Alzheimer's disease (AD) is the major form of dementia in the elderly and is closely related to the toxic effects of microglia sustained activation. In AD, sustained microglial activation triggers impaired synaptic pruning, neuroinflammation, neurotoxicity, and cognitive deficits. Accumulating evidence has demonstrated that aberrant expression of deubiquitinating enzymes is associated with regulating microglia function. Here, we use RNA sequencing to identify a deubiquitinase YOD1 as a regulator of microglial function and AD pathology. Further study showed that YOD1 knockout significantly improved the migration, phagocytosis, and inflammatory response of microglia, thereby improving the cognitive impairment of AD model mice. Through LC-MS/MS analysis combined with Co-IP, we found that Myosin heavy chain 9 (MYH9), a key regulator maintaining microglia homeostasis, is an interacting protein of YOD1. Mechanistically, YOD1 binds to MYH9 and maintains its stability by removing the K48 ubiquitin chain from MYH9, thereby mediating the microglia polarization signaling pathway to mediate microglia homeostasis. Taken together, our study reveals a specific role of microglial YOD1 in mediating microglia homeostasis and AD pathology, which provides a potential strategy for targeting microglia to treat AD.

Sigma-1 receptor (σ ₁R) has become a focus point of drug discovery for central nervous system (CNS) diseases. A series of novel 1-phenylethan-1-one O-(2-aminoethyl) oxime derivatives were synthesized. In vitro biological evaluation led to the identification of 1a, 14a, 15d and 16d as the most high-affinity (K _i < 4 nmol/L) and selective σ ₁R agonists. Among these, 15d, the most metabolically stable derivative exhibited high selectivity for σ ₁R in relation to σ ₂R and 52 other human targets. In addition to low CYP450 inhibition and induction, 15d also exhibited high brain permeability and excellent oral bioavailability. Importantly, 15d demonstrated effective antipsychotic potency, particularly for alleviating negative symptoms and improving cognitive impairment in experimental animal models, both of which are major challenges for schizophrenia treatment. Moreover, 15d produced no significant extrapyramidal symptoms, exhibiting superior pharmacological profiles in relation to current antipsychotic drugs. Mechanistically, 15d inhibited GSK3β and enhanced prefrontal BDNF expression and excitatory synaptic transmission in pyramidal neurons. Collectively, these in vivo proof-of-concept findings provide substantial experimental evidence to demonstrate that modulating σ ₁R represents a potential new therapeutic approach for schizophrenia. The novel chemical entity along with its favorable drug-like and pharmacological profile of 15d renders it a promising candidate for treating schizophrenia.

Objective: To explore the relationship between dietary behavior, dietary frequency and central precocious puberty (CPP) among children in Wuhan, so as to provide a scientific basis for elucidating the possible and related dietary factors leading to precocious puberty. Methods: From February to December 2023, 100 children with CPP and 100 healthy children were collected in Wuhan Children s Hospital for a case control study. Questionnaires and interviews were used to collect demographic information, child lifestyle, as well as dietary behavior and dietary frequency in the past six months. Conditional Logistic regression model was used to analyze the relationship of children s dietary behavior and dietary frequency with CPP. Results: No associations were found between three dietary behavior, including emotional eating, restrictive eating, and external eating, with CPP at baseline ( P >0.05). In the association analysis of dietary frequency and CPP, the risk of CPP was higher in those who consumed red meat >5 times/week ( OR =1.93, 95% CI =1.01-3.68), carbonated beverages ≥4 times/month ( OR =2.70, 95% CI =1.03-7.08), fruit juices ≥4 times/month ( OR =2.31, 95% CI =1.02-5.25), and nutritional supplementation ( OR =2.77, 95% CI =1.47- 5.22 ), whereas the risk of CPP was lower in those who consumed nuts ≥4 times/month ( OR =0.21, 95% CI =0.08-0.57) ( P < 0.05 ). Conclusion Consumption of higher amounts of red meat, high frequency of carbonated and juice based sugary drinks, and nutrient supplementation may be contributing factors to the development of CPP, whereas regular consumption of nuts is less likely to be associated with the development of CPP.

Objective To investigate the efficacy and safety of camrelizumab monoclonal antibody combined with molecular-targeted therapy in elderly patients with unresectable or advanced hepatocellular carcinoma(HCC).Methods A retrospective analysis was performed for the patients with unresectable/advanced HCC who attended six hospitals from January 1,2019 to March 31,2021,and all patients received camrelizumab monoclonal antibody treatment,among whom 84.8%also received targeted therapy.According to the age of the patients,they were divided into elderly group(≥65 years)and non-elderly group(＜65 years).The two groups were assessed in terms of overall survival(OS),progression-free survival(PFS),objective response rate(ORR),disease control rate(DCR),and immune-related adverse events(irAE).The chi-square test or the Fisher's exact test was used for comparison of categorical data between groups;the independent samples t-test was used for comparison of normally distributed continuous data,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups.The Kaplan-Meier method was used for survival analysis,and the log-rank test was used for comparison of survival curves.Univariate and multivariate Cox proportional hazards regression analyses were used to determine the independent influencing factors for PFS and DCR at 6 months.Results A total of 99 HCC patients were enrolled,with 27 in the elderly group and 72 in the non-elderly group.The elderly group had an OS rate of 67.8%,an ORR of 44.4%,and a DCR of 74.1%at 12 months and a median PFS of 6.4(95%confidence interval[CI]:3.0-12.4)months,with no significant differences compared with the non-elderly group(all P＞0.05).The median OS was unavailable for the elderly group,while the non-elderly group had an OS of 18.9(95%CI:13.0-24.8)months;there was no significant difference between the two groups(P=0.485).The univariate and multivariate Cox regression analyses showed that major vascular invasion(MVI)was an independent risk factor for PFS(hazard ratio[HR]=2.603,95%CI:1.136-5.964,P=0.024)and DCR(HR=3.963,95%CI:1.671-9.397,P=0.002)at 6 months,while age,sex,etiology of HBV infection,presence of extrahepatic metastasis,Child-Pugh class B,and alpha-fetoprotein＞400 ng/mL were not associated with PFS or DCR at 6 months.For the elderly group,the incidence rates of any irAE and grade 3/4 irAE were 51.9%and 25.9%,respectively,with no significant differences compared with the non-elderly group(P＞0.05),and skin disease was the most common irAE in both groups(39.4%).Conclusion Camrelizumab monoclonal antibody combined with molecular-targeted therapy has similar efficacy and safety in patients with unresectable/advanced HCC aged≥65 years and those aged＜65 years.MVI is associated with suboptimal response to immunotherapy and poor prognosis.

Objective Sequencing depth is a key parameter in next generation sequencing,which is closely related to the accuracy of sequencing results.Forensic biological evidence examination requires extremely high accuracy.It is crucial to scientifically and reasonably set the sequencing depth analysis threshold for forensic next generation sequencing testing.Methods This study used targeted sequencing data of microhaplotypes from 50 samples with known genotypes.By calculating the accuracy,precision,recall,and F1 score of each locus under various threshold conditions,two types of analysis threshold setting methods,which were based on fixed read count and fixed sequencing depth ratio,were studied extensively.Results The results showed that false positives were observed when the analysis threshold was set at 50×or 100×.When the analysis threshold was set at 200×,false negatives were observed.When the analysis threshold was set at 1.5%,3.0%,or 4.5%,false positives were observed.This study further proposed a third type of analysis threshold setting method,which was based on sequencing depth ratio scatter plots.With this method,no false positive or false negative was observed in the results.This article then explored four factors that lead to significant differences in the sequencing depth of forensic next generation sequencing experiments,compared with the analysis threshold setting method for capillary electrophoresis technology,and discussed the correlation between analysis thresholds and the ability to distinguish mixed DNA.Conclusion Employing the sequencing depth ratio scatter plot method to set analysis threshold has significant application value in next generation sequencing-based forensic genetic marker genotyping.

Objective:To investigate the surgical method of preoperative computed tomography angiography(CTA) combined with color Doppler ultrasonography(CDU) vascular localization assisted by downstream vascularized medial femoral condylar bone flap and its clinical effect in the treatment of refractory scaphoid fracture nonunion.Methods:Retrospective analysis of case data from January 2018 to June 2022 in the Department of Orthopedics at Xingtai General Hospital of North China Medical and Health Group, using a free medial femoral condylar bone flap with descending knee artery as the blood supply to repair old fractures of the scaphoid bone with bone damage. Prior to surgery, CTA combined with CDU technology was used to locate the descending knee artery, and the medial femoral condylar bone flap was designed based on the localization result and bone defects. Follow-up was conducted using outpatient and telephone method. The degree of wrist pain was evaluated using the visual analogue scale (VAS), wrist function was evaluated using the improved Mayo scoring method, the degree of scaphoid arch back deformity was evaluated based on the angle of the scaphoid bone, and stability between the carpal bones was evaluated based on the angle of the scaphoid moon. The normal distribution metric data was represented as Mean±SD, and paired sample t-tests were used to compare the preoperative CDU measurement and the actual intraoperative measurement of the vessel diameter at the origin of the descending knee artery, as well as the distance to the inferior edge of the medial femoral condyle. The VAS score for pain before and after surgery, the Mayo score for wrist function, wrist grip strength, range of motion, scaphoid angle, and scaphoid angle are also compared. Results:A total of 12 patients were enrolled, including 9 males and 3 females; age ranged from 23 to 56 years old, with an average of 36 years old. There were 7 cases of right side fractures, 5 cases of left side fractures, 8 cases of scaphoid waist fractures, and 4 cases of proximal pole fractures. There was no difference between the preoperative CDU location and the actual vessel diameter at the origin of the descending geniculate artery [(1.7±0.5) mm vs. (1.8±0.7) mm] and the distance to the inferior border of the medial femoral condyle [(11.9±2.1) cm vs. (12.1±1.9) cm]( P>0.05). All patients were followed up after the operation, the specific time ranged from 6 months to 2 years, with an average of 13 months. The X-ray films showed that the scaphoid bone healed in all patients after operation, and the average healing time was 15 weeks. After 6 months of follow-up, the VAS scores (3.7±0.9 vs. 0.5±0.1), Mayo scores (46.1±3.8 vs. 86.2±6.1), scapholunar angle [(65.3±4.1)° vs. (47.9±3.5)°], scaphoid angle [(37.1±3.9)° vs. (22.8±2.3)°], wrist flexion and extension range of motion [(79.0±11.7)° vs. (118.5±15.8)°], grip strength [(6.7±4.6) kg vs. (26.1±5.3) kg]were compared before and after surgery, the differences were statistically significant (all P<0.05). Conclusion:The free medial femoral condyle bone flap with a vascular pedicle is one of the ideal method to repair the old navicular fracture with osteonecrosis. The preoperative vascular positioning technique effectively guides the design and cutting of the bone flap and ensures fast and accurate operation during the operation.

Objective To develop an expert consensus on subcutaneous injection for allergen-specific immunotherapy.Methods Relevant domestic and intemational literature was reviewed,and nursing experts who had experiences in subcutaneous injection of allergen-specific immunotherapy were interviewed to form the initial draft of the consensus.A total of 85 experts from 42 hospitals nationwide were invited to participate in discussions.2 rounds of expert consultations,adjustments,revisions,and improvements were made to the initial draft,and an online meeting was held to form the final version of the consensus.The content approved by more than 75％of the expert group is adopted,or it will be discussed or deleted.Results The expert consensus includes operational standards for subcutaneous injection of allergen-specific immunotherapy,identification and management of adverse reactions,and health education.Conclusion The consensus demonstrates strong scientific rigor and practicality,providing guidance for nursing practices in the field of clinical allergology.

Objective:To investigate the surgical method of preoperative computed tomography angiography(CTA) combined with color Doppler ultrasonography(CDU) vascular localization assisted by downstream vascularized medial femoral condylar bone flap and its clinical effect in the treatment of refractory scaphoid fracture nonunion.Methods:Retrospective analysis of case data from January 2018 to June 2022 in the Department of Orthopedics at Xingtai General Hospital of North China Medical and Health Group, using a free medial femoral condylar bone flap with descending knee artery as the blood supply to repair old fractures of the scaphoid bone with bone damage. Prior to surgery, CTA combined with CDU technology was used to locate the descending knee artery, and the medial femoral condylar bone flap was designed based on the localization result and bone defects. Follow-up was conducted using outpatient and telephone method. The degree of wrist pain was evaluated using the visual analogue scale (VAS), wrist function was evaluated using the improved Mayo scoring method, the degree of scaphoid arch back deformity was evaluated based on the angle of the scaphoid bone, and stability between the carpal bones was evaluated based on the angle of the scaphoid moon. The normal distribution metric data was represented as Mean±SD, and paired sample t-tests were used to compare the preoperative CDU measurement and the actual intraoperative measurement of the vessel diameter at the origin of the descending knee artery, as well as the distance to the inferior edge of the medial femoral condyle. The VAS score for pain before and after surgery, the Mayo score for wrist function, wrist grip strength, range of motion, scaphoid angle, and scaphoid angle are also compared. Results:A total of 12 patients were enrolled, including 9 males and 3 females; age ranged from 23 to 56 years old, with an average of 36 years old. There were 7 cases of right side fractures, 5 cases of left side fractures, 8 cases of scaphoid waist fractures, and 4 cases of proximal pole fractures. There was no difference between the preoperative CDU location and the actual vessel diameter at the origin of the descending geniculate artery [(1.7±0.5) mm vs. (1.8±0.7) mm] and the distance to the inferior border of the medial femoral condyle [(11.9±2.1) cm vs. (12.1±1.9) cm]( P>0.05). All patients were followed up after the operation, the specific time ranged from 6 months to 2 years, with an average of 13 months. The X-ray films showed that the scaphoid bone healed in all patients after operation, and the average healing time was 15 weeks. After 6 months of follow-up, the VAS scores (3.7±0.9 vs. 0.5±0.1), Mayo scores (46.1±3.8 vs. 86.2±6.1), scapholunar angle [(65.3±4.1)° vs. (47.9±3.5)°], scaphoid angle [(37.1±3.9)° vs. (22.8±2.3)°], wrist flexion and extension range of motion [(79.0±11.7)° vs. (118.5±15.8)°], grip strength [(6.7±4.6) kg vs. (26.1±5.3) kg]were compared before and after surgery, the differences were statistically significant (all P<0.05). Conclusion:The free medial femoral condyle bone flap with a vascular pedicle is one of the ideal method to repair the old navicular fracture with osteonecrosis. The preoperative vascular positioning technique effectively guides the design and cutting of the bone flap and ensures fast and accurate operation during the operation.

Objective To prepare rabbit polyclonal antibody against mouse IQ and ubiquitin-like domain-containing protein (IQUB) and detect its expression in the mouse testis. Methods Full-length coding sequence of IQUB was inserted into the pET-30a(+) vector to construct pET-30a-IQUB recombinant prokaryotic plasmid. Transformation of pET-30a-IQUB plasmid into E. coli BL21 was performed, and protein expression was induced with isopropyl-beta-D-thiogalactoside (IPTG). The protein was purified through histidine-tagged fusion protein purification column, then denatured by treatment of urea with gradient concentration. New Zealand rabbits were immunized with the denatured protein to produce IQUB polyclonal antibody. Antibody titer was detected by ELISA, and Western blot analysis and immunofluorescence assay were employed to validate the effectiveness and specificity of IQUB antibody. Results pET-30a-IQUB recombinant plasmid was constructed, and protein expression of IQUB was induced successfully with IPTG. The titer of IQUB polyclonal antibody reached 1:1 000 000. The antibody specifically recognized the endogenous IQUB protein of testis in the wild-type adult mouse. IQUB was expressed in spermatogenic cells of different stages. It was localized in the acrosome and flagellum of mature sperms. Conclusion The highly specific rabbit anti-mouse IQUB polyclonal antibody is successfully prepared, which can be used for Western blot and immunofluorescence histochemistry.
Male ; Rabbits ; Animals ; Mice ; Ubiquitins ; Escherichia coli/genetics* ; Isopropyl Thiogalactoside ; Antibodies ; Enzyme-Linked Immunosorbent Assay

Objective To produce rabbit polyclonal antibody against mouse testis expressed 38 (TEX38). Methods Full-length open reading frame sequence of TEX38 was amplified and inserted into the pET-30a-(+) vector to construct pET-30a-TEX38 prokaryotic plasmid. The recombinant plasmid was transformed into E.coli BL21, and expression was induced with isopropyl β-D-thiogalactopyranoside (IPTG). New Zealand white rabbits were immunized with TEX38 protein after purification and denaturation, then TEX38 polyclonal antibodies were collected from rabbit serum samples. ELISA was performed to detect the antibody titer. Western blot and immunofluorescence staining were performed to determine the specificity of TEX38 polyclonal antibodies. Results The pET-30a-TEX38 recombinant plasmid was constructed, and TEX38 prokaryotic protein was expressed and purified successfully. After immunization, the titer of TEX38 antibody reached 1:1 000 000. Western blot analysis and immunofluorescence staining showed that TEX38 was localized in the mouse spermatogenic cells and sperms with a good specificity. Conclusion The rabbit polyclonal antibody against mouse TEX38 is successfully produced, and the expression of TEX38 in mouse spermatogenic cells and sperms is validated.
Male ; Rabbits ; Animals ; Mice ; Testis ; Antibodies ; Enzyme-Linked Immunosorbent Assay ; Immunization ; Spermatozoa ; Escherichia coli