Mutation spectrum of hearing loss patients in Northwest China: Identification of 20 novel variants.

BACKGROUND: Hearing loss (HL) is the most frequent sensory deficit in humans, with strong genetic heterogeneity. The genetic diagnosis of HL is very important to aid treatment decisions and to provide prognostic information and genetic counselling for the patient's family. METHODS: We detected and analysed 362 Chinese non-syndromic HL patients by screening of variants in 15 hot spot mutations. Subsequently, 40 patients underwent further whole-exome sequencing (WES) to determine genetic aetiology. The candidate variants were verified using Sanger sequencing. Twenty-three carrier couples with pathogenic variants or likely pathogenic variants chose to proceed with prenatal diagnosis using Sanger sequencing. RESULTS: Among the 362 HL patients, 102 were assigned a molecular diagnosis with 52 different variants in 22 deafness genes. A total of 41 (11.33%) cases with the biallelic GJB2 (OMIM # 220290) gene mutations were detected, and 21 (5.80%) had biallelic SLC26A4 (OMIM # 605646) mutations. Mitochondrial gene (OMIM # 561000) mutations were detected in seven (1.93%) patients. Twenty of the variants in 15 deafness genes were novel. SOX10 (OMIM # 602229), MYO15A (OMIM # 602666) and WFS1 (OMIM # 606201) were each detected in two patients. Meanwhile, OSBPL2 (OMIM # 606731), RRM2B (OMIM # 604712), OTOG (OMIM # 604487), STRC (OMIM # 606440), PCDH15 (OMIM # 605514), LOXHD1 (OMIM # 613072), CDH23 (OMIM # 605516), TMC1 (OMIM # 606706), CHD7 (OMIM # 608892), DIAPH3 (OMIM # 614567), TBC1D24 (OMIM # 613577), TIMM8A (OMIM # 300356), PTPRQ (OMIM # 603317), SALL1 (OMIM # 602218), and GSDME (OMIM # 608798) were each detected in one patient. In addition, as regards one couple with a heterozygous variant of CDH23 and PCDH15, respectively, prenatal diagnosis results suggest that the foetus had double heterozygous (DH) variants of CDH23 and PCDH15, which has a high risk to cause ID/F type Usher syndrome. CONCLUSION: Our study expanded the spectrum of deafness gene variation, which will contribute to the genetic diagnosis, prenatal diagnosis and the procreation guidance of deaf couple. In addition, the deafness caused by two genes should be paid attention to in the prenatal diagnosis of families with both deaf patients.

A novel microRNA miR-4433a-3p as a potential diagnostic biomarker for lung adenocarcinoma.

Background: Lung adenocarcinoma is one of the leading causes of cancer-related deaths because of the lack of early specific clinical indicators. MicroRNAs (miRNAs) have become the focus in lung cancer diagnosis. Further studies are required to explore miRNA expression in the serum of lung adenocarcinoma patients and their correlation with therapy and analyse specific messenger RNA targets to improve the specificity and sensitivity of early diagnosis. Methods: The Toray 3D-Gene miRNA array was used to compare the expression levels of various miRNAs in the sera of patients with lung adenocarcinoma and healthy volunteers. Highly expressed miRNAs were selected for further analysis. To verify the screening results, serum and pleural fluid samples were analysed using qRT-PCR. Serum levels of the miRNAs and their correlation with the clinical information of patients with lung adenocarcinoma were analysed. The functions of miRNAs were further analysed using the Kyoto Encyclopedia of Gene and Genomes and Gene Ontology databases. Results: Microarray analysis identified 60 and 50 miRNAs with upregulated and downregulated expressions, respectively, in the serum of patients with lung adenocarcinoma compared to those in healthy individuals. Using qRT-qPCR to detection of miRNAs expression in the serum or pleural effusion of patients with early and advanced lung adenocarcinoma, we found that miR-4433a-3p could be used as a diagnostic marker and therapeutic evaluation indicator for lung adenocarcinoma. Serum of miR-4433a-3p levels significantly correlated with the clinical stage. miR-4433a-3p may be more suitable than other tumour markers for the early diagnosis and evaluation of therapeutic effects in lung adenocarcinoma. miR-4433a-3p may affect tumour growth and metastasis by acting on target genes (PIK3CD, UBE2J2, ICMT, PRDM16 and others) and regulating tumour-related signalling pathways (MAPK signal pathway, Ras signalling pathway and others). Conclusion: miR-4433a-3p may serve as a biomarker for the early diagnosis of lung adenocarcinoma and monitoring of therapeutic effects.

Serum bile acid and unsaturated fatty acid profiles of non-alcoholic fatty liver disease in type 2 diabetic patients.

BACKGROUND: The understanding of bile acid (BA) and unsaturated fatty acid (UFA) profiles, as well as their dysregulation, remains elusive in individuals with type 2 diabetes mellitus (T2DM) coexisting with non-alcoholic fatty liver disease (NAFLD). Investigating these metabolites could offer valuable insights into the pathophy-siology of NAFLD in T2DM. AIM: To identify potential metabolite biomarkers capable of distinguishing between NAFLD and T2DM. METHODS: A training model was developed involving 399 participants, comprising 113 healthy controls (HCs), 134 individuals with T2DM without NAFLD, and 152 individuals with T2DM and NAFLD. External validation encompassed 172 participants. NAFLD patients were divided based on liver fibrosis scores. The analytical approach employed univariate testing, orthogonal partial least squares-discriminant analysis, logistic regression, receiver operating characteristic curve analysis, and decision curve analysis to pinpoint and assess the diagnostic value of serum biomarkers. RESULTS: Compared to HCs, both T2DM and NAFLD groups exhibited diminished levels of specific BAs. In UFAs, particular acids exhibited a positive correlation with NAFLD risk in T2DM, while the ω-6:ω-3 UFA ratio demonstrated a negative correlation. Levels of α-linolenic acid and γ-linolenic acid were linked to significant liver fibrosis in NAFLD. The validation cohort substantiated the predictive efficacy of these biomarkers for assessing NAFLD risk in T2DM patients. CONCLUSION: This study underscores the connection between altered BA and UFA profiles and the presence of NAFLD in individuals with T2DM, proposing their potential as biomarkers in the pathogenesis of NAFLD.

[Examination of the relationship between tasks in geriatric health service facilities and benefits of working for older assistant care workers].

Objective　In this study, we clarified the characteristics of tasks performed by older assistant care workers at geriatric health services facilities, by individual characteristics and work status. Additionally, we examined the relationship between the tasks and benefits of working for older assistant care workers.Methods　A self-administered questionnaire survey was provided to 2,571 elderly care facilities employing assistant care workers aged ≥60.　A total of 1,606 responses were obtained, and personal characteristics, working conditions, tasks, and benefits of working for older care assistants were surveyed. Thirteen work tasks were established and categorized into four broad categories; user transfer assistance, facility maintenance, meal-related assistance, and managing and talking to users. Seven benefits of working as care assistants were "contribution to society," "social connection," "purpose of life," "income," "learning from caregiving," "health maintenance and promotion," and "time utilization."Results　Most of the respondents were women and relatively young workers. Among tasks, men, younger workers, and those working >5 days weekly represented a high proportion of those performing user transfer assistance tasks. Tasks on managing and talking to users were not correlated with sex, age, and number of days worked, but those with more flexible work patterns were more likely to be engaged in such tasks. Logistic regression analysis showed that user transfer assistance was correlated with benefits of working, such as "contribution to society," "social connection," and "learning from caregiving." Moreover, facility maintenance was associated with "income," "health maintenance and promotion," and "time utilization" and meal-related assistance was associated with "learning from caregiving." Managing and talking to users was associated with "contribution of society," "social connection," "purpose of life," and "learning from caregiving."Conclusion　The tasks of the workers were associated with individual characteristics and work status. The perception of benefits between the working tasks that involved frequent contact with users and those that did not were considerably different.

Analysis of spinal muscular atrophy carrier screening results in 32,416 pregnant women and 7,231 prepregnant women.

Objectives: Spinal muscular atrophy (SMA) is an autosomal recessive disease that is one of the most common in childhood neuromuscular disorders. Our screenings are more meaningful programs in preventing birth defects, providing a significant resource for healthcare professionals, genetic counselors, and policymakers involved in designing strategies to prevent and manage SMA. Method: We screened 39,647 participants from 2020 to the present by quantitative real-time PCR, including 7,231 pre-pregnancy participants and 32,416 pregnancy participants, to detect the presence of SMN1 gene EX7 and EX8 deletion in the DNA samples provided by the subjects. To validate the accuracy of our findings, we also utilized the Multiplex Ligation-dependent Probe Amplification (MLPA) to confirm the reliability of screening results obtained by quantitative real-time PCR. Result: Among the 39,647 participants who were screened, 726 participants were the carriers of SMN1. The overall carrier rate was calculated to be 1.83% (95% confidence interval: 0.86-2.8%). After undergoing screening, a total of 592 pregnancy carriers were provided with genetic counseling and only 503 of their spouses (84.97, 95% confidence interval: 82.09-87.85%) voluntarily underwent SMA screening. Conclusion: This study provides crucial insights into the prevalence and distribution of SMA carriers among the female population. The identification of 726 asymptomatic carriers highlights the necessity of comprehensive screening programs to identify at-risk individuals and ensure appropriate interventions are in place to minimize the impact of SMA-related conditions.

[Genetic analysis of eighteen patients from Gansu Province with Tetrahydrobiopterin deficiency].

OBJECTIVE: To explore the genetic basis of eighteen patients with Tetrahydrobiopterin deficiency (BH4D) from Gansu Province. METHODS: Eighteen patients diagnosed with BH4D at Gansu Provincial Maternal and Child Health Care Hospital from January 2018 to December 2021 were selected as the study subjects. Whole exome sequencing was carried out, and candidate variants were verified by Sanger sequencing. RESULTS: All of the thirty-six alleles of the eighteen patients were successfully determined by molecular genetic testing. Sixteen patients were found to harbor variants of the PTS gene, and two had harbored variants of the QDPR gene. Ten variants were detected in the PTS gene, with the most common ones being c.259C>T (34.38%) and c.286G>A (15.63%). Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the c.259C>T was classified as a pathogenic variant, whilst the c.286G>A, c.166G>A, c.200C>T, c.272A>G, c.402A>C, c.421G>T, c.84_291A>G and c.317C>T were classified as likely pathogenic variants. A novel c.289_290insCTT variant was classified as likely pathogenic (PM1+PM2_Supporting+PM3+PP3+PP4). The two variants (c.478C>T and c.665C>T) detected in the QDPR gene were both classified as variants of uncertain significance (PM1+PM2_Supporting+PP3+PP4). CONCLUSION: Genetic testing has clarified the pathogenic variants in these BH4D patients, which has enabled timely and accurate clinical intervention and treatment, and provided a reference for genetic counseling and reproductive guidance for their families.

Comparative transcriptome analysis reveals the adaptive mechanisms of halophyte Suaeda dendroides encountering high saline environment.

Suaeda dendroides, a succulent euhalophyte of the Chenopodiaceae family, intermittently spread around northern Xinjiang, China, has the ability to grow and develop in saline and alkali environments. The objective of this study was therefore to investigate the underlying molecular mechanisms of S. dendroides response to high salt conditions. 27 sequencing libraries prepared from low salt (200 mM NaCl) and high salt (800 mM NaCl) treated plants at 5 different stages were sequenced using Illumina Hiseq 2000. A total of 133,107 unigenes were obtained, of which 4,758 were DEGs. The number of DEGs in the high salt group (3,189) was more than the low salt treatment group (733) compared with the control. GO and KEGG analysis of the DEGs at different time points of the high salt treatment group showed that the genes related to cell wall biosynthesis and modification, plant hormone signal transduction, ion homeostasis, organic osmolyte accumulation, and reactive oxygen species (ROS) detoxification were significantly expressed, which indicated that these could be the main mechanisms of S. dendroides acclimate to high salt stress. The study provides a new perspective for understanding the molecular mechanisms of halophytes adapting to high salinity. It also provides a basis for future investigations of key salt-responsive genes in S. dendroides.

Transcriptome Analysis Reveals Key Genes Involved in the Response of Pyrus betuleafolia to Drought and High-Temperature Stress.

Drought and high-temperature stress are the main abiotic stresses that alone or simultaneously affect the yield and quality of pears worldwide. However, studies on the mechanisms of drought or high-temperature resistance in pears remain elusive. Therefore, the molecular responses of Pyrus betuleafolia, the widely used rootstock in pear production, to drought and high temperatures require further study. Here, drought- or high-temperature-resistant seedlings were selected from many Pyrus betuleafolia seedlings. The leaf samples collected before and after drought or high-temperature treatment were used to perform RNA sequencing analysis. For drought treatment, a total of 11,731 differentially expressed genes (DEGs) were identified, including 4444 drought-induced genes and 7287 drought-inhibited genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that these DEGs were more significantly enriched in plant hormone signal transduction, flavonoid biosynthesis, and glutathione metabolism. For high-temperature treatment, 9639 DEGs were identified, including 5493 significantly upregulated genes and 4146 significantly downregulated genes due to high-temperature stress. KEGG analysis showed that brassinosteroid biosynthesis, arginine metabolism, and proline metabolism were the most enriched pathways for high-temperature response. Meanwhile, the common genes that respond to both drought and high-temperature stress were subsequently identified, with a focus on responsive transcription factors, such as MYB, HSF, bZIP, and WRKY. These results reveal potential genes that function in drought or high-temperature resistance. This study provides a theoretical basis and gene resources for the genetic improvement and molecular breeding of pears.

Immune skeletal dysplasia with neurodevelopmental abnormalities caused by a novel variant of EXTL3 gene in a Chinese family.

BACKGROUND: Immune skeletal dysplasia with neurodevelopmental abnormalities (ISDNA) is an extremely rare, autosomal recessive genetic disorder characterized by various skeletal abnormalities, neurodevelopmental deficits, and abnormal immune system function. ISDNA is caused by variation in the exostosin-like 3 (EXTL3) gene, located on chromosome 8p21.2, whose primary function is the biosynthesis of heparan sulfate (HS) skeleton structure. Only a few variations in the EXTL3 gene have been discovered so far. In these years of development, many pathogenic variants in genetic diseases with genetic and phenotypic heterogeneity have been investigated using whole-exome sequencing (WES) technology. METHODS: In this research, a novel EXTL3 variant was first detected in a patient using WES, which was validated from Sanger sequencing in this family. Family history and clinical information were then collected through comprehensive medical examinations and genetic counseling. In silico prediction was then utilized to confirm the pathogenicity of the variant. RESULTS: A novel homozygous variant, NM_001440: c.2015G>A (p.Arg672Gln) in the EXTL3 gene, was identified using WES, which has never been reported before. Sanger sequencing was performed to confirm that the variant segregated with the disease within the family. CONCLUSION: This research identified a novel pathogenic variant in the EXTL3 gene responsible for ISDNA in a Chinese family. It showed the potential diagnostic role of WES in ISDNA, expanded the EXTL3 gene variation spectrum, and demonstrated that the diagnosis of ISDNA using WES is feasible and effective. More comprehensive genetic counseling and precise prenatal diagnosis for the next pregnancy can also be provided to families with genetic disorders.

Receptor-interacting Protein Kinase 2 Is an Immunotherapy Target in Pancreatic Cancer.

Pancreatic ductal adenocarcinoma (PDAC) remains a highly lethal malignancy because of its aggressive nature and the paucity of effective treatment options. Almost all registered drugs have proven ineffective in addressing the needs of patients with PDAC. This is the result of a poor understanding of the unique tumor-immune microenvironment (TME) in PDAC. To identify druggable regulators of immunosuppressive TME, we performed a kinome- and membranome-focused CRISPR screening using orthotopic PDAC models. Our data showed that receptor-interacting protein kinase 2 (RIPK2) is a crucial driver of immune evasion of cytotoxic T-cell killing and that genetic or pharmacologic targeting of RIPK2 sensitizes PDAC to anti-programmed cell death protein 1 (anti-PD-1) immunotherapy, leading to prolonged survival or complete regression. Mechanistic studies revealed that tumor-intrinsic RIPK2 ablation disrupts desmoplastic TME and restores MHC class I (MHC-I) surface levels through eliminating NBR1-mediated autophagy-lysosomal degradation. Our results provide a rationale for a novel combination therapy consisting of RIPK2 inhibition and anti-PD-1 immunotherapy for PDAC. SIGNIFICANCE: PDAC is resistant to almost all available therapies, including immune checkpoint blockade. Through in vivo CRISPR screen, we identified that RIPK2 plays a crucial role in facilitating immune evasion by impeding antigen presentation and cytotoxic T-cell killing. Targeting tumor-intrinsic RIPK2 either genetically or pharmacologically improves PDAC to anti-PD-1 immunotherapy. See related commentary by Liu et al., p. 208 . This article is featured in Selected Articles from This Issue, p. 201.

[Current conditions and challenges of employing older assistant care workers based on geriatric health services facilities' scale].

Objectives　To support care workers, some care facilities employ older individuals as care assistants for peripheral, non-professional tasks. However, the size of the facilities influences their employment needs and support systems for older care workers. Therefore, this study examined the rationale for employing older assistant care workers; the challenges they face; educational systems; and the efforts to facilitate continued employment based on facility size.Methods　Initially, a fax survey was conducted with 3,591 facilities associated with the National Association of Geriatric Health Services Facilities to determine the proportion of older assistant care workers employed. Thereafter, a questionnaire survey was administered to gather more information. Out of 3,591 facilities, 2,170 responded to the survey, and 1,261 responded to the questionnaire. In this study, care assistants aged ≥60 years were classified as older care assistants. The facilities were categorized based on admission capacity into two groups: "small/medium facilities" (≤99 residents) and "large facilities" (≥100 residents). A chi-square test was conducted for data analysis.Results　Of the all geriatric health services facilities, 31.7% employed older assistant care workers. Moreover, the questionnaire survey responses showed they are employed by 687 facilities. Further analysis revealed that larger facilities employed a higher number of older care assistants than small/medium facilities. Small/medium facilities tended to have fewer training systems for older care assistants than larger facilities (30.0% for small/medium facilities vs. 21.6% for large facilities; P=0.014). Furthermore, the work of older care assistants in small/medium facilities was more susceptible to family circumstances than those in larger facilities (15.7% for small/medium facilities vs. 10.2% for large facilities; P=0.033). Compared to small/medium facilities, larger facilities were more likely to state that their employment objective was to reduce the risk of care accidents (19.8% for small/medium facilities vs. 26.3% for large facilities; P=0.046) and provide psychological support, such as periodic interviews, to older care workers to ensure continued employment (24.1% for small/medium facilities vs. for large facilities 37.3%; P<0.001).Conclusion　Larger facilities employeda higher number of older care assistants than small/medium facilities. Furthermore, small/medium facilities were less likely to provide psychological support and had a less structured training system, compared to larger facilities. Additionally, the work of older care assistants in small/medium facilities was more susceptible to family circumstances and other factors, which could affect their work performance. Thus, developing adequate support systems in small/medium facilities is crucial to promoting the employment of older care assistants.

Molecular Dynamics Simulation of the Effect of Defect Size on Magnetostrictive Properties of Low-Dimensional Iron Thin Films.

Defects are an inevitable occurrence during the manufacturing and use of ferromagnetic materials, making it crucial to study the microscopic mechanism of magnetostrictive properties of ferromagnetic materials with defects. This paper conducts molecular dynamics simulations on low-dimensional iron thin films containing hole or crack defects, analyzes and compares the impact of defect size on magnetostrictive properties, and investigates the microscopic mechanism of their effects. The results indicate that the saturation magnetostrictive strains of the defect models do not increase monotonically as the defect size increases. Additionally, it is discovered that the arrangement of atomic magnetic moments in the initial magnetic moment configuration also affects the magnetostrictive properties. When controlling the size of the hole or crack within a certain defect area, it is found that the hole size has less influence on the initial magnetic moment configuration, resulting in a smaller corresponding change in the saturation strain and thus having a lesser impact on the magnetostrictive properties. Conversely, when the crack size changes, the arrangement of the atomic magnetic moments in the initial magnetic moment configuration changes more significantly, resulting in a greater corresponding change in saturation strain, and thus having a greater impact on the magnetostriction performance.

High expression of peroxisomal D-bifunctional protein in cytosol regulates apoptosis and energy metabolism of hepatocellular carcinoma cells via PI3K/AKT pathway.

Peroxisomal D-bifunctional protein (DBP) is an indispensable enzyme of the fatty acid ß-oxidation in the peroxisome of humans. However, the role of DBP in oncogenesis is poorly understood. Our previous studies have demonstrated that DBP overexpression promotes hepatocellular carcinoma (HCC) cell proliferation. In this study, we evaluated the expression of DBP in 75 primary HCC samples using RT-qPCR, immunohistochemistry, and Western blot, as well as its correlation with the prognosis of HCC. In addition, we explored the mechanisms by which DBP promotes HCC cell proliferation. We found that DBP expression was upregulated in HCC tumor tissues, and higher DBP expression was positively correlated with tumor size and TNM stage. Multinomial ordinal logistic regression analysis indicated that lower DBP mRNA level was an independent protective factor of HCC. Notably, DBP was overexpressed in the peroxisome and cytosol and mitochondria of tumor tissue cells. Xenograft tumor growth was promoted by overexpressing DBP outside peroxisome in vivo. Mechanistically, DBP overexpression in cytosol activated the PI3K/AKT signaling axis and promoted HCC cell proliferation by downregulating apoptosis via AKT/FOXO3a/Bim axis. In addition, overexpression of DBP increased glucose uptake and glycogen content via AKT/GSK3ß axis, as well as elevated the activity of mitochondrial respiratory chain complex III to increase ATP content via the mitochondrial translocation of p-GSK3ß in an AKT-dependent manner. Taken together, this study was the first to report the expression of DBP in peroxisome and cytosol, and that the cytosolic DBP has a critical role in the metabolic reprogramming and adaptation of HCC cells, which provides a valuable reference for instituting an HCC treatment plan.

Identification of deep intronic variants of PAH in phenylketonuria using full-length gene sequencing.

BACKGROUND: Phenylketonuria (PKU) is an autosomal recessive congenital metabolic disorder caused by PAH variants. Previously, approximately 5% of PKU patients remained undiagnosed after Sanger sequencing and multiplex ligation-dependent probe amplification. To date, increasing numbers of pathogenic deep intronic variants have been reported in more than 100 disease-associated genes. METHODS: In this study, we performed full-length sequencing of PAH to investigate the deep intronic variants in PAH of PKU patients without definite genetic diagnosis. RESULTS: We identified five deep intronic variants (c.1199+502A>T, c.1065+241C>A, c.706+368T>C, c.706+531>C, and c.706+608A>C). Of these, the c.1199+502A>T variant was found at high frequency and may be a hotspot PAH variant in Chinese PKU. c.706+531T>C and c.706+608A>C are two novel variants that extend the deep intronic variant spectrum of PAH. CONCLUSION: Deep intronic variant pathogenicity analysis can further improve the genetic diagnosis of PKU patients. In silico prediction and minigene analysis are powerful approaches for studying the functions and effects of deep intronic variants. Targeted sequencing after full-length gene amplification is an economical and effective tool for the detection of deep intron variation in genes with small fragments.

[Relationship between the perceived benefits of working and emotional exhaustion among older assistant care workers: A cross-sectional study].

Objectives　This study is intended to clarify the perceived benefits of working among older assistant care workers employed in geriatric health service facilities and examine the relationship between perceived benefits and emotional exhaustion.Methods　We analyzed data from a mail survey of older assistant care workers employed in geriatric health service facilities that the Japan Association of Geriatric Health Services Facilities conducted in 2020. In this survey, those aged ≥60 were defined as older assistant care workers. Responses were obtained from 1,606 older assistant care workers at 599 facilities. The dependent variable was the emotional exhaustion subscale of the Japanese version of the Burnout Questionnaire. The independent variables were the seven benefits of working as an assistant care worker: 1) I can contribute to society; 2) I am connected to society; 3) I have a sense of purpose in life; 4) I am earning the income I expected since I started working as an assistant care worker; 5) I am learning about care; 6) It has maintained and improved my health; and 7) I can use my time effectively. Latent class analysis (LCA) was performed using these seven benefits as independent variables. Multiple regression analysis was performed using emotional exhaustion and the perceived benefits as dependent and independent variables, respectively. The missing values were supplemented by the multiple imputation method.Results　Overall, 1,601 responses were analyzed after excluding 5 respondents who did not answer all the questions. Four patterns were identified from the LCA results: the "benefit-full type," who perceived all benefits; the "benefit-less type," who perceived few benefits; the "benefit-extroverted type," who perceived social contribution and connection as benefits; and the "benefit-introverted type," who perceived health maintenance and improvement and time utilization as benefits. The multiple regression analysis showed that emotional exhaustion scores were higher for the "benefit-extroverted" and "benefit-less" types than for the "benefit-full" type (b=2.465, P<.001 and b=1.931, P<.001, respectively). No difference was found in the case of the "benefit-introverted" type (b=0.050, P=.851).Conclusion　The perceptions of diverse and introverted benefits were associated with lower emotional exhaustion scores among older assistant care workers. Future intervention studies are needed to examine whether obtaining diverse or introverted benefits can decrease the emotional exhaustion score.

[Prenatal diagnosis for a fetus with Walker-Warburg syndrome].

OBJECTIVE: To explore the genetic etiology for a fetus with Walker-Warburg syndrome(WWS). METHODS: A fetus with WWS diagnosed at Gansu Provincial Maternity and Child Health Care Hospital in June 9, 2021 was selected as the study subject. Genomic DNA was extracted from amniotic fluid sample of the fetus and peripheral blood samples from its parents. Trio-Whole exome sequencing (trio-WES) was carried out. Candidate variants were verified by Sanger sequencing. RESULTS: The fetus was found to harbor compound heterozygous variants of the POMT2 gene, namely c.471delC (p.F158Lfs*42) and c.1975C>T (p.R659W), which were respectively inherited from its father and mother. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), they were respectively rated as pathogenic (PVS1+PM2_Supporting+PP4) and likely pathogenic (PM2_Supporting+PM3+PP3_Moderate+PP4). CONCLUSION: Trio-WES may be used for the prenatal diagnosis of WWS. The compound heterozygous variants of the POMT2 gene probably underlay the disorder in this fetus. Above finding has expanded the mutational spectrum of the POMT2 gene and enabled definite diagnosis and genetic counseling for the family.

The spectrum of phenylalanine hydroxylase variants and genotype-phenotype correlation in phenylketonuria patients in Gansu, China.

BACKGROUND: Phenylketonuria (PKU) is a common, congenital, autosomal recessive, metabolic disorder caused by Phenylalanine hydroxylase (PAH) variants. METHODS: 967 PKU patients from Gansu, China were genotyped by Sanger sequencing, multiplex ligation-dependent probe amplification, and whole exome sequencing. We analyzed the variants of PAH exons, their flanking sequences, and introns. RESULTS: The detection of deep intronic variants in PAH gene can significantly improve the genetic diagnostic rate of PKU. The distribution of PAH variants among PKU subtypes may be related to the unique genetic background in Gansu, China. CONCLUSION: The identification of PAH hotspot variants will aid the development of large-scale neonatal genetic screening for PKU. The five new PAH variants found in this study further expand the spectrum of PAH variants. Genotype-phenotype correlation analysis may help predict the prognosis of PKU patients and enable precise treatment regimens to be developed.

New Lidocaine-Based Pharmaceutical Cocrystals: Preparation, Characterization, and Influence of the Racemic vs. Enantiopure Coformer on the Physico-Chemical Properties.

This study describes the preparation, characterization, and influence of the enantiopure vs. racemic coformer on the physico-chemical properties of a pharmaceutical cocrystal. For that purpose, two new 1:1 cocrystals, namely lidocaine:dl-menthol and lidocaine:d-menthol, were prepared. The menthol racemate-based cocrystal was evaluated by means of X-ray diffraction, infrared spectroscopy, Raman, thermal analysis, and solubility experiments. The results were exhaustively compared with the first menthol-based pharmaceutical cocrystal, i.e., lidocaine:l-menthol, discovered in our group 12 years ago. Furthermore, the stable lidocaine/dl-menthol phase diagram has been screened, thoroughly evaluated, and compared to the enantiopure phase diagram. Thus, it has been proven that the racemic vs. enantiopure coformer leads to increased solubility and improved dissolution of lidocaine due to the low stable form induced by menthol molecular disorder in the lidocaine:dl-menthol cocrystal. To date, the 1:1 lidocaine:dl-menthol cocrystal is the third menthol-based pharmaceutical cocrystal, after the 1:1 lidocaine:l-menthol and the 1:2 lopinavir:l-menthol cocrystals reported in 2010 and 2022, respectively. Overall, this study shows promising potential for designing new materials with both improved characteristics and functional properties in the fields of pharmaceutical sciences and crystal engineering.

Judicial and legislative practice and related suggestions on off-label drug use in China.

BACKGROUND: Off-label drug use exists widely in medical practice and is also an area which easily triggers controversy between patients and medical institutions. Previous studies have identified the reasons why off-label drug use long exists. However, there is no multidimensional analysis on real judicial precedents about off-label drug use. This study aimed to investigate the dispute points on off-label drug use based on real cases in China, and proposed suggestions based on newly-leased Physicians Law. METHODS: Our study is a retrospective study with all the 35 judicial precedents on off-label drug use extracted from China Judgments Online from 2014 to 2019. This study mainly used the methods of statistical analysis, inferential analysis, exemplification, literature summarization and comparative analysis. RESULTS: According to the analysis of the 35 precedents of jurisdictions from 11 different aspects, it can be seen that the second-instance and retrial rates of this kind of cases are high, and the disputes between patients and medical institutions are fierce. In judicial practice of off-label drug use, medical institutions are determined whether to bear civil liability according to the constituent elements of medical tort liability: the rate of medical institutions' bearing liability for off-label drug use is not high, and medical institutions are not directly identified as infringing acts and they don't bear tort liability. The clear provisions about off-label drug use in Law of the People's Republic of China on Physicians which was implemented in March 2022 confirm this at the legislative level. CONCLUSIONS: By analyzing the current judicial practice of China's off-label drug use cases, and summarizing the dispute points between medical institution and patients, the constituent elements of tort liability, and the rules of evidence etc., suggestions are proposed to further regulate off-label drug use and promote safe and rational drug use.

PsA inhibits the development of bovine embryos through epigenetic and oxidative stress.

OBJECTIVE: Histone deacetylases (HDACs) are the key regulators involved in the process of embryo development and tumor progression and are often dysregulated in numerous disordered cells, including tumor cells and somatic cell nuclear transfer (SCNT) embryos. Psammaplin A (PsA), a natural small-molecular therapeutic agent, is a potent histone deacetylase inhibitor (HDACi) that alters the regulation of histone. SAMPLES: Approximately 2,400 bovine parthenogenetic (PA) embryos. PROCEDURES: To investigate the effect of PsA on bovine preimplanted embryos, we analyzed the preimplantation development of PA embryos treated with PsA in this study. RESULTS: The blastocyst formation rate of bovine PA embryos decreased sharply with an increase in concentration and duration. Furthermore, the expression of the pluripotency-related gene Nanog was decreased, and the inhibitory effects on histone deacetylases 1 (HDAC1) and DNA methylation transferase 1 (DNMT1) were observed in bovine PA embryos. The acetylation level of histone H3 lysine 9 (H3K9) was enhanced by a PsA treatment of 10 µM for 6 h, while the DNA methylation appeared unchanged. Interestingly, we also found that PsA treatment enhanced the intracellular reactive oxygen species (ROS) generation and decreased the intracellular mitochondrial membrane potential (MMP)- and superoxide dismutase 1 (SOD1)-induced oxidative stress. Our findings improve the understanding of HDAC in embryo development and provide a theoretical basis and reproduction toxicity evaluation for the application of PsA. CLINICAL RELEVANCE: These results indicate that PsA inhibits the development of bovine preimplantation PA embryos, supplying data for the PsA clinical application concentration to avoid reproductive toxicity. In addition, the reproduction toxic effect of PsA may be modulated through increased oxidative stress on the bovine PA embryo, suggesting that PsA in combination with antioxidants, for example, melatonin, might be an effective clinical application strategy.