Objective To evaluate the clinical application of CT-guided coaxial needle biopsy in pancreatic puncture-cutting biopsy via adipose tissue space.Methods The clinical data of 30 patients,who received CT-guided pancreatic coaxial needle biopsy via adipose tissue space at the Pingdu Hospital Area of Affiliated Hospital of Qingdao University of China from June 2021 to April 2024,were retrospectively analyzed.The patients included 20 males and 10 females with a mean age of(65.1±8.7)years.The lesions were located in the pancreatic head(n=7),pancreatic neck(n=4),pancreatic body(n=7),pancreatic tail(n=10)and uncinate process(n=2).Under CT guidance,a 17 G co-axial guiding needle was inserted to establish a puncture pass through adipose tissue space,then an 18 G biopsy needle was used to make tissue sampling.A total of 32 procedures of pancreatic puncture-cutting biopsy were performed,two of these patients received two times of biopsy.After sampling,a self-made blocking agent(a mixed liquid of gelatin sponge particles and snake venom hemagglutinin),was used to occlude the puncture channel so as to prevent complications such as bleeding,pancreatic leakage,needle tract implantation metastasis,etc.Results A total of 32 procedures of pancreatic puncture-cutting biopsy with coaxial guiding needle via adipose tissue space were performed,including two patients who received two times of biopsy.The success rate of puncturing was 100％(32/32),the success rate of sampling was 93.8％(30/32),and the accuracy of sampling was 93.8％(30/32).No serious puncture-related complications occurred in all patients.Conclusion CT-guided pancreatic coaxial needle biopsy via adipose tissue space is clinically safe,accurate in sampling and minimally-invasive.This interventional diagnostic technique is worth applying in clinical practice.

Objective To investigate the incidence and related factors of vitamin D deficiency(VDD) in hospitalized patients with type 2 diabetes mellitus (T2DM). Methods 2139 patients with T2DM who were hospitalized in The Endocrinology Department of Kunshan Hospital of Traditional Chinese Medicine from January 2018 to June 2023 were enrolled in the study. All the subjects were divided into VDD group (n=1301) and T2DM group (n=838),according to the international standard of vitamin D level. General data and biochemical indicators compared between the two groups. According to the interquartile interval of MHR value,it was divided into Q1(MHR≤0.272,n=534),Q2(0.2720.492,n=534)groups.Logistic regression was used to analyze the related factors of VDD in patients with T2DM. Results FPG,HbA1c,TC,TG,LDL-C,MHR,FC-P,HOMA-IR,CTX and proportion of female in VDD group were higher than those in T2DM group (P<0.05),the age and HDL-C in VDD group were lower than those in T2DM group(P<0.05). Multivariate regression analysis showed that sex,age,FPG,HbA1c,TC,TG,LDL-C,FC-P,CTX,MHR were independently assdciated with VDD. Conclusions MHR is a related factor for VDD in patients with T2DM.

Objective:To investigate the clinical phenotype and genetic characteristics of developmental epileptic encephalopathy 18 (DEE18) caused by SZT2 gene variants. Methods:Clinical data of 2 children with SZT2 related DEE18 who visited the Department of Pediatric Neurology, Linyi People′s Hospital in March 2020 and July 2023 were collected. The whole exome sequencing (WES) and Sanger sequencing were applied to verify the child and their parents. SWISS-MODEL software was used to perform protein 3D modeling for the selected SZT2 gene variants. Results:Both of the 2 cases showed severe global developmental delay, epileptic seizures, autism, megacephaly, facial deformity, hypotonia, corpus callosum malformation, persistent cavum septum pellucidum, and slow background activity and focal discharge in video electroencephalography. Case 1 was easy to startle and thin in stature; case 2 had immune deficiency and clustered seizures. WES results showed that case 1 carried a compound heterozygous variant of c.5811G>A (p.W1937X) (paternal) and c.9269delG (p.S3090Ifs *94) (maternal), while case 2 carried a compound heterozygous variant of c.6302A>C(p.H2101P) (paternal) and c.7584dupA (p.E2529Rfs *20) (maternal), the parents of both patients with normal clinical phenotypes. The 4 mutations mentioned above were novel variations that had not yet been reported domestically or internationally. According to the American College of Medical Genetics and Genomics variant classification criteria and guidelines, the p.S3090Ifs *94 variant was interpreted as pathogenic; p.W1937X variant was interpreted as pathogenic; p.E2529Rfs *20 variant was interpreted as likely pathogenic; p.H2101P variant was interpreted as uncertain significance. 3D modeling showed that the variant of p.H2101P resulted in a significant change in the hydrogen bond around the 2 101st amino acid encoded, leading to a decrease in protein stability. The other 3 variants led to early truncation of peptide chain and obvious changes in protein structure. Conclusions:DEE18 caused by SZT2 gene mutation is mainly an autosome recessive genetic disease, and its clinical manifestations include global developmental delay, epileptic seizures, autism, craniofacial malformation, hypotonia, epileptic discharge, corpus callosum malformation, persistent cavum septum pellucidum, shock, small and thin stature, and immune deficiency. Four novel variants related to the SZT2 gene may be the genetic etiology of DEE18 patients in this study.

Objective:To investigate the clinical phenotype and genetic characteristics of infantile epileptic spasm syndrome caused by BRWD3 gene mutation. Methods:Clinical data of a child with BRWD3 related infantile epileptic spasm syndrome who was admitted to Department of Pediatric Neurology of Linyi People′s Hospital on August 2, 2019 were collected and followed up, whole exome sequencing technology and Sanger sequencing were applied to verify the child and his parents, and the pathogenicity of mutation site was analyzed. The studies till June 2023 were searched with keywords of " BRWD3" in both English and Chinese databases of China National Knowledge Infrastructure, Wanfang, Online Mendelian Inheritance in Man, and PubMed. The clinical phenotype and genetic characteristics of patients with BRWD3 related epilepsy were summarized. Results:The patient was a 4 years and 4 months old boy, with a clinical phenotype including severe global development delay, focal seizures (the onset age was 4 months), epileptic spasm (the onset age was 6 months), autism, megacephaly, high forehead as well as hypsarrhythmia. The whole exome sequencing results showed a de novo and frameshift variation c.4318_4319del(p.Q1441Efs*20)(NM_153252) in the BRWD3 gene, and the variation was interpreted as pathogenic (PVS1+PS2+PM2) according to the American College of Medical Genetics and Genomics variant classification criteria and guidelines. A total of 7 English literature articles were retrieved reporting 16 cases of BRWD3 gene related epilepsy in children (including 1 case of infantile epileptic spasm syndrome), and there has been no report in China yet. Totally there were 17 cases of BRWD3 gene related epilepsy including this case. All the cases showed X chromosome dominant inheritance, of whom 15 cases showed minor variations, including 7 missense variations, 3 frameshift variations, 3 splicing variations, 2 nonsense variations, and the remaining 2 cases showed large segment deletions. A total of 15 different variants were found. The phenotypes of the 17 patients mainly included epileptic seizures (17/17), intellectual disability (10/17), motor development disorder (7/17), speech impairment (9/17), megacephaly (8/17), facial malformation (8/17), autism (4/17) and hypotonia (4/17). The common seizure types were found to be focal seizures, occasionally epileptic spasm seizures and tonic seizures. Conclusions:BRWD3 gene variation related epilepsy is an X chromosome dominant genetic disease with a wide clinical phenotype spectrum. BRWD3 gene mutation c.4318_4319del(p.Q1441Efs *20) could cause infantile epileptic spasm syndrome, manifested as severe global developmental delay, epileptic spasm, focal seizures, autism, craniofacial malformation and hypsarrhythmia. This research enriches BRWD3 gene mutation spectrum.

Objective:To analyze the clinical phenotypes and TSC1/TSC2 gene variations in 52 children with tuberous sclerosis complex. Methods:The clinical data of 59 children with tuberous sclerosis complex hospitalized in Linyi People′s Hospital between January 2017 and October 2022 were collected. The analysis of TSC1 and TSC2 gene variations on main family members was performed, and then bioinformatics analysis followed. The positive children were divided into TSC1 gene group and TSC2 gene group, and the difference of clinical characteristics between the two groups was analyzed. Results:Among 59 children, 52 cases were detected TSC1/ TSC2 gene variations (17 cases in the TSC1 gene group and 35 cases in the TSC2 gene group). Of the 52 children, 28 (53.8%) were male, 24 were female (46.2%); 17 (32.7%) were familial cases (10 with TSC1 gene variations and 7 with TSC2 gene variations), 35 (67.3%) were sporadic cases; 46 (88.5%) had hypomelanotic macules, 13 (25.0%) had facial angiofibromas, 5 (9.6%) had shagreen patches, 49 (94.2%) had subependymal nodules/calcifications, 47 (90.4%) had cortical nodules, 2 (3.8%) had subependymal giant cell astrocytomas, 39 (75.0%) had intellectual/developmental disabilities, 49 (94.2%) had epileptic seizures, 8 (15.4%) had cardiac rhabdomyomas, 9 (17.3%) had renal angiomyolipomas, and 4 (7.7%) had retinal hamartomas. Of the 52 children, 49 variations were detected, including 4 large fragment deletion/duplication variations, and 45 point variations; 41 pathogenic variations, 7 likely pathogenic variations, and 1 variation of uncertain significance. In this study, 16 point mutations and 1 large fragment duplication mutation which had not been reported at home and abroad, and 3 high-frequency mutation sites (p.Arg692 *, p.Arg228 *, and p.Arg1200Try) were found. There was a statistically significant difference in the proportion of familial cases [10/17 vs 7/35(20%), χ2=7.838, P=0.005], median onset age of epilepsy [38.0(0.5-134.0) months vs 8.0(0.1-63.0) months, Z=3.506 , P<0.001] and the incidence of developmental retardation/intellectual impairment [8/17 vs 31/35(88.6%), χadj2=8.423, P=0.004] between the TSC1 gene and TSC2 gene groups. Conclusions:Tuberous sclerosis compiex has widespread phenotypes, can affect every body system, especially the skin and nervous system. The pathogenic gene is TSC1/ TSC2. The TSC1 gene group has more familial cases. The TSC2 gene group has an earlier onset age of epilepsy and a higher incidence of developmental retardation/intellectual impairment. In this study, 16 novel point mutations, 1 novel large fragment duplication mutation, and 3 hotspot mutations were identified, expanding the gene variation spectrum of tuberous sclerosis complex.

Objective:To explore the clinical features and genetic etiology of a child with SPONASTRIME dysplasia (SD).Methods:A 9-month-old female who had presented at the Linyi People′s Hospital in August 2022 for short stature was selected as the study subject. Clinical data of the child were collected, and whole exome sequencing (WES) was carried out. Sanger sequencing was used for validating the candidate variants.Results:The child has manifested short stature, mid-face hypoplasia, joint laxity, internal knee rotation, irregularities in the metaphysis of long bones, and flat and concave lumbar vertebrae. WES revealed that she has harbored compound heterozygous variants of the TONSL gene, namely c.3088G>T (p.Glu1030*) and c. 3053G>A (p.Arg1018His), which were inherited from her phenotypically normal parents. Neither variant was reported previously. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c. 3088G>T variant was classified as likely pathogenic (PVS1+ PM2_Supporting), whilst the c. 3053G>A was classified as a variant of uncertain significance (PM2_Supporting+ PM3+ PP3). Conclusion:The c. 3088G>T and c. 3053G>A compound heterozygous variants of the TONSL gene probably underlay the pathogenesis in this patient. Above finding has facilitated the clinical diagnosis and genetic counseling for her family.

Objective:To investigate the clinical phenotype and genetic characteristics of a family with mitochondrial DNA depletion syndrome (MTDPS8B) caused by RRM2B gene mutation. Methods:Data of a family with MTDPS8B admitted to Department of Pediatric Neurology, Linyi People's Hospital in November 2019 were collected. Whole exome sequencing, mitochondrial loop gene sequencing and Sanger sequencing were used for genetic test in the child and family members, and pathogenicity analysis and protein 3D modeling on the mutation sites were conducted. The clinical phenotype and genetic characteristics of the family members were summarized.Results:Six subjects in these 8 individuals from the three generations of this family underwent genetic test. Four subjects had c.627G>A(p.M209I) (NM_001172477) heterozygous variation in RRM2B gene on the chromosome 8, and the variation was highly conserved; and no report on this variation has been found in domestic and foreign databases yet. Protein 3D modeling found that this variation may affect protein stability and function. The proband (male) and two older sisters carried the same variant had MTDPS8B, manifested as gastrointestinal dysfunction and progressive decline in motor function and intelligence, while the mother carried the same variant only had intellectual disability; the father and eldest sister did not carry the mutation and had normal clinical phenotype; the maternal grandparents had normal clinical phenotype and had passed away without genetic test; variation and disease were isolated in this family. The variation was interpreted as pathogenic (PM1+PM2+PP1+PP3) according to the American College of Medical Genetics and Genomics variant classification criteria and guidelines. Conclusion:The c.627G>A variant can cause RRM2B gene-related disease, and family members carried the same variants vary in clinical severity.

Objective To investigate the incidence and related factors of vitamin D deficiency(VDD) in hospitalized patients with type 2 diabetes mellitus (T2DM). Methods 2139 patients with T2DM who were hospitalized in The Endocrinology Department of Kunshan Hospital of Traditional Chinese Medicine from January 2018 to June 2023 were enrolled in the study. All the subjects were divided into VDD group (n=1301) and T2DM group (n=838),according to the international standard of vitamin D level. General data and biochemical indicators compared between the two groups. According to the interquartile interval of MHR value,it was divided into Q1(MHR≤0.272,n=534),Q2(0.2720.492,n=534)groups.Logistic regression was used to analyze the related factors of VDD in patients with T2DM. Results FPG,HbA1c,TC,TG,LDL-C,MHR,FC-P,HOMA-IR,CTX and proportion of female in VDD group were higher than those in T2DM group (P<0.05),the age and HDL-C in VDD group were lower than those in T2DM group(P<0.05). Multivariate regression analysis showed that sex,age,FPG,HbA1c,TC,TG,LDL-C,FC-P,CTX,MHR were independently assdciated with VDD. Conclusions MHR is a related factor for VDD in patients with T2DM.

Background The human body is usually exposed to a variety of heavy metals at the same time, and different types and concentrations of heavy metals may have complex interactions during their absorption and metabolism in the human body. Seminal fructose is an important energy source for sperm movement. A large number of studies have shown that metal exposure may impair semen quality, and seminal fructose is an important factor affecting male reproduction, so it is necessary to investigate the relationship between mixed heavy metal exposure and seminal fructose to explore the mechanism of semen quality damage caused by metal exposure. Objective To understand the status of common heavy metal exposure in men of childbearing age in Puyang City, Henan Province, and to study the relationship between mixed exposure to heavy metals and seminal fructose, as well as potential interactions among heavy metals. Methods Volunteers were recruited from the Puyang Maternal and Child Health Hospital Reproductive Center for a cross-sectional survey on general demographic characteristics, smoking, alcohol consumption, and other information. Semen samples were collected to detect 12 metals such as vanadium (V), manganese (Mn), cobalt (Co), nickel (Ni), zinc (Zn), selenium (Se), silver (Ag), cadmium (Cd), barium (Ba), thallium (Tl), iron (Fe), and lead (Pb) in seminal plasma and seminal fructose. After correcting for selected confounding factors, a Bayesian kernel machine regression (BKMR) model was used to evaluate the impact of seminal plasma heavy metal mixed exposure and its interactions on seminal fructose. Results A total of 825 adult males were enrolled. The concentrations in M (P₂₅, P₇₅) of V, Mn, Co, Ni, Zn, Se, Ag, Cd, Ba, Tl, Fe, and Pb in seminal plasma were 0.39 (0.28, 0.54), 12.31 (8.92, 17.52), 0.26 (0.18, 0.38), 5.15 (3.32, 8.64), 182159.80 (121847.80, 199144.50), 13.61 (10.55, 17.68), 0.03 (0.02, 0.04), 0.34 (0.27, 0.46), 8.64 (5.94, 13.43), 0.06 (0.05, 0.08), 168.74 (114.17, 259.45), and 1.69 (1.15, 2.36) μg·L⁻¹ respectively. The Spearman correlation results indicated that there was a negative correlation between V, Mn, Co, Zn, Se, Ba, Tl, or Fe in seminal plasma and seminal fructose (P<0.05), and the values of r (95%CI) were −0.044 (−0.087, −0.001), −0.129 (−0.171, −0.087), −0.055 (−0.099, −0.012), −0.099 (−0.143, −0.056), −0.053 (−0.097, −0.010), −0.068 (−0.111, −0.025), −0.095 (−0.138, −0.052), and −0.082 (−0.125, −0.039), respectively. The results of multiple linear regression indicated that there was a negative correlation between the exposure level of Cd, Mn, Zn, Ag, Ba, Tl, or Fe in seminal plasma and seminal fructose (P<0.05), the values of associated β (95%CI) were −0.551 (−0.956, −0.147), −0.315 (−0.419, −0.212), −0.187 (−0.272, −0.103), −0.161 (−0.301, −0.021), −0.188 (−0.314, −0.062), −1.159 (−2.170, −0.147), and −0.153 (−0.230, −0.076), respectively. The BKMR model analysis showed that seminal fructose level decreased with the increase of plasma metal mixed exposure concentration. Compared with all metal exposure at P₅₀, the seminal fructose level decreased by 0.2374 units when all metal exposure was at P₇₅. Seminal plasma Zn [posterior inclusion probabilities (PIPs)=1.0000] had the strongest effect on seminal fructose, followed by Mn (PIPs=0.5872), Se (PIPs=0.5656), and Ba (PIPs=0.5398). The univariate exposure-response curve showed a negative approximate linear correlations between Ba or Mn and seminal fructose, a positive linear correlation between Se and seminal fructose, and an approximate inverted U-shaped association between Zn and seminal fructose. No significant interaction between studied metals was found. Conclusion Mixed metal exposure may lead to decrease of seminal fructose, in which Zn, Mn, Se, and Ba may play an important role. Mn and Zn exposure may reduce the level of seminal fructose, Se may increase the level of seminal fructose, and there may be a threshold effect between Zn exposure and seminal fructose level. No interaction between different metals on seminal fructose is found.

OBJECTIVE: To explore the clinical features and genetic etiology of a child with Multiple congenital malformations-hypotonia-epilepsy syndrome type 3 (MCAHS3) and provide prenatal diagnosis for her parents. METHODS: A female child who had presented at Linyi People's Hospital on 27 July 2022 for recurrent convulsions for over 4 years was selected as the study subject. Clinical data of the child were collected. Peripheral blood samples were taken from the child and her parents and subjected for whole exome sequencing (WES). Candidate variants were verified by Sanger sequencing. Prenatal diagnosis was carried out on amniotic fluid sample at 18 weeks' gestation. Bioinformatic software was used to analyze the pathogenicity of the protein model for the variant loci. RESULTS: The child was a 4-year-old female with frequent seizures, peculiar facial appearance, hypotonia and severe developmental delay. Genetic analysis revealed that she has harbored compound heterozygous variants of the PIGT gene, namely c.1126del (p.H376Tfs*56) and c.1285G>C (p.E429Q), which were respectively inherited from her mother and father. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.1126del (p.H376Tfs*56) variant was predicted to be pathogenic (PVS1+PM2_Supporting+PM4), and c.1285G>C (p.E429Q) variant was predicted to be likely pathogenic (PM2_Supporting+PM3+PM4). Prenatal diagnosis suggested that the fetus also harbored the same compound heterozygous variants, and the pregnancy was terminated with induced labor. CONCLUSION The c.1126del (p.H376Tfs*56) and c.1285G>C (p.E429Q) compound heterozygous variants of the PIGT gene probably underlay the MCAHS3 in this patient, and prenatal diagnosis has prevented birth of further affected child in this family.
Humans ; Female ; Child ; Pregnancy ; Child, Preschool ; Muscle Hypotonia/genetics* ; Prenatal Diagnosis ; Computational Biology ; Epileptic Syndromes ; Facies