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 investigate the relationship between serum interacting protein kinase (RIPK)-3 levels and mycoplasma pneumoniae pneumonia (MPP) correlation between disease severity and lung function in children.Methods:A case-control study was conducted. A total of 147 children with MPP admitted to the Children's Hospital Affiliated to Xuzhou Medical University from April 2023 to March 2024 were selected, including 76 cases of mild MPP (mild MPP group) and 71 cases of severe MPP (severe MPP group). Additionally, 36 healthy children who underwent physical examinations during the same period were selected as the healthy control group. The levels of serum RIPK3, C-reactive protein (CRP), lactate dehydrogenase (LDH), and D-dimer of each group were compared, and the correlations between each index and the severity of MPP in children were analyzed. The lung function indicators, including the percentage of peak expiratory flow (PEF% pred) and the percentage of forced expiratory volume in one second (FEV1% pred), were compared between groups, and the correlations between these indicators and the serum RIPK3 levels in children with MPP were analyzed. Measurement data with normal distribution was expressed as " xˉ±s", one-way ANOVA was used on comparison between groups, LSD-t test was used on pairwise comparison.Measurement data with skewed distribution were expressed as M( Q1, Q3). The Kruskal-Wallis H test was used for comparisons among multiple groups, and the Dunn test was used for pairwise comparisons. Counting data was expressed as case(%), χ2 test was used on comparison between groups. The relationships between serum RIPK3, CRP, LDH, D-dimer and the severity of MPP in children were analyzed by Spearman correlation analysis. The relationships between serum RIPK3 levels and PEF% pred, FEV1% pred were analyzed by Pearson correlation analysis. Results:The levels of serum RIPK3, CRP, LDH and D-dimer in children with mild MPP and severe MPP were all higher than those in the healthy control group [(329.67±59.69), (406.77±47.47) ng/L vs. (103.16±15.93) ng/L, (13.49±4.76), (37.76±18.94) mg/L vs. (1.59±0.81) mg/L, (253.12±50.96), (501.96±167.55) U/L vs. (186.06±45.49) U/L, (0.90±0.19), (3.01±1.46) mg/L vs. (0.29±0.12) mg/L], and the levels in the severe MPP group were all higher than those in the mild MPP group (all P < 0.05). The levels of serum RIPK3, CRP, LDH and D-dimer in children with MPP were positively correlated with the severity of the disease (r values were 0.59, 0.76, 0.83, 0.67, all P < 0.001). The levels of PEF% pred and FEV1% pred in children with mild MPP and severe MPP were all lower than those in the healthy control group [(72.65±7.67)%, (59.93±10.17)% vs. (98.54±4.81)%, (73.79±6.56)%, (60.56±10.54)% vs. (98.08±4.65)%], and the levels in the severe MPP group were all lower than those in the mild MPP group (all P < 0.05). The level of serum RIPK3 in children with MPP was negatively correlated with the lung function indicators PEF% pred and FEV1% pred (r values were -0.78, -0.77, all P < 0.001). Conclusions:The serum RIPK3 level in children with MPP is positively correlated with the severity of their condition and negatively correlated with the lung function indicators PEF% pred and FEV1% pred.

Objective: To investigate the impact of milk and egg supplementation on body composition and bone mineral density of rural primary school students in Yunnan Province, so as to provide a reference for developing targeted nutritional intervention strategies. Methods: In December 2023, a cluster sampling method was adopted to select students from grades one to three in four primary schools each from Jinggu and Shidian countys of Yunnan Province, as the intervention group (662 students). Additionally, two boarding primary schools were selected from each county based on the principle of matching scale and student numbers as the control group (455 students). Starting from April 2023, the intervention group received 200 mL milk and 50 g eggs during the break on school days for 8 months, while the control group maintained their usual diet behavior. Body composition was measured by using bioelectrical impedance analysis, and distal radial bone mineral density was assessed via dual energy X-ray absorptiometry in April and December 2023. The intervention effects were analyzed by using a difference in-differences approach. Results: The final measurements of body fat percentage, skeletal muscle mass and fat free mass of the intervention group and the control group of primary school students were significantly higher than the baseline values, and the net effect of milk and egg intervention on these body composition indicators was not statistically significant ( P >0.05, both before and after adjustment). In contrast, bone mineral density increased significantly by 0.02 g/cm 2 in the intervention group. The net intervention effect on bone mineral density was statistically significant ( β=0.02, 95%CI =0.00-0.04), and remained significant after model adjustment ( β=0.02, 95%CI =0.00-0.04) (both P < 0.05). Subgroup analysis showed statistically significant effects of the intervention among girls ( β=0.02, 95%CI =0.00-0.04), day students ( β=0.04, 95%CI =0.01-0.07), and students with normal nutritional status ( β=0.02, 95%CI =0.00-0.04) (all P <0.05). No significant effect of milk and egg supplementation was observed on body composition indicators (all P <0.05). Conclusions Milk and egg supplementation can improve bone mineral density among rural primary school students in Yunnan Province. It is recommended that rural school aged children should increase intake of milk and eggs to support growth and development.

Objective:To investigate the relationship between serum interacting protein kinase (RIPK)-3 levels and mycoplasma pneumoniae pneumonia (MPP) correlation between disease severity and lung function in children.Methods:A case-control study was conducted. A total of 147 children with MPP admitted to the Children's Hospital Affiliated to Xuzhou Medical University from April 2023 to March 2024 were selected, including 76 cases of mild MPP (mild MPP group) and 71 cases of severe MPP (severe MPP group). Additionally, 36 healthy children who underwent physical examinations during the same period were selected as the healthy control group. The levels of serum RIPK3, C-reactive protein (CRP), lactate dehydrogenase (LDH), and D-dimer of each group were compared, and the correlations between each index and the severity of MPP in children were analyzed. The lung function indicators, including the percentage of peak expiratory flow (PEF% pred) and the percentage of forced expiratory volume in one second (FEV1% pred), were compared between groups, and the correlations between these indicators and the serum RIPK3 levels in children with MPP were analyzed. Measurement data with normal distribution was expressed as " xˉ±s", one-way ANOVA was used on comparison between groups, LSD-t test was used on pairwise comparison.Measurement data with skewed distribution were expressed as M( Q1, Q3). The Kruskal-Wallis H test was used for comparisons among multiple groups, and the Dunn test was used for pairwise comparisons. Counting data was expressed as case(%), χ2 test was used on comparison between groups. The relationships between serum RIPK3, CRP, LDH, D-dimer and the severity of MPP in children were analyzed by Spearman correlation analysis. The relationships between serum RIPK3 levels and PEF% pred, FEV1% pred were analyzed by Pearson correlation analysis. Results:The levels of serum RIPK3, CRP, LDH and D-dimer in children with mild MPP and severe MPP were all higher than those in the healthy control group [(329.67±59.69), (406.77±47.47) ng/L vs. (103.16±15.93) ng/L, (13.49±4.76), (37.76±18.94) mg/L vs. (1.59±0.81) mg/L, (253.12±50.96), (501.96±167.55) U/L vs. (186.06±45.49) U/L, (0.90±0.19), (3.01±1.46) mg/L vs. (0.29±0.12) mg/L], and the levels in the severe MPP group were all higher than those in the mild MPP group (all P < 0.05). The levels of serum RIPK3, CRP, LDH and D-dimer in children with MPP were positively correlated with the severity of the disease (r values were 0.59, 0.76, 0.83, 0.67, all P < 0.001). The levels of PEF% pred and FEV1% pred in children with mild MPP and severe MPP were all lower than those in the healthy control group [(72.65±7.67)%, (59.93±10.17)% vs. (98.54±4.81)%, (73.79±6.56)%, (60.56±10.54)% vs. (98.08±4.65)%], and the levels in the severe MPP group were all lower than those in the mild MPP group (all P < 0.05). The level of serum RIPK3 in children with MPP was negatively correlated with the lung function indicators PEF% pred and FEV1% pred (r values were -0.78, -0.77, all P < 0.001). Conclusions:The serum RIPK3 level in children with MPP is positively correlated with the severity of their condition and negatively correlated with the lung function indicators PEF% pred and FEV1% pred.

Objective To explore the molecular mechanism of Gasdermin B(GSDMB)regulating the fate of intestinal epithelial cells.Methods The human GSDMB plasmid was overexpressed into two human intestinal epithelial cell lines(NCM460 and HT-29 cells)and human colon-derived organoids.Western blotting was used to confirm the efficiency of electroporation.Cell counting kit(CCK8),cell apoptosis,and cell cycle by flow cytometry were performed to analyze the effect of GSDMB overexpression on cell function.Transcriptome sequencing was used to analyze the downstream effector molecules of GSDMB.T test was used to compare the data between the two groups.Results The overexpression of GSDMB protein in the two intestinal epithelial cell lines was successfully reconstructed.The absorbance value(A)of human intestinal epithelial cells overexpressing GSDMB protein[NCM460 cells:(1.17±0.01),HT-29 cells:(0.96±0.06)]was significantly lower than that of blank control cells[NCM460 cells:(1.67±0.12),HT-29 cells:(1.24±0.07)](t=7.24 and 5.46,P＜0.05).The number of apoptotic cells in the GSDMB overexpression group[NCM460 cells:(12.03±1.55),HT-29 cells:(29.30±4.48)]was significantly higher than that in the blank group[NCM460 cells:(4.96±1.74),HT-29 cells:(6.95±3.42)](t=5.26 and 6.97,P＜0.05).Cell cycle analysis showed that the ratio of cells at G0/G1 phase in the GSDMB overexpression group[NCM460 cells:(47.98±5.28)％,HT-29 cells:(38.04±3.45)％]was significantly lower than that in the control group[NCM460 cells:(59.54±3.90)％,HT-29 cells:(63.81±1.76)％](t=3.05 and 11.53,P＜0.05).Transcriptome sequencing results showed that the dual specificity phosphatase 4 and 6(DUSP4 and DUSP6)genes were significantly upregulated after GSDMB protein expression.Fluorescence quantitative PCR results confirmed that the relative expression levels of DUSP4(2.45±0.15)and DUSP6(4.34±0.22)in intestinal epithelial cells transfected with GSDMB were significantly higher than those in the control group(1.06±0.05 and 1.01±0.02)(t=15.08 and 26.52,P＜0.05).After GSDMB-expressing NCM460 cells were treated with the DUSP inhibitor BCI,the BCI treatment group had a significantly increased expression level of p-ERK compared to the control group[(1.14±0.17)vs.(0.58±0.12)](t=5.42,P=0.002);the A value(1.84±0.07)and G0/G1 phase ratio(59.83±2.17)％in the BCI treatment group were significantly higher than those in the non-treatment group[(1.52±0.10)and(52.10±2.23)％],and the number of apoptosis in the BCI treated group(7.60±0.56)was significantly lower than that in the untreated group(12.57±1.00)(t=4.71,4.31,7.52,P＜0.05).TUNEL staining in human colon organoids showed a significant increase in apoptotic cells,and the relative expression level of DUSP6 protein(0.85±0.09)was significantly higher than that of the control group(0.21±0.04),accompanied by a decrease in p-ERK levels[(0.83±0.18)vs.(0.19±0.06)],with statistical significance(t=11.95,P＜0.001;t=6.56,P＜0.001).Conclusion GSDMB may inhibit cell proliferation,induce cell cycle arrest,and promote apoptosis by upregulating dual specificity phosphatase DUSP6-mediated ERK phosphorylation,thus affecting the fate of intestinal epithelial cells.

Regional anatomy teaching not only requires students to deal with the basic knowledge of human body including the level, location and adjacent relationship, but also to understand the clinical application of anatomical structure. Based on the four aspects of field anatomy, simulated surgery, clinical application lectures and CBL teaching, this study formulated a suitable assessment method to reconstruct the teaching system of regional anatomy relying on the improvement of the laboratory environment and the teacher team, aiming at cultivating students' clinical practice ability as the core and building a new regional anatomy course to meet the teaching needs of the new era.

OBJECTIVE: To determine the carrier rate for 21 inherited metabolic diseases among a Chinese population of childbearing age. METHODS: A total of 897 unrelated healthy individuals (including 143 couples) were recruited, and DNA was extracted from their peripheral blood samples. Whole exome sequencing (WES) was carried out to screen potential variants among 54 genes associated with 21 inherited metabolic diseases. Pathogenic and likely pathogenic variants and unreported loss-of-function variants were analyzed. RESULTS: One hundred fourty types of pathogenic/likely pathogenic variants (with an overall number of 183) and unreported loss-of-function variants were detected, which yield a frequency of 0.20 per capita. A husband and wife were both found to carry pathogenic variants of the SLC25A13 gene and have given birth to a healthy baby with the aid of preimplantation genetic diagnosis. The detected variants have involved 40 genes, with the most common ones including ATP7B, SLC25A13, PAH, CBS and MMACHC. Based on the Hardy-Weinberg equilibrium, the incidence of the 21 inherited metabolic diseases in the population was approximately 1/1100, with the five diseases with higher incidence including citrullinemia, methylmalonic acidemia, Wilson disease, glycogen storage disease, and phenylketonuria. CONCLUSION This study has preliminarily determined the carrier rate and incidence of 21 inherited metabolic diseases among a Chinese population of childbearing age, which has provided valuable information for the design of neonatal screening program for inherited metabolic diseases. Pre-conception carrier screening can provide an important measure for the prevention of transmission of Mendelian disorders in the population.
Asians/genetics* ; China ; Exome ; Female ; Humans ; Infant, Newborn ; Metabolic Diseases/genetics* ; Mitochondrial Membrane Transport Proteins/genetics* ; Oxidoreductases/genetics* ; Whole Exome Sequencing

Objective:To explore the strategy of preimplantation genetic testing for monogenic disorders (PGT-M) with variants of uncertain significance (VUS).Methods:Monogenic disorder couples who carried VUS and sought fertility counseling between 2018 and 2020 in Reproductive and Genetic Hospital of CITIC-Xiangya were recruited in this study. The pathogenicity of VUS was reanalyzed according to the Standards and Guidelines for the Interpretation of Sequence Variants released by the American College of Medical Genetics and Genomics (ACMG) and the Bayesian Classification. Those VUSs were reclassified as "pathogenic/likely pathogenic variants (P/LP)", "likely pathogenic VUS", "variants of uncertain significance", or "likely benign VUS". PGT-M was applied to families with VUS upgraded as "P/LP" or "likely pathogenic VUS" under the principle of couples fully voluntary and understanding the risks. We also followed up the developmental status of fetuses and the health condition of the born children.Results:1) A total of 25 variants were detected in 16 families with monogenic disorders, including 1 P, 3 LP, and 21 VUS. After reanalysis, 11 VUS and 7 VUS were upgraded as LP (52.4%) and "likely pathogenic VUS" (33.3%), respectively. Two VUS were still reclassified as "variants of uncertain significance"(9.5%), and 1 VUS was reclassified as "likely benign VUS" (4.8%). 2) PGT-M was implemented for 14 families with monogenic disorders, including 9 families with VUS upgraded as LP, 2 families with one LP/P and one "likely pathogenic VUS", and 3 families with only "likely pathogenic VUS". 3) Twelve healthy babies were born after PGT-M. Following up was done according to the onset age of diseases: 8 offsprings did not show the symptoms as probands, and 4 offsprings had not yet reached the age of onset and need continuous follow-up.Conclusion:It is necessary to actively search for new evidence and reanalyze the pathogenicity of VUS according to ACMG guidelines before PGT-M. Under fully informed consent of the patients, PGT-M can be carried out for VUS reclassified as "P/LP" and "likely pathogenic VUS", to reduce the risk of recurrence.

Objective:To explore the strategy of preimplantation genetic testing for monogenic disorders (PGT-M) with variants of uncertain significance (VUS).Methods:Monogenic disorder couples who carried VUS and sought fertility counseling between 2018 and 2020 in Reproductive and Genetic Hospital of CITIC-Xiangya were recruited in this study. The pathogenicity of VUS was reanalyzed according to the Standards and Guidelines for the Interpretation of Sequence Variants released by the American College of Medical Genetics and Genomics (ACMG) and the Bayesian Classification. Those VUSs were reclassified as "pathogenic/likely pathogenic variants (P/LP)", "likely pathogenic VUS", "variants of uncertain significance", or "likely benign VUS". PGT-M was applied to families with VUS upgraded as "P/LP" or "likely pathogenic VUS" under the principle of couples fully voluntary and understanding the risks. We also followed up the developmental status of fetuses and the health condition of the born children.Results:1) A total of 25 variants were detected in 16 families with monogenic disorders, including 1 P, 3 LP, and 21 VUS. After reanalysis, 11 VUS and 7 VUS were upgraded as LP (52.4%) and "likely pathogenic VUS" (33.3%), respectively. Two VUS were still reclassified as "variants of uncertain significance"(9.5%), and 1 VUS was reclassified as "likely benign VUS" (4.8%). 2) PGT-M was implemented for 14 families with monogenic disorders, including 9 families with VUS upgraded as LP, 2 families with one LP/P and one "likely pathogenic VUS", and 3 families with only "likely pathogenic VUS". 3) Twelve healthy babies were born after PGT-M. Following up was done according to the onset age of diseases: 8 offsprings did not show the symptoms as probands, and 4 offsprings had not yet reached the age of onset and need continuous follow-up.Conclusion:It is necessary to actively search for new evidence and reanalyze the pathogenicity of VUS according to ACMG guidelines before PGT-M. Under fully informed consent of the patients, PGT-M can be carried out for VUS reclassified as "P/LP" and "likely pathogenic VUS", to reduce the risk of recurrence.

Objective To analyze the drug consumption of the joint logistics support force during field training in Yunnan, summarize the drug use characteristics of medical staff, and analyze the drug use rules, so as to provide the basis for modular drug support. Methods The basic information and drug use of the personnel in an institution of the joint logistics support force during the resident training period from April to June 2020 were collected and sorted out. The drug use indicators were calculated by using the defined daily dose (DDD) as the unit, including: daily drug dose (DDDs), daily drug dose cost (DDDc), drug use intensity (DUI), drug utilization rate (DUR) and drug ranking ratio (R) were used to describe drug use characteristics. Results During the resident training period, the amount of musculoskeletal system drugs was large, and the number of users was large. The dosage of cardiovascular system drugs, digestive system drugs and hormone drugs was large, but the number of users was small. Sensory organ drugs, dermatological drugs, heat-clearing agents (Chinese patent medicine) and respiratory system drugs were small in dosages, but the number of users was large. Ranked by DDDs, the top 3 drugs with the highest dosage were Yunnan Baiyao plaster, Yunnan Baiyao Band-Aid and Loratadine Tablets. Ranked by DDDc, the largest daily cost was Budesonide and Formoterol Fumarate Powder for Inhalation. As age increases, the intensity of drug use increased; The intensity of drug use in women was greater than that in men. The intensity of drug use was different for different categories personnel. Conclusion Musculoskeletal system drugs are important drugs for field training. The task personnel should formulate a list of medicines for basic diseases based on their own medication for basic diseases and the duration of the task, and submit it to the medicinal material guarantee department to ensure the carrying drugs. The factors affecting modular drug support in field training include: task type, region, solar term, duration and personnel composition, etc.