OBJECTIVES: The purinergic receptor P2X2 (P2RX2) encodes an ATP-gated ion channel permeable to Na⁺, K⁺, and especially Ca²⁺. Loss-of-function mutations in P2RX2 are known to cause autosomal dominant nonsyndromic deafness 41 (DFNA41), which manifests as high-frequency hearing loss, accelerated presbycusis, and increased susceptibility to noise-induced damage. Zebrafish, owing to their small size, rapid development, high fecundity, transparent embryos, and high gene conservation with humans, provide an ideal model for studying human diseases and developmental mechanisms. This study aims to generate a p2rx2 knockout zebrafish model using CRISPR/Cas9 gene editing system to investigate the effect of p2rx2 deficiency on the auditory system, providing a basis for understanding P2RX2-related hearing loss and developing gene therapy strategies. METHODS: Two CRISPR targets (sgRNA1 and sgRNA2) spaced 47 bp apart were designed within the zebrafish p2rx2 gene. Synthesized sgRNAs and Cas9 protein were microinjected into single-cell stage Tübingen (TU)-strain zebrafish embryos. PCR and gel electrophoresis verified editing efficiency at 36 hours post-fertilization (hpf). Surviving embryos were raised to adulthood (F0), tail-clipped, genotyped, and screened for positive mosaics. F1 heterozygotes were generated by outcrossing, and F2 homozygous mutants were obtained by intercrossing. Polymerase chain reaction (PCR) combined with sequencing verified mutation type and heritability. At 5 days post-fertilization (dpf), YO-PRO-1 staining was used to examine hair cell morphology and count in lateral line neuromasts and the otolith region. Auditory evoked potential (AEP) thresholds at 600, 800, 1 000, and 2 000 Hz were measured in nine 4-month-old wild type and mutant zebrafish per group. RESULTS: A stable p2rx2 knockout zebrafish line was successfully established. Sequencing revealed a 66 bp insertion at the first target site introducing a premature stop codon (TAA), leading to early termination of protein translation and loss of function. Embryos developed normally with no gross malformations. At 5 dpf, mutants exhibited significantly reduced hair cell density in the otolith region compared with wild type, although lateral line neuromasts were unaffected. AEP testing showed significantly elevated auditory thresholds at all 4 frequencies in homozygous mutants compared with wild type (all P<0.001), indicating reduced hearing sensitivity. CONCLUSIONS We successfully generated a p2rx2 loss-of-function zebrafish model using CRISPR/Cas9 technology. p2rx2 deficiency caused hair cell defects in the otolith region and increased auditory thresholds across frequencies, indicating its key role in maintaining zebrafish auditory hair cell function and hearing perception. The phenotype's restriction to the otolith region suggests tissue-specific roles of p2rx2 in sensory organs. This model provides a valuable tool for elucidating the molecular mechanisms of P2RX2-related hearing loss and for screening otoprotective drugs and developing gene therapies.
Animals ; Zebrafish/genetics* ; Receptors, Purinergic P2X2/deficiency* ; CRISPR-Cas Systems/genetics* ; Gene Knockout Techniques ; Phenotype ; Zebrafish Proteins/genetics* ; Disease Models, Animal

Objective The quality evaluation of 20 batches of Ganoderma was carried out by the combination of DNA identification and physicochemical analysis.Methods The 20 batches of Ganoderma were identified by DNA extraction,polymerase chain reaction(PCR)amplification,purification,sequencing,and inputting the Chinese medicines into DNA barcode identification system.Then the content determination of marker components was carried out.Results Ganoderma of No.1,2,3,4,5,6,9,10,11,12,13,14,15,16,18 and 20 belonged to Ganoderma lucidum(Leyss.ex Fr.)Karst.in the Pharmacopoeia,while No.7,8,17 and 19 did not belong to Ganoderma lucidum(Leyss.ex Fr.)Karst.in the Pharmacopoeia.Content determination was conducted on 16 batches of Ganoderma lucidum(Leyss.ex Fr.)Karst.,in which 15 batches were up to the standards of Pharmacopoeia,the decreasing sequence of contents was as follows:No.2,1,5,10,18,3,4,16,15,11,13,14,6,9,and 12,and No.20 was failed to meet the standards,with a content of 0.38％.Conclusion The combination of DNA molecular identification and physicochemical analysis can be used for rapid quality evaluation of the authenticity and superiority of Ganoderma.

Objective:To explore the clinical risk factors and pregnancy outcomes in women with hyperglycemia in pregnancy (HIP) complicated with hypertensive disorders of pregnancy (HDP).Methods:This retrospective cohort study included 43 973 singleton live births delivered at the Obstetrics and Gynecology Hospital of Fudan University between September 2017 and December 2022. Participants were categorized into four groups: HH group (HIP with HDP, n=1 011), HIP group ( n=5 469), HDP group ( n=3 486), and control group ( n=34 007). Baseline characteristics and perinatal outcomes were compared using the Chi-square test (or Fisher's exact test). Logistic regression identified risk factors and adverse outcome risks and analyze the impact of HH on neonatal weight. Results:HH accounted for 15.6% (1 011/6 480) of HIP cases, with an overall incidence of 2.3% (1 011/43 973). HIP and HDP were strongly correlated [ OR=1.803, 95% CI: 1.672-1.945]. Advanced maternal age (≥35 years at estimated due date), primiparity, and pre-pregnancy overweight/obesity (body mass index ≥24 kg/m2) were independent risk factors for HH [ OR (95% CI): 1.305 (1.113-1.529), 1.845 (1.545-2.203), and 2.316 (1.981-2.718), respectively]. Compared to the HIP group, the HH group had significantly higher risks of preterm birth [10.3% (104/1 011) vs. 6.3% (344/5 469), OR=1.627 (95% CI:1.280-2.068)], cesarean delivery [57.0% (576/1 011) vs. 41.9% (2 289/5 469), OR=1.701 (95% CI:1.474-1.963)], and neonatal birth weight < P10 [13.9% (141/1 011) vs. 9.0% (494/5 469), OR=1.668 (95% CI:1.336-2.083)]. Stratified analysis revealed a 73.4% increased risk of birth weight < P10 in the gestational diabetes mellitus A1 with HDP subgroup ( aOR=1.734, 95% CI: 1.416-2.125). Conclusions:Advanced age, pre-pregnancy overweight/obesity, and primiparity are risk factors for HH. Compared to isolated HIP, HH is associated with elevated risks of preterm birth, cesarean delivery, and abnormal neonatal birth weight, though the impact on birth weight may vary by HIP subtype.

Objective:To explore the clinical risk factors and pregnancy outcomes in women with hyperglycemia in pregnancy (HIP) complicated with hypertensive disorders of pregnancy (HDP).Methods:This retrospective cohort study included 43 973 singleton live births delivered at the Obstetrics and Gynecology Hospital of Fudan University between September 2017 and December 2022. Participants were categorized into four groups: HH group (HIP with HDP, n=1 011), HIP group ( n=5 469), HDP group ( n=3 486), and control group ( n=34 007). Baseline characteristics and perinatal outcomes were compared using the Chi-square test (or Fisher's exact test). Logistic regression identified risk factors and adverse outcome risks and analyze the impact of HH on neonatal weight. Results:HH accounted for 15.6% (1 011/6 480) of HIP cases, with an overall incidence of 2.3% (1 011/43 973). HIP and HDP were strongly correlated [ OR=1.803, 95% CI: 1.672-1.945]. Advanced maternal age (≥35 years at estimated due date), primiparity, and pre-pregnancy overweight/obesity (body mass index ≥24 kg/m2) were independent risk factors for HH [ OR (95% CI): 1.305 (1.113-1.529), 1.845 (1.545-2.203), and 2.316 (1.981-2.718), respectively]. Compared to the HIP group, the HH group had significantly higher risks of preterm birth [10.3% (104/1 011) vs. 6.3% (344/5 469), OR=1.627 (95% CI:1.280-2.068)], cesarean delivery [57.0% (576/1 011) vs. 41.9% (2 289/5 469), OR=1.701 (95% CI:1.474-1.963)], and neonatal birth weight < P10 [13.9% (141/1 011) vs. 9.0% (494/5 469), OR=1.668 (95% CI:1.336-2.083)]. Stratified analysis revealed a 73.4% increased risk of birth weight < P10 in the gestational diabetes mellitus A1 with HDP subgroup ( aOR=1.734, 95% CI: 1.416-2.125). Conclusions:Advanced age, pre-pregnancy overweight/obesity, and primiparity are risk factors for HH. Compared to isolated HIP, HH is associated with elevated risks of preterm birth, cesarean delivery, and abnormal neonatal birth weight, though the impact on birth weight may vary by HIP subtype.

Objective To explore the correlations between serum soluble growth stimulation gene 2 protein (sST2), brain natriuretic peptide (BNP) and the degree of carotid atherosclerosis in patients with acute ischemic stroke. Methods A total of 205 patients with acute ischemic stroke were selected as study group, and were divided into mild group with 100 cases, moderate group with 70 cases, and severe group with 35 cases according to the severity of carotid atherosclerosis. Another 150 healthy people who received physical examination during the same period were selected as control group. Serum levels of sST2 and BNP were compared among all groups. The correlations of sST2 and BNP levels with the degree of carotid atherosclerosis were analyzed. The influencing factors of different degree of carotid atherosclerosis in patients with acute ischemic stroke were analyzed. The predictive value of sST2 combined with BNP on the degree of carotid atherosclerosis in patients with acute ischemic stroke was analyzed. Results The serum sST2 level of the study group was significantly lower than that of the control group, and the serum BNP level was significantly higher than that of the control group (P < 0.05). The sST2 levels in moderate and severe groups were significantly lower, BNP levels were significantly higher than that in the mild group (P < 0.05). BNP level in the severe group was significantly higher, sST2 level was significantly lower than that in the moderate group (P < 0.05). The degree of carotid atherosclerosis in patients with acute ischemic stroke was correlated with age, history of hypertension, history of smoking, history of diabetes, levels of total cholesterol (TC), C-reactive protein (CRP), homocysteine (Hcy) and low density lipoprotein (LDL) (P < 0.05). Age, hypertension history, smoking history, diabetes history, TC, CRP, Hcy, LDL, BNP and sST2 were the influencing factors for degree of carotid atherosclerosis (P < 0.05). Serum BNP level was positively correlated with the degree of carotid atherosclerosis, while sST2 level was negatively correlated with the degree of carotid atherosclerosis (P < 0.001). The area under the curve (AUC) for the combined prediction of sST2 and BNP was significantly greater than that for the individual prediction of sST2 and BNP (P < 0.001). Conclusion Clinical observation of the changes in serum levels of sST2 and BNP in patients with acute ischemic stroke can effectively assess the progression of the disease, which is beneficial for improving the severity of carotid atherosclerosis in patients. The combined assessment of sST2 and BNP shows good diagnostic performance for evaluating the severity of carotid atherosclerosis in patients with acute ischemic stroke.

Objective:To investigate the effects of customized dental-bone supported osteotomy guide plate in bilateral sagittal split osteotomy.Methods:24 patients(48 sides)with maxillofacial deformity underwent BSSO were included.The maxillofacial region of all patients was scanned by CT,the plaster dental models were scanned using laser surface scanner,and the 3D models were established.The osteotomy guide plates of the inner horizontal and anterior sagittal bone incision of mandible ramus were manufactured by digital technology.All splits underwent operation with(27 side)and without(21 sides)the osteotomy guide plate respectively by the same doc-tor,and the time for the inner horizontal and anterior sagittal bone incision of mandible ramus was recorded.Postoperative CT scan was performed to evaluate the surgical effects according to the lingual split scale(LSS)classification.Results:The wound in all patients healed well and no serious complication was observed.The time for the inner horizontal and anterior sagittal bone incision of mandible ramus in plate group and no plate group was(125.67±2.23)s and(141.15±3.69)s respectively(P＜0.05).The probability of mandi-ble splitting according to Hunsuck standard osteotomy line increased from 42.86％to 66.67％,and the probability of osteotomy line passing through mandibular nerve canal decreased from 33.33％to 7.41％,by using osteotomy guide plate.In addition,LSS4 type of osteotomy line was avoided by using osteotomy guide plates.Data analysis showed that the split pattern of sagittal split osteotomy of the mandibular ramus was influenced by the application of osteotomy guide plates(P＜0.05).Conclusion:The customized dental-bone supported osteotomy guide plate is effective in the completion of the operation and reducing time consuming and surgical complication in BSSO.

Objective:To investigate the current situation of medical social worker supervision in Shanghai, for reference to promote the high-quality development of medical social work.Methods:From June to July 2022, a questionnaire survey was conducted on the in-service medical social workers in all medical institutions with medical social work departments or posts in Shanghai. The questionnaire mainly included demographic information, current status of supervision implementation, and effectiveness of supervision. The data were analyzed descriptively with t test for comparison between groups and the Pearson test was for correlation analysis. Results:A total of 99 medical social workers were included in this study, 65 had received supervision, and medical institutions where 58 people located had established the supervision system. The average scores of actual and expected supervisory support received by medical social workers were 3.71 and 4.20 respectively, and the mean burnout level score was 32.91. The effect of establishing a supervision institutions on burnout was statistically significant ( P<0.05) and actual access to supervision support was negatively associated with burnout ( P<0.05). Conclusions:The overall situation of medical social work supervision in Shanghai was positive, but the demand for supervision was not fully satisfied; Supervision had a positive impact on reducing the burnout level. It is suggested that medical social work should further strengthen the cultivation of supervisory talents, promote the construction of supervisory systems, and improve the quality of supervision.

BackgroundHome-based intervention occupies a prominent place in the treatment of autism spectrum disorder （ASD） in children， whereas previous studies on the effect of home-based early intensive behavioral intervention on verbal ability of children with ASD are somewhat inadequate. ObjectiveTo study the effects of intensive family behavioral of intervention based on Verbal Behavior Milestones Assessment and Placement Program （VB-MAPP） on the language ability of children with ASD， so as to provide references for the development of family intervention strategies for children with ASD. MethodsChildren aged 2 to 3 years old who attended the Children's Rehabilitation Department of the Second Affiliated Hospital of Shantou University Medical College from September 2020 to December 2021 and met the Diagnostic and Statistical Manual of Mental Disorders， fifth edition （DSM-5） diagnostic criteria for ASD were selected as the study objects. A non-randomized concurrent control trial was conducted to compare a study group （n=24） receiving home-based early intensive behavioral intervention using VB-MAPP with a control group （n=14） receiving training from other special institutions or early childhood education institutions. The intervention lasted for 6 months in both groups. Before and after the intervention， the VB-MAPP milestone assessment was performed in the two groups， and the VB-MAPP milestone score， mand， trac and listener responds were selected as the study indicators. Then the intervention effect was compared between two groups， and the multiple linear regression was performed to screen the related influencing factors. ResultsAfter intervention， the total milestone assessment score， mand， tact and listener responds scores of study group were higher than those of control group， with statistical difference （Z=-4.339~-2.195， P<0.05 or 0.01）. Multiple linear regression analysis denoted that the average weekly hours of home-based intervention in the first three months had certain effect on listener responds （β=1.029， P<0.05）. ConclusionApplication of home-based early intensive behavior intervention using VB-MAPP may contribute to the improvement of verbal abilities such as mand， tact and listener responds in children with ASD. ［Funded by 2020 Guangdong Provincial Science and Technology Innovation Strategy Special Fund （number， 20200601）］

The blood brain barrier (BBB) is a physical and metabolic barrier that maintains central nervous system homeostasis and protects brain tissues from potentially hazardous circulating substances. This article reviews the biological characteristics of caludin-5 during cerebral ischemia, its role in BBB integrity and permeability, as well as the research progress of related drug therapy based on calludin-5.

Objective This study focused on the application regularity of medicinal and dietary substances (MDS) of traditional Chinese medicine (TCM) diet therapy during rehabilitation, in order to help patients with Corona Virus Disease 2019 (COVID-19) reduce sequelae and improve their life quality. Methods The official websites of the national and provincial health committees, the website of the National Administration of Traditional Chinese Medicine, the China BioMedical Literature Database (CBM), China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP), and Wanfang Database were used to search the keywords, such as “coronavirus” “novel coronavirus pneumonia” “COVID-19” “protocol” “guideline” “consensus” and “rehabilitation period”. The search time was from the establishment of databases to July 31, 2022. The prevention and control protocols of various provinces and cities were manually supplemented and screened out. The information on the frequency, property, flavor, meridian tropism, and efficacy of MDS was collected for association rule analysis through the Apriori algorithm. Hierarchical cluster analysis was performed using the Euclidean distance and longest distance. Results A total of 18 protocols were screened out, including 56 lists of TCM diet therapy, and 47 kinds of MDS with a frequency of 132 times during the rehabilitation of COVID-19. Among them, six lists of diet therapy were collected from national websites, 26 from local government websites, and 24 from social and academic institution websites. The intended population can be divided into seven categories including normal recovery, lung-spleen Qi deficiency, deficiency of both Qi and Yin, spleen-stomach weakness, deficiency of Yang Qi, kidney Qi deficiency, and blood deficiency. Shanyao (Dioscoreae Rhizoma) and Lianzi (Nelumbinis Semen), followed by Dazao (Jujubae Fructus) were used most commonly in MDS, with mainly flat property, sweet flavor, and spleen and lung meridians in meridian tropism. Besides, deficiency-tonifying drugs were commonly used in MDS. Through association rule analysis, 12 groups of association MDS pairs were obtained. The pair of Yiyiren (Coicis Semen) and Chenpi (Citri Reticulatae Pericarpium) had the highest Lift value, and Yiyiren (Coicis Semen) was used most frequently in the MDS category for eliminating pathogenic factors. The results of complex network analysis showed that the core MDS were Yiyiren (Coicis Semen), Shanyao (Dioscoreae Rhizoma), Huangqi (Astragali Radix), Fuling (Poria), and Dazao (Jujubae Fructus). Three core categories were classified by cluster analysis, including the category of strengthening spleen, nourishing kidney, and grasping Qi, the category of removing phlegm, abating panting, and regulating Qi, and the category of strengthening the middle-energizer and reinforcing Qi. Conclusion Based on the TCM theory, most patients during the rehabilitation of COVID-19 are in a state of lingering pathogens due to deficient vital Qi. TCM diet therapy is based on the principle of “giving both reinforcing and reducing treatment”, and the MDS combinations focus on both reinforcing the health Qi and eliminating pathogenic factors. The diet therapy mainly uses the MDS with flat property and sweet flavor, which belongs to deficiency-tonifying drugs, adding suitable MDS of pathogen-eliminating drugs according to different situations. The ultimate goal is to promote lung inflammation absorption, improve pulmonary fibrosis, increase immunity, reduce the occurrence of sequelae, and improve life quality.