Visual electrophysiology measurements have become a routine method of functional examination in ophthalmology. Small animal visual electrophysiology is an important tool for exploring the functionality of the visual system in small animals, finding widespread applications in neuroscience and drug research and development. This guide aims to offer a standardized operational guide for the operation of visual electrophysiological norms in small animals to ensure the accuracy and repeatability of the test. The study emphasizes different types of visual electrophysiological tests, such as electroretinogram(ERG)and visual evoked potential(VEP), evoked in small animals, and their application in different disease models. Detailed descriptions are provided regarding the selection and preparation of experimental animals, including the requirements of animal species, anesthesia methods and test environment. In terms of operational procedures, this guide highlights the correct electrode placement, the selection of stimulus parameters, and the key steps for signal acquisition and processing. According to different animal models, the corresponding operation suggestions were provided, and the troubleshooting methods of common problems were introduced. Beyond fundamental operations, this guide also focuses on the interpretation and reporting of test results. It explains various types of electrophysiological waveforms. In summary, this operational specification for small animal visual electrophysiology provides a comprehensive and detailed framework for researchers to ensure the standardization and reliability of tests. By following these guidelines, researchers can effectively utilize small animal visual electrophysiology techniques to gain insight into the function and abnormalities of the visual system.

Fundus photography in small animals holds great significance for studying alterations in the structure and function of retinal blood vessels. However, the absence of uniform operating practices can lead to inconsistencies and incomparability of data. To address this issue, this study aims to develop a standard operation guide for fundus photography in small animals. It will provide researchers with standardized operation procedures and accurate data analysis methods to ensure high accuracy and reproducibility in data collection and analysis. The adoption of these guidelines enables a high level of data consistency while providing more precise results than traditional methods. This guideline bridges the gap in the lack of standard operating norms for fundus photography in small animals, offering researchers a reliable operational framework. This guideline not only helps to improve the accuracy and comparability of data, but also provides strong support for retinal vascular research in clinical practice.

Objective To investigate the neuroprotective effect of Huangpu Tongqiao Capsule(HPTQ)in a rat model of Wilson disease(WD)and explore the underlying mechanisms.Methods SD rat models of WD were established by feeding of copper-supplemented chow diet and drinking water for 12 weeks,and starting from the 9th week,the rats were treated with low-,moderate-and high-dose HPTQ,penicillamine,or normal saline by gavage on a daily basis for 3 weeks.Copper levels in the liver and 24-h urine of the rats were detected,and their learning and memory abilities were evaluated using Morris water maze test.HE staining was used to observe morphological changes of CA1 region neurons in the hippocampus,and neuronal apoptosis was detected with TUNEL staining.Hippocampal expressions of endoplasmic reticulum stress(ERS)-mediated apoptosis pathway-related proteins GRP78,CHOP,caspase-12,cleaved caspase-9,and cleaved caspase-3 at both the mRNA and protein levels were detected using RT-qPCR,immunofluorescence assay or Western blotting.Results Compared with normal control rats,the rat models with copper overload-induced WD exhibited significantly increased copper levels in both the liver and 24-h urine,impaired learning and memory abilities,obvious hippocampal neuronal damage in the CA1 region and increased TUNEL-positive neurons(P<0.01),with also lowered mRNA and protein expressions of GRP78,CHOP,caspase-12,cleaved caspase-9,and cleaved caspase-3 in the hippocampus(all P<0.01).Treatments with HPTQ and penicillamine significantly lowered copper level in the liver but increased urinary copper level,improved learning and memory ability,alleviated neuronal damage and apoptosis in the hippocampus,and decreased hippocampal expressions of GRP78,CHOP,caspase-12,cleaved caspase-9,and cleaved caspase-3 in the rat models(P<0.01 or 0.05).Conclusion HPTQ Capsule has neuroprotective effects in rat models of WD possibly by inhibiting ERS-mediated apoptosis pathway.

Objective To investigate the neuroprotective effect of Huangpu Tongqiao Capsule(HPTQ)in a rat model of Wilson disease(WD)and explore the underlying mechanisms.Methods SD rat models of WD were established by feeding of copper-supplemented chow diet and drinking water for 12 weeks,and starting from the 9th week,the rats were treated with low-,moderate-and high-dose HPTQ,penicillamine,or normal saline by gavage on a daily basis for 3 weeks.Copper levels in the liver and 24-h urine of the rats were detected,and their learning and memory abilities were evaluated using Morris water maze test.HE staining was used to observe morphological changes of CA1 region neurons in the hippocampus,and neuronal apoptosis was detected with TUNEL staining.Hippocampal expressions of endoplasmic reticulum stress(ERS)-mediated apoptosis pathway-related proteins GRP78,CHOP,caspase-12,cleaved caspase-9,and cleaved caspase-3 at both the mRNA and protein levels were detected using RT-qPCR,immunofluorescence assay or Western blotting.Results Compared with normal control rats,the rat models with copper overload-induced WD exhibited significantly increased copper levels in both the liver and 24-h urine,impaired learning and memory abilities,obvious hippocampal neuronal damage in the CA1 region and increased TUNEL-positive neurons(P<0.01),with also lowered mRNA and protein expressions of GRP78,CHOP,caspase-12,cleaved caspase-9,and cleaved caspase-3 in the hippocampus(all P<0.01).Treatments with HPTQ and penicillamine significantly lowered copper level in the liver but increased urinary copper level,improved learning and memory ability,alleviated neuronal damage and apoptosis in the hippocampus,and decreased hippocampal expressions of GRP78,CHOP,caspase-12,cleaved caspase-9,and cleaved caspase-3 in the rat models(P<0.01 or 0.05).Conclusion HPTQ Capsule has neuroprotective effects in rat models of WD possibly by inhibiting ERS-mediated apoptosis pathway.

convalescents, and to screen for broad-spectrum neutralizing antibodies against the SARS-CoV-2 RBD. Methods Using biotinylated RBD as a molecular probe, flow cytometry was employed to perform single-cell sorting of B cells from peripheral blood mononuclear cells (PBMCs) of convalescents. The obtained B cells were lysed and subjected to reverse transcription, followed by nested PCR amplification of the heavy and light chains of antibodies was conducted using random primers. The amplified products were cloned into corresponding expression vectors, and the respective matched heavy-light chain plasmids were co-transfected into 293F cells for expression. Monoclonal antibodies were then purified using Protein A column chromatography. Neutralization experiments were conducted with the wild-type (WT) pseudovirus, and antibodies with IC50<0.1 μg/mL were selected for further testing of neutralizing breadth and potency against the wild-type (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and currently prevalent pseudovirus strains (XBB, BA.5, BF.7). Results A total of 21 RBD-specific monoclonal B cells were obtained from two recovered patients, resulting in the isolation of 13 pairs of antibody light/heavy chains. Nine antibodies were successfully expressed, with P1-A1, P1-B6, and P1-B9 exhibiting IC50 values below 0.1 μg/mL against the pseudovirus of the wild-type strain (WT). Specifically, P1-B6 effectively neutralized the wild-type strain (WT), Beta variant (B.1.351), and Delta variant (B.1.617.2), with IC50 values reaching 0.01 μg/mL. P1-B9 demonstrated effective neutralization against the wild-type strain (WT), Beta variant (B.1.351), Delta variant (B.1.617.2), and Gamma variant (P.1) pseudoviruses, with IC50 values of 0.42 μg/mL, 0.63 μg/mL, 0.28 μg/mL, and 2.50 μg/mL, respectively. Additionally, P1-B6 exhibited good neutralization against BA.5 and BF.7 pseudoviruses, with IC50 values of 0.06 μg/mL and 0.09 μg/mL, respectively. Conclusions Infection with the SARS-CoV-2 WT strain can induce the generation of neutralizing antibodies with broad-spectrum activity. Generating these broadly neutralizing antibodies does not require an excessively high somatic hypermutation. The obtained antibodies can be used as candidates for SARS-CoV-2 diagnosis and prevention.

Epilepsy is a recurrent, transient seizure disorder of the nervous system that affects the intellectual development, life and work, and psychological health of patients. People with epilepsy worldwide experience great suffering. Stressful stimuli such as infection, mental stress, and sleep deprivation are important triggers of epilepsy, and chronic stressful stimuli can lead to frequent seizures and comorbidities. The hypothalamic-pituitary-adrenal (HPA) axis is the most important system involved in the body’s stress response, and dysfunction thereof is thought to be associated with core epilepsy symptoms and related psychopathology. This article explores the intrinsic relationships of corticotropin-releasing hormone, adrenocorticotropic hormone, and glucocorticoids with epilepsy in order to reveal the role of the HPA axis in the pathogenesis of epilepsy. We hope that this information will yield future possible directions and ideas for fully understanding the pathogenesis of epilepsy and developing antiepileptic drugs.

Objective: To investigate the role of methylation of placental glucocorticoid response gene in the association between pregnancy-related anxiety in the third trimester and birth outcomes. Methods: Based on a prospective cohort study, singleton live births and their mothers from the Ma'anshan Birth Cohort Study (MABC) were included as participants in this study. The maternal pregnancy-related anxiety symptoms in the third trimester of pregnancy were evaluated by using the Pregnancy-related Anxiety Questionnaire. The neonatal birth outcomes were collected from medical records. The placental tissues from 300 pregnant women with pregnancy-related anxiety and 300 without pregnancy-related anxiety were collected to detect the methylation of FKBP5, NR3C1 and HSD11B2 genes using the Methyl Target approach. The methylation factors were extracted by exploratory factor analysis. Linear regression or logistic regression models were used to analyze the association between pregnancy-related anxiety in the third trimester, methylation factor scores, and birth outcomes. The mediating role of methylation factors in the association between pregnancy-related anxiety in the third trimester and birth outcomes was analyzed by using the Process procedure. Results: The mean age of 2 833 pregnant women was (26.60±3.60) years old. After adjusting for confounding factors, pregnancy-related anxiety in the third trimester increased the risk of small-for-gestational-age (OR=1.32, 95%CI:1.00-1.74). A total of 5 methylation factors were extracted, and the factor 5 was loaded with FKBP5 CpGs 18-21. Pregnancy-related anxiety in the third trimester was negatively correlated with the factor 5 (β=-0.24,95%CI:-0.44--0.05). The factor 5 was positively correlated with the gestational age (β=0.17, 95%CI:0.06-0.27). In addition, the factor 2 (β=0.02,95%CI:0.00-0.04) and factor 3 (β=0.03,95%CI:0.01-0.05) were positively correlated with 5-min Apgar score after delivery. However, this study did not found the mediating role of the scores of the factor characterized by FKBP5 in the relationship between pregnancy-related anxiety and birth outcomes. Conclusion: Pregnancy-related anxiety in the third trimester may reduce the methylation level of FKBP5 CpGs 18-21 in placental tissues and is associated with the risk of small-for-gestational-age.
Infant, Newborn ; Pregnancy ; Female ; Humans ; Young Adult ; Adult ; Pregnancy Trimester, Third ; Placenta ; Glucocorticoids/metabolism* ; Cohort Studies ; Prospective Studies ; Methylation ; Factor V/metabolism* ; Anxiety/genetics*

Abstract: Objective　To investigate the fundus arteriosclerosis and its influencing factors in HIV/AIDS patients after long-term highly active antiretroviral therapy (HAART). Methods　The clinical basic data and fundus examination data of 203 HIV/AIDS patients before and after HAART in the Fourth People 's Hospital of Nanning from January 2020 to June 2022 were collected to evaluate the occurrence of fundus arteriosclerosis and analyze its influencing factors. Results　Of the 203 HIV/AIDS patients, 159 patients developed fundus arteriosclerosis, with an incidence of 78.33%, including 33 patients with grade Ⅰ(20.75%), 87 patients with grade Ⅱ (54.72%), 28 patients with grade Ⅲ(17.61%), and 11 patients (6.92%) with Grade Ⅳ. Before HAART, there was no significant difference in CD4+T lymphocyte count, CD8+T lymphocyte count, viral load, white blood cell count, platelet count, hemoglobin, serum creatinine, blood urea nitrogen, triacylglycerol, total cholesterol, fasting blood glucose, alanine aminotransferase, aspartate aminotransferase and serum total bilirubin between the atherosclerosis group and normal group (P>0.05). After 6 months of HAART, CD8+T lymphocyte count, triacylglycerol and fasting blood glucose in atherosclerosis group were significantly higher than those in normal group (P<0.05). In the stratified comparison of CD4+ and CD8+ lymphocyte counts after 6 months of HAART, the proportion of patients with CD4+ lymphocyte count (CD4+)<200 (cells/μL) in the atherosclerosis group was significantly higher than that in the normal group; the proportion of patients with CD4+ lymphocyte count (CD4+)≥500 (cells/μL) was significantly lower than that in the normal group; the proportion of patients with CD8+ lymphocyte count CD8+≥ 800/μL was significantly higher than that in the normal group (all P<0.05). Binary logistic regression analysis showed that opportunistic infection, HIV course, CD4+T, CD8+T lymphocyte count after HAART and triglyceride were independent risk factors for ocular fundus atherosclerosis in HIV/AIDS patients (all P<0.05). Conclusions　The incidence of ocular fundus arteriosclerosis is high in HIV/AIDS patients. More than 4 years of HIV course, combined opportunistic infection, Low CD4+T lymphocyte count after 6 months of HAART, high CD8+T lymphocyte count and high triglyceride level are independent risk factors for ocular fundus arteriosclerosis in HIV/AIDS patients. Fundus screening should be performed before and after HAART in such population, HAART program should be formulated for the risk of cardiovascular disease, and risk management of cardiovascular disease should be strengthened during treatment to improve patient outcomes.

Abstract: Objective　To construct SARS-CoV-2 receptor binding domain molecular probe for monoclonal memory B cell sorting and obtain RBD specific neutralizing antibodies from peripheral blood mononuclear cells (PBMCs) of COVID-19 convalescents by single-cell sorting. Methods　The SARS-CoV-2 RBD sequence was downloaded from GenBank, and the Avi tag and 6-histidine tags were added at the C-terminal. After codon optimization, it was chemically synthesized, cloned into the pDRVI1.0 vector, expressed after transfection of 293F cells, and biotinylated consequently. RBD-specific B cells were sorted out with this probe1 from the PBMCs of convalescents recovered from COVID-19. After B cells were lysed, the variable regions of heavy chain and light chain were amplified, cloned into the antibody expression vector, and transfected into 293F cells to express the antibody. Then the antibody was purified from the supernatant using protein A column and SARS-CoV-2 pseudovirus was used to test their neutralizing activity. Results　RBD-Avi probe was produced and successfully biotinylated sequentially with an efficiency of 30%-50%. Western blot analysis revealed that the biotinylated probe was recognized by the antibodies purified from COVID-19 convalescent plasma. Using this probe, 7 and 16 RBD-specific memory B cells were successfully isolated from the PBMCs of two convalescent individuals, accounting for 0.24% and 0.17% of the total cell population, respectively. After amplifying the variable regions of antibody heavy and light chains from the lysed B cells, 7 and 12 pairs of antibody heavy-light chains were obtained. A total of 16 antibodies were expressed in the convalescent individuals, and most of the purified antibodies showed neutralizing activity against the pseudovirus, with IC50 values of 6 antibodies below 1 μg/mL. The IC50 values of XJ-A9 and SCF-F1 against the wild-type pseudovirus were 0.07 μg/mL and 0.35 μg/mL, respectively. Conclusion　The SARS-CoV-2 RBD molecular probe constructed in this study has good antigenicity, and the isolated antibodies present neutralizing activity against wild-type SARS-CoV-2 pseudovirus.