ObjectiveTo investigate the awareness and clinical practice of guidelines for the prevention and treatment of chronic hepatitis B （2022 edition） among clinicians. MethodsFrom July 19 to December 31， 2024， a self-designed electronic questionnaire was distributed via the WeChat mini program to collect related data from 1 588 clinicians nationwide， including their awareness and practice based on 18 questions regarding testing and referral， diagnosis and treatment， and follow-up. ResultsAmong all respondents， only 350 clinicians correctly understood all the updated key points of antiviral indications and treatment for special populations in the 2022 edition of guidelines for the prevention and treatment of chronic hepatitis B， with an overall awareness rate of 22.0%. Only 20% ‍—‍ ‍40% of the patients with positive HBV DNA and an age of >30 years receive antiviral therapy， while 80% ‍—‍ ‍100% of the patients with positive HBV DNA and a family history of hepatitis B cirrhosis or hepatocellular carcinoma receive antiviral therapy. The median follow-up rates at 1 year， 3 years， and 5 years were 67.5% 57.5% and 47.5%，respectively， showing a trend of gradual reduction， which might be associated with the influencing factors such as insufficient time for follow-up management by clinicians， insufficient awareness of the disease among patients， and poor adherence to follow-up. ConclusionThere is a gap between the awareness and practice of guidelines for the prevention and treatment of chronic hepatitis B （2022 edition） among clinicians. It is recommended to further strengthen training and focus on the whole process of “detection， diagnosis， treatment， and management” for patients with chronic hepatitis B in healthcare institutions， in order to promote the implementation of the guidelines.

Objective: To explore the longitudinal associations between dietary macronutrients and lung function in schoolaged children, so as to provide the nutritional research evidence for promoting children s lung health. Methods: In November 2021, two primary schools located in Chengdu, Sichuan Province were selected from the Southwest China Childhood Nutrition and Growth (SCCNG) cohort by a stratified cluster random sampling method, enrolling a total of 1 112 school aged children aged 8 to 13 years. At baseline, the dietary and sociodemographic characteristics of the children were assessed. One year later, the forced vital capacity (FVC) of the children was measured and converted into Z scores (FVC- Z ), while the vital capacity index (VCI) was also calculated. Generalized linear regression analysis was employed to examine the associations between dietary macronutrients and lung function, considering interactions with gender and age, followed by stratified analysis. Results: After adjusting for confounding factors, the analysis results of the generalized linear regression model showed that the carbohydrate energy ratio was negatively correlated with FVC- Z ( β =-0.02) and VCI ( β =-0.16), while the fat energy ratio showed a positive correlation with FVC- Z ( β =0.03) and VCI ( β =0.23) ( P <0.05). The protein energy ratio was positively correlated with FVC- Z ( β =0.09) and VCI ( β =0.60) specifically in girls ( P <0.05). Additionally, there was an interaction effect of age on the associations between macronutrients and lung function ( P <0.01); in children aged 8-9 and 10-11, the carbohydrate energy supply ratio was negatively correlated with FVC- Z ( β =-0.04, -0.03) and VCI ( β =-0.29, -0.21), and fat energy supply ratio was positively correlated with FVC- Z ( β =0.07, 0.05) and VCI ( β =0.46, 0.32) ( P <0.05). Conclusions There are age and sex differences in the association of dietary macronutrients with lung function, with a low carbohydrate, high fat diet promoting lung function in children. Additionally, protein intake appears to have a positive influence on the lung function of girls. The early school age period may represent a critical window for dietary interventions aimed at promoting lung health.

Objective:To establish a fluorescent quantitative RT-PCR assay based on N_sgRNA of SARS-CoV-2 and preliminarily apply it on real samples.Methods:Recombinant plasmid, specific primers and probes of N_sgRNA were designed and synthesized based on Wuhan-Hu-1/2019_MN908947 and synthesis mechanism of subgenomic RNA (sgRNA). Using recombinant plasmid as amplification templates, the optimal reaction conditions and reaction system were screened according to the Ct value, fluorescence intensity, and shape of amplification curve and was evaluated for sensitivity, reproducibility, and specificity. Meanwhile, the possibility of practical application of the method was explored by testing 172 clinical samples and 256 municipal wastewater samples. Results:A qRT-PCR assay for N_sgRNA in SARS-CoV-2 was initially established. The detection limit of the assay was 20 copies/mL, and the variation coefficients of in-batch (0.002%~0.767%) and batch to batch repetition (0.016%~0.752%) were less than 1%. Only N_sgRNA recombinant plasmid was detected in the specificity assay. So the method is more highly sensitive, specific and reproducible. The RatiosgRNA/ gRNA of clinical samples HK.3, EG.5.1, JN.1 and their sub-lineages and their corresponding urban sewage samples in epidemic period were significantly different ( P<0.05). There is a strong correlation between the median of RatiosgRNA/ gRNA in clinical samples and sewage samples in the same period (correlation coefficient r=1.000, P=0.010). Conclusions:In this study, a qRT-PCR method for detecting N_sgRNA of SARS-CoV-2 was established and the method has the characteristics of higher sensitivity, stronger specificity and better repeatability, and it can be used to detect SARS-CoV-2 infectivity.

Objective:To investigate the relationship between screen time and content, and the mental health status of adolescents. The findings will inform the formulation of targeted intervention policies to enhance adolescent mental health.Methods:Between September and November 2023, 5 197 students from 64 junior high, senior high, and vocational schools across 13 districts in Wuhan were recruited, using the stratified whole-cluster random sampling to investigate their screen behavior and mental health status. Mental health status was measured using the Mental Health Inventory for Chinese Middle School Students (MMHI-60). A generalized additive model was used to explore the nonlinear association between screen time and mental health status. Additionally, a mixed-effects model was utilized to explore the dose-response associations between average daily total screen time, screen time for different content types, and adolescents' mental health status and the impact of the proportion of different screen contents on mental health outcomes.Results:The age of the participants was (14.40±1.48) years, with 56.07% being boys. The MMHI-60 score averaged 1.73±0.70. The M( Q1,Q3) for daily total screen time was 50.00 (0.00,128.57) minutes. The M( Q1,Q3) for screen time dedicated to gaming, studying, socializing, and watching videos were 0.00 (0.00, 20.00), 8.57 (1.64, 44.50), 4.28 (0.00, 30.00), and 0.00 (0.00, 25.71) minutes, respectively. A non-linear association was observed between average daily screen time and adolescent mental health problem score, 0-1 hour of daily screen time was beneficial for adolescent mental, compared to no screen time. However, screen time exceeding 1 hour was detrimental, with the negative impact increasing alongside screen time duration. When total daily screen time was held constant, the proportion of time spent on gaming ( β=0.14, 95% CI: 0.05-0.23, P=0.003) and video ( β=0.21, 95% CI: 0.09-0.28, P<0.001) was positively correlated with mental health problems, whereas the proportion of time spent on studying was negatively correlated with mental health problems ( β=-0.17, 95% CI: -0.24 - -0.11, P<0.001). Conclusions:Moderate screen time is advantageous for adolescent mental health. However, it is crucial to minimize the proportion of screen time dedicated to video and gaming activities to mitigate potential adverse effects.

Objective To construct a Gabra3 gene knockout cell model and explore transcriptomic and proteomic alterations in murine neuronal cells,in order to investigate the molecular mechanisms underlying the increased depth of slow-wave sleep observed following Gabra3 deletion.Methods Multiple sgRNA sequences were designed,and the CRISPR/Cas9 system was used to knock out the Gabra3 gene in the murine GT1-7 neuronal cell line.Gene sequencing was performed to assess knockout efficiency,and TA cloning was used to validate the knockout results.Protein immunoblotting experiments were conducted to confirm the knockout,while cell proliferation assays were used to validate the knockout phenotype.Total RNA and protein were extracted for transcriptomic and proteomic sequencing,respectively.A range of bioinformatics analyses was conducted to assess the functional consequences of Gabra3 knockout in GT1-7 neuronal cells.Results Following Gabra3 gene knockout,pathways related to cortisol and aldosterone synthesis and secretion,as well as circadian rhythm,were significantly enriched.Three key genes,BMP2,GLI2,and DLL1,were identified.Proteomic profiling revealed widespread disturbances in protein expression following Gabra3 knockout.Conclusion Gabra3 gene knockout may increase slow-wave sleep depth by modulating the expression of hormone secretion-related genes and altering circadian regulatory pathways.

Objective:To evaluate the efficacy and long-term outcomes of fludarabine, cyclophosphamide, and rituximab (FCR) in treatment-na?ve patients with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) .Methods:Clinical data from 68 CLL/SLL patients treated with FCR at Jiangsu Province Hospital (August 2008–May 2021) were retrospectively analyzed to assess efficacy, safety, and survival outcomes.Results:Among 68 patients [46 males, 22 females; median age 55 (47, 60) years], 13.1% (8/61) had a complex karyotype, 32.3% (20/62) had immunoglobulin heavy variable region mutated (IGHV-M) type, 6.6% (4/61) had del (17p), and 14.8% (8/54) had del (11q). Patients received a median of 6 (4, 6) FCR cycles. The overall response rate was 88.2% (60/68), including 47.0% (32/68) complete remissions. Over a median follow-up of 82 (59, 98) months, 66.2% (45/68) experienced disease progression. Median progression-free survival was 56 (21, 123) months, while median overall survival was not reached. The 5- and 10-year PFS rates were 42.6% (95% CI: 31.9–56.8% ) and 28.7% (95% CI: 19.0–43.4% ), respectively. Poor PFS was associated with del (17p) ( HR=5.04, 95% CI: 1.72–14.74, P=0.003), del (11q) ( HR=5.27, 95% CI: 2.11–13.15, P<0.001), IGHV unmutated (IGHV-UM) ( HR=4.11, 95% CI: 1.72–9.79, P=0.001), complex karyotype (CK) ( HR=3.53, 95% CI: 1.58–7.85, P=0.002), β 2-microglobulin >3.5 mg/L ( HR=2.87, 95% CI: 1.37–6.01, P=0.005). In multivariate analysis, IGHV-UM remained an independent predictor of PFS ( HR=8.63, 95% CI: 1.09–68.40, P=0.042). Sixteen patients with IGHV-M and lacking del (17p) or CK had a median PFS of 123 (58,123) months and a 5-year PFS rate of 70.7% (95% CI: 49.7–99.1% ), reaching a plateau after 5 years with no recurrences by 10 years. Common grade 3–4 adverse events included hematologic toxicity (44.1%, 30/68), infection (36.7%, 25/68), and liver dysfunction (4.4%, 3/68). Among 25 patients receiving single-agent BTK inhibitors after FCR progression, median follow-up was 45 (26, 64) months; 36% (9/25) experienced disease progression, with a median PFS time of 55 (27, 55) months. Conclusion:First-line FCR provides durable long-term benefits for patients with IGHV-M CLL without del (17p) or CK.

Alzheimer's disease (AD) is a neurodegenerative disease with clinical hallmarks of progressive cognitive impairment. Synergistic effects of the Aβ-Tau cascade reaction are tightly implicated in AD pathology, and microglial NLRP3 inflammasome activation drives neuronal tauopathy. However, the underlying mechanism of how Aβ mediates NLRP3 inflammasome remains unclear. Herein, we determined that oligomeric Aβ (o-Aβ) bound to microglial Kv2.1 and promoted Kv2.1-dependent potassium efflux to activate NLRP3 inflammasome resulting in neuronal tauopathy by using Kv2.1 inhibitor drofenine (Dfe) as a probe. The underlying mechanism has been intensively investigated by assays with Kv2.1 knockdown in vitro (si-Kv2.1) and in vivo (AAV-ePHP-si-Kv2.1). Dfe deprived o-Aβ of its capability to promote microglial NLRP3 inflammasome activation and neuronal Tau hyperphosphorylation by inhibiting the Kv2.1/JNK/NF-κB pathway while improving the cognitive impairment of 5×FAD-AD model mice. Our results have highly addressed that the Kv2.1 channel is required for o-Aβ-driven microglial NLRP3 inflammasome activation and neuronal tauopathy in AD model mice and highlighted that Dfe as a Kv2.1 inhibitor shows potential in the treatment of AD.

Objective To explore the correlation between changes in gut microbiota and the occurrence of chronic renal failure(CRF)in elderly patients,providing theoretical evidence for the prevention and treatment of CRF in the elderly.Methods A total of 57 elderly patients with CRF admitted to Department of Nephrology,Jilin People's Hospital from June 2021 to June 2023 were selected as observation group,while 65 healthy individuals who underwent physical examinations at the same hospital during the same period were chosen as control group.Baseline data,gut microbiota composition,and biochemical indicators were compared between two groups.Results The levels of γ-glutamyl transferase,creatinine,urea,uric acid,N-terminal pro-brain natriuretic peptide,parathyroid hormone,hypersensitive C-reactive protein,and serum phosphorus were higher in observation group than those in control group,whereas serum calcium was lower;all differences were statistically significant(P＜0.05).Firmicutes and Bacteroidetes were dominant in two groups,accounting for 49.22％and 43.12％in observation group and 49.47％and 42.01％in control group,respectively.At the genus level,Bacteroides predominated,comprising 8.69％and 9.68％of the total,respectively.Multivariate Logistic regression analysis showed that Actinobacteria(95％CI:0.015-0.391,P=0.002),Lactobacillus(95％CI:0.224-0.577,P＜0.001),Bifidobacterium(95％CI:0.150-0.933,P=0.035),and Streptococcus(95％CI:0.256-0.687,P-0.001)were negatively associated with the occurrence of CRF in the elderly,whereas Clostridium(95％CI:2.176-15.047,P＜0.001)and Enterococcus(95％CI:1.579-7.224,P=0.002)were positively associated.Conclusion Changes in gut microbiota are closely related to the development of CRF in the elderly.Clinically,attention should be paid to the cultivation and identification of gut microbiota in this population,in order to provide guidance for the diagnosis and treatment of intestinal flora disorders in patients with CRF,guide probiotic application,and formulate microecological regulation programs.

BACKGROUND:The cervical facet joint,as an important anatomical structure of the posterior column of the cervical spine,plays an important role in neck activity,stress transmission,and maintaining cervical stability. In recent years,anatomical and biomechanical studies have shown that asymmetry of cervical facet joints can cause degeneration of facet joints,which may be the main cause of cervical spine degeneration in young people. Existing research is mostly focused on adults,and there are also reports on preschool and school-age children in China,while there are few reports on the morphological parameters of cervical facet joints in adolescents.OBJECTIVE:Through three-dimensional reconstruction of the cervical facet joints in adolescents,measuring their relevant morphological parameters,and comparing them with those in children and adults,we explored the age-related changes in the morphological development of cervical facet joints,providing a theoretical basis for the diagnosis,treatment,and prevention of cervical spondylosis arising from cervical facet joints.METHODS:A total of 62 adolescents aged 13-18 years were selected to undergo spiral CT scan of cervical vertebrae and 3D reconstruction,requiring no bone destruction,tumor,deformity,or fracture,no changes in vertebrae morphology and structure,no previous spinal operations. The guardian's informed consent to the experimental protocol was obtained. By age group,group A was 13-14 years old;group B was 15-16 years old;group C was 17-18 years old. Thecorrelation morphometry and statistical analysis of C2-C7 facet joints were performed in adolescents of each group.RESULTS AND CONCLUSION:(1) In three groups of subjects,the facet joint surface heights and widths displayed decreasing and increasing trends in relation to the change of vertebra order. The facet joint surfaces on the inferior surface showed larger height and width compared to the corresponding indicators on the superior surface. (2) The intra-articular height of the articular process was lowest in C5 among the three groups of ages,and it showed a positive correlation with age. (3) Among the three groups,the gaps between the articular surfaces of the joints in C4-5 of group A,C3-4 of group B,and C4-5 of group C weresignificantly larger than the rest of the gaps in each group. Except for C4-5,there were no significant differences between the two groups. Except for C2-3,the remaining gaps between the vertebrae in group C were significantly larger than those in the two groups. (4) It is indicated that the morphology of the cervical facet joint surface gradually transitions from circular to elliptical as the vertebral order increases. In inter-group comparison,facet joint surface height is significantly affected by age compared to facet joint surface width. The area of the lower facet joint surface of each segment is greater than that of the upper facet joint surface,with only significant differences in the shape and area of C4-5 and C5-6. In addition,the minimum height of the facet joint is located at C5,and the significantly widened gap between the facet joint surfaces is mainly located at C3-4 and C4-5. Therefore,cervical instability often occurs at the mid-level.

Objective To clarify the active ingredients and the potential molecular mechanism of Guben Qushi Huayu Prescription in treating psoriasis recurrence.Methods An ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS)was applied to analyze the whole formula and the constituents absorbed into blood of Guben Qushi Huayu Prescription,and molecular docking technology was used to study the binding affinity of the constituents absorbed into blood with psoriasis-related immunomodulatory proteins such as CD69 and CD103 proteins.Results Mass spectrometry analysis identified 21 active ingredients such as paeoniflorin in Guben Qushi Huayu Prescription,including several known anti-inflammatory and immunomodulatory compounds.Analysis of the constituents absorbed into blood identified 11 ingredients,including paeoniflorin,that may affect the course of psoriasis through blood circulation.Molecular docking studies revealed that the constituents absorbed into blood,including astilbin,isoastilbin,chlorogenic acid,neochlorogenic acid,cryptochlorogenic acid,helicine,paeoniflorin,ononin,all had high binding affinities with CD69 and CD103 proteins.Conclusion This research reveals the main active ingredients of Guben Qushi Huayu Prescription and their potential mechanism for regulating the recurrence of psoriasis by mass spectrometry and molecular docking technology,contributing to providing scientific basis for further pharmacological research and clinical application.