ObjectiveTo explore the mechanism by which the Shenfu Xiongze prescription regulates autophagy in rats with myocardial remodeling through the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） signaling pathway. MethodsA rat model of myocardial remodeling induced by isoprenaline （ISO） was established. Rats were divided into the blank group，the model group，the low-，medium-， and high-dose groups of Shenfu Xiongze prescription，and the captopril group， 6 rats in each group. Except for the blank group，the rat model of myocardial remodeling was established in the other groups by intraperitoneal injection of 2.5 mg·kg^-1 ISO for 3 consecutive weeks. At the same time of modeling， the low-，medium-， and high-dose groups of Shenfu Xiongze prescription were administered the corresponding doses of Shenfu Xiongze prescription solution （8.4，16.8，and 33.6 g·kg^-1），and the captopril group was administered captopril solution （25 mg·kg^-1）. As for the blank group and the model group， the same volume of normal saline was given. The treatment was continued for 3 weeks. Echocardiography was used to observe the cardiac structure and function，and the heart weight index was detected. Masson staining and hematoxylin-eosin （HE） staining were used to observe the pathological morphology changes of myocardial tissue. The levels of interleukin-6 （IL-6） and B-type natriuretic peptide （BNP） in serum were detected by enzyme-linked immunosorbent assay （ELISA）. The expression of type Ⅰ collagen （Collagen Ⅰ），type Ⅲ collagen （Collagen Ⅲ），and microtubule-associated protein 1 light chain 3 （LC3） proteins in myocardial tissue was determined by immunohistochemistry. Autophagy was observed by transmission electron microscopy. The mRNA expression of Collagen Ⅰ，Collagen Ⅲ，α-smooth muscle actin （α-SMA），LC3，yeast Atg6 homolog protein （Beclin-1），AMPK，and mTOR in myocardial tissue was detected by quantitative real-time polymerase chain reaction （real-time PCR）. The protein expression of Collagen Ⅰ，α-SMA，transforming growth factor-β₁ （TGF-β₁），LC3，Beclin-1，p62， phosphorylation（p）-AMPK，p-mTOR，AMPK，and mTOR was detected by Western blot. ResultsCompared with the normal group，rats in the model group exhibited significantly decreased values of ejection fraction （EF） and left ventricular fractional shortening （FS） （P<0.01）， significantly increased values of left ventricular end-diastolic diameter （LVIDd） and left ventricular end-systolic diameter （LVIDs） （P<0.01）. Additionally， the model group also showed increased degrees of inflammatory infiltration and fibrosis of myocardial tissue， significantly elevated levels of serum IL-6 and BNP （P<0.01）， significantly increased mRNA and protein levels of Collagen Ⅰ，Collagen Ⅲ，α-SMA，and mTOR （P<0.01），and markedly decreased mRNA and protein levels of LC3，Beclin-1，and AMPK （P<0.05，P<0.01）. Compared with the model group， the low-，medium-， and high-dose groups of Shenfu Xiongze prescription presented significantly elevated EF and FS values （P<0.01） and lowered LVIDd and LVIDs （P<0.05）. In these groups， the inflammation and fibrosis were alleviated significantly. They also exhibited decreased serum levels of IL-6 and BNP （P<0.01）， significantly reduced protein expression of Collagen Ⅰ， α-SMA， TGF-β₁， p62， and p-mTOR （P<0.01）， significantly decreased mRNA expression of Collagen Ⅰ， Collagen Ⅲ， α-SMA， and mTOR （P<0.01）， and significantly increased mRNA and protein levels of LC3， Beclin-1， and AMPK （P<0.05，P<0.01）. ConclusionThe Shenfu Xiongze prescription can improve the myocardial remodeling induced by ISO in rats by regulating the autophagy level，enhance cardiac function，and reduce inflammatory and fibrotic levels. This effect may be achieved through the AMPK/mTOR signaling pathway.

Diabetic retinopathy （DR） is a microvascular complication of diabetes and one of its most common complications. Prolonged hyperglycemia induces oxidative stress， inflammatory responses， apoptosis， and pathological angiogenesis， ultimately disrupting the blood-retinal barrier（BRB） and leading to visual impairment or even blindness. Recent studies show that the nuclear factor erythroid 2-related factor 2 （Nrf2） signaling pathway plays an important role in the development of DR's pathological changes. Meanwhile， Chinese herbal monomers have been shown to modulate the Nrf2 signaling pathway， thereby intervening in the development of DR. In terms of inhibiting oxidative stress， saponin compounds such as platycodin-D and ginsenoside Rb1 downregulate the expression of malondialdehyde （MDA）， thereby ameliorating retinal oxidative stress. Flavonoids such as total flavonoids from Pueraria lobata flower and puerarin upregulate the expression of superoxide dismutase （SOD） and glutathione peroxidase （GPx）， effectively clearing lipid peroxides. Regarding the suppression of inflammation， phenolic compounds like resveratrol and chlorogenic acid inhibit the nuclear factor kappa B （NF-κB） pathway， reducing the release of tumor necrosis factor-alpha （TNF-α） and mitigating inflammatory responses. In the context of inhibiting apoptosis， polysaccharides such as Polygonatum sibiricum polysaccharide and Angelica sinensis polysaccharide downregulate the expression of the pro-apoptotic protein Bcl-2-associated X protein （Bax） and suppress the activity of the executioner Caspase-3， thereby reducing the apoptosis rate. As for the inhibition of neovascularization， compounds including bilobalide and physcion significantly decrease the protein expression of vascular endothelial growth factor （VEGF）， leading to a reduction in retinal pathological angiogenesis. Furthermore， Chinese herbal compound prescriptions such as Tongluo Zhujing pills， Yiqi Huoxue Yangyin decoction， Qiming granules， and Danlou tablets can also intervene in the onset and progression of DR through the mechanisms described above. In summary， both Chinese herbal monomers and Chinese herbal compound prescriptions can modulate the Nrf2 signaling pathway to inhibit oxidative stress， alleviate inflammation， and participate in maintaining BRB integrity， suppressing retinal neovascularization， and preventing neurodegeneration， thereby delaying the progression of DR. Therefore， this paper reviews and summarizes recent studies at home and abroad on how traditional Chinese medicine （TCM） works to treat DR， and the relationship between the Nrf2 pathway and DR. It aims to provide research ideas for preventing and treating DR.

Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals ; Swine ; Antibodies, Neutralizing/immunology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Immunoglobulin Fab Fragments/immunology* ; Antibodies, Viral/immunology* ; Picornaviridae/immunology* ; Humans ; HEK293 Cells ; Swine Diseases/diagnosis* ; Picornaviridae Infections/diagnosis*

Hepatic fibrosis is a chronic pathological condition characterized by hepatic stellate cell activation and excessive deposition of extracellular matrix， which would progress to liver cirrhosis and even hepatocellular carcinoma. Therefore， reversal of hepatic fibrosis is of great importance for improving quality of life and prolonging survival time. Currently， various therapeutic drugs for hepatic fibrosis have entered the stage of clinical trial. This article reviews the research advances in therapeutic drugs for hepatic fibrosis in the recent years， in order to provide insights into the treatment of hepatic fibrosis and future research directions for drugs.

Objective:To explore the correlations of sleep quality and architecture with heart rate variability (HRV) in patients with stenoses of vertebrobasilar artery system and internal carotid artery system.Methods:A retrospective study was performed; 72 patients with stenosis or occlusion of the head and neck arteries (not resulting in cerebral infarction) admitted to Department of Neurology, Second Affiliated Hospital of Zhengzhou University from June 2023 to June 2024 were chosen, including 33 patients with moderate-to-severe stenosis or occlusion of the vertebrobasilar system (VB group) and 39 patients with moderate-to-severe stenosis or occlusion of the internal carotid artery system (ICA group). Pittsburgh sleep quality index (PSQI) and polysomnography (PSG) were used to evaluate the sleep quality and architecture, respectively; and 24-hour ambulatory electrocardiogram was used to assess the HRV. Differences in PSQI score, PSG and HRV parameters between the two groups were compared; partial correlation analysis was used to explore the correlations of HRV parameters with PSQI scores and PSG parameters; multivariate linear regression was used to analyze the independent influencing factors for HRV.Results:(1) Compared with the ICA group, the VB group exhibited significantly higher PSQI scores, spontaneous arousal index (SAI), ratio of time of stage 1 non-rapid eye movement sleep/total sleep time (T N1/T t), and apnea-hypopnea index (AHI), while significantly lower ratio of time of rapid eye movement sleep/total sleep time (T R/T t), spindle wave density in stage 2 non-rapid eye movement sleep (N2), lowest blood oxygen saturation, standard deviation of normal to normal intervals (SDNN) of all sinus beats, low-frequency power (LF), and high-frequency power (HF, P<0.05). (2) In both VB group and ICA group, SDNN was negatively correlated with PSQI score ( r=-0.461, P=0.020; r=-0.378, P=0.036). In the VB group, SDNN was negatively correlated with T N1/T t ( r=-0.467, P=0.019) and SAI ( r=-0.551, P=0.004), and positively correlated with ratio of time of stage 3 non-rapid eye movement sleep/total sleep time (T N3/T t, r=0.686, P<0.001) and spindle wave density in N2 ( r=0.518, P=0.008); LF and HF were negatively correlated with SAI ( r=-0.481, P=0.015; r=-0.564, P=0.003). In the ICA group, HF was negatively correlated with spindle wave density in N2 ( r=-0.369; P=0.041). (3) Multivariate linear regression results indicated that T N3/T t (β=0.348, P=0.018), SAI (β=-0.330, P=0.018), and spindle wave density in N2 (β=0.286, P=0.013) were independent influencing factors for Ln_SDNN in patients with moderate-to-severe stenosis or occlusion of the vertebrobasilar system. Conclusion:Patients with stenosis or occlusion of the vertebrobasilar system exhibit poorer subjective sleep quality, increased light sleep, heightened arousal, and reduced sleep stability compared with those with stenosis or occlusion of the internal carotid artery system, which may be caused by the imbalance of autonomic nerve function.

Objective:To investigate the prevalence of sarcopenia and potential influencing factors in middle-aged and elderly populations in Zhejiang Province.Methods:Data were obtained from Zhejiang Provincial Household Economic Status Survey, a cross-sectional survey was condcuted in middle-aged and olde adults selected through multi-stage sampling in three cities in Zhejiang (Huzhou, Jiaxing and Shaoxing) in July 2023. A total of 3 019 study participants, average age 62.3 years old, 53.5% men, were included according to the inclusion and exclusion criteria. Sarcopenia screening was conducted by using the questionnaire with five sarcopenia related-items. Univariable and multivariable logistic regression analysis was used to identify the factors associated with sarcopenia.Results:The prevalence of sarcopenia in the middle-aged and old study participants was 4.47%. Significant differences were observed between the participants with or without sarcopenia in terms of age, educational level, BMI, alcohol consumption status, diet habit, physical activity level, sleep quality, number of chronic diseases, childhood socioeconomic status, adulthood community socioeconomic status, muscle strength, walking assistance, ability to stand from seat, ability to climb stairs, and fall frequency ( P<0.05). Multivariable logistic regression analysis revealed that old age (≥75 years: OR=2.82, 95% CI: 1.60-4.97), low body weight ( OR=1.96, 95% CI: 1.06-3.62), unhealthy diet habit ( OR=1.57, 95% CI: 1.01-2.46), physical inactivity ( OR=5.80, 95% CI: 3.09-10.88), poor or very poor sleep quality ( OR=1.65, 95% CI:1.23-2.41), number of chronic diseases (1 chronic disease: OR=1.84, 95% CI: 1.08-3.14; 2 chronic diseases: OR=3.22, 95% CI: 1.81-5.71; 3 or more chronic diseases: OR=3.74, 95% CI: 2.11-6.65), poor childhood socioeconomic status ( OR=2.98, 95% CI: 1.23-7.20), and poor adulthood community socioeconomic status ( OR=3.87, 95% CI: 1.63-9.17) were significant risk factors for sarcopenia. Conclusions:The prevalence of sarcopenia was relatively low in middle-aged and old population in Zhejiang. Age, BMI, unhealthy diet, physical activity level, sleep quality, number of chronic diseases, childhood socioeconomic status, and adulthood community socioeconomic status were identified as significant influencing factors.

Objective:To investigate the association between blood pressure variability (BPV) and all-cause mortality and cardiovascular events in patients undergoing continuous ambulatory peritoneal dialysis (CAPD), and provide reference for clinical management in CAPD patients.Methods:This retrospective cohort study included patients who received CAPD at Guangdong Provincial People's Hospital between May 1, 2010, and July 31, 2023. Baseline and clinical data of the patients were collected. Coefficient of variation of systolic blood pressure (CVSBP) was used to assess BPV. The patients were divided into CVSBP T1, CVSBP T2 and CVSBP T3 groups based on CVSBP tertiles, and the differences among the three groups were compared. Diastolic blood pressure and mean arterial pressure were used to further assess BPV and sensitivity analysis was conducted. The primary endpoint was the composite outcome of all-cause mortality and cardiovascular events. Kaplan-Meier survival curve and Cox regression analysis were used to analyze the association between CVSBP and the primary endpoint.Results:A total of 358 CAPD patients were included, with age of (43.6±13.3) years, and 197 males (55.0%). The proportion of males, proportion of smoking, baseline blood urea nitrogen, serum creatinine and serum albumin in CVSBP T2 (9.08%≤CVSBP<12.55%, n=120) group and CVSBP T3 (CVSBP≥12.55%, n=119) group were lower than those in CVSBP T1 group (CVSBP<9.08%, n=119), and baseline systolic blood pressure, residual kidney Kt/V and total Kt/V were higher than those in CVSBP T1 group, with statistically significant difference among the three groups (all P<0.05). During follow-up of 37(23, 76) months, 49 patients (13.7%) experienced the composite endpoint events, including 12 patients (3.4%) of all-cause deaths and 42 patients (11.7%) of cardiovascular events. Kaplan-Meier survival analysis showed that the incidence of composite endpoint events in CVSBP T3 group was higher than that in CVSBP T1 group and CVSBP T2 group, but the difference was not statistically significant (Log-rank χ2=3.795, P=0.150). Multivariate Cox regression analysis showed that, after adjusting for age, sex, diabetes, baseline systolic blood pressure, residual renal function, and serum albumin, as a continuous variable, CVSBP was not associated with the risk of composite outcome in CAPD patients ( HR=1.058, 95% CI 0.985?1.135, P=0.122); as a categorical variable, with CVSBP T1 group as reference, CVSBP T2 group and CVSBP T3 group were not associated with the risk of composite outcome ( HR=1.222, 95% CI 0.471?3.167, P=0.681; HR=1.827, 95% CI 0.737?4.530, P=0.193). The sensitivity analysis showed that increased variability of diastolic blood pressure ( HR=1.162, 95% CI 1.063?1.270, P=0.021) and increased variability of mean arterial pressure ( HR=1.114, 95% CI 1.030?1.204, P=0.007) were correlated with higher risk of composite outcome in CPAD patients. Conclusions:Systolic blood pressure variability during follow-up is not associated with risk of composite outcome of all-cause mortality and cardiovascular events in CAPD patients. Increased variability of diastolic blood pressure and increased variability of mean arterial pressure are associated with a higher risk of composite outcome in CPAD patients. Interventions to reduce BPV may be helpful to improve the long-term prognosis of CAPD patients.

This study aimed to identify,characterize,and functionally analyze the CYP450 gene fam-ily within Hyalomma asiaticum unfed nymphs(HasiUN)under regular physiological conditions,laying the groundwork for further study into the biosynthesis and metabolism of exogenous sub-stances associated with the CYP450 gene in HasiUN.Through our methods,we identified and characterized HasiUNCYP450 genes.We then annotated them using the gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG).This was achieved with the help of bioinforma-tics,based on the transcriptome of 2-week-old HasiUN.Further on,we conducted a preliminary a-nalysis of tissue expression,distribution,and potential function with qRT-PCR and network inter-action predictions.Our analyses showed that 87 HasiUN CYP450 genes,ranging in size from 1089 to 1692 bp,encoded predominantly basic,hydrophilic,and unstable protein sequences.These se-quences had nearly identical numbers of transmembrane regions,both with and without a signal peptide.Phylogenetic and domain analyses indicated that HasiUN CYP450,having a complete CYP450 domain,were distributed among various clans,with most members found in clan 3.CYP20 clans,present in 13 species from 2 families and 7 genera,required either one-host or three-hosts to undergo their full life cycle.MEME analysis results hint at different clans having variations in the amount and combination of motifs.Expression level analysis showed that HasiUN CYP450 was expressed at different levels,with the highest distribution of normally expressed genes in clan 3 and low expression of all mito clan genes.Functional annotation revealed HasiUN CYP450 key as-sociation with biological processes,metabolism,and organismal systems according to the GO and KEGG annotations.Distribution results showed that HasiUN CYP20a1 was expressed in specific tissues,including salivary glands and ovaries.Functional analysis points to CYP20a1's role in UV-induced toxic metabolism,endocrine disruptor metabolism,and hazardous substance metabolism in the nervous system.Our research presents the first analysis of HasiUN CYP450 gene's biological characteristics and expression pattern under normal physiological conditions.Additionally,investi-gations into the distribution and function of a new category,HasiUN CYP20a1,were embarked upon.These findings establish a theoretical foundation for continued functional study of HasiUN CYP45 gene.

Objective:To observe the high-risk histopathological feature (HRF) and their correlation with prognosis in children with intraocular retinoblastoma (RB) in the intraocular stage after failed eyepreserving treatment and enucleation surgery.Methods:A single-center retrospective case study. From August 2018 to January 2023, 64 children (64 eyes) with advanced intraocular RB who were diagnosed in Department of Ophthalmology of Beijing Children's Hospital and underwent enucleation surgery after failed eye-preserving treatment were included in the study. The median follow-up time was 51 months. The gender of the children patients, the age of visit and enucleation, International Intraocular Retinoblastoma Classification (IIRC), the initial chemotherapy regimen (hereinafter referred to as "chemotherapy"), the time of enucleation surgery, pathological results, post-enucleation treatment methods and prognosis were collected. The Mann-Whitney U test was used for comparison between groups. Survival analysis was performed using the Kaplan-Meier method, and the log-rank test was used for comparison between groups. Results:Among 64 cases and 64 eyes, 37 were male and 27 were female. The age of seeking medical treatment was 20 (11-31) months. The age at which the surgery was performed was 29 (16-40) months. The number of eyes in IIRC stage D and E was 16 and 48 respectively. The initial chemotherapy regimens simply applied (hereinafter referred to as "alone") intravenous systemic chemotherapy (IVC) and ophthalmic artery infusion chemotherapy (IAC) in 40 cases and 11 cases, 13 cases of IVC+IAC. All patients with positive HRF received systemic adjuvant chemotherapy after surgery. There were 37 eyes (57.8%, 37/64) positive for HRF. There was no statistically significant difference in the positive rate of HRF between children in IIRC stage D and stage E ( χ2=0.021, P=0.884). Among the 37 eyes with HRF, the numbers of eyes with extensive choroidal invasion, posterior lamina cribrosa optic nerve invasion, scleral invasion and optic nerve stump involvement were 17 (45.9%, 17/37), 16 (43.2%, 16/37), 3 (8.1%, 3/37) and 3 (8.1%, 3/37), respectively. During the follow-up period, there were 5 cases (7.8%, 5/64) of extraocular metastasis of the tumor and death, all of which were stage E and had HRF. Among them, the initial treatment plan was IAC for 4 cases, one case of IVC. The survival rates of children among the IVC, IAC or IVC+IAC regimens were 97.5% (39/40), 63.6% (7/11), and 100.0% (13/13), respectively. The comparison of survival rates among different chemotherapy regimens showed statistically significant differences ( χ2=14.233, P<0.001). The results of survival analysis showed that the cumulative survival rate of those with extensive choroidal invasion, posterior lamina cribrosa optic nerve infiltration, and those who received IAC was significantly lower than that of those without extensive choroidal invasion, posterior lamina cribrosa optic nerve infiltration, and those who received IVC+IAC and IVC ( P<0.05). Conclusions:Eye-preserving treatment for children with advanced intraocular RB may increase the positive rate of HRF and the risk of extraocular metastasis. The IVC+IAC eye-protecting treatment plan can improve the survival rate of children.

Objective:To investigate the clinical and pathological significance of the DICER1 mutation in follicular thyroid carcinoma (FTC).Methods:Sixty-eight cases of primary FTC resected between 2009 and 2023 were retrieved from The Affiliated Hospital of Qingdao University, Qingdao, China. Sanger sequencing was performed to identify DICER1 and TERT promoter mutations in all cases. Cases with DICER1 or TERT promoter mutations were subject to additional examination of potential mutations in KRAS, HRAS, and NRAS. The clinical and pathological features of DICER1-mutant FTCs were then analyzed. The relationship between DICER1 mutations and TERT-promoter/RAS mutations was also assessed.Results:DICER1 mutations were detected in 16 of the 68 FTC cases (23.5%), with 11 near E1813 at exon 25, 6 near D1709 at exon 24, and 1 in the splice region of exon 25. Two cases harbored two (distinct) mutations. All patients with DICER1-mutant FTC were female. Compared with patients with DICER1-wild-type FTC, those with DICER1-mutant were much younger, and had a higher proportion of minimally invasive subtype. Nine FTCs with DICER1 mutations were subject to further sequencing on adjacent non-cancerous tissues or lymph node tissues, but no mutations were detected. TERT-promoter or RAS hotspot mutations were not identified in any of the DICER1-mutant cases. However, TERT-promoter mutation was found in 6 DICER1-wild-type cases (8.8%, 6/68), with 3 cases also having RAS hotspot mutations and exhibiting highly aggressive biological behaviors.Conclusion:DICER1 mutations may occur in FTCs and appear mutually exclusive with RAS and TERT-promoter mutations, warranting further study as RAS-like mutations.