OBJECTIVE: To observe the effects of acupuncture on podocyte autophagy and long non-coding RNA SOX2 overlapping transcript (LncRNA SOX2OT)/mammalian target of rapamycin C1 (mTORC1)/Unc-51-like kinase 1 (ULK1) pathway in rats with diabetic kidney disease (DKD), and to explore the mechanism by which acupuncture reduces urinary protein. METHODS: A total of 40 SPF-grade male Sprague-Dawley rats were randomly divided into a control group (n=10) and a modeling group (n=30). The DKD model was established by feeding a high-fat, high-sugar diet combined with intraperitoneal injection of streptozotocin (STZ) in the modeling group. Twenty rats with successful DKD model were randomly divided into a model group (n=10) and an acupuncture group (n=10). The acupuncture group received "spleen and stomach-regulating" acupuncture at bilateral "Zusanli" (ST36), "Fenglong" (ST40), "Yinlingquan" (SP9), and "Zhongwan" (CV12), 30 min per session, once daily, five times per week, for four weeks. The general condition, fasting blood glucose (FBG), 2-hour postprandial glucose (2hPG), serum creatinine (SCr), blood urea nitrogen (BUN), 24-hour urinary protein quantification, and urine albumin-to-creatinine ratio (UACR) were compared before and after the intervention. After intervention, urinary podocyte injury marker SPON2 was measured by ELISA. Podocyte autophagosomes and glomerular basement membrane ultrastructure in renal tissue were observed via transmission electron microscopy. Podocyte apoptosis was assessed by TUNEL staining. The protein expression of microtubule-associated protein 1 light chain 3Ⅱ (LC3-Ⅱ), mTORC1, ULK1, Beclin-1, and p62 in renal tissue was detected by Western blot. LncRNA SOX2OT expression in renal tissue was measured by real-time PCR. RESULTS: After the intervention, compared with the control group, the model group exhibited increased food and water intake, increased urine output, weight loss, and loose stools; compared with the model group, the food and water intake, urine volume, and loose stools were improved in the acupuncture group. Compared with the control group, FBG, 2hPG, SCr, BUN, 24-hour urinary protein quantification, UACR, and urinary SPON2 were all higher in the model group (P<0.01); compared with the model group, the FBG, 2hPG, SCr, BUN, 24-hour urinary protein quantification, UACR, and urinary SPON2 were all lower in the acupuncture group (P<0.01). Compared with the control group, the model group showed reduced podocyte autophagosomes and thickened glomerular basement membrane; compared with the model group, the acupuncture group had increased podocyte autophagosomes and less thickened basement membrane. Compared with the control group, the podocyte apoptosis index (AI) was higher in the model group (P<0.01); compared with the model group, the AI was lower in the acupuncture group (P<0.01). Compared with the control group, the expression of ULK1, Beclin-1, and LC3-Ⅱ proteins was lower, and the expression of mTORC1 and p62 proteins was higher in the model group (P<0.01). Compared with the model group, the expression of ULK1, Beclin-1, and LC3-Ⅱ proteins was higher, and the expression of mTORC1 and p62 proteins was lower in the acupuncture group (P<0.01). Compared with the control group, the LncRNA SOX2OT expression was lower in the model group (P<0.01). Compared with the model group, LncRNA SOX2OT expression was higher in the acupuncture group (P<0.01). CONCLUSION The "spleen and stomach-regulating" acupuncture method could improve renal function in DKD rats, reduce blood glucose and urinary protein excretion, alleviate podocyte injury, and enhance podocyte autophagy. The mechanism may be related to modulation of the renal LncRNA SOX2OT/mTORC1/ULK1 pathway.
Animals ; Podocytes/cytology* ; Diabetic Nephropathies/physiopathology* ; Rats, Sprague-Dawley ; Male ; Rats ; Mechanistic Target of Rapamycin Complex 1/genetics* ; Autophagy ; Acupuncture Therapy ; Autophagy-Related Protein-1 Homolog/genetics* ; RNA, Long Noncoding/metabolism* ; Humans ; Signal Transduction

Objective To investigate the current status of self-management in adult patients with epilepsy and re-lated influencing factors.Methods Epilepsy Self-Management Scale(ESMS)was used to evaluate the self-management abilities of 180 patients with epilepsy in a district of Shanghai,China,and the impact of related factors on the self-management abilities of patients was analyzed,including demographic factors(sex,age,marital status,and educational level,etc.)and clinical factors(age of onset,course of disease,seizure frequency,and types of medication,etc.).Results The total score of self-management was(102.62±6.11)points for the patients with epilepsy.There was a sig-nificant difference in self-management score between the patients with different occupations(F=2.950,P<0.05)and be-tween the patients with different family incomes(F=7.298,P<0.05).The patients who missed fewer doses of antiepilep-tic drugs had a higher self-management score(F=35.948,P<0.05),and there was a significant difference in self-management score between the patients with different follow-up frequencies(F=15.619,P<0.05).Medication manage-ment showed the strongest correlation with safety management(r=0.435,P<0.05),while safety management showed the lowest correlation with seizure management score(r=-0.152,P<0.05).The multiple linear regression analysis showed that medication compliance and follow-up frequency in specialized care were influencing factors for total self-management score.Conclusion The patients with epilepsy have poor self-management abilities,and improving employment rate,in-creasing family income,enhancing medication compliance,and increasing follow-up frequency in specialized care have a certain clinical significance in improving the self-management abilities of patients.

Objective:To identify and observe disease-causing gene variants and clinical phenotypes in a Han Chinese family with Leber congenital amaurosis (LCA).Methods:A retrospective study. A patient with LCA10 and his parents who had presented at Department of Ophthalmology of Henan Provincial People's Hospital on May 2022 were selected as the study subject. Detailed medical and family histories were recorded, fundus photography and flash electroretinogram (F-ERG) were performed. Peripheral venous blood samples (3 ml) of the proband and his parents were collected to extract whole genomic DNA, then whole exome sequencing (WES) and mitochondrial DNA (mtDNA) sequencing were carried out for the proband to determine the disease-causing gene and variants. All variants were annotated by bioinformatics analysis. According to the American College of Medical Genetics and Genomics (ACMG) guidelines, the pathogenicity of all detected variants were evaluated. Candidate variants were verified by Sanger sequencing, and in vitro minigene assay were performed to evaluate the impact of the missense variant with insufficient evidence on mRNA splicing.Results:The proband, male, 7-month-old, presented with an inability to follow light or objects, eye poking, photophobia, nystagmus, partial loss of retinal pigment epithelium around the fovea of the macula. At the age of 2 years old, F-ERG revealed severe reduction, elongation, or even no waveform of a-wave and b-wave in both eyes. No obvious abnormality was found in the clinical phenotype of his parents. The result of WES revealed that the proband carried two variants in exon 40 and exon 2 of CEP290, a frameshift variant c.5515_5518del (p.Glu1839Lysfs*11) (V1) and a novel missense variant c.74C>T (p.Ala25Val) (V2), respectively. The result of mitochondrial DNA sequencing was negative. Sanger sequencing confirmed that the heterozygous frameshift variant was inherited from his father and the heterozygous novel missense variant was inherited from his mother, which constituted compound heterozygous variants. In vitro minigene splicing assay confirmed that V2 created a new splicing donor at exon 2, leading to the in-frame deletion of 30bp fragment during transcription and loss of 10 amino acid residues in the protein. The two variants were pathogenic (V1) and likely pathogenic (V2) based on ACMG guidelines, respectively. Conclusions:The c.5515_5518del and novel c.74C>T compound heterozygous variants of the CEP290 gene probably are the cause of LCA10 in this family, which lead to the production of a truncated protein and aberrant splicing of pre-mRNA, respectively. LCA is characterized by early onset, severe impairment of visual function, and a wide range of disease-causing variations.

Objective:To explore the training mode of continuing education for clinical engineers, and provide reference for the training of clinical engineers.Methods:A total of 31 clinical engineers were selected as the two-stage training mode combined with short-term centralized intensive basic training and standardized post professional training. Comparative analysis of assessment scores before and after training was performed, and the satisfaction of comprehensive effect and post competency improvement after training was evaluated. SPSS 19.0 was used for t-test. Results:The assessment scores after training (85.06±7.31) were significantly higher than those before (69.55±6.74) training ( P = 0.001). The comprehensive training effect of clinical engineers and the satisfaction rate of post competency improvement reached a high level, which were 70.97% (22/31) and 83.87% (26/31), respectively. Conclusion:The two-stage training mode combined with short-term centralized basic intensive training and standardized post professional training can not only effectively improve the theoretical knowledge level, practical ability and post competency of clinical engineers, but also shorten the centralized training time, which is conducive to solving the practical problems that affect the continuing education training program due to the lack of human resources of clinical engineers.

Objective:To compare the consistency and difference of non-mydriatic two-field 45° ultra-wide field Optos and Clarus500 fundus imaging in a large-scale diabetic retinopathy (DR) screening.Methods:A diagnostic methodology study. From November 2020 to August 2021, 526 eyes of 277 patients with type 2 diabetes who diagnosed in Department of Ophthalmology, Henan Provincial People's Hospital were included in the study. Among them, there were 175 males with 328 eyes and 102 females with 198 eyes; the age was 53±10 years old. The same experienced technician performed the non-mydriatic dual-field 45° fundus imaging and the non-mydriatic ultra-wide-angle imaging system Optos, Clarus500 single-field fundus imaging examination on the patient on the same day, and obtained the dual-field 45° fundus image and Optos, Clarus500 single-field fundus image. The Optos and Clarus500 single-field fundus images in the same area as the dual-field 45° fundus image were captured by Photoshop software, and the Optos and Clarus500 dual-field fundus images were obtained. Subsequently, two experienced ophthalmologists performed interpretation and DR grading of the 5 groups of images, respectively. Images with inconsistent grading results were interpreted by a third ophthalmologist and used as the final grading result. In order to avoid the mydriatic dual-field 45° imaging interpretation results as the standard, the consistency and detection rate difference of the two ultra-wide-angle imaging systems in the rapid DR screening results were evaluated. The weighted Kappa ( κ) test was used to analyze the consistency of DR diagnosis between dual-field 45° fundus imaging and Optos and Clarus500 fundus imaging; χ2 test was used to compare the detection rates of DR between different imaging systems. Results:Compared with the dual-field 45° fundus image, the Clarus500 single-field had a higher DR detection rate ( χ2=24.965, P<0.001), and the Optos dual-field fundus image had a lower DR detection rate ( χ2=49.559, P<0.001). Compared with the DR detection rate of dual-field 45° fundus image, Optos single-field fundus image, Clarus500 double-field fundus image had no significant difference ( χ2=2.572, 0.649; P=0.109, 0.421). Compared with Optos, Clarus500 single-field and dual-field fundus images DR detection rate, the difference was statistically significant ( χ2=43.214, 61.216; P<0.001). Consistency assessment of DR grading results: dual-field 45° fundus images and Clarus500 dual-field fundus images ( κ value=0.932, 95% confidence interval ( CI) 0.907-0.956) were highly consistent; dual-field 45° fundus images and Optos single-field fundus images [ κ value=0.474, 95% CI 0.417-0.532], Optos dual-field fundus image ( κ value=0.495, 95% CI 0.438-0.551), Optos dual-field fundus image ( κ value=0.495, 95% CI 0.438-0.551) and Clarus500 dual-field fundus image ( κ value=0.452, 95% CI 0.395-0.506) were moderately consistent; dual-field 45°fundus images and Clarus500 single-field fundus images ( κ value=0.354, 95% CI 0.303-0.403) and Optos single-field fundus images and Clarus500 single-field fundus images ( κ value=0.347, 95% CI 0.287-0.393) showed general agreement. Conclusions:Compared with Optos dual-field fundus image, dual-field 45°fundus image and Clarus500 dual-field fundus image have high consistency in the grading results of DR rapid screening. Compared with Optos single-field fundus image, the detection rate of the DR of Clarus500 single-field fundus image is higher.

Objective: To explore the relationship between anthropometric indices and the incident risk of hypertension, compare novel anthropometric indices with traditional indices in hypertension prediction and establish hypertension prediction models among elderly Chinese. Methods: A total of 27 009 retirees from the Dongfeng Motor Corporation were recruited at baseline in 2008 and the first follow-up was conducted in 2013. After the exclusion of participants less than 60 years old, participants with hypertension, coronary heart disease, stroke, cancer, and those with missing data, a total of 6 784 elderly participants were enrolled in this study. A multivariate logistic regression model was used to analyze the relationship between traditional anthropometric indices, body mass index (BMI), waist circumference (WC), waist-to-height ratio (WHtR), novel anthropometric indices, visceral adiposity index (VAI), body shape index (ABSI), body roundness index (BRI), and the incident risk of hypertension. Meanwhile, a multivariate logistic regression model was used to establish hypertension prediction models. Receiver operating characteristic (ROC) curve was applied to compare the prediction ability of different models. Results: A total of 1 787 incident cases of hypertension were identified, with the incidence of hypertension about 27.59%. Significant positive associations were detected between BMI, WC, WHtR, VAI, BRI and the incident risk of hypertension after adjusting for potential confounders (all P values<0.05). In men, the OR (95%CI) (upper tertile vs lower tertile) was 1.45 (1.22-1.73) for VAI, and 1.86 (1.55-2.23) for BRI. In women, the OR (95% CI) (upper tertile vs lower tertile) was 1.55 (1.22-1.96) for VAI, and 1.60 (1.27-2.01) for BRI. For ABSI, no significant association was observed in either men (OR (95%CI): 1.07 (0.90-1.28)) or women (OR (95%CI): 1.03 (0.82-1.29), both P values >0.05). The basic hypertension prediction model included age, drinking (only in men), education status (only in men), systolic blood pressure (SBP), diastolic blood pressure (DBP), and fasting blood glucose levels. Based on the basic prediction model, BMI (in men: AUC=0.697) and BRI (only in men: AUC=0.696) improved area under ROC curve (AUC) significantly (P<0.05). BMI was the strongest predictor in both men (AUC=0.697) and women (AUC=0.685) in the extended model. Conclusion Significant positive associations were detected between BMI, WC, WHtR, VAI, BRI and the incident risk of hypertension among elderly Chinese. BMI was the strongest predictor in hypertension prediction model compared with other anthropometric indices.

Objective To explore the application of bumetanide to inhibition of tumor cell proliferation.Methods In different cell lines, the expression of natrium,kalium, chloride cotransporter 1 ( NKCC1) was detected by Western blotting while the proliferation of different tumor cells was examined by CCK-8 kit.Results The target protein NKCC1 expression in lung cancer cell line ( A549 ) and colorectal cancer cell line ( HCT116 ) was significantly higher than that in chronic myelogenous leukemia cell line (K562), esophageal cancer cell line (Eca109), cervical carcinoma cell line (HeLa), T lymphocytic leukemia cell line (Jurkat) and breast cancer cell line (MCF7).IC50 Values of bumetanide were significantly lower in A549 and HCT116 than in K562, Eca109,HeLa,Jurkat and MCF7.Furthermore, the inhibiory rate and the target protein expression level were positively correlated.Conclusion Bumetanide can inhibit tumor cell proliferation and NKCC1 can serve as a potential target of anticancer drugs.

Objective Subarachnoid hemorrhage ( SAH) is a devastating disease with high fatality and morbidity and micro-glia/macrophage ( M/M) plays a vital role in SAH brain injury with complicated pathophysiological mechanism .This study was to ob-serve the effect of Nrf2 deficiency on M/M activation and M1 polarization after subarachnoid hemorrhage in mice . Meth ods We col-lected 70 wild-type ( WT) ICR mice and 35 Nrf2-knockout ( KO) mice to establish the SAH model by injecting fresh autologous blood into pre-chiasmatic cistern.WT mice were arranged into four groups: sham operation group, post operative day 1 (POD1) group, POD3 group and POD5 group.Then WT mice and Nrf2 Nrf2-knockout mice were divided into sham operation WT group , sham opera-tion KO group, SAH WT group and SAH KO group.Western blotting (WB) and immunohistochemistry (IHC) were applied to observe the activation and proliferation of M/M after SAH on WT mice .Difference in activation and M 1 polarization were observed by detecting Iba1 expression in WB and CD 16/32 +Iba1 +cells in immunofluorescence between WT and KO mice . Results Gray scale values of Iba1 expression by WB in WT mice are 0.491 ±0.039, 0.657 ± 0.069, 0.930 ±0.046 and 0.926 ±0.046;average optical intensity values of Iba1 expression by IHC in WT mice are 0.412 ±0.122, 0.625 ±0.135, 0.963 ±0.213 and 0.978 ±0.224.The data indica-ted that Iba1 expression increased in SAH KO group in comparison to SAH WT group on 1, 3, 5 day after SAH (P<0.05).Moreover, Nrf2 deficiency promoted the activation and polarization of M /M by increased Iba1 protein expression and CD16/32 +Iba1 +cells after SAH ( P<0.05). Conclusion SAH induces M/M activation and proliferation in mice, and Nrf2 deficiency promotes the activa-tion, proliferation and M1 polarization after SAH .

Objective To evaluate the potential involvement of melatonin in the activation of nuclear factor-E2-related factor 2-antioxidant response element (Nrf2-ARE) signaling pathway and anti-oxidative stress in an experimental model of traumatic brain injury (TBI).Methods One hundred and sixteen ICR male mice,aged 6-8 weeks,were randomly divided into sham-operated group,model group,vehicle group and melatonin treatment group (n=29);mice in the later three groups were established TBI models by Flierl improved free-fall method;0,1,2,3 and 4 h after that,mice in the melatonin treatment group were performed intraperitoneal injection of melatonin (10 mg/kg,3 mg/mL diluted in normal saline),and those in the vehicle group were given the same volume of normal saline.Twenty-four h after TBI,the neuronal degeneration was investigated by Fluoro-Jade C (FJC) staining and brain water content was measured by wet-to-dry weight ratio method;colorimetric method was employed to detect the malondialdehyde (MDA),superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels;Western blotting was engaged to analyze the protein content of nuclear Nrf2,cytoplasmic Nrf2,NADPH quinine oxidoreductase-1 (NQO1) and heme oxidase-1 (HO-1);real-time quantitative-PCR was employed to evaluate the mRNA content of HO-1 and NQO-1;electrophoresis mobility shift assay (EMSA) was performed to measure the Nrf2 DNA binding activity.Results As compared with the sham-operated group,the other three groups had significantly increased number of cortical degenerative neurons,water content,MDA content,nuclear Nrf2 expression,and nuclear HO-1 and NQO-1 protein and mRNA expressions,and had significantly lower levels of cytoplasmic Nrf2 and GPx.As compared with the model group and vehicle group,the melatonin treatment group had significantly decreased number of cortical degenerative neurons,water content,MDA content and nuclear Nrf2 expression,and had significantly increased cytoplasmic Nrf2 level,GPx content and nuclear HO-1 and NQO-1 protein and mRNA expressions (P＜0.05).The SOD content in the sham-operated group and melatonin treatment group was significantly higher than that in the model group and vehicle group (P＜0.05).EMSA showed that the Nrf2 DNA binding activity increased accordingly in the order of sham-operated group,model group,vehicle group and melatonin treatment group.Conclusion The melatonin can improve the anti-oxidative stress neuroprotection following TBI,which may be associated with the activation of Nrf2-ARE signaling pathway.

Objective Subarachnoid hemorrhage ( SAH) is a devastating disease with a high mortality.This study was to in-vestigate the effect of Nrf2 on secondary brain injury following SAH and its action mechanism in mice. Methods SAH models were established in wild-type ( WT) and Nrf2 knockout ( KO) ICR male mice by injecting fresh blood drawn from the femoral artery into the pre-chiasmatic cistern.The animals were divided into four groups, WT sham, WT SAH, KO sham, and KO SAH.At 24 hours after modeling, the expression levels of malondialdehyde ( MDA) , GSH/GSSG, TNF-αand IL-1β, the volume of brain water, and content of Evans blue were measured, the activity scores obtained, and cerebral vasospasm of the anterior and middle cerebral arteries ( ACA and MCA) detected. Results At 24 hours, the expressions of MDA, TNF-α, and IL-1βwere (3.299 ±0.335), (1.187 ± 0.436), and (59.330 ±21.787) mg/g in the WT sham group, (4.339 ±0.328), (2.432 ±0.434), and (121.584 ±21.675) mg/g in the WT SAH group, (3.488 ±0.634), (1.170 ±0.312), and (58.497 ±15.608) mg/g in the KO sham group, and (5.335 ±0.499), (3.132 ±0.548), and (171.117 ±50.479) mg/g in the KO SAH group, markedly increased in the SAH groups as compared with the sham controls (P<0.05), while the GSH/GSSG levels were significantly higher in the former two groups than in the latter (0.553 ±0.100 and 0.375 ±0.068 vs 0.714 ±0.091, 0.761 ±0.114, P<0.01).The contents of brain water and Evans blue were (0.784 ±0.005) and (7.055 ±1.046) μg/g in the WT sham group, (0.808 ±0.004) and (7.230 ±1.192) μg/g in the WT SAH group, (0.784 ±0.004) and (9.620 ±1.290) μg/g in the KO sham group, and (0.819 ±0.004) and (11.628 ±1.040)μg/g in the KO SAH group, remarkably increased in the SAH groups in comparison with the sham groups (P<0.05).The apoptosis rate 8.916 and 82.100 ±6.870 vs 70.833 ±8.750 and 51.767 ±13.006), ACA radius/wall thickness value (13.885 ±3.360 and 14.212 ±3.2545 vs 8.024 ±2.780 and 6.861 ±2.702), MCA radius/wall thickness value (18.648 ±2.893 and 19.435 ±2.775 vs 6.337 ±3.993 and 5.107 ±3.805), and activity score (2.733 ±0.450 and 2.767 ±0.430 vs 1.967 ±0.928 and 1.433 ±0.679) (all P<0.01). Conclusion Nrf2 knockout increases oxidative stress and inflammatory reaction following SAH and consequently aggravates secondary brain injury.Nrf2 has a protective effect against SAH-induced brain injury.