Objective:To investigate the effects of fluoride exposure on kidney injury in rats and the sirtuin 3 (SIRT3)-fork head protein O3a (FOXO3a)-tensin homolog induced putative kinase 1 (PINK1)/E3 ubiquitin ligase (PARKIN) pathway.Methods:Twenty-four 4-week-old SD rats (clean grade, body mass 100 - 150 g) were selected and divided into three groups according to the randomized numeric table: control group, low fluoride group, and high fluoride group, with eight rats in each group (half male and half female). The control group was given free access to tap water (fluoride ion concentration < 0.5 mg/L), while the low fluoride and high fluoride groups were given free access to tap water and sodium fluoride solutions with fluoride ion concentrations of 5.0 and 50.0 mg/L, respectively, for a period of 180 days. The formation of dental fluorosis in rats was observed and recorded, and the femur, urine and blood samples of rats were collected to measure bone fluoride, urinary fluoride, and blood fluoride levels, and to detect kidney function related indicators (serum uric acid, creatinine, and urea nitrogen contents). Morphological changes of renal tissues stained with hematoxylin-eosin (HE) were observed under a light microscope. Real-time fluorescence quantitative PCR (qRT-PCR) and Western blotting were used to detect the mRNA and protein expression levels of renal SIRT3, FOXO3a, PINK1, PARKIN, microtubule associated protein 1 light chain 3 (LC3), autophagy receptor protein (P62), respectively.Results:Seven and one rats in the low and high fluoride groups were found to haveⅠdegree dental fluorosis, while zero and seven rats were found to haveⅡdegree dental fluorosis. Compared with the control group, rats in the low and high fluoride groups had higher levels of bone fluoride (μg/g: 1.18 ± 0.06, 2.16 ± 0.07 vs 0.52 ± 0.05), urinary fluoride (mg/L: 4.43 ± 0.11, 7.46 ± 0.09 vs 2.58 ± 0.14), blood fluoride (μg/ml: 0.77 ± 0.06, 1.68 ± 0.10 vs 0.52 ± 0.08), serum uric acid (μg/ml: 61.01 ± 4.17, 103.92 ± 5.43 vs 28.68 ± 2.91), creatinine (μg/ml: 74.82 ± 9.61, 132.05 ± 5.35 vs 22.38 ± 4.11), and urea nitrogen (μg/ml: 13.36 ± 1.27, 14.55 ± 0.34 vs 0.29 ± 0.07, P < 0.05). Under the light microscope, the kidneys of the control group showed tight and orderly arrangement of renal tubules and glomerular cells, with complete and clear cell contours. The low fluoride group was similar to the control group and no significant abnormalities were observed. The high fluoride group showed abnormal glomerular structure and atrophy, with some areas of renal tubules showing epithelial cell edema and unclear intercellular boundaries. The results of qRT-PCR assay showed that compared with the control group, the low and high fluoride groups had lower mRNA expression levels of SIRT3 (0.82 ± 0.03, 0.58 ± 0.02 vs 1.00 ± 0.08), P62 (0.75 ± 0.07, 0.28 ± 0.09 vs 1.00 ± 0.07, P < 0.05), and higher mRNA expression levels of FOXO3a (1.35 ± 0.04, 3.01 ± 0.23 vs 1.00 ± 0.08), PINK1 (1.58 ± 0.09, 3.28 ± 0.09 vs 1.00 ± 0.07), PARKIN (1.51 ± 0.04, 1.67 ± 0.10 vs 1.00 ± 0.05), LC3 (1.74 ± 0.07, 2.38 ± 0.18 vs 1.00 ± 0.08, P < 0.05). The results of Western blotting showed that compared with the control group, the low and high fluoride groups had lower protein expression levels of SIRT3 (0.91 ± 0.01, 0.55 ± 0.03 vs 1.00 ± 0.01), P62 (0.94 ± 0.27, 0.66 ± 0.38 vs 1.00 ± 0.19, P < 0.05), and higher protein expression levels of FOXO3a (1.14 ± 0.03, 1.22 ± 0.05 vs 1.00 ± 0.02), PINK1 (1.46 ± 0.03, 1.56 ± 0.03 vs 1.00 ± 0.05), PARKIN (1.98 ± 0.02, 2.33 ± 0.11 vs 1.00 ± 0.06), LC3 (4.10 ± 0.58, 4.93 ± 0.33 vs 1.00 ± 0.13, P < 0.05). Conclusion:Exposure to fluoride can cause renal tissue injury in rats, with downregulation of SIRT3 and P62 expression levels, and upregulation of FOXO3a, PINK1, PARKIN, and LC3 expression levels.

OBJECTIVE: To explore the clinical and genetic characteristics of eight children with Primary hypertrophic cardiomyopathy (HCM). METHODS: Eight children with HCM admitted to the Department of Cardiology of Henan Children's Hospital from January 2018 to December 2021 were selected as the study subjects. Clinical data of the children were collected. Whole exome sequencing was carried out on two children, and trio whole exome sequencing was carried out on the remainder 6 children. Sanger sequencing was used to verify the candidate variants in the children and their parents, and the pathogenicity of the variants was evaluated based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). RESULTS: The patients had included 5 males and 3 females, with their ages ranging from 5 to 13 years old. The average age of diagnosis was (7.87 ± 4.8) years old, and the cardiac phenotype showed non-obstructive HCM in all of the patients. WES has identified variants of the MYH7 gene in 4 children, including c.2155C>T (p.Arg719Trp), c.1208G>A (p.Arg403Gln), c.1358G>A (p.Arg453His), and c.1498G>A (p.Glu500Lys). Based on the guidelines from the ACMG, the first 3 variants were classified as pathogenic, while c.1498G>A (p.Glu500Lys) was classified as likely pathogenic (PM1+PM2_Supporting+PM6+PP3), which was also unreported previously. The remaining four children had all harbored maternal variants, including MYL2: c.173G>A (p.Arg58Gln; classified as pathogenic), TPM1: c.574G>A (p.Glu192Lys) and ACTC1: c.301G>A (p.Glu101Lys)(both were classified as likely pathogenic), and MYBPC3: c.146T>G (p.Ile49Ser; classified as variant of uncertain significance). Seven children were treated with 0.5 ~ 3 mg/(kg·d) propranolol, and their symptoms had improved significantly. They were followed up until September 30, 2022 without further cardiac event. CONCLUSION Genetic testing can clarify the molecular basis for unexplained cardiomyopathy and provide a basis for clinical diagnosis and genetic counseling. Discovery of the c.1498G>A (p.Glu500Lys) variant has also expanded the spectrum of MYH7 gene mutations underlying HCM.
Female ; Male ; Humans ; Child ; Child, Preschool ; Adolescent ; Cytoskeletal Proteins ; Family ; Genetic Counseling ; Genetic Testing ; Cardiomyopathy, Hypertrophic/genetics*

Objective:To investigate the association between relative mitochondrial DNA copy number (mtDNA-CN) and coal-burning-borne endemic fluorosis (abbreviated as coal-burning-borne fluorosis).Methods:From June 2018 to March 2019, using cross-sectional study, 482 patients with coal-burning-borne fluorosis were selected as the case group in Bijie City, a typical coal-burning-borne fluorosis area of Guizhou Province; meanwhile, 212 healthy individuals from Changshun County, a non-coal-burning-borne fluorosis area in Guizhou Province, were selected as the control group. Questionnaire survey and physical examination were used to collect general condition such as basic information and living habits of the two groups, peripheral venous blood samples were collected, and real-time fluorescence quantitative PCR was used to detect the relative mtDNA-CN in peripheral blood. The correlation between relative mtDNA-CN and coal-burning-borne fluorosis was analyzed by binary and unordered multi-class logistic regression.Results:There were significant differences in the body mass index (BMI), and the distribution of gender rario, marital status and education level between the control group and the case group ( t = 7.91, χ 2 = 5.11, 13.33, 34.32, P < 0.05). The relative mtDNA-CN in the control group was higher than that in the case group [median (quartile): 202 (138, 292) vs 131 (96, 217), Z = - 7.80, P < 0.001]. The results of binary logistic regression analysis [odds ratio (95% confidence interval)] showed that educational level [primary school: 0.572 (0.377 - 0.868), junior high school and above: 0.292 (0.174 - 0.493)], relative mtDNA-CN [131 - < 217: 0.265 (0.144 - 0.488), ≥217: 0.183 (0.100 - 0.335)] and BMI [1.222 (1.142 - 1.307)] were the influencing factors for the risk of coal-burning-borne fluorosis( P < 0.05). In subgroups with different BMI and educational levels, the relative mtDNA-CN was significantly negatively correlated with the risk of coal-burning-borne fluorosis( Ptrend < 0.05), and there was no interaction between mtDNA-CN and BMI and educational levels ( Pinteraction > 0.05). The results of unordered multi-class logistic regression analysis showed that the relative mtDNA-CN were significantly negatively correlated with the risk of dental fluorosis and skeletal fluorosis ( Ptrend < 0.05). Conclusion:The higher the relative mtDNA-CN, the lower the risk of coal-burning-borne fluorosis, suggesting that mtDNA-CN may be a potential biomarker of coal-burning fluorosis.

Objective:To explore the relationship between children's intelligence and urinary fluoride in Suojia Township of Miao, Yi and Hui Nationalities (referred to as Suojia Township), a coal-burning-borne endemic fluorosis area in Guizhou Province.Methods:In April 2019, 173 children aged 10 to 13 years old were selected from three schools in Suojia Township. According to whether they had dental fluorosis, the children were divided into case group ( n = 104) and control group ( n = 69). Middle segment urine samples of the children were collected and urinary fluoride level was determined by the method of ion-selective electrode. Combined Raven's Test-the Rural in China (CRT-RC2) was used for children's intelligence quotient (IQ) test. Linear regression analysis was used to observe the association between urinary fluoride and IQ, and the results were expressed by regression coefficient ( β) and 95% confidence interval ( CI). Results:Urinary fluoride level of case group was higher than that of control group [(2.14 ± 1.78) vs (1.53 ± 0.98) mg/L], and IQ was lower than that of control group [(92.33 ± 11.68) vs (100.38 ± 11.87) points], and the differences were statistically significant ( t = 2.58, 4.41, P < 0.05). The linear regression equation of urinary fluoride ( X) and IQ ( Y) of case group was Y = 96.99 - 2.86 X. For every 1 mg/L increase in urinary fluoride level, IQ decreased by 2.86 points ( β = - 2.86, 95% CI: - 5.48 - - 0.24). Conclusion:Long-term exposure to fluoride pollution from coal burning may damage children's intelligence, and children's IQ decreases with increase of fluoride level in urine.

Objective:To investigate the relationship between gene polymorphism of osteoprotegerin (OPG) and coal-burning endemic fluorosis in Guizhou Province.Methods:In 2018 and 2019, a case-control study was conducted in Bijie City, a typical coal-burning endemic fluorosis area in Guizhou Province, and 260 cases of coal-burning endemic fluorosis patients were selected as case group. According to the "Diagnostic Criteria for Endemic Skeletal Fluorosis" (WS 192-2008), the case group was divided into severe group (130 cases) and mild group (130 cases). At the same time, 130 cases without dental fluorosis and skeletal fluorosis symptoms were chosen as control group in Changshun County, a non-coal-burning endemic fluorosis area in Guizhou Province. Whole blood genomic DNA was extracted, and real-time fluorescence quantitative PCR with TaqMan-MGB probe was utilized to type the OPG gene rs2460985, rs2073618, rs6469804 and rs6993813 four single nucleotide polymorphism (SNP) loci of all samples, and genetic model analysis was performed to compare the frequency distribution of its alleles, genotypes and constructed haplotypes in control, mild and severe groups. Results:By Hardy-Weinberg equilibrium test, the genotype frequencies of the four SNP loci all reached genetic balance in control, mild and severe groups ( P > 0.05). There was a statistically significant difference in the genotype frequency of OPG gene rs6469804 locus among the three groups (χ 2 = 10.615, P < 0.05), and the difference in the genotype frequency of this locus between the control group and the severe group was statistically significant (χ 2 = 6.784, P < 0.05). The results of genetic model analysis showed that in comparison between the control group and the severe group, the optimal genetic model of rs6469804 locus was the overdominant genetic model, the frequency distribution of genotypes AA + GG and AG in the control group and the severe group was statistically significant [odds ratio ( OR) = 1.94, 95% confidence interval ( CI): 1.16 - 3.23, P < 0.05], genotype AG was a risk factor for coal-burning endemic fluorosis. In comparison between the control group and the mild group, the optimal genetic model of rs2073618 locus was the recessive genetic model, the frequency distribution of genotypes GG + GC and CC in the control group and the mild group was statistically significant ( OR = 3.17, 95% CI: 1.08 - 9.30, P < 0.05), genotype CC was a risk factor for coal-burning endemic fluorosis. In comparison between the control group and the mild group, haplotypes C-C-G-T and T-G-A-C were risk factors for coal-burning endemic fluorosis (adjusted OR = 2.41, 1.98, 95% CI: 1.29 - 4.50, 1.22 - 3.23, P < 0.05); in comparison between the control group and the severe group, haplotype T-G-A-C was a risk factor for coal-burning endemic fluorosis (adjusted OR = 1.87, 95% CI: 1.14 - 3.07, P < 0.05). Conclusion:OPG gene rs6469804 locus genotype AG and rs2073618 locus genotype CC may be risk factors for coal-burning endemic fluorosis.

Objective:To understand the current situation of children's fluorosis in the coal-burning-borne endemic fluorosis areas (abbreviated as coal-burning fluorosis) in Suojia Miao, Yi and Hui Township (Suojia Township for short) in Liuzhi Tequ, Guizhou Province, and to provide scientific basis for formulating prevention and control strategies and measures.Methods:In 2019, the cluster sampling method was adopted to select children aged 8-12 years old from 6 primary schools in Suojia Township, Liuzhi Tequ, Guizhou Province to conduct a questionnaire survey to collect basic information, and perform dental fluorosis examination and indexing in accordance with the "Diagnosis of Dental Fluorosis" standards. Immediate urine samples were collected from children in April and October, and urinary fluoride content was determined by ion selective electrode method.Results:A total of 1 381 children aged 8-12 years old were investigated, aged (9.84 ± 1.38) years old, including 679 boys and 702 girls. A total of 625 children with dental fluorosis were detected, and the detection rate was 45.26%; the dental fluorosis index was 1.00, and the prevalence intensity was moderate; the main score of dental fluorosis was extremely mild, accounting for 37.00% (511/1 381). The detection rates of dental fluorosis in children aged 8 to 12 years old were 35.10% (106/302), 35.83% (115/321), 47.96% (129/269), 55.23% (153/277), and 57.55% (122/212), respectively; the difference between different ages was statistically significant (χ 2 = 48.949, P < 0.01), and the detection rate of dental fluorosis in children increased with age(χ 2trend = 45.254, P < 0.01).The detection rates of dental fluorosis in boys and girls were 43.59% (296/679) and 46.87% (329/702), respectively, and there was no significant difference between different genders (χ 2 = 1.492, P > 0.05). In April and October, 123 and 107 urine samples of children aged 8-12 years old were tested. The geometric mean of urinary fluoride was 1.55 and 0.47 mg/L, respectively. The urinary fluoride level in April was higher than the normal range (< 1.40 mg/L). Conclusions:Suojia Township in Liuzhi Tequ of Guizhou Province is still a fluorosis area, and there is a big difference in urinary fluorine level in different months, which indicates that the residents in this area may have intermittent high fluorine intake, and prevention and control of endemic fluorosis should be further strengthened.

Objective: To assess the diagnostic performance of contrast-enhanced spectral mammography (CESM) in suspected breast lesions. Methods: A total of 97 patients with suspected breast cancer identified by clinical examination or screening underwent two-views CESM examination on the basis of digital breast tomosynthesis (DBT) combined with full-field digital mammography (FFDM), and they were finally confirmed by biopsy or pathology. Three senior radiologists analyzed images, including lesion visibility, lesion characteristics, enhancement type, degree of enhancement, BIRDS classification, etc. Finally, based on the pathology, we compared the CESM+DBT+FFDM and DBT+FFDM two models according to sensitivity, specificity and ROC for diagnostic performance. Results: There were a total of 120 lesions. Eighty-nine lesions were malignant, 31 benign; CESM was not enhanced in 2 cases, mild enhancement was performed in 22 cases, moderately intensive in 15 cases, highly intensive in 81 cases, and 2 cases were not enhanced; mass-enhanced in 96 cases, including ring-enhanced in 12 cases, 22 cases of non-mass type. The sensitivities of the combination of CESM and not combination of CESM were 91.0% and 80.9%, respectively, and the specificities were 93.5% and 87.1%, respectively. The area under the ROC curve of combination of CESM was higher than the without combination of CESM (0.923 and 0.900, P<0.05), The difference was statistically significant. Conclusion For suspicious lesions, CESM examination can improve the diagnosis accuracy of breast cancer.

OBJECTIVETo study the distribution of haplotypes of Y-chromosomal short tandem repeats (Y-STR) loci among three ethnic minorities from Guizhou, China.

METHODSTwenty four Y-STR loci of 174 unrelated males were amplified with a Microreader(TM)24Y Direct ID System kit. Capillary electrophoresis was carried out on an ABI 3100 Genetic Analyzer, and the data was analyzed with GeneMapper software.

RESULTSSeventy six haplotypes were identified for the 24 Y-STR loci among the three ethnic minorities, including 13 from the Qiangs, 35 from the Manchurians, and 28 from the Shes, with the corresponding Haplotype Diversity (HD) being 0.7327, 0.9578, and 0.9344. Genetic distance between the Shes and Qiangs was relatively close, whilst that for Manchurians was relatively far.

CONCLUSIONAnalysis of the genetic characteristics and relationship of the three ethnic minorities from Guizhou can provide a reference for the study of their origin, evolution and patrilineal fusion.

Objective To evaluate the diagnostic performance of digital breast tomosynthesis (DBT) breast X-ray photography image texture characteristics based deep learning classification model on differentiating malignant masses. Methods Retrospectively collected 132 cases with simplex breast lesions (89 benign lesions and 43 malignant lesions) which were confirmed by pathology and DBT during January 2016 to December 2016 in Nanfang Hospital. DBT was performed before biopsy and surgery. Image of cranio-caudal view (CC) and medio-lateral oblique (MLO) were captured. The lesion area was segmented to acquire ROI by ITK-SNAP software. Then the processed images were input into MATLAB R2015b to establish a feature model for extracting texture features. The characteristics with high correlation was analyzed from Fisher score and one sample t test. We built up support vector machine (SVM) classification model based on extracted texture and added neural network model (CNN) for deep learning classification model. We randomly assigned collected cases into training group and validation group. The diagnosis of benign and malignant lesions were served as the reference. The efficiency was evaluated by ROC classification model. Result We extracted 82 texture characteristics from 132 images of leisure (132 images of CC and 132 images of MLO) by establishing deep learning classification model of breast lesions. We randomly chose and combined characteristics from 15 texture characteristics with statistical significance, then differentiated benign and malignant by SVM classification model. After 50 iterations on each combination of characteristics, the average diagnostic efficacy was compared to obtained the one with higher efficacy. Nine of CC and 8 of MLO was selected. The result showed that the sensitivity, specificity, accuracy and area under curve (AUC) of the model to differentiate simplex breast lesions for CC were 0.68, 0.77, 0.74 and 0.74, for MLO were 0.71, 0.71, 0.71 and 0.76. Conclusions MLO has better diagnostic performance for the diagnosis than CC. The deep learning classification model on breast lesions which was built upon DBT image texture characteristics on MLO could differentiate malignant masses effectively.

Objective To investigate the effects of comprehensive pelvic floor rehabilitation program on pelvic floor muscle strength, female sexual function index (FSFI) score and sexual satisfaction scale for women (SSS-W) score in patients with perimenopausal female sexual dysfunction. Methods The clinical data of 120 patients with perimenopausal female sexual dysfunction were retrospectively analyzed, and all the patients were treated with estrogen and progesterone replacement therapy. Among the patients, 80 patients were treated with Kegel training program (control group), and 80 patients were treated with Kegel training program combined with comprehensive pelvic floor rehabilitation program (treatment group). The pelvic floor muscle strength, FSFI score, SSS-W score, incidence of algopareunia and sexual life frequency were compared between 2 groups. Results The pelvic floor muscle strength after treatment in treatment group was significantly better than that in control group. The Ⅲ,ⅣandⅤgrade of pelvic floor muscle strength in treatment group was in 5, 42 and 33 cases, and that in control group was in 21, 47 and 12 cases, and there was statistical difference (P<0.05). The FSFI scores and SSS-W scores after treatment in treatment group were significantly higher than those in control group:(32.24 ± 3.70) scores vs. (28.51 ± 2.95) scores and (139.80 ± 16.17) scores vs. (122.33 ± 13.75) scores, and there were statistical differences (P<0.05). The incidence of algopareunia after treatment in treatment group was significantly lower than that in control group:10.00%(8/80) vs. 55.00%(44/80), and there was statistical difference (P<0.05). The sexual life frequency after treatment in treatment group was significantly better than that in control group, the 0, 1, 2 and≥3 times/month of sexual life frequency in treatment group was in 0, 8, 20 and 52 cases, and that in control group was in 5, 34, 24 and 17 cases, and there was statistical difference (P<0.05). Conclusions Comprehensive pelvic floor rehabilitation program in patients with perimenopausal female sexual dysfunction can efficiently promote the recovery process of pelvic floor muscle strength, improve FSFI score and SSS-W score, and increase the degree of sexual satisfaction.