This study explored the effect of the Mycobacterium tuberculosis(Mtb)PE/PPE family protein PE_PGRS37 on the growth of Mycobacterium smegmatis(Ms)and macrophage autophagy during Mtb infection.The pe_pgrs37 gene was amplified from Mtb genome through PCR,and the recombinant vector pAIN-PE_PGRS37 was successfully constructed through homologous recombi-nation.pAIN-PE_PGRS37 and pAIN were integrated into Ms through electroshock to construct pAIN-PGRS37/Ms and pAIN/Ms re-combinant bacteria.Western blotting indicated that the PE_PGRS37 protein was correctly expressed in pAIN-PE_PGRS37/Ms.The re-combinant bacteria were inoculated in 7H9/7H10 medium,and their colony morphology and growth curves were observed.No signifi-cant difference in colony morphology was observed between pAIN-PE_PGRS37/Ms and pAIN/Ms.The growth rate significantly in-creased between 10 and 16 h,and a plateau was reached at 26 h.After infection of U937 cells with pAIN-PE_PGRS37/Ms and pAIN/Ms,macrophage autophagy flow was detected with western blotting and immunofluorescence.In the pAIN-PE_PGRS37/Ms-infected group,compared with the pAIN/Ms-infected group,macrophage LC3-II and p62 protein expression was significantly up-regulated(P<0.001)and inhibited autophagosome and lysosome fusion.The intracellular survival of the recombinant bacteria was detected through colony counting,and pAIN-PE_PGRS37/Ms showed significantly greater survival in macrophages at 12 h,24 h,and 48 h than pAIN/Ms(P<0.05).Our results suggested that PE_PGRS37 protein promotes Mycobacterium survival in macrophages by blocking macro-phage autophagy flow,thus inhibiting macrophage autophagy.

Objective:To explore the value of ultrasound elastography (USE) in evaluating the stability and prognosis of carotid plaque (CP) in patients with cerebral infarction(CI).Methods:A total of 94 patients with CI admitted to Dongying District People′s Hospital of Dongying City from January 2022 to December 2023 were retrospectively selected as the study objects, all of whom were treated with carotid artery dissection. The presence of CP was confirmed by pathological examination. The head and neck of patients were examined by conventional ultrasound and USE before surgery, respectively, and the diagnostic value of the two methods on CP stability was compared. The difference of strain value and strain rate among different plaque types was compared, and the correlation between different plaque types and the score of ESSEN Stroke Risk Scale (ESRS) was analyzed.Results:The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of conventional ultrasonic detection of CP were 75.00%, 71.43%, 74.00%, 87.10% and 52.63%; and of USE were 90.28%, 89.29%, 90.00%, 95.59%, 78.13%. The strain values and strain rates of stable plaques were lower than those of vulnerable plaques : (0.73 ± 0.11) × 10 3 vs. (2.42 ± 0.57) × 10 3, (6.57 ± 0.84) s -1 vs. (9.36 ± 2.55) s -1, there were statistical differences ( P<0.01). The ESRS scores of patients with vulnerable plaques were mostly ≥3 scores, and those with stable plaques were mostly<3 scores, there was statistical differences ( P<0.01). The scores of National Institutes of Health Stroke Scale (NIHSS) and modified Rankin Scale (mRS) 3 months after the onset of stable plaque were lower than those of vulnerable plaque patients: (26.80 ± 8.47) scores vs. (34.67 ± 8.98) scores, (3.97 ± 0.84) scores vs. (4.55 ± 1.61) scores, there were statistical differences ( P<0.01 or <0.05). Conclusions:USE has significant advantages in detecting CP stability in CI patients, which can efficiently and accurately assess the risk of plaque and provide important information for clinical use.

This study explored the effect of the Mycobacterium tuberculosis(Mtb)PE/PPE family protein PE_PGRS37 on the growth of Mycobacterium smegmatis(Ms)and macrophage autophagy during Mtb infection.The pe_pgrs37 gene was amplified from Mtb genome through PCR,and the recombinant vector pAIN-PE_PGRS37 was successfully constructed through homologous recombi-nation.pAIN-PE_PGRS37 and pAIN were integrated into Ms through electroshock to construct pAIN-PGRS37/Ms and pAIN/Ms re-combinant bacteria.Western blotting indicated that the PE_PGRS37 protein was correctly expressed in pAIN-PE_PGRS37/Ms.The re-combinant bacteria were inoculated in 7H9/7H10 medium,and their colony morphology and growth curves were observed.No signifi-cant difference in colony morphology was observed between pAIN-PE_PGRS37/Ms and pAIN/Ms.The growth rate significantly in-creased between 10 and 16 h,and a plateau was reached at 26 h.After infection of U937 cells with pAIN-PE_PGRS37/Ms and pAIN/Ms,macrophage autophagy flow was detected with western blotting and immunofluorescence.In the pAIN-PE_PGRS37/Ms-infected group,compared with the pAIN/Ms-infected group,macrophage LC3-II and p62 protein expression was significantly up-regulated(P<0.001)and inhibited autophagosome and lysosome fusion.The intracellular survival of the recombinant bacteria was detected through colony counting,and pAIN-PE_PGRS37/Ms showed significantly greater survival in macrophages at 12 h,24 h,and 48 h than pAIN/Ms(P<0.05).Our results suggested that PE_PGRS37 protein promotes Mycobacterium survival in macrophages by blocking macro-phage autophagy flow,thus inhibiting macrophage autophagy.

Objective:To validate the usefulness of copy number variation sequencing (CNV-seq) in detecting the deletion/duplication of the DMD gene in Duchenne muscular dystrophy (DMD)/Becker muscular dystrophy (BMD) patients. Methods:One hundred and seventy-seven cases who visited the Department of Medical Genetics, Affiliated Hospital of Kunming University of Science and Technology/the First People′s Hospital of Yunnan Province from April 2018 to November 2023 were collected. All patients had previously accepted multiplex ligation-dependent probe amplification (MLPA) to detect the deletion/duplication of the DMD gene, including 90 cases of normal control with a negative result of MLPA and 87 cases with the deletion or duplication of the DMD gene (61 cases of DMD and 26 cases of BMD). CNV-seq was performed in a single-blind manner to detect DMD gene deletion or duplication for all of 177 cases to obtain the detection efficiency of CNV-seq in comparison with MLPA. Results:Comparing to MLPA, CNV-seq had a coincidence rate of 88.7% (157/177) for detecting DMD gene deletion/duplication, with a sensitivity of 77.0% (67/87), a specificity and a positive predictive value of both 100.0% (90/90 and 67/67, respectively), a negative predictive value of 81.8% (90/110), and a Kappa value of 0.773. Of the 87 patients with the deletion or duplication of the DMD gene, CNV-seq detected 67 cases with DMD gene deletion/duplication, including 62 cases with deletion and 5 cases with duplication, with fragment ranging from 150 to 750 kb. While CNV-seq missed 23.0% (20/87) of positive cases, mainly due to the involved fragments spanning only 1 to 4 exons, and with a variation size less than 50 kb, below the resolution (100 kb) of CNV-seq. The detection rate of CNV-seq in BMD cases (84.6%, 22/26) was a little higher than that in DMD cases (73.8%, 45/61), but there was no significant difference between 2 subgroups ( χ2=1.211, P=0.271). The results of CNV-seq in normal controls were all negative, and consistent with the results of MLPA. Conclusion:CNV-seq can detect 77.0% (67/87) of deletion/duplication of the DMD gene in patients with DMD/BMD, while the deletion/duplication less than 100 kb may be inevitably unidentified, therefore it is recommended as an assistant screening technique in prenatal diagnosis for DMD gene deletion or duplication.

Objective:To explore the value of ultrasound elastography (USE) in evaluating the stability and prognosis of carotid plaque (CP) in patients with cerebral infarction(CI).Methods:A total of 94 patients with CI admitted to Dongying District People′s Hospital of Dongying City from January 2022 to December 2023 were retrospectively selected as the study objects, all of whom were treated with carotid artery dissection. The presence of CP was confirmed by pathological examination. The head and neck of patients were examined by conventional ultrasound and USE before surgery, respectively, and the diagnostic value of the two methods on CP stability was compared. The difference of strain value and strain rate among different plaque types was compared, and the correlation between different plaque types and the score of ESSEN Stroke Risk Scale (ESRS) was analyzed.Results:The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of conventional ultrasonic detection of CP were 75.00%, 71.43%, 74.00%, 87.10% and 52.63%; and of USE were 90.28%, 89.29%, 90.00%, 95.59%, 78.13%. The strain values and strain rates of stable plaques were lower than those of vulnerable plaques : (0.73 ± 0.11) × 10 3 vs. (2.42 ± 0.57) × 10 3, (6.57 ± 0.84) s -1 vs. (9.36 ± 2.55) s -1, there were statistical differences ( P<0.01). The ESRS scores of patients with vulnerable plaques were mostly ≥3 scores, and those with stable plaques were mostly<3 scores, there was statistical differences ( P<0.01). The scores of National Institutes of Health Stroke Scale (NIHSS) and modified Rankin Scale (mRS) 3 months after the onset of stable plaque were lower than those of vulnerable plaque patients: (26.80 ± 8.47) scores vs. (34.67 ± 8.98) scores, (3.97 ± 0.84) scores vs. (4.55 ± 1.61) scores, there were statistical differences ( P<0.01 or <0.05). Conclusions:USE has significant advantages in detecting CP stability in CI patients, which can efficiently and accurately assess the risk of plaque and provide important information for clinical use.

Objective:To validate the usefulness of copy number variation sequencing (CNV-seq) in detecting the deletion/duplication of the DMD gene in Duchenne muscular dystrophy (DMD)/Becker muscular dystrophy (BMD) patients. Methods:One hundred and seventy-seven cases who visited the Department of Medical Genetics, Affiliated Hospital of Kunming University of Science and Technology/the First People′s Hospital of Yunnan Province from April 2018 to November 2023 were collected. All patients had previously accepted multiplex ligation-dependent probe amplification (MLPA) to detect the deletion/duplication of the DMD gene, including 90 cases of normal control with a negative result of MLPA and 87 cases with the deletion or duplication of the DMD gene (61 cases of DMD and 26 cases of BMD). CNV-seq was performed in a single-blind manner to detect DMD gene deletion or duplication for all of 177 cases to obtain the detection efficiency of CNV-seq in comparison with MLPA. Results:Comparing to MLPA, CNV-seq had a coincidence rate of 88.7% (157/177) for detecting DMD gene deletion/duplication, with a sensitivity of 77.0% (67/87), a specificity and a positive predictive value of both 100.0% (90/90 and 67/67, respectively), a negative predictive value of 81.8% (90/110), and a Kappa value of 0.773. Of the 87 patients with the deletion or duplication of the DMD gene, CNV-seq detected 67 cases with DMD gene deletion/duplication, including 62 cases with deletion and 5 cases with duplication, with fragment ranging from 150 to 750 kb. While CNV-seq missed 23.0% (20/87) of positive cases, mainly due to the involved fragments spanning only 1 to 4 exons, and with a variation size less than 50 kb, below the resolution (100 kb) of CNV-seq. The detection rate of CNV-seq in BMD cases (84.6%, 22/26) was a little higher than that in DMD cases (73.8%, 45/61), but there was no significant difference between 2 subgroups ( χ2=1.211, P=0.271). The results of CNV-seq in normal controls were all negative, and consistent with the results of MLPA. Conclusion:CNV-seq can detect 77.0% (67/87) of deletion/duplication of the DMD gene in patients with DMD/BMD, while the deletion/duplication less than 100 kb may be inevitably unidentified, therefore it is recommended as an assistant screening technique in prenatal diagnosis for DMD gene deletion or duplication.

OBJECTIVE: To perform carrier screening for spinal muscular atrophy (SMA) among 3049 reproductive-age individuals from Yunnan region and determine the copy number of survival motor neuron (SMN) gene and carrier frequencies. METHODS: Multiplex ligation-dependent probe amplification (MLPA) was used to determine the copy number of exon 7 of SMN1 and SMN2 genes and identify those with a single copy of SMN1 gene. Prenatal diagnosis was performed for couples whom were both found to be SMA carriers. RESULTS: In total 62 SMA carriers were identified among the 3049 subjects, which yielded a carrier frequency of 1 in 49 (2.03%). No statistical difference was found in the carrier frequency between males and females (1.91% vs. 2.30%, P>0.05). Respectively, 1.3% (41/3049) and 0.69% (21/3049) of the carriers were caused by heterozygous deletion and conversion of the SMN1 gene. The average copy number for SMN1 alleles was 1.99. Two couples were found to be both as SMA carriers, for whom the birth of an affected fetus was avoided by prenatal diagnosis. CONCLUSION No difference was found in the carrier frequency of SMA-related mutations between the two genders in Yunnan region, which was in keeping to an autosomal recessive inheritance pattern. Determination of the carrier frequency for SMA and SMN gene variants may provide a basis for genetic counseling and prenatal diagnosis for the disease.
China ; Female ; Genetic Carrier Screening ; Genetic Counseling ; Genetic Variation ; Heterozygote ; Humans ; Male ; Muscular Atrophy, Spinal ; genetics ; Pregnancy ; Prenatal Diagnosis ; Survival of Motor Neuron 1 Protein ; genetics ; Survival of Motor Neuron 2 Protein ; genetics

Objective To explore the construction, application advantages and values of the ECG network system under Wise Information Technology.Methods A total of 240 patients enrolled in Department of Neurology, the First Affiliated Hospital of Wenzhou Medical University from March 2016 to May 2016 were randomly divided into two groups: conventional ECG group (control group) and network ECG group (experimental group), with 120 cases respectively. The ECG test time and satisfaction of patients and nurses were compared between two groups.Results Compared with the conventional ECG group, patients in network ECG group took less time to have an ECG examination, which decreased from an average of 33.32 min to 9.59 min. And the time required for nurses decreased from an average of 10.27 min to 4.82 min. The differences were statistically significant (P<0.001). Patients' satisfaction increased from 89.8% to 99.5%, while medical staff's satisfaction increased from 91.5% to 99.3%. The differences were statistically significant (P<0.001). Conclusions Network ECG examination greatly reduces the waste of human resources and saves operating costs, enhances the health care staff satisfaction and is worth further promotion in the clinical application.