Objective: To define the mechanism by which swine TRIM25 restricts vesicular stomatitis virus replication. Methods: Porcine 3D4/21 cells with TRIM25 overexpression or knockdown were infected with vesicular stomatitis virus. Viral replication was quantified by immunoblotting, quantitative reverse transcription polymerase chain reaction, and 50% tissue culture infectious dose assays. Type I interferon signaling was assessed by transcript quantification, interferon-beta and interferon-stimulated response element reporter assays, and co-immunoprecipitation.Viral RNA binding was tested by RNA immunoprecipitation. Results: TRIM25 overexpression reduced viral RNA and infectious titers, whereas TRIM25 knockdown increased replication (p < 0.01). TRIM25 increased interferon-beta and interferon-stimulated gene expression and enhanced interferon-beta and interferonstimulated response element promoter activity (p < 0.01). Mechanistically, TRIM25 promoted Lys63-linked ubiquitination of RIG-I and increased phosphorylation of TANK-binding kinase 1 and interferon regulatory factor 3. TRIM25 also bound vesicular stomatitis virus genomic RNA, and binding required the C-terminal region. Conclusions and Relevance: Porcine TRIM25 restricts vesicular stomatitis virus replication by amplifying type I interferon signaling and directly binding viral RNA.

OBJECTIVE To explore the molecular characteristics of small colony variants(SCVs)of Staphylococcus aureus isolates so as to provide theoretical bases for clinical control of the persistent and recurrent infections in-duced by the SCVs.METHODS The clinical blood specimens that were collected from the First Affiliated Hospital of Traditional Chinese Medicine University of Guangzhou were cultured to acquire 1 strain of SCV S.aureus S2.The colonial morphology,growth rate,catalase,plasma-coagulase,production capability of biofilm and antimi-crobial susceptibility of the strain SCVs were observed,the growth curve and autolysis curve were drawn,the whole genome sequencing was performed for the S2,the phylogenetic evolution of the genome and the single nu-cleotide polymorphism(SNP)were analyzed.RESULTS The S2 was identified as S.aureus,with the catalase and plasma-coagulase tested positive.The strain was resistant to various types of antibiotics,including macrolides like erythromycin and azithromycin,fluroquinolones such as levofloxacin and ciprofloxacin,and penicillin.The forma-tion of pigment was reduced,the formation of hemolytic rings was low;the growth rate of the strain was slower than that of ATCC 25923(quality control strain)and ZC1(the normal control strain),the autolytic activity of the strain was higher than that of the ATCC 25923 and ZC1,and the biofilm production of the strain was remarkably less than that of the ATCC 29213(P＜0.05).The whole genome sequencing demonstrated that the molecular typ-ing of the S2 was ST239-t030 clone,which harbored a variety of drug resistance genes and virulence genes,and the menB gene had missense mutation.CONCLUSION The formation of SCVs may be associated with the inhibited synthesis of menadione caused by the missense mutation of menB gene.

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:Sarcopenia obesity is characterized by the coexistence of sarcopenia and obesity with a continuously increasing prevalence.Aerobic exercise can alleviate the progression of sarcopenic obesity,but the overall metabolic changes in skeletal muscle after exercise are still unclear.OBJECTIVE:To investigate the effects of exercise on various aspects of the energy metabolic pathways in the skeletal muscle of mice with sarcopenic obesity.METHODS:Male C57BL/6J mice were randomly divided into normal control(normal diet)and model(high-fat diet)groups.After 12 weeks of feeding,sarcopenic obesity model mice were screened by body mass and behavior assessments.The sarcopenia obesity mice were then divided into the sedentary and exercise groups.Mice in the exercise group were subjected to treadmill training on the basis of a high-fat diet.After 8 weeks of intervention,body mass,lipid metabolism,muscle volume of calf muscle group of the hind limb,skeletal muscle morphology,and expressions of energy metabolic pathway-related genes were detected.RESULTS AND CONCLUSION:Compared with the normal control group,the levels of triglycerides,total cholesterol,and free fatty acid in the sedentary group were significantly increased,along with significantly increased lipid droplets in skeletal muscle(P<0.05).Compared with the sedentary group,all of the above indicators in the exercise group showed a significant decreasing trend(P<0.05).Compared with the normal control group,grip strength and fatigue latency time,muscle volume,and fiber cross-sectional area were significantly decreased in the sedentary group,but the mRNA expression of Atrogin-1 and Murf-1 was elevated(P<0.05).Compared with the sedentary group,grip strength and fatigue latency time,muscle volume,and fiber cross-sectional area were significantly improved in the exercise group(P<0.05),while the mRNA expression of Atrogin-1 and Murf-1 was reduced(P<0.05).Compared with the normal control group,the mRNA expression of intramuscular transcription factors Pparα and Pgc-1α was decreased in the sedentary group(P<0.05),the mRNA expression of fatty acid synthesis-related enzymes Srebp1c and Fasn was elevated(P<0.05),and the mRNA expression of the β-oxidation system Cpt1β,Acox1,Acox3 and fatty acid metabolism Arf1 and Plin3 was decreased(P<0.05).Compared with the sedentary group,the abnormal expression of the above genes was significantly reversed in the exercise group(P<0.05).To conclude,aerobic exercise can alleviate lipid deposition and improve muscle quality and strength of sarcopenic obesity mice by regulating the expression of genes in the intramuscular energy metabolism network.

Objectives:To compare the incision exposure time,and accuracy rates of puncture and screw placement of open screw placement with robot-assisted percutaneous needle puncture technique with tradition-al robot-assisted open screw placement,and to explore the learning curve of such technique.Methods:The clinical data of 53 patients who underwent robot-assisted open pedicle screw fixation surgery between August 2020 and December 2023 were retrospectively analyzed.According to the surgical methods,21 patients were divided into observation group(treated with robot-assisted pedicle screw placement with percutaneous needle puncture technique),and the other 32 patients were divided into control group(treated with traditional robot-assisted open pedicle screw placement).There were no statistically significant differences in gender,age,and disease type between the two groups of patients(P＞0.05).The total operation time,incision exposure time,blood loss,accuracy rate of puncture,and accuracy rate of screw placement,as well as complications,were recorded.The patients in the observation group were divided into three groups(A,B and C)according to the order of operation,with 7 cases in each group,and the total operation time,robot operation time,accuracy rate of puncture,accuracy rate of screw placement,and complications in each group were compared.ln curve regression analysis was used to analyze the trend of operation time with the number of operations.Results:There was no statistical difference between the observation group and control group in the total operation time(265.5±108.8min vs 309.2±154.9min,P＞0.05).The incision exposure time in the observation group(189.00±94.74min)was statistically less than that in the control group(309.2±154.9min)(P＜0.05).In the control group,the incision was exposed immediately at the onset of surgery,so the incision exposure time was the same as the total operation time.The accuracy rate of puncture between the observation group and control group showed no statistical difference(94.2％vs 95.0％,P＞0.05).The accuracy rates of screw placement of both groups were 100％.There was no statistical difference in blood loss between the observation group and con-trol group(343.8±207.0mL vs 480.6±392.0mL,P＞0.05).There were 1 cases of incision infection in the obser-vation group and 3 cases in the control group,with no statistical difference(P＞0.05).The total operation time of groups A,B and C was 350.9±92.8min,263.7±116.2min and,181.9±25.8min respectively,and the robot operation time was 90.1±25.1min,73.9±12.1min and,65.4±9.5min respectively.The differences between the three groups were statistically significant(P＜0.05).The results of the ln curve regression analysis showed that the total operation time[y=-71.324ln(x)+419.603]and the robot operation time[y=-15.757ln(x)+110.527]de-creased significantly with the increase of the number of cases and reached a relatively stable level in the 8th case.There was no significant difference in the accuracy rate of puncture,accuracy rate of screw placement and the incidence of complications between groups A,B and C(P＞0.05).Conclusions:Robot-assisted percuta-neous needle puncture technique reduces the incision exposure time and maintains a high screw placement accuracy.With the increase of the number of surgical cases,the total operation time and robot operation time decrease,and the learning curve becomes stable fast.

Objective:To explore the effects three of anesthetics(tribromoethanol,isoflurane,and pentobarbital so-dium)on the outcome of mice with post-stroke dysphagia(PSD)induced by photothrombosis(PT)method,and to e-valuate which anesthetic is more suitable for the preparation of this model.Methods:Sixty-six male C57BL/6J mice were divided into Tribromoethanol group,Isoflurane group,Pentobarbital sodium group and Sham group.The post-stroke dysphagia model was established by PT.Before and 5 min after modeling,a laser speckle imager was used to measure the regional cerebral blood flow(rCBF)decrease rate of mice and record the wake-up time of mice.Forty-eight hours after modeling,the mortality rate of PSD mice in three groups was recorded and the rCBF decrease rate was meas-ured again.The neurological function of mice was evaluated using the neurological deficit score,the water intake of mice was recorded using the 4-min drinking test,the infarct volume ratio was measured using the TTC staining method,and the swallowing counts induced by water administration was recorded using the multichannel physiological recorder MP160 and the myoelectric area of the swallowing muscle was calculated.Results:There was no statistical difference in the percentage of decrease in rCBF,infarct volume ratio,neurological deficit score,water intake,swallowing counts,and myoelectric area of swallowing muscle among the three groups of PSD mice 48 h after modeling(P＞0.05).Com-pared with the Tribromoethanol group and the Pentobarbital sodium group,the rCBF of the mice in the Isoflurane group decreased rapidly within 5 min(P＜0.05),and the mortality rate of the mice was lower and the awakening time was shorter.(P＜0.05).Conclusion:The use of different anesthesia will affect the mortality rate,wake-up time and the downward trend of rCBF within 48 h after modeling of PSD mice.Among the three anesthetics,isoflurane is more suit-able as an anesthetic for the PSD mouse model.

OBJECTIVE To explore the molecular characteristics of small colony variants(SCVs)of Staphylococcus aureus isolates so as to provide theoretical bases for clinical control of the persistent and recurrent infections in-duced by the SCVs.METHODS The clinical blood specimens that were collected from the First Affiliated Hospital of Traditional Chinese Medicine University of Guangzhou were cultured to acquire 1 strain of SCV S.aureus S2.The colonial morphology,growth rate,catalase,plasma-coagulase,production capability of biofilm and antimi-crobial susceptibility of the strain SCVs were observed,the growth curve and autolysis curve were drawn,the whole genome sequencing was performed for the S2,the phylogenetic evolution of the genome and the single nu-cleotide polymorphism(SNP)were analyzed.RESULTS The S2 was identified as S.aureus,with the catalase and plasma-coagulase tested positive.The strain was resistant to various types of antibiotics,including macrolides like erythromycin and azithromycin,fluroquinolones such as levofloxacin and ciprofloxacin,and penicillin.The forma-tion of pigment was reduced,the formation of hemolytic rings was low;the growth rate of the strain was slower than that of ATCC 25923(quality control strain)and ZC1(the normal control strain),the autolytic activity of the strain was higher than that of the ATCC 25923 and ZC1,and the biofilm production of the strain was remarkably less than that of the ATCC 29213(P＜0.05).The whole genome sequencing demonstrated that the molecular typ-ing of the S2 was ST239-t030 clone,which harbored a variety of drug resistance genes and virulence genes,and the menB gene had missense mutation.CONCLUSION The formation of SCVs may be associated with the inhibited synthesis of menadione caused by the missense mutation of menB gene.

Objective:To explore the effects three of anesthetics(tribromoethanol,isoflurane,and pentobarbital so-dium)on the outcome of mice with post-stroke dysphagia(PSD)induced by photothrombosis(PT)method,and to e-valuate which anesthetic is more suitable for the preparation of this model.Methods:Sixty-six male C57BL/6J mice were divided into Tribromoethanol group,Isoflurane group,Pentobarbital sodium group and Sham group.The post-stroke dysphagia model was established by PT.Before and 5 min after modeling,a laser speckle imager was used to measure the regional cerebral blood flow(rCBF)decrease rate of mice and record the wake-up time of mice.Forty-eight hours after modeling,the mortality rate of PSD mice in three groups was recorded and the rCBF decrease rate was meas-ured again.The neurological function of mice was evaluated using the neurological deficit score,the water intake of mice was recorded using the 4-min drinking test,the infarct volume ratio was measured using the TTC staining method,and the swallowing counts induced by water administration was recorded using the multichannel physiological recorder MP160 and the myoelectric area of the swallowing muscle was calculated.Results:There was no statistical difference in the percentage of decrease in rCBF,infarct volume ratio,neurological deficit score,water intake,swallowing counts,and myoelectric area of swallowing muscle among the three groups of PSD mice 48 h after modeling(P＞0.05).Com-pared with the Tribromoethanol group and the Pentobarbital sodium group,the rCBF of the mice in the Isoflurane group decreased rapidly within 5 min(P＜0.05),and the mortality rate of the mice was lower and the awakening time was shorter.(P＜0.05).Conclusion:The use of different anesthesia will affect the mortality rate,wake-up time and the downward trend of rCBF within 48 h after modeling of PSD mice.Among the three anesthetics,isoflurane is more suit-able as an anesthetic for the PSD mouse model.

BACKGROUND:Sarcopenia obesity is characterized by the coexistence of sarcopenia and obesity with a continuously increasing prevalence.Aerobic exercise can alleviate the progression of sarcopenic obesity,but the overall metabolic changes in skeletal muscle after exercise are still unclear.OBJECTIVE:To investigate the effects of exercise on various aspects of the energy metabolic pathways in the skeletal muscle of mice with sarcopenic obesity.METHODS:Male C57BL/6J mice were randomly divided into normal control(normal diet)and model(high-fat diet)groups.After 12 weeks of feeding,sarcopenic obesity model mice were screened by body mass and behavior assessments.The sarcopenia obesity mice were then divided into the sedentary and exercise groups.Mice in the exercise group were subjected to treadmill training on the basis of a high-fat diet.After 8 weeks of intervention,body mass,lipid metabolism,muscle volume of calf muscle group of the hind limb,skeletal muscle morphology,and expressions of energy metabolic pathway-related genes were detected.RESULTS AND CONCLUSION:Compared with the normal control group,the levels of triglycerides,total cholesterol,and free fatty acid in the sedentary group were significantly increased,along with significantly increased lipid droplets in skeletal muscle(P<0.05).Compared with the sedentary group,all of the above indicators in the exercise group showed a significant decreasing trend(P<0.05).Compared with the normal control group,grip strength and fatigue latency time,muscle volume,and fiber cross-sectional area were significantly decreased in the sedentary group,but the mRNA expression of Atrogin-1 and Murf-1 was elevated(P<0.05).Compared with the sedentary group,grip strength and fatigue latency time,muscle volume,and fiber cross-sectional area were significantly improved in the exercise group(P<0.05),while the mRNA expression of Atrogin-1 and Murf-1 was reduced(P<0.05).Compared with the normal control group,the mRNA expression of intramuscular transcription factors Pparα and Pgc-1α was decreased in the sedentary group(P<0.05),the mRNA expression of fatty acid synthesis-related enzymes Srebp1c and Fasn was elevated(P<0.05),and the mRNA expression of the β-oxidation system Cpt1β,Acox1,Acox3 and fatty acid metabolism Arf1 and Plin3 was decreased(P<0.05).Compared with the sedentary group,the abnormal expression of the above genes was significantly reversed in the exercise group(P<0.05).To conclude,aerobic exercise can alleviate lipid deposition and improve muscle quality and strength of sarcopenic obesity mice by regulating the expression of genes in the intramuscular energy metabolism network.

Adverse pregnancy outcome (APO) refers to any pregnancy outcome other than a normal live birth, mainly including preeclampsia, fetal growth restriction, recurrent pregnancy loss, etc., which seriously threaten the health of both the mother and the fetus. Most APOs originate from abnormal uterine endometrial microenvironment before pregnancy. Once a woman confirms pregnancy, intervening in the uterine endometrium at this stage is often too late. The uterine endometrium, as a key site for embryo implantation and development, its abnormal microenvironment can lead to APO. Therefore, in-depth exploration of the relationship between the pre-pregnancy uterine endometrial microenvironment and APO is helpful for clarifying the mechanism of APO occurrence and providing key ideas and methods for early prevention and treatment. This article reviews the latest research on the mechanism of action between uterine endometrial microenvironment and APO, systematically summarizing the research progress and main conclusions in this field from several aspects such as immune imbalance, inflammatory response, abnormal energy metabolism, and senescence of uterine endometrial stromal cells.