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.

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.

Objective:To investigate the expression and clinical significance of non coding RNA GATA binding protein 6 antisense RNA1 (lncRNA GATA6-AS1) and GATA binding protein 6 messenger RNA (GATA6 mRNA) in ovarian cancer.Methods:A prospective study was conducted on 98 patients with ovarian cancer treated at the Second Hospital of Tianjin Medical University, from January 2022 to January 2025, and 98 patients with benign ovarian lesions during the same period were selected as the control group. Real-time quantitative polymerase chain reaction (qPCR) was used to detect the serum and tissue expression levels of lncRNA GATA6-AS1 and GATA6 mRNA in patients with ovarian cancer and benign ovarian lesions. Pearson analysis was performed to assess the correlation between serum lncRNA GATA6-AS1 and GATA6 mRNA levels in patients with ovarian cancer. Receiver operating characteristic (ROC) curves were plotted to analyze the diagnostic value of serum lncRNA GATA6-AS1 and GATA6 mRNA levels in patients with ovarian cancer.Results:Compared with the control group, the serum levels of lncRNA GATA6-AS1 and GATA6 mRNA in the study group were decreased: 0.76 ± 0.18 vs. 1.00 ± 0.22, 0.75 ± 0.19 vs. 1.01 ± 0.21, with statistically significant differences ( P<0.05). Pearson analysis revealed a positive correlation between serum lncRNA GATA6-AS1 and GATA6 mRNA levels in the study group ( r = 0.61, P<0.05). Compared with benign ovarian lesion tissues, the expression levels of lncRNA GATA6-AS1 and GATA6 mRNA in ovarian cancer tissues were reduced: 0.65 ± 0.15 vs. 1.01 ± 0.20, 0.59 ± 0.12 vs. 1.01 ± 0.24, with statistically significant differences ( P<0.05). In serous ovarian cancer tissues, the expression levels of lncRNA GATA6-AS1 and GATA6 mRNA were lower than those in mucinous ovarian cancer tissues: 0.61 ± 0.10 vs. 0.77 ± 0.16, 0.54 ± 0.11 vs. 0.74 ± 0.14, with statistically significant differences ( P<0.05). The expression of lncRNA GATA6-AS1 and GATA6 mRNA in ovarian cancer tissues was associated with tumor size, International Federation of Gynecology and Obstetrics stage, differentiation degree, lymph node metastasis, peritoneal metastasis, and ascites ( P<0.05). The ROC curve indicated that the combined diagnosis of serum lncRNA GATA6-AS1 and GATA6 mRNA for ovarian cancer had an area under the curve (AUC) of 0.905, which was significantly higher than that of lncRNA GATA6-AS1 alone ( Z = 2.45, P = 0.014) and GATA6 mRNA alone ( Z = 2.16, P = 0.031). Conclusions:The lncRNA GATA6-AS1 and GATA6 mRNA in serum and tissue of ovarian cancer patients decrease, showing a positive correlation. The joint of the two has high diagnostic value for ovarian cancer.

Objective:To analyze the application value of ultrasound Thyroid Imaging Reporting and Data System (TI-RADS) classification in the differential diagnosis of thyroid nodules and its correlation with patients' midkine, thymidine kinase 1, and thyroid function indicators.Methods:This study was a prospective study. A total of 108 patients with thyroid nodules (120 nodules) who visited Jinan 2 nd People's Hospital from December 2022 to February 2024 were included. Ultrasound examinations were conducted to complete the TI-RADS classification, and serum levels of midkine (MK) and thymidine kinase 1 (TK1), as well as thyroid function indicators, were measured. Based on the patients' pathological results, the consistency between the TI-RADS diagnostic results and the pathological findings was analyzed. The diagnostic efficacy of TI-RADS classification was calculated. The incidence of malignant nodules across various TI-RADS categories was assessed, and the differences in serum levels of MK, TK1, and thyroid function indicators between patients with benign and malignant nodules were compared. Additionally, a correlation analysis was performed to analyze the correlation between the TI-RADS classification of thyroid nodules and the serum levels of MK, TK1, and thyroid function indicators. Results:Pathological examination results revealed 38 malignant nodules and 82 benign nodules. The consistency between ultrasound TI-RADS classification diagnoses and pathological results was good ( Kappa = 0.90, P < 0.001), with diagnostic sensitivity, specificity, and accuracy of 86.84% (33/38), 97.56% (80/82), and 94.17% (113/120), respectively. Among the ultrasound TI-RADS classifications, the malignant rates for category Ⅳb and category Ⅴ thyroid nodules were the highest, at 92.00% (23/25) and 100.00% (10/10), respectively. Serum levels of MK, TK1, and thyroid-stimulating hormone (TSH) were significantly higher in patients with malignant thyroid nodules than in those with benign thyroid nodules (all P < 0.05). Correlation analysis indicated that TI-RADS classification was positively correlated with serum levels of MK, TK1, and TSH ( r = 0.56, 0.60, 0.52, all P < 0.05). Conclusions:Ultrasound TI-RADS classification can be used to differentiate the nature of thyroid nodules, demonstrating high accuracy with low risks of missed or misdiagnoses. It is also closely related to indicators such as MK, TK1, and TSH, providing a basis for the detection of thyroid cancer.

Objective This review aims to investigate the application of eye tracking(ET)in patients with speech impairment in the ICU.The review summarizes the current status and prospects of ET technology applications.Methods Following the scoping reviews framework,a systematic search was conducted across Web of Science,PubMed,Cochrane Library,Embase,CINAHL,CNKI,Wanfang Data,Chinese Medical Database,and Chinese biomedical database.The search covered publications from the inception of each database to October 29th,2024.The included studies were comprehensively analyzed.Results A total of 12 pieces of the literature were included,including 4 prospective cohort studies,4 experimental studies,3 prospective observational studies,and 1 randomized controlled trial.The application population of ET in the ICU mainly includes patients on mechanical ventilation,those at high risk of delirium,patients with spinal cord injury,etc.The types of integrated ET system equipment are mainly head-mounted and fixed;the types of ET involve gaze,blinking,etc.;the functions include standardized scale assessment,free interaction,and eye-movement model recognition.The main outcome indicators of the research are feasibility,physical symptoms and social-psychological status.Conclusion ET is applicable to a specific group of ICU patients with aphasia,and has shown good feasibility and effectiveness in the expression of patients'basic needs,self-assessment of symptoms and improvement of psychosocial status.

Microglia activation-mediated neuroinflammatory responses serve as a critical pathological ba-sis for the development and progression of various brain diseases.The role of circular RNAs(circRNAs)in the regulation of neuroinflammation is increasingly being recognized.This study aimed to investigate the effect and molecular mechanisms of targeted inhibition of circular RNA Homeodomain Interacting Pro-tein Kinase 3(HIPK3)(circHIPK3)on lipopolysaccharide(LPS)-induced microglial polarization in BV2 cells.The results showed that LPS stimulation significantly induced polarization of BV2 cells towards the pro-inflammatory M1 phenotype and upregulated circHIPK3 expression(P＜0.01).Engineered extra-cellular vesicles(EVs)with rabies viral glycoprotein(RVG)loaded with circHIPK3 siRNA(RVG-EVs-sicHIPK3)were successfully constructed.Transmission electron microscopy(TEM)revealed their typi-cal EV morphology.nanoparticle tracking analysis(NTA)indicated a peak particle size of 70 nmn.And Western blotting analysis confirmed the expression of characteristic membrane marker proteins.Treatment with RVG-EVs-sicHIPK3 significantly suppressed the LPS-induced elevation of inflammatory cytokines(TNF-α,IL-6,IL-1β)in the supernatant and reduced the expression of M1 phenotypic marker proteins(CD16 and CD86)(P＜0.01).Concurrently,RVG-EVs-sicHIPK3 increased the number of mitophago-somes within cells,upregulated the ratio of the autophagy-related proteins LC3-Ⅱ/LC3-I(P＜0.01),and downregulated the expression of the autophagy-related protein p62 and mitochondrial-specific proteins(TOMM20 and TIMM23)(P＜0.01).The mitophagy inhibitor Mdivi-1 significantly reversed the RVG-EVs-sicHIPK3-mediated downregulation of inflammatory cytokine levels,M1 marker proteins,and mito-chondrial protein expression(P＜0.01).This study demonstrates that inhibiting circHIPK3 reduces LPS-induced microglial polarization towards the M1 phenotype.The protective mechanism is closely associated with enhanced mitophagic flux and the promotion of damaged mitochondrial clearance.

Objective:To analyze the application value of ultrasound Thyroid Imaging Reporting and Data System (TI-RADS) classification in the differential diagnosis of thyroid nodules and its correlation with patients' midkine, thymidine kinase 1, and thyroid function indicators.Methods:This study was a prospective study. A total of 108 patients with thyroid nodules (120 nodules) who visited Jinan 2 nd People's Hospital from December 2022 to February 2024 were included. Ultrasound examinations were conducted to complete the TI-RADS classification, and serum levels of midkine (MK) and thymidine kinase 1 (TK1), as well as thyroid function indicators, were measured. Based on the patients' pathological results, the consistency between the TI-RADS diagnostic results and the pathological findings was analyzed. The diagnostic efficacy of TI-RADS classification was calculated. The incidence of malignant nodules across various TI-RADS categories was assessed, and the differences in serum levels of MK, TK1, and thyroid function indicators between patients with benign and malignant nodules were compared. Additionally, a correlation analysis was performed to analyze the correlation between the TI-RADS classification of thyroid nodules and the serum levels of MK, TK1, and thyroid function indicators. Results:Pathological examination results revealed 38 malignant nodules and 82 benign nodules. The consistency between ultrasound TI-RADS classification diagnoses and pathological results was good ( Kappa = 0.90, P < 0.001), with diagnostic sensitivity, specificity, and accuracy of 86.84% (33/38), 97.56% (80/82), and 94.17% (113/120), respectively. Among the ultrasound TI-RADS classifications, the malignant rates for category Ⅳb and category Ⅴ thyroid nodules were the highest, at 92.00% (23/25) and 100.00% (10/10), respectively. Serum levels of MK, TK1, and thyroid-stimulating hormone (TSH) were significantly higher in patients with malignant thyroid nodules than in those with benign thyroid nodules (all P < 0.05). Correlation analysis indicated that TI-RADS classification was positively correlated with serum levels of MK, TK1, and TSH ( r = 0.56, 0.60, 0.52, all P < 0.05). Conclusions:Ultrasound TI-RADS classification can be used to differentiate the nature of thyroid nodules, demonstrating high accuracy with low risks of missed or misdiagnoses. It is also closely related to indicators such as MK, TK1, and TSH, providing a basis for the detection of thyroid cancer.

Objective:To analyze the correlation of bone mineral density (BMD) , serum transforming growth factor β1 (TGF- β1) , and retinol-binding protein 4 (RBP4) with osteoporosis in gestational diabetes mellitus (GDM) . Methods:A total of 180 GDM patients admitted to Department of Gynecology, Second Hospital of Tianjin Medical University from Jan. 2022 to Jan. 2024 were included as the observation group, and 30 healthy pregnant women undergoing examination at the same time were included into the control group. BMD [stiffness index (SI) , broadband ultrasound attenuation (BUA) , speed of sound (SOS) ], serum TGF-β1, and RBP4 levels were compared between the two groups; GDM women with complicated with osteoporosis were included in the osteoporosis subgroup, and pregnant women without osteoporosis were included in the non-osteoporosis subgroup. BMD, serum TGF- β1 and RBP4 levels were compared between the two groups, and the relationship among bone mineral density and serum TGF- β1, RBP4 was analyzed by Pearson correlation. Receiver operating characteristic curve (ROC) curve was drawn to evaluate the diagnostic value of serum TGF- β1 and RBP4 levels in GDM complicated with osteoporosis. Results:The observation group had significantly higher levels of serum TGF- β1 and RBP4 than the control group, while lower BUA, SOS and SI ( t=99.04, 28.48, 4.10, 3.54, 6.29, P < 0.05) ; Among 180 pregnant women with GDM, 38 cases had osteoporosis and 142 cases did not have osteoporosis. The osteoporosis subgroup had significantly higher serum TGF- β1 and RBP4 levels than the other subgroup, while BMD indexes such as BUA, SOS and SI were significantly lower ( t=3.35, 3.48, 3.77, 2.85, 3.41, P < 0.05) ; Pearson correlation analysis showed that BMD indexes, such as BUA, SOS and SI, were significantly correlated with serum TGF- β1 and RBP4 in pregnant women with GDM ( r=-0.61, 0.58, -0.60, -0.58, -0.60, -0.63, P<0.05) ; The area under the curve (AUC) of TGF- β1, RBP4 and combined detection of GDM women with osteoporosis was 0.572, 0.653 and 0.659, respectively. Conclusions:In GDM women complicated with osteoporosis, the levels of serum TGF- β1 and RBP4 increase significantly, and BMD decreases significantly. Serum TGF- β1 and RBP4 in GDM women are closely related to BMD index, which can provide early diagnosis basis for GDM complicated with osteoporosis.

Objective To explore the characteristics and influencing factors of complicated intra-abdominal infection in ICU patients after abdominal surgery.Methods A retrospective study was performed on ICU patients(n=92,observation group)developing complicated intra-abdominal infection after abdominal surgery versus patients(n=104,control group)without complicated intra-abdominal infection after abdominal surgery in our hospital from January 2020 to December 2023.The characteristics of patients who developed complicated intra-abdominal infections were analyzed and the relevant influencing factors were identified using multivariate Logistic regression analysis.Results A total of 124 strains of pathogenic bacteria were isolated and identified in 92 infected patients,with 64.52％(80/124)of Gram-negative strains comprising mainly 29.03％(36/124)of Escherichia coli,25.81％(32/124)of Gram-positive strains comprising mainly 12.90％(16/124)of Enterococcus faecalis,and 9.68％(12/124)of fungi comprising mainly 6.45％(8/124)of Candida albicans.Multivariate Logistic regression results showed that preoperative underlying disease,surgical methods,duration of surgery,invasive procedures,antibiotic use,and length of ICU stay,enteral nutrition were the influencing factors of complicated intra-abdominal infection in ICU patients after abdominal surgery(P＜0.0 5).Conclusion Patients with complicated intra-abdominal infection are infected with a variety of pathogenic bacteria,predominantly Gram-negative.The patient's preoperative underlying disease,surgical approach,duration of surgery,invasive procedures,use of antibiotics,length of ICU stay,and early enteral nutrition all affect the risk of complicated intra-abdominal infection in ICU patients after abdominal surgery.

ObjectiveTo investigate the liver histopathological features of HBeAg-negative patients in the indeterminate phase of low-viral-load chronic hepatitis B virus （HBV） infection. MethodsA total of 271 patients with low-viral-load HBeAg-negative chronic HBV infection who underwent liver biopsy in Department of Infectious Diseases， Affiliated Hospital of Yan’an University， from September 2013 to June 2021 were enrolled as subjects， and the degree of liver injury was compared between patients based on age， sex， presence or absence of the family history of hepatitis B， HBsAg， and alanine aminotransferase （ALT） level. The chi-square test was used for comparison of categorical data between two groups. ResultsAmong the 271 patients with HBeAg-negative chronic HBV infection， 86 patients （31.73%） grade≥A2 liver inflammatory activity， 72 （26.57%） had a liver fibrosis stage of ive， and 112 （41.33%） had moderate or severe liver histological injury. The proportion of patients with grade≥A2 liver inflammatory activity in the patients with ALT>20 U/L was significantly higher than that in the patients with ALT≤20 U/L （χ²=3.938， P=0.047）. There were no significant differences in the proportion of patients with grade≥A2 liver inflammatory activity between the patients with different ages， sexes， family history of hepatitis B， HBsAg levels （all P>0.05），there were no significant differences in the proportion of patients with a liver fibrosis stage of ≥F2 between the patients with different ages， sexes， family history of hepatitis B， HBsAg， and ALT levels （all P>0.05）， and the stratified analysis of patients aged≤30 years and patients without the family history of hepatitis B showed no statistical significance between groups （all P>0.05）. There was no significant difference in the degree of liver histological injury between the patients with different ages， sexes， family history of hepatitis B， HBsAg， and ALT levels （all P>0.05）. ConclusionSignificant liver injury is observed in more than 40% of the patients with low-viral-load HBeAg-negative chronic HBV infection， and there is no significant difference in the degree of liver histological injury between the patients with different ages， sexes， family history of hepatitis B， HBsAg， and ALT levels. Even for the patients aged≤30 years who deny the family history of hepatitis B， there is still a considerable proportion of patients with liver injury， which should be taken seriously by clinicians.