Objective To seek an optimal method for the separation,culture of mouse embryonic germ cells (EGCs) in vitro,and to observe the influence of Activated Schwann cells (ASCs)-derived neurotrophins on the differentiation capability of mouse EGCs into neurogenic cells.Methods The gonadal ridges and a few abdominal tissues of the 11-day postcoitum (dpc) mouse embryos were isolated and disaggregated by 0.125％ trypsin-0.02％ EDTA,followed by culture of the mouse EGCs on mouse embryonic fibroblast (MEF) feeders.Monoclonal formation of the mouse EGCs was observed,and the staining of stage specificity embryo antigen-1 (SSEA-1),alkaline phosphatase (AKP),periodic acid-Schiff staining (PAS) were applied to identify the mouse EGCs.Two groups were divided as followed:mouse EGCs+basic medium (control group) and mouse EGCs+ASCs (experimental group).Immunofluorescence (NeuN,MBP,GFAP)analysis was used to evaluate the neurogenic differentiation of mouse EGCs and then to calculate the statistical positive rates of cell staining.All experimental results were analyzed statistically.Results (1) Identification ofmouse EGCs:Mouse EGCs were characterized by a dome-shaped colony containing a large nucleus and a relatively small amount of cytoplasm.All mouse EGCs were positive staining of SSEA-1,AKP,and PAS;(2)The neural induction of mouse EGCs:After one week induction,there were few round or oval cells with long axon-like processes migrating from the edge of the EGCs clones.3 weeks later,the neurogenic-like cells increased quickly.The results of immunofluorescence (NeuN,MBP,GFAP)staining demonstrated that mouse EGCs could differentiate into neurogenic cells under the influence of ASCs.The positive rate of cell staining was significant.Conclusion In this study,a simple,economical method was applied to successfully separate the mouse EGCs in vitro; mouse EGCs can differentiate into neurogenic cells under the influence of ASCs-derived neurotrophins.

BACKGROUND: Emerging studies have focused on cell transplantation. Schwann cells (SCs) can secrete various neurotrophic factors and improve local environment around injury. Plenty of documents have demonstrated that SCs could promote functional recovery following spinal injury. Many transplanting methods are available for treating spinal cord injury, and the intravenous cell transplantation is profitable for easy operation and avoidance of additional trauma. OBJECTIVE: To investigate the effects of intravenous transplantation of SCs on spinal cord injury in rats. METHODS: The bilateral sciatic nerves of Wistar rats were separated in vitro, cultured by tissue clot method, identified by S-100 and labeled by Hoechst33342. Sixty rat models with T10 spinal cord injury were prepared using impactor model- II type weight drop apparatus. Then the injured rats were randomly divided into 3 groups: blank control, DMEM control and SCs transplantation groups. No treatment was performed in the blank control group. Totally 1 mL DMEM and or SCs was injected into rats of DMEM control and SCs transplantation groups by tail vein respectively. Basso Beattie Bresnahan (B6B) scores were performed at 1 day before and 1, 3 days, 1 week and weekly after operation. The migration of transplanted SCs was observed at 2 weeks and 4 after transplantation. The expressions of glial fibrillary acidic protein (GFAP) and neuron specific enolase (NSE) were detected by haematoxylin-eosin staining and immune-fluorescence staining.RESULTS AND CONCLUSION: The purity of SCs reached 95%. Hoechst33342 positive cells were observed throughout the injured and the nearby region of spinal cord at 1, 2, and 4 weeks after transplantation. The statistical difference of BBB score among the SCs transplantation, blank control, and the DMEM control groups displayed at 4 weeks after transplantation (P < 0.05), and the BBB scores of the SCs transplantation were higher than other groups. Haematoxylin-eosin staining showed the cavity formed in each group at 8 weeks after transplantation, but the area of SCs transplantation was smaller than that of the blank control and DMEM control groups. The immunofluorescence staining indicated that the expression of GFAP were more intense in the blank control group and DMEM control than SCs transplantation (P < 0.05), while the expression of NSE was more intense in SCs transplantation than other groups (P< 0.05). It implied that intravenous transplantation of SCs promotes regeneration of axon and improves neurological functions after spinal cord injury in rats.

BACKGROUND:Schwann cells can secrete various neurotrophic factors,and promote functional recovery of injured spinal cord.However,xenogenic Schwann cells transplantation may induce autoimmune response.Moreover,local transplantation results in secondary injury.Vein transplantation may reached injury site passing the blood spinal cord barrier,but the treatment concentration is not effective.OBJECTIVE:To investigate the therapeutic effects of transplantation of autologous activated Schwann Cells(AASCs)via subarachnoid space on spinal cord injury(SCI)in rats.METHODS:A total of 66 rats were used to establish SCI models,and the model rats were randomly divided into 3 groups.The unilateral saphenous nerves of rats were ligated directly for 1 week to activate Schwann cells,but inactivated and model control groups were not subjected to nerve ligation.1 cm nerve was excised from distal end of each group,and Schwann cells were isolated and cultured by tissue mass method.The AASCs,autologous Schwann cells(ASCs)were injected with corresponding Hoechst33342-labeted SCs suspension,but the model control group was injected with DMEM injection.The basso beattie bresnahan(BBB)score and footprint analysis,as well as HE and GFAP immunohistochemistry staining were performed to evaluate functional recovery of rat hind limbs.RESULTS AND CONCLUSION:On 4 weeks after injury,BBB scores of AASCs were significantly superior to the other groups (P＜0.05).Two weeks after transplantation,some SCs migrated to injured spinal cord.Compared with ASCs group,the center distance of forward and hind feet and extorsion angle of the third toe of hind limb were significantly reduced in the AASCs group at 5 weeks(P＜0.05),the glial scar area was significantly decreased at 13 weeks(P＜0.05),and the cavity area of injured region was signiflcentJy diminished(P＜0.05).Results show that AASCs transplantation via subarachnoid space promoted functional recovery after SCI in rats.

Background/Aims: Anti-phospholipase A2 receptor (PLA2R) autoantibody is the main biomarker of idiopathic membranous nephropathy (IMN). We aimed to find a new cutoff value of anti-PLA2R for patients with IMN and to explore the relevance between this antibody and baseline clinical parameters. Methods: A total of 670 subjects including 374 IMN cases and 296 non-IMN controls were included between January 2017 and January 2020. All clinical parameters were collected at the time of renal biopsy. The levels of anti-PLA2R were detected by a commercial enzyme-linked immunosorbent assay (ELISA) kit. The optimal cutoff value was calculated by a receiver operating characteristic curve and compared in diagnostic efficiency. Results: The optimal cutoff value of anti-PLA2R for IMN was 7.45 RU/mL with the highest Youden index, and the corresponding sensitivity, specificity, positive predictive value and negative predictive value were 80.75%, 97.97%, 98.05% and 80.11%, respectively. Anti-PLA2R levels in IMN patients demonstrated a significant positive correlation with serum creatinine and 24-hour urinary protein, while they showed a negative correlation with serum albumin and estimated glomerular filtration rate. Conclusions The recommended cutoff value of anti-PLA2R is 7.45 RU/mL using ELISA detection for distinguishing IMN from non-IMN nephropathy. The level of anti-PLA2R is related to baseline renal function in IMN. This new threshold can improve the diagnostic efficiency and facilitate early diagnosis of IMN.

OBJECTIVE: To valuate test results of normal hearing persons with different ages using disyllabic mandarin speech test materials (MSTMs). Obtaining speech recognition threshold (SRT) and P-I function of different ages as clinical reference of hearing recovery and individual's ability to perceive and process speech. METHOD: One hundred and twenty subjects with normal hearing who speak mandarin well in their daily lives were enrolled in this study and divided into four groups (18-30, 31-40, 41-50 and 51-60 years old). Nine lists of disyllabic mandarin speech test materials with equal difficulty were utilized to test each age group. RESULT: There are good agreement between SRT and mean PTA thresholds (at 0.5, 1.0, 2.0, and 4.0 kHz) in each age group. PTA thresholds have little change as age increasing in age group 18-30, 31-40 and 41-50 (P > 0.05). PTA threshold of age group 51-60 increases more apparently compared with the other three groups (P < 0.01). SRT thresholds of each age group increase as age increasing (P < 0.05) and SRT threshold of age group 51-60 increases more apparently (P < 0.01). Slopes of P-I function in each age group are 5.8%/dB, 4.7%/dB, 3.8%/dB, 2.9%/dB respectively. CONCLUSION Nine lists of disyllabic MSTMs were used to test normal hearing persons in different ages and SRT and P-I function of four different age groups were obtained. As age increases, SRT increases and slope of P-1 functions decreases. PTA and SRT thresholds of age group 51-60 increase more apparently. The SRT and P-I functions provide reference data of normal hearing for utilizing of disyllabic mandarin speech test materials clinically.
Adolescent ; Adult ; Female ; Humans ; Language ; Male ; Middle Aged ; Phonetics ; Reference Values ; Speech Reception Threshold Test ; Young Adult

Lung cancer is characterized by high incidence and mortality rates and invasiveness, and its occurrence and development are influenced by various factors. Mitochondria, as ubiquitous organelles in the human body, regulate cellular processes, such as metabolism, signal transduction, oxidative stress, and genomic instability, thereby affecting the initiation and progression of lung cancer. This article summarizes the recent research progress on mitochondrial-targeted drugs, mitochondrial transfer, and mitochondrial gene therapy for lung cancer treatment. This work also discusses the principles and prospects of mitochondrial therapy to provide new insights for lung cancer treatment.

Objective To investigate the correlation between homocysteine (Hcy) and cerebrovascular hemodynamic accumulative scores in primary hypertension patients. Methods A cross-sectional survey was conducted in 2 767 patients with essential hypertension who were simultaneously tested for serum Hcy and cerebral vascular function in the health management/physical examination center in Chongqing General Hospital from October 2015 to March 2018. The prevalence of hyperhomocysteinemia (HHcy) was also explored. Differences between cerebrovascular hemodynamic accumulative scores and its abnormal rate among different Hcy levels were evaluated using the analysis of variance and χ2tests, and logistic regression was used to analyze the correlation between Hcy and cerebrovascular hemodynamic accumulative scores. Results The median level of Hcy in primary hypertension was 11.8 (9.3-15.0) μmol/L. HHcy prevalence was 25.15% (27.01% in men and 19.80% in women), which was higher in men than women (χ2=14.576, P<0.001) and was increasing with age (P<0.001). The proportion of stroke, proportion of taking hypotensive medications, age, fasting plasma glucose, systolic pressure, pulse pressure, and Hcy were significantly higher in the abnormal score (<75 points) group (P<0.001) than in the normal score (≥75 points) group. The average cerebrovascular hemodynamic accumulative score was 86.99±16.10 points. The score in the highest quartile of Hcy (77.91±16.10) was significantly lower than that in other quartiles. The abnormal score rate (<75 points) was 15.25% and was increasing with the Hcy level (χ2=13.986, P<0.001). Logistic regression showed that Hcy in the second, third, and highest quartiles observed in abnormal scores was, respectively, 1.913-fold, 2.045-fold, and 7.497-fold higher than that in the lowest quartile after adjusting the confounding factors. Conclusion Hcy may be an independent risk factor for abnormal cerebrovascular hemodynamic accumulative scores in primary hypertension. Cerebrovascular dysfunction should be closely monitored when Hcy was higher than 15 μmol/L.

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