OBJECTIVE: To explore the genetic etiology of 46 Chinese pedigrees affected with Hereditary multiple exostoses (HME) and provide genetic counseling and reproductive intervention. METHODS: Whole-exome sequencing and Sanger sequencing were carried out on 87 patients from the 46 pedigrees to analyze the variants of EXT1 and EXT2 genes. Pathogenicity of the variants was assessed based on the guidelines from the American College of Medical Genetics and Genomics and Association for Molecular Pathology (ACMG/AMP). Prenatal diagnosis and preimplantation genetic testing (PGT) were provided for couples with identified pathogenic mutations. This study was approved by the Medical Ethics Committee of the hospital (Ethics No.: LL-SC-SG-2014-010). RESULTS: In total 17 and 22 pathogenic variants were respectively identified in the EXT1 and EXT2 genes, among which 5 EXT1 and 12 EXT2 variants were unreported previously. Three patients with no family history were found to harbor de novo variants of the EXT1 gene. Twenty nine couples had opted for PGT or underwent prenatal diagnosis following natural conception, and 17 healthy babies were born. CONCLUSION This study has clarified the genetic etiology of 45 HME pedigrees and identified 17 novel variants, which has enriched the mutational spectrum of the EXT1 and EXT2 genes. Reproductive intervention through PGT and prenatal diagnosis have prevented the recurrence of HME in these families.
Humans ; Female ; Male ; Pedigree ; Exostoses, Multiple Hereditary/diagnosis* ; N-Acetylglucosaminyltransferases/genetics* ; Adult ; Exostosin 1 ; Asian People/genetics* ; Genetic Testing ; Exostosin 2 ; Mutation ; China ; Prenatal Diagnosis ; Pregnancy ; Genetic Counseling ; Preimplantation Diagnosis ; Exome Sequencing ; East Asian People

Objective To explore the effect of back-pushing manipulation on motor ability and skele-tal muscle mitochondrial function in rats with chronic fatigue syndrome(CFS).Methods Twenty-four male Sprague-Dawley rats were randomly divided into a blank group of 8 and a modeling group of 16.The CFS rat model was established using forced weight-bearing swimming combined with chronic stress stimulation for 21 days.After successful modeling,the modeling group was further divided into the modeling control(MC)group and the back-pushing manipulation(BPM)group,each of 8.In the back-pushing manipulation group,all rats were given daily 20-minute back pushing for 14 consecu-tive days after modeling.Then,all groups were recorded semi-quantitative scores of general conditions(SQS-GC),body mass,and exhaustion swimming time(EST).Moreover,the grasping ability of limbs was assessed by using the mesh screen test(MST).After the intervention,the tissue of the erector spinae muscle was taken to detect the adenosine triphosphate(ATP)content using the biochemi-cal method,and the activities of mitochondrial respiratory chain complexes Ⅰ to Ⅳ were detected by using the spectrophotometric method.Results After modeling,compared with the blank group,the SQS-GC of the MC group and BPM groups were higher(P<0.01),while EST was shorter and the MST score was lower(P<0.01 or P<0.05).However,after intervention,compared with the MC group,the SQS-GC of the BPM group was lower(P<0.01),while EST and the MST score was higher(P<0.01 or P<0.05).Compared with the blank group,the ATP content of the modelling group was significantly lower(P<0.01),and the activities of mitochondrial respiratory chain complexes I,II,III,and IV de-creased significantly(P<0.01).However,after intervention,all the above values of the BPM group in-creased significantly,compared with the MC group(P<0.01 or P<0.05).Conclusion The back-pushing manipulation can improve the mitochondrial function and energy metabolism,as well as the exercise ca-pacity,and alleviate fatigue of CFS rats,which may be related to the improvement of the activities of mitochondrial respiratory chain complexes I,II,III,and IV,and the increase of ATP content in the skeletal muscles.

Objective To explore the hearing characteristics and early screening methods of hearing loss in diabetes mellitus(DM)patients.Methods From September 2022 to November 2023,800 DM patients in the Department of Endocrinology in People's Liberation Army General Hospital of Southern Theater Command were enrolled.According to the results of electrical audiometry,they were divided into a normal hearing DM group(n=214)and a hearing loss group(n=586).Binary logistic regression was used to analyze the influencing factors of hearing loss in DM patients.Results Age,homeostasis model assessment of insulin resistance(HOMA-IR),serum creatinine(Scr),urinary albumin/creatinine ratio(UACR)DM duration,hearing loss value,and proportion of DPN,DR,DKD,hypertension,coronary heart disease,perceived hearing loss,DPOAE abnormality and ABR abnormality in hearing loss group was higher than that in normal hearing group(P<0.05).DBP,homeostasis model insulin sensitivity index and estimated glomerular filtration rate(eGFR)were lower than those in normal hearing group(P<0.05).Logistic regression analysis showed that DM duration(OR 0.987,95%CI 0.987～1.002,P=0.001),eGFR(OR 0.974,95%CI 0.964～0.983,P=0.000),and hypertension(OR 0.659,95%CI 0.443～0.980,P=0.040)and coronary heart disease(OR 0.488,95%CI 0.246～0.968,P=0.040)were the influencing factors of hearing loss in DM patients.Conclusions Hearing loss is the complications of DM and is related to factors such as the DM duration.Given the insidious onset of hearing loss,DM patients should undergo a combined hearing screening using multiple methods in the early stage.

The diagnostic value of Mycobacterium tuberculosis(MTB)complex and rifampicin resistance test kits(fluorescence PCR method)in detecting for MTB complex and rifampicin resistancein sputum samples was evaluated.A total of 271 patients with suspected tuberculosis were prospectively and consecutively enrolled at Hunan Chest Hospital between April 1,2024,and November 30,2024.Of these,229 patients were confirmed to have tuberculosis,whereas 42 patients were not-tuberculosis samples were col-lected from all patients and subjected to fluorescence PCR,Xpert MTB/RIF(abbreviated as Xpert),and MGIT 960 culture and drug sensitivity testing.Clinical diagnosis and MTB culture results served as reference standards for TB diagnosis,whereas phenotypic drug susceptibility testing and Xpert served as reference standards to for assessment of rifampicin resistance.The sensitivity,specificity,positive predictive value,and negative predictive value of the fluorescence PCR method were analyzed.Kappa tests were performed to analyze the concordance between detection techniques.With clinical diagnosis as the gold standard,the sensitivity and specificity of the fluorescence PCR method for the diagnosis of TB were 65.1%(149/229)and 97.6%(41/42),and the consistency test for the fluo-rescence PCR and Xpert methods showed high consistency(Kappa value=0.993).With the MGIT 960 liquid culture as the reference standard,the positive detection rate of the fluorescence PCR method for the detection of patients with positive cultures was 91.9%(102/111,95%CI:85.2%-96.2%),and the positive detection rate for 147 patients with sputum culture-negative TB was 27.9%(41/147,95%CI:21.3%-35.6%).With the phenotypic drug sensitivity test as the gold standard,the sensitivity and specificity of the fluo-rescence PCR method for the detection of rifampicin resistance were 100.0%(31/31)and 96.6%(28/29)respectively,and the consis-tency between tests was high(Kappa value=0.967).With Xpert as the gold standard,the sensitivity and specificity of fluorescence PCR for the detection of rifampicin resistance were 95.8%(46/48)and 99.0%(99/100),respectively,and the consistency between tests was high(Kappa value=0.953).Finally,samples with rifampicin resistance detected with the fluorescence PCR method had a clear rpoB gene mutation type according to one-generation sequencing.In conclusion,the fluorescence PCR method showed high sen-sitivity in detecting MTB complex groups and rifampicin resistance,and had high concordance with Xpert.Therefore,this technique can serve a rapid test for TB diagnosis to increase the rate of positive TB pathology and detection of rifampicin resistance.This method is particularly suitable for use in lower-income countries and economically disadvantaged grassroots communities.

Objective To explore the effect of back-pushing manipulation on motor ability and skele-tal muscle mitochondrial function in rats with chronic fatigue syndrome(CFS).Methods Twenty-four male Sprague-Dawley rats were randomly divided into a blank group of 8 and a modeling group of 16.The CFS rat model was established using forced weight-bearing swimming combined with chronic stress stimulation for 21 days.After successful modeling,the modeling group was further divided into the modeling control(MC)group and the back-pushing manipulation(BPM)group,each of 8.In the back-pushing manipulation group,all rats were given daily 20-minute back pushing for 14 consecu-tive days after modeling.Then,all groups were recorded semi-quantitative scores of general conditions(SQS-GC),body mass,and exhaustion swimming time(EST).Moreover,the grasping ability of limbs was assessed by using the mesh screen test(MST).After the intervention,the tissue of the erector spinae muscle was taken to detect the adenosine triphosphate(ATP)content using the biochemi-cal method,and the activities of mitochondrial respiratory chain complexes Ⅰ to Ⅳ were detected by using the spectrophotometric method.Results After modeling,compared with the blank group,the SQS-GC of the MC group and BPM groups were higher(P<0.01),while EST was shorter and the MST score was lower(P<0.01 or P<0.05).However,after intervention,compared with the MC group,the SQS-GC of the BPM group was lower(P<0.01),while EST and the MST score was higher(P<0.01 or P<0.05).Compared with the blank group,the ATP content of the modelling group was significantly lower(P<0.01),and the activities of mitochondrial respiratory chain complexes I,II,III,and IV de-creased significantly(P<0.01).However,after intervention,all the above values of the BPM group in-creased significantly,compared with the MC group(P<0.01 or P<0.05).Conclusion The back-pushing manipulation can improve the mitochondrial function and energy metabolism,as well as the exercise ca-pacity,and alleviate fatigue of CFS rats,which may be related to the improvement of the activities of mitochondrial respiratory chain complexes I,II,III,and IV,and the increase of ATP content in the skeletal muscles.

Objective To explore the hearing characteristics and early screening methods of hearing loss in diabetes mellitus(DM)patients.Methods From September 2022 to November 2023,800 DM patients in the Department of Endocrinology in People's Liberation Army General Hospital of Southern Theater Command were enrolled.According to the results of electrical audiometry,they were divided into a normal hearing DM group(n=214)and a hearing loss group(n=586).Binary logistic regression was used to analyze the influencing factors of hearing loss in DM patients.Results Age,homeostasis model assessment of insulin resistance(HOMA-IR),serum creatinine(Scr),urinary albumin/creatinine ratio(UACR)DM duration,hearing loss value,and proportion of DPN,DR,DKD,hypertension,coronary heart disease,perceived hearing loss,DPOAE abnormality and ABR abnormality in hearing loss group was higher than that in normal hearing group(P<0.05).DBP,homeostasis model insulin sensitivity index and estimated glomerular filtration rate(eGFR)were lower than those in normal hearing group(P<0.05).Logistic regression analysis showed that DM duration(OR 0.987,95%CI 0.987～1.002,P=0.001),eGFR(OR 0.974,95%CI 0.964～0.983,P=0.000),and hypertension(OR 0.659,95%CI 0.443～0.980,P=0.040)and coronary heart disease(OR 0.488,95%CI 0.246～0.968,P=0.040)were the influencing factors of hearing loss in DM patients.Conclusions Hearing loss is the complications of DM and is related to factors such as the DM duration.Given the insidious onset of hearing loss,DM patients should undergo a combined hearing screening using multiple methods in the early stage.

The diagnostic value of Mycobacterium tuberculosis(MTB)complex and rifampicin resistance test kits(fluorescence PCR method)in detecting for MTB complex and rifampicin resistancein sputum samples was evaluated.A total of 271 patients with suspected tuberculosis were prospectively and consecutively enrolled at Hunan Chest Hospital between April 1,2024,and November 30,2024.Of these,229 patients were confirmed to have tuberculosis,whereas 42 patients were not-tuberculosis samples were col-lected from all patients and subjected to fluorescence PCR,Xpert MTB/RIF(abbreviated as Xpert),and MGIT 960 culture and drug sensitivity testing.Clinical diagnosis and MTB culture results served as reference standards for TB diagnosis,whereas phenotypic drug susceptibility testing and Xpert served as reference standards to for assessment of rifampicin resistance.The sensitivity,specificity,positive predictive value,and negative predictive value of the fluorescence PCR method were analyzed.Kappa tests were performed to analyze the concordance between detection techniques.With clinical diagnosis as the gold standard,the sensitivity and specificity of the fluorescence PCR method for the diagnosis of TB were 65.1%(149/229)and 97.6%(41/42),and the consistency test for the fluo-rescence PCR and Xpert methods showed high consistency(Kappa value=0.993).With the MGIT 960 liquid culture as the reference standard,the positive detection rate of the fluorescence PCR method for the detection of patients with positive cultures was 91.9%(102/111,95%CI:85.2%-96.2%),and the positive detection rate for 147 patients with sputum culture-negative TB was 27.9%(41/147,95%CI:21.3%-35.6%).With the phenotypic drug sensitivity test as the gold standard,the sensitivity and specificity of the fluo-rescence PCR method for the detection of rifampicin resistance were 100.0%(31/31)and 96.6%(28/29)respectively,and the consis-tency between tests was high(Kappa value=0.967).With Xpert as the gold standard,the sensitivity and specificity of fluorescence PCR for the detection of rifampicin resistance were 95.8%(46/48)and 99.0%(99/100),respectively,and the consistency between tests was high(Kappa value=0.953).Finally,samples with rifampicin resistance detected with the fluorescence PCR method had a clear rpoB gene mutation type according to one-generation sequencing.In conclusion,the fluorescence PCR method showed high sen-sitivity in detecting MTB complex groups and rifampicin resistance,and had high concordance with Xpert.Therefore,this technique can serve a rapid test for TB diagnosis to increase the rate of positive TB pathology and detection of rifampicin resistance.This method is particularly suitable for use in lower-income countries and economically disadvantaged grassroots communities.

Objective:To investigate the correlation between hearing loss and microvascular complications in diabetes.Methods:This cross-sectional study conducted the data from 572 patients with diabetes hospitalized in the Endocrinology Department of the General Hospital of Southern Theater Command from September 2022 to July 2023. All participants underwent electrical audiometry and acoustic immittance in the ENY department. Based on the audiometric results, participants were categorized into normal hearing group and hearing loss group. Additionally, 572 non-diabetic patients from the outpatient department were enrolled as the non-diabetic group. The general information and laboratory results were collected and compared using t test, rank sum test or χ2 test. Binary logistic regression analysis was used to evaluate the association of diabetic hearing loss with diabetic kidney disease(DKD), diabetic retinopathy (DR), and diabetic peripheral neuropathy (DPN). Results:Among 572 patients with diabetes, 429 suffered from hearing loss and 143 were normal. χ2 test showed significant differences in combined DKD and DPN between two groups, but not in DR. Multivariate binary logistic regression analysis identified DKD and DPN as risk factors for hearing loss, but no correlation was found with DR. Conclusion:Diabetic patients with DKD or DPN should be monitored for potential hearing loss. Early screening and treatment are crucial to prevent severe hearing impairment.

Gene-knockout technology is increasingly used as a powerful tool for establishing animal models of hyperuricemia(HUA).HUA gene-knockout animal models are not only helpful in revealing the molecular mechanisms of uric acid metabolism but are also of great value for evaluating potential therapeutic strategies.In this paper,the application of gene-knockout technology in HUA animal models is discussed in detail by reviewing the domestic and foreign literature,focusing on the knockout of urate oxidase,glucose transporter 9,and ATP-binding cassette transporter G2.The review provides a reference and guidance for the further establishment of HUA animal models by gene knockout technology.

Objective:To use pan-genomics and subtractive proteomics techniques to screen the new antibacterial targets from the Staphylococcus aureus genome,and to lay the foundation for the development of anti-Staphylococcus aureus drugs.Methods:The genome sequencing data of 50 strains with sequencing level Complete were collected by searching the whole genome sequencing data in the National Center for Biotechnology Information(NCBI)Database with Staphylococcus aureus as the keyword;BPGA tool was used to conduct the pan-genomics analysis on the genomic data to obtain the core genes of Staphylococcus aureus;subtractive proteomics technique was used to screen the potential antibacterial targets from the core genes.These potential antibacterial targets were used as the receptors;LibDock software was used to screen the potential anti-Staphylococcus aureus drugs from the US Food and Drug Administration(FDA)-approved drug library;molecular docking technology was used to analyze the binding abilities of the drugs and targets.Results:There were 14 379 gene families in the 50 Staphylococcus aureus genomes,of which 1 620 were the core genes.The subtractive proteomics analysis results showed that tyrosine autokinase 1335 was the potential anti-Staphylococcus aureus target.LibDock software screened out nine compounds,including balofloxacin,tenofovir disoproxil fumarate,and adefovir,that may exert anti-Staphylococcus aureus effects on this target protein.The molecular docking results showed there was good binding abilities between the targets and the compounds.Conclusion:Tyrosine autokinase may be the potential target for antii-Staphylococcus aureus.