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.

The cardiovascular-kidney-metabolic syndrome (CKM) is a systemic disease caused by the interaction and mutual influence of cardiovascular diseases, chronic kidney disease, and metabolic abnormalities. It severely impairs patients' quality of life, increases the risk of death, and has become a serious global public health issue. Sodium-glucose transporter 2 inhibitor (SGLT2i) is a novel class of oral hypoglycemic agent with both cardiorenal protection and metabolic regulation effects. It has become an emerging strategy to prevent the onset, slow the progression, and improve the prognosis of CKM. This article provides a comprehensive review of the current applications of SGLT2i in CKM and their potential mechanisms for ameliorating metabolic risk factors and conferring cardio-renal protection, thereby offering a theoretical foundation for the clinical use of SGLT2i in patients with this newly defined critical clinical condition.

Objective: To investigate independent risk factors for lower extremity amputation (LEA) in hospitalized patients with diabetic foot ulcers ( DFUs) . Methods: A multicenter retrospective analysis was conducted on the clinical data of 329 DFUs hospitalized patients with diabetic foot ulcers from four general hospitals across the na⁃tion. A multivariate Logistic regression model was constructed , and prediction analysis was performed using R 4. 2. 1 . The discriminative ability of the model was assessed using receiver operating characteristic curves , while calibration accuracy and clinical applicability were evaluated via calibration curves and decision curve analysis. Results : The study revealed that patients with higher education backgrounds showed lower disease severity (Wagnergrade) (Z = - 4. 331 , P < 0. 05) . A history of amputation , pre⁃existing lower extremity vascular disease , abnormal dorsalis pedis artery pulsation , and a history of coronary heart disease were significantly associated with the severity of DFUs , resulting in higher Wagner scores (P < 0. 05) . In the amputation prognosis analysis , prolonged duration of diabetes and elevated white blood cell count were positively correlated with amputation risk ( both P < 0. 01) .Multivariable regression identified non⁃higher education , low hemoglobin levels , decreased total cholesterol , and abnormally elevated platelet counts as independent risk factors for high Wagner grades ( ≥ grade 3 ) ( all P <0. 05) . The integrated predictive model incorporating these factors demonstrated strong discriminative performance ,with an area under curve of 0. 880 (95% CI: 0. 801 - 0. 960) . The calibration curve slope approached the ideal value , and decision curve analysis confirmed the model ′s clinical net benefit within a threshold probability range of 10% - 65% . Conclusion Lower education level , poor baseline nutritional status , infection , hypercoagulability ,and underlying vascular diseases collectively constitute key factors contributing to elevated amputation risk in DFUs patients. The developed predictive model exhibits high accuracy and may assist clinicians in formulating individual⁃ized intervention strategies.

The cardiovascular-kidney-metabolic syndrome (CKM) is a systemic disease caused by the interaction and mutual influence of cardiovascular diseases, chronic kidney disease, and metabolic abnormalities. It severely impairs patients' quality of life, increases the risk of death, and has become a serious global public health issue. Sodium-glucose transporter 2 inhibitor (SGLT2i) is a novel class of oral hypoglycemic agent with both cardiorenal protection and metabolic regulation effects. It has become an emerging strategy to prevent the onset, slow the progression, and improve the prognosis of CKM. This article provides a comprehensive review of the current applications of SGLT2i in CKM and their potential mechanisms for ameliorating metabolic risk factors and conferring cardio-renal protection, thereby offering a theoretical foundation for the clinical use of SGLT2i in patients with this newly defined critical clinical condition.

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 investigate the relationship between intramuscular adipose tissue index (IATI) calculated from computed tomography images at transverse process of the first lumbar and all-cause mortality in maintenance dialysis patients, and to provide a reference for improving the prognosis in these patients.Methods:It was a multicenter retrospective cohort study. The clinical data of patients who received maintenance hemodialysis or peritoneal dialysis treatment from January 1, 2017 to December 31, 2019 in 4 grade Ⅲ hospitals including Zhongda Hospital Affiliated to Southeast University, Taizhou People's Hospital Affiliated to Nanjing Medical University, Affiliated Hospital of Yangzhou University, and the Third Affiliated Hospital of Soochow University were retrospectively collected. IATI was calculated by low attenuation muscle (LAM) density/skeletal muscle density. The receiver-operating characteristic curve was used to determine the optimal cut-off value of IATI, and the patients were divided into high IATI group and low IATI group according to the optimal cut-off value. The differences of baseline clinical data and measurement parameters of the first lumbar level between the two groups were compared. The follow-up ended on December 23, 2022. The endpoint event was defined as all-cause mortality within 3 years. Kaplan-Meier survival curve and log-rank test were used to analyze the survival rates and the differences between the two groups. Multivariate Cox regression analysis models were used to analyze the association between IATI and the risk of all-cause mortality in maintenance dialysis patients. Multivariate logistic regression analysis model was used to analyze the influencing factors of high IATI.Results:A total of 478 patients were eligibly recruited in this study, with age of (53.55±13.19) years old and 319 (66.7%) males, including 365 (76.4%) hemodialysis patients and 113 (23.6%) peritoneal dialysis patients. There were 376 (78.7%) patients in low IATI (<0.42) group and 102 (21.3%) patients in high IATI (≥0.42) group. The proportion of age ≥ 60 years old ( χ2=24.746, P<0.001), proportion of diabetes mellitus ( χ2=5.570, P=0.018), fasting blood glucose ( t=-2.145, P=0.032), LAM density ( t=-3.735, P<0.001), LAM index ( t=-7.072, P<0.001), and LAM area/skeletal muscle area ratio ( Z=-9.630, P<0.001) in high IATI group were all higher than those in low IATI group, while proportion of males ( χ2=11.116, P<0.001), serum albumin ( Z=2.708, P=0.007) and skeletal muscle density ( t=12.380, P<0.001) were lower than those in low IATI group. Kaplan-Meier survival analysis showed that the 3-years overall survival rate of low IATI group was significantly higher than that in high IATI group (Log-rank χ2=19.188, P<0.001). Multivariate Cox regression analysis showed that IATI<0.42 [<0.42/≥0.42, HR(95% CI): 0.50 (0.31-0.83), P=0.007] was an independent protective factor of all-cause mortality, and age ≥60 years old [ HR (95% CI): 2.61 (1.60-4.23), P<0.001], diabetes mellitus [ HR (95% CI): 1.71 (1.06-2.78), P=0.029] and high blood neutrophil/lymphocyte ratio [ HR (95% CI): 1.04 (1.00-1.07), P=0.049] were the independent risk factors of all-cause mortality in maintenance dialysis patients. Stepwise Cox regression analysis showed that IATI<0.42 was still an independent protective factor of all-cause mortality in maintenance dialysis patients [<0.42/≥0.42, HR (95% CI): 0.45 (0.27-0.76), P=0.003]. Multivariate logistic regression analysis showed that low skeletal muscle density [ OR (95% CI): 0.84 (0.81-0.88), P<0.001] and high serum triglyceride [ OR (95% CI): 1.39 (1.07-1.82), P=0.015] were the independent influencing factors of IATI≥0.42. Conclusion:IATI<0.42 of the first lumbar level is an independent protective factor of all-cause mortality in maintenance dialysis patients. Localized myosteatosis within high-quality skeletal muscle may reduce the risk of all-cause mortality in these patients.

Immunotherapy,represented by immune checkpoint inhibitors,may lead to immune-related adverse events(irAEs)while demonstrating favorable efficacy in patients with malignant tumors.Many domestic and international guidelines or consensus have been established to assist clinicians in effectively managing the majority of irAEs.However,further exploration about irAEs is required regarding the definition and determination,temporal patterns,individual variances,utilization of hormones and immunosuppressants,correlation between irAEs and therapeutic outcomes,immune reactivation,and special populations.The identification and management of severe,refractory,and multiple irAEs necessitate additional solutions.This paper aims to clarify 10 prominent issues concerning irAEs individually and provide assistance for clinicians.

Immunotherapy,represented by immune checkpoint inhibitors(ICIs),has significantly changed the treatment status of most patients with malignant tumors.In order to regulate and guide the immunotherapy of cancer patients in China,the Chinese Society of Clinical Oncology(CSCO)has updated the Immune Checkpoint Inhibitor Clinical Practice Guidelines every year since 2020,which has been widely praised.The Version 2024 released this year has been significantly updated in terms of content,which can be summarized in five keywords as:"addition","deletion","upgrade","downgrade"and"definiteness",with the characteristic of a large overall update range,the continuous advancement of immunotherapy,continuous enrichment of combination strategies,more first-line treatment options,and excellent performance of several drugs.This article provides a point-to-point interpretation of major updates in the new guideline.

Objective In cases and events of mixed STR typing of aborted fetus,two methods for calculating paternity index(PI)of suspected biological fathers are proposed,which could be useful for theoretical reference for parental identification including mixed STR typing.Methods Depending on whether the fetal genotypes can be identified,the simple PI calculation method and the PI calculation method of deduced biological paternal genes when the fetal genotypes cannot be identified are proposed.Results The simple PI calculation method is to indentify the fetal genotypes first and then calculate according to the standard triplet.The PI calculation method of deduced biological paternal genes is to deduce all the possible genotypes of biological fathers conforming to Mendel's law(inference)without considering the ratio of peak height and peak area in mixed typing,and then calculate the parental index separately,taking the minimum value as the parental index of the locus.Conclusion When mixture ratio of fetus in the mixed typing of aborted tissue MR≥0.43,the accuracy of separation is very high and the simple PI calculation method can be accurate,so it is recommended.If 0.05≤MR＜0.43,it is suggested to use the calculation method of deduced biological paternal genes,which can avoid misjudgment of irrelevant persons to the greatest extent.If MR＜0.05,there's a high risk of fetal allele loss,we should not perform a paternity test on the mixed spot.Since the cumulative parental index calculated by deduced the biological paternal genes is usually lower than the value calculated by dividing the fetal genotype,the CPI may be lower than 10 000 when fewer loci are identified,and then more genetic markers should be detected.

We retrospectively analyzed the clinical data of seven patients (four men and three women) with primary hyperoxaluria (PH) type 1 (PH1) in the Department of Nephrology of Zhongda Hospital, Southeast University from January 2018 to October 2023. The mean age at disease onset was 32.1 (range: 26-42) years. The mean age at diagnosis was 40.6 (range: 28-51) years. All patients initially had kidney stones, and three patients were found to have renal insufficiency at the time of disease onset. Among them, two patients underwent hemodialysis immediately. Symptoms at the first visit included bone pain ( n=7), joint pain or deformity ( n=5), fatigue ( n=5), hypotension ( n=3), and subcutaneous nodules ( n=2). Four patients had a family history of PH. All patients had varying degrees of anemia (60-114 g/L), significant hypoalbuminemia (16.5-32.1 g/L), and hypercoagulable state (D-dimer: 2 230-12 781 μg/L). Seven patients received maintenance hemodialysis; their mean age was 37.7 (range: 26-50) years. The mean duration from disease onset to hemodialysis was 5.6 (range: 0-20) years. Five patients repeatedly experienced dialysis access dysfunction. Three patients underwent kidney transplantation before a diagnosis was made, and all transplanted kidneys lost function due to oxalate deposition. The mean follow-up duration was 14.43 (range: 4-38) months. Unfortunately, one patient died. All seven patients underwent computed tomography of the abdomen. All patients suffered skeletal abnormalities, bilateral nephrolithiasis, and nephrocalcinosis. Six patients carried AGXT gene mutations, including four compound heterozygous mutations and two pure homozygous mutations.The mutation sites included: c.823-824dup.AG (p.S275Rfs*38)(exon 8), c.815-816ins.GA (p.S275Rfs*38)(exon 8), c.595G>A (p.G199S) (exon 5), c.32C>G (p.P11R) (exon 1), and c.638C>T (p.A213V)(exon 6). According to the American College of Medical Genetics and Genomics guidelines, two loci were identified as likely pathogenic variants, seven were identified as pathogenic variants, and one locus was identified as having uncertain significance. In addition, patients 1 and 4 underwent skin biopsy, patient 2 underwent renal transplant biopsy, and patient 3 underwent bone marrow biopsy. Interestingly, significant oxalate deposition was found in the tissues. Therefore, PH1 is a rare autosomal recessive inherited disease. This study not only enhanced the understanding of the clinical characteristics of PH1 patients but also had great significance in early diagnosis and treatment of the disease.