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 incidence and risk factors of open gingival embrasures (OGEs) in the incisor region after treatment with clear aligners in adult non-extraction patients and provide a reference for preventing the occurrence of an open gingival wedge gap in the incisal area after orthodontic treatment. Methods: This study has been reviewed and approved by the institutional medical ethics committee, and informed consent was obtained from the patients. A total of 125 adult patients with malocclusion who completed clear aligner treatment at Hefei Stomatological Hospital from September 2022 to December 2024 were selected as the study subjects. Based on the presence or absence of OGEs in the incisor region observed in frontal intraoral photographs taken immediately after treatment completion, the patients were divided into a normal group and an OGE group. Clinical data, including intraoral photographs, digital models, and cone-beam computed tomography before and after treatment, were analyzed. Measurements such as incisor overlap and rotation, crown morphology, number of attachments, and interproximal enamel reduction (IPR) were recorded and analyzed. Results: The incidence of OGEs between the maxillary and mandibular central incisors after clear aligner treatment in adult patients was 28.8% and 39.2%, respectively. No statistically significant differences were observed between the normal and OGE groups in terms of sex, Angle's classification, gingival biotype, overbite, overjet, IPR amount, age, treatment duration, tooth axis angulation, or horizontal movement distance of mandibular central incisors before and after treatment (P 0.05). However, significant differences were found in the number of attachments, anteroposterior distance between mesial incisal angles, distance from the interproximal contact point (ICP) to the alveolar bone crest (ABC) (ICP-ABC), horizontal distance between mesial cementoenamel junction (CEJ) of two adjacent central incisors (CEJ-CEJ) and labial alveolar bone thickness (P 0.05). IPR amount and mandibular incisor intrusion were significantly associated with the severity of OGEs (P 0.05). Regression analysis revealed that the number of attachments, anteroposterior distance between mesial incisal angles, ICP-ABC distance, and CEJ-CEJ horizontal distance were significantly correlated with the occurrence of OGEs. Conclusion The incidence of open gingival embrasures in the mandibular central incisor region is relatively high among adult patients treated with clear aligners. The number of attachments (n = 2), the anteroposterior distance between the mesio-incisal angles, the distance from the tooth contact point to the alveolar bone crest, and the horizontal distance between adjacent cementoenamel junctions have been identified as risk factors for the development of open gingival embrasures upon completion of orthodontic treatment.

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 explore the effects of lipoxygenase(LOX)on severe fever with thrombocytopenia syndrome virus(SFTSV)in human umbilical vein endothelial cells(HUVECs).Methods The CCK-8 assay was used to assess the cytotoxicity of LOX in HUVECs.Real-time quantitative PCR(qPCR)was adopted to detect the replication of viral RNA in cells.The infection rate of SFTSV to HUVECs was observed via indirect immunofluorescence assay(IFA).The expressions of intracellular viral NP protein and autophagy related proteins microtubule-associated protein 1 light chain 3(LC3)and adenosine 5'-monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)were detected via Western blotting.Lipid metabolomics was used to analyze the differences in expressions of intracellular metabolites in the SFTSV-infected Huh 7 with or without LOX treatment.Results The results of CCK-8 showed no obvious cytotoxicity to HUVECs with LOX≤8 mg/mL.With the increase of LOX concentrations,the viral RNA level,viral infection rate,and expressions of SFTSV nucleoprotein(NP)protein were gradually decreased,respectively.The autophagy reaction was activated in SFTSV-infected HUVECs,evidenced by the increased expression level of LC3 Ⅱ protein.Compared with the untreated group,LOX treatment resulted in decreased levels of LC3 Ⅱ and AMPK phosphorylation,and enhanced phosphorylation of mTOR in SFTSV-infected HUVEC.Conclusion LOX negatively regulates autophagy by inhibiting AMPK/mTOR signaling pathway,thereby inhibiting SFTSV replication.

ObjectiveTo investigate the association of SH2B1 rs7359397 （C>T） polymorphism with the progression to hepatic fibrosis in the elderly patients with metabolic dysfunction-associated steatotic liver disease （MASLD） in Beijing， China， and to provide an important genetic basis for the precise subtyping， prognostic evaluation， and individualized treatment of elderly MASLD patients in China. MethodsA total of 505 elderly patients （aged ≥65 years） who participated in regular physical examination in Mentougou Kuangshan Hospital of Beijing Jingmei Group General Hospital from November 2020 to September 2021 and were diagnosed with MASLD by abdominal ultrasound were enrolled as MASLD group， and 381 elderly population who underwent physical examination in the same community hospital during the same period of time and were not found to have MASLD by abdominal ultrasound were enrolled as control group. FibroScan was used to measure liver fat content and determine fibrosis stage. The 96-well microfluidic chip technique was used to identify SH2B1 rs7359397 polymorphism. The independent-samples t test was used for comparison of normally distributed continuous data between the two groups， and the chi-square test or the adjusted chi-square test was used for comparison of categorical data between the two groups. Univariate and multivariate Logistic regression analyses were used to identify the independent predictive factors for MASLD and its comorbidities. ResultsCompared with the control group， the MASLD group had a significantly younger age and significantly higher levels of waist circumference， hip circumference， waist-hip ratio， body mass index （BMI）， alanine aminotransferase， aspartate aminotransferase， triglyceride， platelet count， and fibrosis-4 （FIB-4） index， as well as a significantly lower level of high-density lipoprotein cholesterol （all P<0.05）. Among the 381 patients in the control group， 264 （69.29%） had genotype CC and 117 （30.71%） had genotype CT+TT， while among the 505 patients in the MASLD group， 317 （62.77%） had genotype CC and 188 （37.23%） had genotype CT+TT， suggesting that the MASLD group had a significantly higher proportion of patients with genotype CT+TT compared with the control group （χ²=4.09， P=0.043）. In the MASLD group， compared with the genotype CC group， the genotype CT+TT group had a significantly lower proportion of patients with FIB-4 ≥2 or atherosclerotic cardiovascular diseases （P<0.05）. The multivariate Logistic regression analysis showed that after adjustment for age， sex， and BMI， carrying T allele was a protective factor against progressive hepatic fibrosis （odds ratio ［OR］=0.481， 95% confidence interval ［CI］： 0.249 — 0.929， P=0.029）. In the subgroups of comorbidities with hypertension， metabolic syndrome， and obesity， genotype CT+TT was associated with a significant reduction in the risk of progressive hepatic fibrosis （hypertension： OR=0.27， 95%CI：0.09 — 0.77， P=0.014； metabolic syndrome： OR=0.30， 95%CI： 0.11 — 0.79， P=0.015； obesity： OR=0.11， 95%CI： 0.03 — 0.48， P=0.003）. After adjustment for age， sex， and BMI， in the patients with MASLD， the patients with genotype CT+TT had a significant reduction in the prevalence rate of atherosclerotic cardiovascular diseases compared with those with genotype CC （OR=0.506， 95%CI：0.336 — 0.761， P=0.001）. ConclusionSH2B1 rs7359397 （C>T） polymorphism is associated with the reduction in the risk of hepatic fibrosis and atherosclerotic cardiovascular diseases in MASLD patients.

Objective:To evaluate the clinical rehabilitation effect of external counterpulsation(ECP)on patients with coronary heart disease(CHD)by meta-analysis.Methods:We searched the databases of CNKI,WanFang,VIP,CBM,PubMed,Web of Science,Cochrane Library and Embase for randomized controlled trials(RCTs)and prospective cohort studies upon rehabilitative effect of ECP on CHD patients before May 2024.And meta-analysis was conducted to calculate the pooled MD and 95％CI using the random(P＜0.5 or I2≥50％)or fixed effect models(other situations).Results:13 eligible literatures were finally included in the meta-analysis.Compared with participants in control group,those in trial group had significant higher left ventricular ejection fraction(MD=4.15,95％CI 2.55～5.76,P＜0.001),stroke volume(MD=9.11,95％CI 7.59～10.64,P＜0.001),peak oxygen uptake(MD=5.42,95％CI 2.53～8.32,P＜0.001),6-min walking distance(MD=31.14,95％CI 24.89～37.40,P＜0.001),metabolic equiv-alent(MD=0.58,95％CI 0.45～0.71,P＜0.001),exercise duration time(SMD=0.77,95％CI 0.55～0.99,P＜0.001),oxygen pulse(MD=0.88,95％CI 0.68～1.09,P＜0.001),and significant lower left ventricular end-diastolic diameter(MD=-3.19,95％CI-5.20～-2.61,P＜0.001).Conclusion:This study showed that ECP could effectively improve heart function,exercise capacity and tolerance of CHD patients.

Objective To explore the effects and mechanisms of Changpu Yujin Tang(CPYJT)on the NOD-like receptor thermal protein domain associated protein 3/cysteinyl aspartate specific proteinase-1/Gasdermin D(NLRP3/Caspase-1/GSDMD)signaling pathway-mediated neuroinflammation in rats with Tourette syndrome(TS).Methods SPF-grade male SD rats were randomly divided into the Control and TS groups.After successful modeling in the TS group,the rats were randomly divided into the Model,Tiapride,and CPYJT groups,and were treated with the corresponding drugs for 4 weeks.After the treatment,the rats' behavior was scored,H & E staining was used to observe pathological changes in the striatum,ELISA was used to measure the content of IL-1β and IL-18,RT-qPCR was used to detect the expression of NLRP3 and ASC mRNA,and Western blot was used to detect the expression of NLRP3,ASC,Caspase-1,Cleaved-Caspase-1,GSDMD,and GSDMD-NT proteins.Results Compared with the Control group,the Model group showed increased scores of stereotyped and motor behaviors(P＜0.01),pathological changes in the striatal tissue,increased content of IL-1β and IL-18(P＜0.01),and upregulated expression of NLRP3,ASC mRNA,and NLRP3,ASC,Caspase-1,Cleaved-Caspase-1,GSDMD,and GSDMD-NT proteins(P＜0.01).Compared with the Model group,the Tiapride group and the CPYJT group showed decreased scores of stereotyped and motor behaviors(P＜0.01),improved pathological damage in the striatal tissue,reduced content of IL-1β and IL-18(P＜0.01),and inhibited expression of NLRP3,ASC mRNA,and NLRP3,ASC,Caspase-1,Cleaved-Caspase-1,GSDMD,and GSDMD-NT proteins(P＜0.01).Conclusion The therapeutic effect of CPYJT on TS is related to the inhibition of the neuroinflammatory response mediated by the NLRP3/Caspase-1/GSDMD signaling pathway.

Objective:To analyze the symptoms,diagnosis and treatment of upper urinary tract calculi patients combined with mild and moderate benign prostatic hyperplasia(BPH)after ureteral stent implantation.Methods:One hundred and six BPH pa-tients who were hospitalized for upper urinary tract calculi and had ureteral stents retained from January 2019 to December 2022 were selected and divided into 2 weeks group and 4 weeks group according to the time of removal of ureteral stents after surgery.Their gener-al clinical data were analyzed and compared.International Prostatic Symptom Scale(IPSS),postoperative ureteral Stent Symptom Questionnaire(USSQ),and incidence of adverse events after ureteral stent removal were recorded before and after removal.Results:The scores of IPSS were significantly increased in all patients,and symptoms in urinary tract had improved significantly after discharge(P＜0.05).Compared with the 2 weeks group,the USSQ score of the 4 weeks group was significantly increased(P＜0.05).And no significant adverse event was observed in the 2 weeks group after the removal of ureteral sten.Conclusion:IPSS score and USSQ score increased significantly during stent implantation in BPH patients with lithiasis.And complications increased sig-nificantly over time.Following thorough clinical assessment,early ureteral stent removal demonstrates both safety and efficacy,repre-senting an optimal therapeutic approach in selected cases.

Objective::To evaluate the early pregnancy loss (EPL) rates in women with and without low molecular weight heparin (LMWH) treatment during early pregnancy.Methods::A retrospective, non-randomized study was conducted at Guangzhou Women and Children’s Medical Center between June 2019 and March 2022, involving women diagnosed with polycystic ovary syndrome (PCOS). All participants conceived following standard preconception care and voluntarily chose either the control group or the LMWH intervention group during the first month of pregnancy. The intervention was administered throughout the entire first trimester. Early and final pregnancy outcomes were recorded, with a particular focus on EPL rates. In addition, venous blood samples and clinical data were collected to compare hormonal profiles, blood lipid levels, and anthropometric parameters between the two groups. Statistical analyses included the two-tailed unpaired Student’s t-test, Mann–Whitney U test, Chi-square test, and Kaplan–Meier survival analysis. A value of P < 0.050 was considered statistically significant. Results::Thirty-eight women in the LMWH group and 102 women in the control group were included. The EPL rates in the LMWH and control groups were 5.3% (2/38) and 26.5% (27/102), respectively ( χ2 = 7.582, P = 0.006, odds ratio ( OR) = 0.154, 95% confidence interval ( CI): 0.035-0.685). The age ( P = 0.005), PCOS subtype ( P = 0.012), and levels of total cholesterol ( P = 0.003), and high-density lipoprotein ( P = 0.018) were significantly different between these two groups. Continued follow-up was performed to observe the long-time effects of LMWH treatment in early pregnancy. Seventy-three patients were successfully delivered, 23 patients in the LMWH group and 50 patients in the control group. There was no significant difference between the LMWH and control groups in gestation length, bleeding during delivery, birth weight, gender of the newborn, or mode of delivery. In addition, Kaplan-Meier curve analysis revealed that LMWH treatment may decrease the risk of EPL in PCOS patients in the first trimester ( χ2 = 4.144, P = 0.040). Conclusion::LMWH treatment during early pregnancy may reduce the EPL rate in women with PCOS.

The endoplasmic reticulum is an important organelle responsible for the synthesis,folding and processing of proteins.When the endoplasmic reticulum protein folding ability is affect-ed,internal unfolded proteins or misfolded proteins accumulate in the endoplasmic reticulum accumulation,leading to endoplasmic reticulum structural dysfunction resulting in endoplasmic reticu-lum stress.Autophagy is the process by which cells selectively remove stress endoplasmic reticulum and error proteins.Under stress,the endoplasmic reticulum can maintain normal physiolog-ical functions through unfolded protein responses and autophagy.Postmenopausal osteoporosis is mainly due to an imbalance be-tween bone resorption by osteoclasts and bone formation by oste-oblasts.Endoplasmic reticulum stress and autophagy both modu-late bone cells status with consequences for bone homeostasis.This article provides a review of the progress of research on en-doplasmic reticulum stress and autophagy interaction in the path-ogenesis of postmenopausal osteoporosis.