1.Effect of fluoride exposure on endoplasmic reticulum-mitochondrial calcium transfer and apoptosis in primary nerve cells
Yongheng LU ; Shuang ZHU ; Feiyan ZHAO ; Fujun AI ; Yanjie LIU ; Yangting DONG ; Zhizhong GUAN ; Na WEI
Chinese Journal of Tissue Engineering Research 2026;30(1):111-119
BACKGROUND:Previous studies have found that neuronal damage caused by continuous excessive fluoride exposure is related to Ca2+overload,but the mechanism of Ca2+flow conversion between intracellular calcium stores and cell apoptosis damage is still unclear.OBJECTIVE:To investigate the effect of fluoride exposure on Ca2+transport channel proteins and apoptosis levels in the mitochondria-associated endoplasmic reticulum membrane of primary cultured neural cells.METHODS:Primary nerve cells of neonatal SD rats were cultured in vitro and identified by immunofluorescence staining with neuronal nucleus-specific antibody up to day 7.The nerve cells were divided into control group(containing 0 mmol/L sodium fluoride),low fluoride group(containing 0.5 mmol/L sodium fluoride),and high fluoride group(containing 1 mmol/L sodium fluoride).The cell morphological changes were observed by light microscope 24 hours after fluorine exposure.The expression levels of apoptosis-related protein BAX/BCL-2 and calcium transfer-related pathways VDAC1,GRP 75,and IP3R were detected using western blot assay.The expression levels of VDAC1,GRP 75,and IP3R mRNA were detected by RT-PCR.Ca2+levels were detected by Rhood-2AM Ca2+probe.Mitochondrial membrane potential detection kit was used to detect the change in mitochondrial membrane potential.The level of apoptosis was determined by flow cytometry and TUNEL staining.RESULTS AND CONCLUSION:(1)The purity of neurons cultured on day 7 had been determined to be over 90%,with few impurities,good growth status,and tight cell network connections,meeting the requirements of subsequent experiments.(2)Compared with the control group,growth of neural cell clusters in the low-fluoride group and the high-fluoride group increased;the processes were broken;the cell body was rounded,and the connection network between cells was destroyed.Compared with the low-fluoride group,the cell damage changes in the high-fluoride group were more obvious.(3)Compared with the control group,the protein expressions of VDAC1,GRP75,and IP3R were increased in the low-fluoride group and the high-fluoride group(P<0.05),and the ratio of apoptosis-related protein BAX/BCL-2 was increased(P<0.05).Compared with the control group,the expression of VDAC1 and GRP75 mRNA in the low-fluoride group was significantly increased(P<0.05);the expression levels of VDAC1,GRP75,and IP3R mRNA in the high-fluoride group were significantly increased(P<0.01).(4)The level of cell apoptosis increased significantly after fluoride exposure,and the high-fluoride group was significantly higher than the control and low-fluoride groups(P<0.01).(5)After fluoride exposure,the concentration of mitochondrial Ca2+in nerve cells increased significantly(P<0.05),the mitochondrial membrane potential decreased(P<0.01),and the degree of damage in the high-fluoride group was more obvious(P<0.05).The results show that fluoride exposure impairs the morphological structure of primary neural cells,resulting in upregulation of Ca2+transfer pathway protein expression between the endoplasmic reticulum and mitochondria,mitochondrial Ca2+overload,mitochondrial damage,and increased levels of apoptosis.
2.Effect of fluoride exposure on endoplasmic reticulum-mitochondrial calcium transfer and apoptosis in primary nerve cells
Yongheng LU ; Shuang ZHU ; Feiyan ZHAO ; Fujun AI ; Yanjie LIU ; Yangting DONG ; Zhizhong GUAN ; Na WEI
Chinese Journal of Tissue Engineering Research 2026;30(1):111-119
BACKGROUND:Previous studies have found that neuronal damage caused by continuous excessive fluoride exposure is related to Ca2+overload,but the mechanism of Ca2+flow conversion between intracellular calcium stores and cell apoptosis damage is still unclear.OBJECTIVE:To investigate the effect of fluoride exposure on Ca2+transport channel proteins and apoptosis levels in the mitochondria-associated endoplasmic reticulum membrane of primary cultured neural cells.METHODS:Primary nerve cells of neonatal SD rats were cultured in vitro and identified by immunofluorescence staining with neuronal nucleus-specific antibody up to day 7.The nerve cells were divided into control group(containing 0 mmol/L sodium fluoride),low fluoride group(containing 0.5 mmol/L sodium fluoride),and high fluoride group(containing 1 mmol/L sodium fluoride).The cell morphological changes were observed by light microscope 24 hours after fluorine exposure.The expression levels of apoptosis-related protein BAX/BCL-2 and calcium transfer-related pathways VDAC1,GRP 75,and IP3R were detected using western blot assay.The expression levels of VDAC1,GRP 75,and IP3R mRNA were detected by RT-PCR.Ca2+levels were detected by Rhood-2AM Ca2+probe.Mitochondrial membrane potential detection kit was used to detect the change in mitochondrial membrane potential.The level of apoptosis was determined by flow cytometry and TUNEL staining.RESULTS AND CONCLUSION:(1)The purity of neurons cultured on day 7 had been determined to be over 90%,with few impurities,good growth status,and tight cell network connections,meeting the requirements of subsequent experiments.(2)Compared with the control group,growth of neural cell clusters in the low-fluoride group and the high-fluoride group increased;the processes were broken;the cell body was rounded,and the connection network between cells was destroyed.Compared with the low-fluoride group,the cell damage changes in the high-fluoride group were more obvious.(3)Compared with the control group,the protein expressions of VDAC1,GRP75,and IP3R were increased in the low-fluoride group and the high-fluoride group(P<0.05),and the ratio of apoptosis-related protein BAX/BCL-2 was increased(P<0.05).Compared with the control group,the expression of VDAC1 and GRP75 mRNA in the low-fluoride group was significantly increased(P<0.05);the expression levels of VDAC1,GRP75,and IP3R mRNA in the high-fluoride group were significantly increased(P<0.01).(4)The level of cell apoptosis increased significantly after fluoride exposure,and the high-fluoride group was significantly higher than the control and low-fluoride groups(P<0.01).(5)After fluoride exposure,the concentration of mitochondrial Ca2+in nerve cells increased significantly(P<0.05),the mitochondrial membrane potential decreased(P<0.01),and the degree of damage in the high-fluoride group was more obvious(P<0.05).The results show that fluoride exposure impairs the morphological structure of primary neural cells,resulting in upregulation of Ca2+transfer pathway protein expression between the endoplasmic reticulum and mitochondria,mitochondrial Ca2+overload,mitochondrial damage,and increased levels of apoptosis.
3.Alcohol-related liver disease-associated acute-on-chronic liver failure: Mechanisms, dynamic assessment, and precise management
Na AI ; Huaining CUI ; Xiaohui FANG ; Tao LIU ; Yanhang GAO
Journal of Clinical Hepatology 2026;42(8):1755-1760
Acute-on-chronic liver failure (ACLF) is a high-mortality syndrome that develops during the acute deterioration of chronic liver disease and is characterized by multiple organ dysfunction driven by systemic inflammation. Due to gut microbiota dysbiosis, endotoxin translocation, and amplified inflammatory responses, alcohol-related liver disease-associated ACLF (ALD-ACLF) is more pronounced and is often accompanied by a high risk of secondary infection and extrahepatic organ involvement, thereby exhibiting distinct etiological and clinical characteristics. This article systematically reviews the advances in the definitional boundaries, pathophysiological mechanisms, dynamic early warning, and prognostic assessment of ALD-ACLF, elaborates on risk stratification and key issues in clinical management, and discusses the potential impact of the dual-hit phenotype of metabolic dysfunction and alcohol-related liver disease on risk stratification in ALD-ACLF. This article also highlights the need to further strengthen etiology-oriented early identification, dynamic stratification, control of predisposing factors, and earlier intervention, in order to provide a reference for the precise assessment and individualized intervention of these patients.
4.Analysis of the occurrence and risk factors of microperforations in surgical gloves used in dermatovenereology surgeries
Dan-li TANG ; Wei-na ZHANG ; Yan-yan NIU ; Ai-xiu SHI ; Fei HAN
Journal of Regional Anatomy and Operative Surgery 2025;34(5):444-447
Objective To clarify the occurrence,location distribution,and risk factors of microperforations in surgical gloves used in dermatovenereology surgeries.Methods A total of 898 sterilized surgical gloves worn by right-handed medical staff during dermatovenereology surgeries in Suqian Hospital of Jiangsu Province Hospital from May 2022 to April 2024 were selected as the research objects.The occurrence and location distribution of microperforations in all sterilized surgical gloves after surgery were collected.Univariate analysis and binary Logistic regression analysis were conducted on the factors that might lead to the occurrence of microperforations.Results Among the 898 gloves selected in this study,61 gloves(6.79%)had microperforations;the incidence of microperforations in the gloves worn on the left hand was significantly higher than that in the gloves worn on the right hand(P<0.05);microperforations were prone to occur on the palmar surfaces of the index finger and thumb of the gloves.The results of univariate and binary Logistic regression analyses showed that the use of special instruments,surgery duration≥60 minutes,ingrown nails surgery,and worn by the chief surgeon were the risk factors for the occur-rence of microperforations in sterilized surgical gloves(OR>1,P<0.05),while wearing double-layer gloves was the protective factor to avoid the occurrence of microperforations(OR<1,P<0.05).Conclusion The sterilized surgical gloves are more likely to occur microperforations if involved special instruments in surgery,surgery duration≥60 minutes,and ingrown nails surgery,and worn by the chief surgeon,while wearing double-layer gloves can reduce the incidence of microperforations.
5.Analysis of the occurrence and risk factors of microperforations in surgical gloves used in dermatovenereology surgeries
Dan-li TANG ; Wei-na ZHANG ; Yan-yan NIU ; Ai-xiu SHI ; Fei HAN
Journal of Regional Anatomy and Operative Surgery 2025;34(5):444-447
Objective To clarify the occurrence,location distribution,and risk factors of microperforations in surgical gloves used in dermatovenereology surgeries.Methods A total of 898 sterilized surgical gloves worn by right-handed medical staff during dermatovenereology surgeries in Suqian Hospital of Jiangsu Province Hospital from May 2022 to April 2024 were selected as the research objects.The occurrence and location distribution of microperforations in all sterilized surgical gloves after surgery were collected.Univariate analysis and binary Logistic regression analysis were conducted on the factors that might lead to the occurrence of microperforations.Results Among the 898 gloves selected in this study,61 gloves(6.79%)had microperforations;the incidence of microperforations in the gloves worn on the left hand was significantly higher than that in the gloves worn on the right hand(P<0.05);microperforations were prone to occur on the palmar surfaces of the index finger and thumb of the gloves.The results of univariate and binary Logistic regression analyses showed that the use of special instruments,surgery duration≥60 minutes,ingrown nails surgery,and worn by the chief surgeon were the risk factors for the occur-rence of microperforations in sterilized surgical gloves(OR>1,P<0.05),while wearing double-layer gloves was the protective factor to avoid the occurrence of microperforations(OR<1,P<0.05).Conclusion The sterilized surgical gloves are more likely to occur microperforations if involved special instruments in surgery,surgery duration≥60 minutes,and ingrown nails surgery,and worn by the chief surgeon,while wearing double-layer gloves can reduce the incidence of microperforations.
6.Successful Management of Refractory Chylothorax in Preterm Infants Using Hypertonic Glucose Pleurodesis
Young Seok DO ; Euiseok JUNG ; Sung Hyeon PARK ; Jeong Min LEE ; Ha Na LEE ; Jiyoon JEONG ; Soo Hyun KIM ; Byong Sop LEE ; Ki Soo KIM ; Ellen Ai-Rhan KIM
Neonatal Medicine 2024;31(3):73-79
Neonatal chylothorax is a potentially fatal respiratory condition caused by a congenital or traumatic etiology. Conventional therapies, such as fasting, total parenteral nutrition, and intravenous octreotide, are generally successful in such cases; however, more invasive therapeutic measures, such as pleurodesis, should be considered in refractory cases. This case report presents two preterm infants with refractory chylothorax who were non-responsive to conventional treatment but were successfully managed using hypertonic glucose pleurodesis. The first case was that of a female infant born at 24+5 weeks of gestation (585 g) and diagnosed with postsurgical chylothorax at 68 days of life. Even after the initiation of fasting and intravenous octreotide administration, pleural drainage did not reduce. Therefore, the patient underwent three intermittent procedures of 50% glucose pleurodesis, which resulted in the resolution of the chylothorax and subsequent chest tube removal after 37 days. The second case was a female infant born at 34+6 weeks (3,040 g), who was diagnosed with congenital chylothorax immediately after birth. Fasting and intravenous octreotide failed to show any clinical effects; therefore, the patient underwent pleurodesis for 3 consecutive days. After the procedure, the amount of pleural drainage substantially decreased, and the chest tube was removed after 14 days. In both cases, a temporal relation between pleurodesis and chylothorax resolution was observed, suggesting that hypertonic glucose pleurodesis may be an effective and safe alternative for treating refractory chylothorax in preterm infants with minimal side effects. Further studies are needed to establish the optimal protocol for this procedure and to compare its efficacy with that of other pleurodesis agents.
7.Successful Management of Refractory Chylothorax in Preterm Infants Using Hypertonic Glucose Pleurodesis
Young Seok DO ; Euiseok JUNG ; Sung Hyeon PARK ; Jeong Min LEE ; Ha Na LEE ; Jiyoon JEONG ; Soo Hyun KIM ; Byong Sop LEE ; Ki Soo KIM ; Ellen Ai-Rhan KIM
Neonatal Medicine 2024;31(3):73-79
Neonatal chylothorax is a potentially fatal respiratory condition caused by a congenital or traumatic etiology. Conventional therapies, such as fasting, total parenteral nutrition, and intravenous octreotide, are generally successful in such cases; however, more invasive therapeutic measures, such as pleurodesis, should be considered in refractory cases. This case report presents two preterm infants with refractory chylothorax who were non-responsive to conventional treatment but were successfully managed using hypertonic glucose pleurodesis. The first case was that of a female infant born at 24+5 weeks of gestation (585 g) and diagnosed with postsurgical chylothorax at 68 days of life. Even after the initiation of fasting and intravenous octreotide administration, pleural drainage did not reduce. Therefore, the patient underwent three intermittent procedures of 50% glucose pleurodesis, which resulted in the resolution of the chylothorax and subsequent chest tube removal after 37 days. The second case was a female infant born at 34+6 weeks (3,040 g), who was diagnosed with congenital chylothorax immediately after birth. Fasting and intravenous octreotide failed to show any clinical effects; therefore, the patient underwent pleurodesis for 3 consecutive days. After the procedure, the amount of pleural drainage substantially decreased, and the chest tube was removed after 14 days. In both cases, a temporal relation between pleurodesis and chylothorax resolution was observed, suggesting that hypertonic glucose pleurodesis may be an effective and safe alternative for treating refractory chylothorax in preterm infants with minimal side effects. Further studies are needed to establish the optimal protocol for this procedure and to compare its efficacy with that of other pleurodesis agents.
8. Effects of sodium pyruvate on the morphology, structure and function of erythrocytes stored in vitro in type 2 diabetes rats
Yu BAI ; Na YAO ; Xiaoyi XIE ; Mandi WU ; Yinghui CUI ; Jianrong GUO ; Yongbin CHI
Chinese Journal of Clinical Pharmacology and Therapeutics 2024;29(2):139-145
AIM: To observe the effect of RBC preservation solution with sodium pyruvate on the morphology, structure and function of RBC stored in vitro in type 2 diabetes rats. METHODS: Thirty SPF male SD rats, were randomly divided into 3 groups (n=10): non-T2DM conventional RBC preservation solution (group A), T2DM conventional RBC preservation solution (group B) and T2DM sodium pyruvate RBC preservation solution (group C). The leukoreduced RBC from the tail vein and stored for 0 d (T0), 7 d (T1), 14 d (T2), 21 d (T3) and 28 d (T4) to detect the morphology, structure and the contents of 2, 3-DPG, reactive oxygen species (ROS), malondialdehyde (MDA) and lactic acid (LA) of RBC in group A, B and C. The RBC stored for 14 days in vitro were labeled with PKH26, and its survival rate were tested in vivo at 1, 4, 10 and 16 hours after intravenous infusion. RESULTS: At T0, the RBC morphology of group A was intact, which was better than that of group B and group C. With the extension of storage time, the morphology of RBC in each group gradually transformed into a spindle-spherical shape. Compared with group A, the incidence of acanthocytes in group B and group C was higher, and the incidence of acanthocytes in group C was lower than that in group B. Compared with group A, the content of 2, 3-DPG in group B and group C decreased, while ROS and MDA increased at different time points (P<0.05). The content of 2,3-DPG in group C was higher than that in group B (P<0.05), and the contents of ROS and MDA were lower than those in group B (P<0.05). LA content in group B was higher than that in group A and group C (P<0.05). At T2-T4, the LA content in group C was lower than that in group A (P<0.05). The survival rate of RBC in group A was higher than that in group B and C, and the survival rate of RBC in group B was lower than that in group C (P<0.05). CONCLUSION: Sodium pyruvate added RBC preservation solution has a certain protective effect on RBC stored in vitro in type 2 diabetic rats, and its mechanism may be related to its antioxidant effect.
9.Research progress in regulatory mechanism and traditional Chinese medicine intervention of circular RNA for coronary atherosclerotic heart disease
Lan-Tian HU ; Xue-Na XIE ; Yu-Ying WANG ; Mei LIU ; Hong-Ai GUO ; Rong YUAN ; Qi-Qi XIN ; Yu MIAO ; Wei-Hong CONG
Chinese Pharmacological Bulletin 2024;40(11):2014-2019
Coronary atherosclerotic heart disease(CHD)is an ischemic cardiovascular condition caused by the narrowing or blockage of the vascular lumen due to coronary atherosclerosis.Clinically,it presents as angina pectoris,heart failure,or sud-den cardiac death,and stands as one of the primary causes of mortality among both urban and rural populations in China.Cir-cRNA,classified as non-coding RNAs,can function as upstream regulatory molecules for miRNA or RNA-binding proteins.They actively participate in various pathological processes associated with CHD,including endothelial cell dysfunction,smooth mus-cle cell migration,macrophage-derived foam cell formation,an-giogenesis,myocardial injury,and repair,as well as post-in-farction heart failure.The expression pattern of these molecules is highly specific to the illness and tissue,indicating their poten-tial as therapeutic targets for disease management and as biomar-kers.Furthermore,they also open up new avenues for drug tar-get development in the field of traditional Chinese medicine.This article aims to provide an overview of the recent research progress on circRNA in the regulation of coronary heart disease,as well as the mechanisms involved in traditional Chinese medi-cine.It serves as a valuable reference for future research on cor-onary heart disease.
10.Efficacy of bronchoalveolar lavage combined with prone positioning in children with Mycoplasma pneumoniae pneumonia and atelectasis:a prospective randomized controlled study
Ri-Na SA ; Ai-Qiong WANG ; You-Han GAO ; Xiao-An LI ; Hugejiletu
Chinese Journal of Contemporary Pediatrics 2024;26(1):31-36
Objective To study the efficacy of bronchoalveolar lavage(BAL)combined with prone positioning in children with Mycoplasma pneumoniae pneumonia(MPP)and atelectasis and its effect on pulmonary function.Methods A prospective study was conducted on 94 children with MPP and atelectasis who were hospitalized in Ordos Central Hospital of Inner Mongolia from November 2020 to May 2023.The children were randomly divided into a treatment group and a control group,with 47 children in each group.The children in the treatment group were given conventional treatment,BAL,and prone positioning,and those in the control group were given conventional treatment and BAL.The two groups were compared in terms of fever,pulmonary signs,length of hospital stay,lung recruitment,and improvement in pulmonary function.Results Compared with the control group,the treatment group had significantly shorter time to improvement in pulmonary signs and length of hospital stay and a significantly higher rate of lung recruitment on day 7 of hospitalization,on the day of discharge,and at 1 week after discharge(P<0.05).Compared with the control group,the treatment group had significantly higher levels of forced vital capacity(FVC)as a percentage of the predicted value,forced expiratory volume(FEV)in 1 second as a percentage of the predicted value,ratio of FEV in 1 second to FVC,forced expiratory flow at 50%of FVC as a percentage of the predicted value,forced expiratory flow at 75%of FVC as a percentage of the predicted value,and maximal mid-expiratory flow as a percentage of the predicted value on the day of discharge and at 1 week after discharge(P<0.05).There was no significant difference in the time for body temperature to return to normal between the two groups(P>0.05).Conclusions In the treatment of children with MPP and atelectasis,BAL combined with prone positioning can help to shorten the time to improvement in pulmonary signs and the length of hospital stay and promote lung recruitment and improvement in pulmonary function.

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