ObjectiveTo investigate the mechanism by which Mingshi prescription regulates the retinal melanopsin-dopamine （Opn4-DA） axis in myopic mice to inhibit endoplasmic reticulum （ER） stress in the retina and sclera， thereby delaying axial elongation associated with myopia. MethodsSixty 4-week-old male SPF-grade C57BL/6J mice were randomly divided into a normal group， a form-deprived myopia group （FDM group）， an intrinsically photosensitive retinal ganglion cells ablation group （ipRGCs group）， a Mingshi Prescription group （MSF group， 5.2 g·kg^-1）， and an ipRGCs + MSF group （5.2 g·kg^-1）. Except for the normal group， all other groups underwent FDM modeling. Additionally， the ipRGCs and ipRGCs + MSF groups received retinal ipRGC ablation. Three weeks after modeling， the MSF and ipRGCs + MSF groups were administered Mingshi prescription via continuous gavage for six weeks. After refraction and axial length were measured in all mice， eyeballs were collected along with retinal and scleral tissues. Pathological and morphological changes in the retina， choroid， and sclera were observed using periodic acid-Schiff （PAS） staining. Western blot was employed to detect the relative protein expression levels of dopamine D1 receptor （DRD1）， C/EBP homologous protein （CHOP）， and glucose-regulated protein 78 （GRP78） in the retina， and CHOP and GRP78 in the sclera. Real-time PCR was used to detect the relative mRNA expression of Opn4， CHOP， and GRP78 in the retina， and CHOP and GRP78 in the sclera. Immunofluorescence staining （IF） was performed to detect the expression of Opn4 and DRD1 in retinal tissues. ResultsCompared with the normal group， the FDM group showed a significant myopic shift in refraction （P<0.05） and a significant increase in axial length （P<0.05）. The retinal layers were thinner， the number of ganglion cells was reduced， and collagen fibers in the sclera were loosely arranged with evident gaps. Opn4 and DRD1 protein and mRNA expression in the retina were significantly decreased （P<0.05）， while CHOP and GRP78 protein and mRNA expression in both retinal and scleral tissues were significantly increased （P<0.05）. Compared with the FDM group， the ipRGCs group exhibited further increases in myopic refraction and axial length （P<0.05）， more pronounced thinning and looseness in the retinal， choroidal， and scleral layers， lower expression of Opn4 and DRD1 protein and mRNA in the retina （P<0.05）， and higher expression of CHOP and GRP78 protein and mRNA in the retina and sclera （P<0.05）. Compared with the FDM group， the MSF group showed significantly reduced refractive error and axial length （P<0.05）， with improved cellular number， arrangement， and thickness in ocular tissues， increased Opn4 and DRD1 protein and mRNA expression in the retina （P<0.05）， and reduced CHOP and GRP78 protein and mRNA expression in both retina and sclera （P<0.05）. Similarly， the ipRGCs + MSF group showed significant improvements in terms of the above items compared with the ipRGCs group （P<0.05）. ConclusionMingshi Prescription delays myopic axial elongation and refractive progression by regulating the Opn4-DA axis in the retina of myopic mice， thereby inhibiting ER stress in the retina and sclera. This intervention promotes Qi and blood nourishment of the eyes， softens the fascia， and restores ocular rhythm.

Objective:To investigate long-term auditory changes and characteristics of Alport syndrome（AS） patients with different degrees of renal injury. Methods:Retrospectively analyzing clinical data of patients diagnosed AS from January 2007 to September 2022, including renal pathology, genetic detection and hearing examination. A long-term follow-up focusing on hearing and renal function was conducted. Results:This study included 70 AS patients, of which 33（25 males, 8 females, aged 3.4-27.8 years） were followed up, resulting in a loss rate of 52.9%.The follow-up period ranged from 1.1to 15.8 years, with 16 patients followed-up for over 10 years. During the follow-up, 10 patients presenting with hearing abnormalities at the time of diagnosis of AS had progressive hearing loss, and 3 patients with new hearing abnormalities were followed up, which appeared at 5-6 years of disease course. All of which were sensorineural deafness. While only 3 patients with hearing abnormalities among 13 patients received hearing aid intervention. Of these patients,7 developed end-stage renal disease（ESRD）, predominantly males （6/7）. The rate of long-term hearing loss was significantly different between ESRD group and non-ESRD group（P=0.013）. There was no correlation between the progression of renal disease and long-term hearing level（P>0.05）. kidney biopsies from 28 patients revealed varying degrees of podocyte lesion and uneven thickness of basement membrane. The severity of podocyte lesion was correlated with the rate of long-term hearing loss（P=0.048）, and there was no correlation with the severity of hearing loss（P>0.05）. Among 11 cases, theCOL4A5mutationwas most common （8 out of 11）, but there was no significant correlation between the mutation type and hearing phenotype（P>0.05）. Conclusion:AS patients exhibit progressive hearing loss with significant heterogeneity over the long-term.. THearing loss is more likely to occur 5-6 years into the disease course. Hearing abnormalities are closely related to renal disease status, kidney tissue pathology, and gene mutations, emphasizing the need for vigilant long-term hearing follow-up and early intervention.
Male ; Child ; Female ; Humans ; Nephritis, Hereditary/pathology* ; Retrospective Studies ; Kidney ; Deafness ; Hearing Loss/genetics* ; Kidney Failure, Chronic/pathology* ; Mutation

Objective:To summarize initial experience of applying nanopore third-generation sequencing detection method (nanopore sequencing) for genetic diagnosis of non-classical 21 hydroxylase deficiency (NC 21-OHD), and to explore its performance and application prospects.Methods:Clinical data of the two NC 21-OHD patients, who were hospitalized at the First Affiliated Hospital of Zhengzhou University in May 2019, were collected. Peripheral venous blood was collected and genome DNA extracted. Genetic variants was detected by nanopore sequencing and underwent bioinformatic analysis. Pathogenetic mutations in CYP21A2 gene were validated with PCR-sanger sequencing in the two patients and their parents.Results:The average reads length and sequence depth in the patient one was 12, 792 bp and 27.19×. The average reads length and sequence depth in the patient two was 13, 123 bp and 21.34×. Compound variants of c.293-13C>G/c.844G>T (p.Val282Leu) and c.332_339delGAGACTAC (p.Gly111Valfs)/c.844G>T (p.Val282Leu) were detected in these two patients, which were consistent with clinical phenotype of NC 21-OHD. Further analysis showed that c.293-13C>G mutation was inherited from her father and c.844G>T (p.Val282Leu) mutation was inherited from her mother for the patient one. The c.844G>T (p.Val282Leu) mutation was inherited from her father and c.332_339delGAGACTAC (p.Gly111Valfs) mutation from her mother.Conclusions:The heterozygous mutations in CYP21A2 gene are the cause of NC 21-OHD in these two patients. Nanopore sequencing technique is a reliable new detection method for patients with NC 21-OHD.

Objective:To investigate the effect of long non-coding RNA (lncRNA) human histocompatibility leukocyte antigen complex P5 (HCP5) on the neuronal injury induced by oxygen glucose deprivation/reoxygenation (OGD/R), and to analyze its potential mechanism.Methods:SH-SY5Y cells were divided into control group (CON group, normal medium culture), model group (Model group, OGD/R), interference control group (si-NC group, OGD/R after HCP5 small interfering RNA negative control (si-NC)), HCP5 interference group (si-HCP5 group, OGD/R after HCP5 small interfering RNA (si-HCP5)), HCP5 interference+ inhibitor control group (si-HCP5+ anti-NC group, OGD/R after transfection of si-HCP5, miR-525-5p inhibitor negative control (anti-NC)), HCP5 interference+ miR-525-5p inhibitor group (si-HCP5+ anti-miR-525-5p group, OGD/R after transfection of si-HCP5, miR-525-5p inhibitor). qRT-PCR was used to detect the expression of lncRNA HCP5 and miR-525-5p in cells.The activity of SH-SY5Y cells was detected by MTT.The level of reactive oxygen species (ROS) in the cells was detected by fluorescent probe of dichlorofluorescein diacetate (DCFH-DA). The apoptosis of SH-SY5Y cells was detected by flow cytometry.Western blot was used to detect the expression of BTG2, Bcl-2 related X protein (Bax), B lymphocyte tumor 2 (Bcl-2) and cleaved caspase-3 protein.SPSS 25.0 software was used to analyze the data, and one-way ANOVA was used for comparison between multiple groups, and SNK- q test was used for further comparison between two groups. Results:There were statistically significant differences in lncRNA HCP5, miR-525-5p RNA levels and BTG2 protein expression levels among the 6 groups ( F=28.853, 59.241, 13.731, all P<0.001). Compared with the CON group, the Model group had higher level of lncRNA HCP5, lower level of miR-525-5p, and higher level of BTG2 protein (all P<0.05). Compared with the Model group, the si-HCP5 group had lower level of lncRNA HCP5, higher level of miR-525-5p, and lower level of BTG2 protein (all P<0.05). Compared with the si-HCP5+ anti-NC group, the si-HCP5+ anti-miR-525-5p group had higher level of lncRNA HCP5, lower level of miR-525-5p, and higher level of BTG2 protein (all P<0.05). There were statistically significant differences in cell activity and ROS levels among the six groups of cells ( F=16.180, 59.950, both P<0.001). The cell activity of the Model group was lower than that of the CON group (0.33±0.12, 0.63±0.11) ( P<0.05), and the ROS level was higher than that of the CON group (224.62±23.27, 100.00±0.00) ( P<0.05). The cell activity of the si-HCP5+ anti-miR-525-5p group was lower than that of the si-HCP5+ anti-NC group (0.38±0.08, 0.58±0.08) ( P<0.05), and the ROS level was higher than that of the si-HCP5+ anti-NC group (207.83±19.39, 135.27±14.36) ( P<0.05). There were statistically significant differences in the apoptosis rate and expression levels of apoptotic proteins Bcl-2, Bax, and cleared Caspase-3 among the six groups of cells ( F=27.994, 29.660, 45.000, 52.983, all P<0.001). There were no statistically significant difference in Bax, Bcl-2, cleared Caspase-3 protein levels, and apoptosis rate in SH-SY5Y cells between the Model group and the si-NC group, as well as between the si-HCP5 group and the si-HCP5+ anti-NC group (all P>0.05). Compared with the CON group, the apoptosis rate, levels of Bax and cleared Casase-3 protein in the Model group were significantly upregulated (all P<0.05), while the Bcl-2 protein level was significantly downregulated ( P<0.05). Compared with the Model group and si-NC group, the si-HCP5 group showed significant downregulation of cell apoptosis rate and levels of Bax and cleared Caspase-3 protein (all P<0.05), while the Bcl-2 protein level was upregulated ( P<0.05). Compared with the si-HCP5 group and si-HCP5+ anti-NC group, the si-HCP5+ anti-miR-525-5p group showed significant upregulation of cell apoptosis rate and levels of Bax and cleared Caspase-3 protein levels (all P<0.05), and significant downregulation of Bcl-2 protein levels ( P<0.05). Conclusion:lncRNA HCP5 may inhibit the expression of BTG2 by targeting up-regulation of miR-525-5p, thus leading to apoptosis of nerve cells in OGD/R models.

Objective:To investigate the risk factors for bleeding and thrombosis during extracorporeal membrane oxygenation (ECMO) therapy in critically ill patients and determine the best predictors of coagulation-related complications.Methods:A retrospective analysis was performed on patients who received ECMO for respiratory or circulatory failure at Xiangya Hospital of Central South University from January 2020 to December 2022. The outcome was whether bleeding or thrombosis occurred from 24 h after ECMO insertion to before weaning. The differences in demographic characteristics, weaning conditions, prognosis, routine blood tests, organ function, coagulation and blood product transfusion of each group were compared. Logistic regression analysis was used to evaluate the risk factors for bleeding and thrombosis, and ROC curve evaluation was used to assess their capacity to predict complications.Results:A total of 61 patients with ECMO were enrolled, with 21 cases of bleeding and 14 cases of thrombosis during ECMO. Compared with the nonbleeding group, the activated partial thromboplastin time, thromboplastin time (TT), and transfusions of frozen plasma and red blood cells were higher in the bleeding group (all P<0.05). Compared with the nonthrombotic group, the increase in body weight, D-dimer (DD), fibrinogen degradation product (FDP), and improvement of arterial oxygen partial pressure (ΔPO 2) within 24 h were significantly higher in the thrombotic group (all P<0.05). Logistic regression analysis revealed that TT ( OR=1.039, 95% CI: 1.006~1.072, P=0.018) and frozen plasma transfusion volume ( OR=1.046, 95% CI: 1.010-1.083, P=0.012) were risk factors for bleeding events. FDP ( OR=1.030, 95% CI: 1.009-1.051, P=0.005), DD ( OR=1.181, 95% CI: 1.044-1.336, P=0.008), and ΔPO 2 ( OR=1.007, 95% CI: 1.002-1.012, P=0.006) were risk factors for thrombosis. According to ROC curve analysis, the AUCs of TT, frozen plasma transfusion, and combined indexes for predicting bleeding were 0.712, 0.690, and 0.816, respectively. The combined indices had a cut-off value of 0.273, a sensitivity of 75.61%, and a specificity of 80.00%. The AUCs of FDP, DD, ΔPO 2, and combined FDP with ΔPO 2 for predicting thrombosis were 0.778, 0.748, 0.786, and 0.868, respectively. The cut-off value of the combined index was 0.157, the sensitivity was 68.09%, and the specificity was 92.86%. Conclusions:TT combined with frozen plasma transfusion volume predicted bleeding optimally, while FDP plus ΔPO 2 predicted thrombotic events better during ECMO treatment in critically ill patients.

【Objective】 To analyze the risk factors of postpartum stress urinary incontinence (SUI) and to establish a nomogram model. 【Methods】 A total of 278 puerpera who gave birth at our hospital during Dec.2018 and Aug.2020 were selected as the modeling group, and 132 puerpera who gave birth during Sep.2020 and Sep.2021 were involved in the verification group. Factors affecting postpartum SUI were identified with univariate and multivariate logistic regression, and a nomogram prediction model was constructed with R software. The predictive effectiveness and discrimination of the model were assessed, and the decision curve analysis (DCA) was drawn to evaluate the clinical application value of the model. 【Results】 A total of 84 cases (30.22%) in the modeling group developed SUI 2 months after delivery. Fetal weight, delivery method, maternal age, mobility (Δhy) and rotation Angle (Δβ) were factors affecting postpartum SUI (P<0.05). Multivariate logistic regression analysis showed that increased fetal weight, normal delivery, increased Δhy, and increased Δβ were independent risk factors of postpartum SUI (P<0.05). The constructed nomogram fitted well. The H-L fit curve of the modeling group and verification group were (χ²=7.514, P=0.312) and (χ²=6.157, P=0.267), respectively. The area under the receiver operating characteristic curve of the modeling group and verification group were 0.815 and 0.760, respectively, indicating high specificity and consistency. DCA indicated that when the high-risk threshold probability of the model was between 0.06-0.80, the nomogram model had a high clinical value. 【Conclusion】 Increased fetal weight, normal delivery, increased Δhy and elevated Δβ are independent risk factors that affect postpartum SUI. The nomogram model constructed has good predictive effectiveness and discrimination, and high clinical application value.

OBJECTIVE To investigated the anti-de-pressant effects of the fruit Areca catechu L.(ACL)and elucidated its potential underlying mechanism using a rat model of chronic unpredictable mild stress(CUMS).METHODS CUMS was induced in rats to establish a depression animal model for 28 d.According to the baseline sucrose preference,the male rats were divided into six different groups.They were treated with parox-etine hydrochloride,ACL,and water once a day until the behavioral tests were performed.The levels of corticoste-rone(CORT),malondialdehyde(MDA),catalase(CAT),and total superoxide dismutase(T-SOD)in serum were de-tected using a commercial kit,and the concentrations of 5-hydroxytryptamine(5-HT)and dopamine(DA)mono-amine neurotransmitters in the brain tissues were detect-ed by liquid chromatography-tandem mass spectrometry.Doublecortin(DCX)expression in the hippocampal den-tate gyrus(DG)was determined by immunofluorescence,and the relative abundance of brain-derived neurotrophic factor(BDNF),TrkB,PI3K,p-Akt/Akt,PSD-95,and p-GSK-3β/GSK-3β of brain tissues were assayed by West-ern blotting.RESULTS ACL markedly increased sucrose preference,decreased the immobility time,and short-ened the feeding latency of CUMS-induced rats.CUMS induction resulted in marked changes in the contents of the monoamine neurotransmitters(5-HT and DA)in the hippocampus and cortex of brain tissues and the levels of CORT,MDA,CAT,and T-SOD in serum,whereas ACL administration alleviated these considerable changes.ACL promoted DCX expression in DG and increased the protein levels of BDNF,TrkB,PI3K,p-Akt/Akt,PSD-95,and p-GSK-3β/GSK-3β in the brains of CUMS-induced rats.CONCLUSION Our results indicated that ACL may improve depression-like behaviors in CUMS-induced rats by decreasing the hyperfunction and oxidative stress of the hypothalamic-pituitary-adrenal axis,stimulating hippo-campal neurogenesis,and activating the BDNF signaling pathway.

Centenarians are known as the "model population" in the study of healthy aging. Understanding their characteristics of health and longevity and its related influencing factors would facilitate the exploration of the possible path to achieve health and longevity under the background of population aging in China. In recent years, the population based observational studies, including China Hainan Centenarian Cohort Study (CHCCS), have found the main health characteristics of centenarians and their relationship with quality of life. This paper summarizes the relevant research results from CHCCS in recent years, and compare them with the results of similar domestic studies. The main health indicators include blood pressure, blood lipid, blood glucose, hemoglobin, body measurement and renal function. The health outcomes including diet, nutrition, health-related quality of life, activities of daily living, geriatric syndrome. The results are expected to provide epidemiological evidence for the development of healthy aging policies.

Objective:To explore the association between parathyroid hormone (PTH) and all-cause death in the centenarians in Hainan Province.Methods:The study was based on China Hainan Centenarian Cohort Study, a total of 1 002 centenarians were included in the baseline survey, the follow up was conducted to collect the information about their death outcome. The association between baseline PTH level and all-cause death was analyzed by Cox proportional risk regression model using continuous variables, dichotomous variables, and quad variables as independent variables of the centenarians.Results:As of May 31 st, 2021, a total of 522 centenarians had died in the follow up period, the median of the follow up time was 4.16 (1.31,5.04) years, and the mortality rate was 52.10%. After fully adjusting the covariates, the multiple Cox proportional hazards regression analyses showed that the risk of all-cause death increased by 3.7% for every 10 pg/ml increase in PTH value, the HR was 1.037 (95% CI: 1.007-1.067) in all the centenarians. The risk of death in the centenarians in the highest PTH group was 1.458 (95% CI: 1.131-1.878) times higher than that in the lowest PTH group. Conclusions:Higher parathyroid hormone level was associated with the elevated risk for all-cause death in the centenarians in Hainan. PTH can be used as a reference index for the prediction of the death risk of long-lived elderly in community.

Objective:To examine the association between depression and all-cause death in centenarians in Hainan Province.Methods:Based on China Hainan Centenarian Cohort Study, 1 002 centenarians with integral data were included in the baseline survey, follow up was conducted to collect the information about their survival status and death outcome. Depression was evaluated by the 15-item Geriatric Depression Scale (GDS-15) and based on the evaluation scores, the mental status of the centenarians was classified as: severe depressive disorder (MDD; score ≥10), minor depressive disorder (MnDD; score 6-9), and normal status (score ≤5). Multivariable-adjusted Cox proportional hazards regression models was used to estimate the hazard ratios ( HRs) and 95% CIs. Results:As of May 31 st, 2021, a total of 522 deaths had been reported, with a mortality density of 152.39/1 000 person-years and a median follow up time M( Q1, Q3) of 4.16 (1.31, 5.04) years, in the centenarians. After adjusting the factors of socio-demographic characteristics and lifestyle, multivariate Cox regression analysis showed that the risk of all-cause mortality increased by 3.1% for every 1-point increase in GDS-15 score ( HR=1.031, 95% CI: 1.002-1.061). Compared with centenarians with normal status, the risk of mortality in the centenarians with MDD increased by 35.9% ( HR=1.359, 95% CI: 1.023-1.806), the centenarians with MDD had an average 1.94 years of life lost. Among the female centenarians, the risk of mortality in MnDD group and MDD group increased by 29.3% ( HR=1.293, 95% CI: 1.053-1.590) and 42.3% ( HR=1.423, 95% CI: 1.047-1.935) compared with normal group, those with MDD had an average 1.71 years of life lost. Conclusions:Depression was related to the increased risk of all-cause death and life lost in the centenarians in Hainan.