively alleviate depressive symptoms while minimizing side effects.Curcumin,a natural polyphenolic compound derived from the rhizome of turmeric,demonstrates considerable potential in the prevention and treatment of depres-sion,owing to its diverse array of biological activities.In recent years,numerous studies have investigated the use of curcumin for the treatment of depression.This article aims to provide a comprehensive review of the mechanisms of action underlying curcumin's efficacy in treating depression.Specifically,it focu-ses on its ability to improve neurotransmitter imbalances,restore neural plasticity,alleviate neural damage,mitigate dysfunction of the hypothalamic-pituitary-adrenal(HPA)axis,regulate in-flammatory factors and neuroinflammatory signaling pathways,and inhibit oxidative stress.This review is intended to offer in-sights and methodological references for basic research on curcu-min,as well as for the development of novel therapeutic agents for the treatment of depression.

Depression is a prevalent mental and emotional disor-der that often results in significant emotional disturbances,cog-nitive dysfunction,and memory impairments.It is characterized by a high incidence rate,a substantial disability burden,and limited therapeutic efficacy.Currently,the long-term use of medications for the treatment of depression can result in a range of adverse reactions,highlighting the urgent need to explore no-vel approaches that can effectively alleviate depressive symptoms while minimizing side effects.Curcumin,a natural polyphenolic compound derived from the rhizome of turmeric,demonstrates considerable potential in the prevention and treatment of depres-sion,owing to its diverse array of biological activities.In recent years,numerous studies have investigated the use of curcumin for the treatment of depression.This article aims to provide a comprehensive review of the mechanisms of action underlying curcumin's efficacy in treating depression.Specifically,it focu-ses on its ability to improve neurotransmitter imbalances,restore neural plasticity,alleviate neural damage,mitigate dysfunction of the hypothalamic-pituitary-adrenal(HPA)axis,regulate in-flammatory factors and neuroinflammatory signaling pathways,and inhibit oxidative stress.This review is intended to offer in-sights and methodological references for basic research on curcu-min,as well as for the development of novel therapeutic agents for the treatment of depression.

Objective To explore the effect of life reconstruction training on anxiety and depression among individuals with chronic spinal cord injury(SCI)and their caregivers,and to analyze the influencing factors.Methods A total of 216 SCI patients and 79 caregivers who participated in life reconstruction training were recruited from the Hope House for SCI individuals in Shanghai Yangzhi Rehabilitation Hospital from May,2017 to Octo-ber,2019.They were assessed with Self-Rating Anxiety Scale(SAS)and Self-Rating Depression Scale(SDS)be-fore and after intervention.Results After intervention,the scores of SAS and SDS decreased(P<0.05)in both SCI patients and their caregivers.SAS scores were more likely to reduce in SCI patients who suffered anxiety at admission(OR=2.315,95%CI 1.328 to 4.033,P=0.003)and their caregivers(OR=3.632,95%CI 1.270 to 10.390,P=0.016),as well as non-agricultural-registered SCI patients(OR=1.908,95%CI 1.074 to 3.391,P=0.028).Being unmarried,having an injury duration≥21 years,injury due to disease,and having depression at admission significantly promoted the reduction in SDS scores among SCI patients(P<0.05).Caregivers with a junior-high-school education or less were more likely to experience reductions in scores of SAS and SDS compared with those with higher education(P<0.05).Caregivers with five to 15 years of caregiving experience achieved more anxiety reduction(OR=7.155,95%CI 1.806 to 28.342,P=0.005).Conclusion Life reconstruction training is effective on anxiety and depression among individuals with chronic SCI and their caregivers.It is recommended to prioritize anxiety interventions for patients with rural household registra-tion in China,along with depression management for the married/divorced/widowed individuals,with<21 years since injury,and trauma-induced lesions.For caregivers,it is needed to focus to anxiety and depression in better-educated individuals,as well as anxiety in those with either<5 or≥15 years of caregiving experience.