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.

ObjectiveTo evaluate the clinical efficacy of Huayu Tongluo Formula （化瘀通络方， HTF） in patients with mid-stage diabetic kidney disease of blood stasis syndrome and explore its potential mechanisms. MethodsA multi-center， randomized， double-blind， placebo-controlled clinical trial was conducted. Ninety patients of mid-stage diabetic kidney disease of blood stasis syndrome were divided into a control group of 46 cases and a treatment group of 44 cases. Both groups received conventional western medicine treatment， the treatment group additionally taking HTF， while the control group taking a placebo of the formula. The treatment was administered once daily for 24 weeks. The primary outcomes included 24-hour urine total protein （24 h-UTP）， serum albumin （Alb）， glycated hemoglobin （HbA1c）， and serum creatinine （Scr）.The secondary outcomes included changes in levels of endothelin-1 （ET-1）， nitric oxide （NO）， vascular endothelial growth factor （VEGF）， and traditional Chinese medicine （TCM） syndrome scores before and after treatment. Clinical efficacy was evaluated based on TCM syndrome scores and overall disease outcomes. Adverse reactions and endpoint events were recorded. ResultsIn the treatment group after treatment， 24 h-UTP， ET-1， and VEGF levels significantly decreased （P＜0.05）， Alb and NO levels significantly increased （P＜0.05）； while the TCM syndrome scores for edema， lumbar pain， numbness of limbs， dark purple lips， dark purple tongue or purpura， and thin， rough pulse all significantly decreased （P＜0.05）. In the control group， no significant changes were observed in any of the indicators after treatment （P＞0.05）.Compared with the control group， the treatment group showed significant reductions in 24 h-UTP， ET-1， and VEGF levels， and increases in Alb and NO levels （P＜0.05）. The TCM syndrome scores for edema， lumbar pain， dark purple tongue or purpura， and thin， rough pulse were all lower in the treatment group than in the control group （P＜0.05）. The total effective rate of TCM syndrome in the treatment group was 59.09% （26/44）， and the overall clinical effective rate was 45.45% （20/44）. In the control group， these rates were 15.22% （7/46） and 8.7% （4/46）， respectively， with the treatment group showing significantly better outcomes （P＜0.05）. A total of 7 adverse events occurred across both groups， with no significant difference （P＞0.05）. No endpoint events occurred during the study. ConclusionOn the basis of conventional treatment of Western medicine， HTF can further reduce urinary protein levels and improve clinical symptoms in patients with mid-stage diabetic kidney disease of blood stasis syndrome. The mechanism may be related to its effects on endothelial function.

Exosomal circular RNA(circRNAs)are pivotal in cancer biology,and tumor pathophysiology.These stable,non-coding RNAs encapsulated in exosomes participated in cancer progression,tumor growth,metas-tasis,drug sensitivity and the tumor microenvironment(TME).Their presence in bodily fluids positions them as potential non-invasive biomarkers,revealing the molecular dynamics of cancers.Research in exosomal circRNAs is reshaping our understanding of neoplastic intercellular communication.Exploiting the natural properties of exosomes for targeted drug delivery and disrupting circRNA-mediated pro-tumorigenic signaling can develop new treatment modalities.Therefore,ongoing exploration of exoso-mal circRNAs in cancer research is poised to revolutionize clinical management of cancer.This emerging field offers hope for significant breakthroughs in cancer care.This review underscores the critical role of exosomal circRNAs in cancer biology and drug resistance,highlighting their potential as non-invasive biomarkers and therapeutic targets that could transform the clinical management of cancer.

Objective:To compare the effects of different automatic normal tissue objective(NTO)weights(WNTO)of Eclipse planning system on intensity modulated radiation therapy(IMRT)of fixed field for abdominal tumor.Methods:Twenty patients with rectal cancer and twenty patients with cervical cancer who received radiotherapy in The Second People's Hospital of Huludao from September 2022 to January 2024 were selected.5 groups of different plans[WNTO20,40,60,80 and 100]were respectively formulated for each patient.Among of them,the WNTO20 plan was control group,and other 4 groups were respectively WNTO40 group,WNTO60 group,WNTO80 group and WNTO100 group.Then,the differences in dosimetry and monitor units(MU)among 5 groups of plans were compared.Results:In the WNTO 80 and 100 groups of patients with rectal cancer,the conformation index(CI)values of target regions of them were respectively 1.02±0.04 and 1.00±0.03,all of which approached to 1,and the CI values of two groups were lower than that of control group,and the differences were statistically significant(t=15.986,18.422,P<0.05).The homogeneity index(HI)of WNTO 80 and 100 groups were respectively 0.09±0.12 and 0.10±0.14,which were higher than that of control group,and the differences were statistically significant(t=-1.371,-1.463,P<0.05).The 52Gy dose volume(Vmax52)of small intestine of the two groups were respectively(4.48±14.49)and(4.77±10.47)cm3,which were higher than that of control group,and the differences were statistically significant(t=-1.360,-2.005,P<0.05).The 30 Gy dose volume percentage(V30 Gy)values of the right femoral heads of the two groups were respectively(12.28±4.57)%and(10.96±4.17)%,which were lower than that of control group,and the differences were statistically significant(t=6.893,6.75,P<0.05).The 30 Gy dose volume percentage(V30 Gy)values of the left femoral head of the two groups were respectively(11.57±4.41)%and(10.17±3.78)%,which were lower than that of control group,and the differences were statistically significant(t=6.782,5.963,P<0.05).The MU value of WNTO100 group was(1254±93)MU,which was higher than that of control group,and the difference was statistically significant(t=-3.741,P<0.05).In the WNTO 80 and 100 groups of patients with cervical cancer,the CI values of target regions of them were respectively 1.03±0.32 and 1.02±0.03,all of which approached to 1,and the CI values of the two groups were lower than those of control group,and the differences were statistically significant(t=20.069,19.475,P<0.05).The HI values of them were respectively 0.07±0.05 and 0.08±0.01,which were higher than that of control group,and the differences were statistically significant(t=-12.445,-19.478,P<0.05).The V30 Gy values of the right femoral heads of the two groups were respectively(16.11±4.71)%and(14.90±4.56)%,which were lower than that of control group,and the differences were statistically significant(t=7.875,8.020,P<0.05).The V30 Gy values of the left femoral head of the two groups were respectively(15.21±3.71)%and(13.93±3.77)%,which were lower than that of control group,and the differences were statistical significance(t=7.617,7.436,P<0.05).The Vmax52 of small intestine of the two groups were respectively(0.82±1.10)and(4.47±8.81)cm3,which were higher than that of control group,and the difference was statistically significant(t=-2.985,-2.388,P<0.05).Conclusion:The WNTO80 is recommended for IMRT plan of radiotherapy on abdominal tumor,which can more quickly and better meet the assessment conditions of IMRT plan.

Liquiritigenin(LG)is a flavone of pharmacological importance,however,its application potential is severely limited due to its poor water solubility.LG could be disassociated slightly in water to form phenolate anion,therefore,better solubilization effect is expected by inclusion with cationic cyclodextrins(CCDs).In this work,four kinds of CCDs modified with amino groups at the primary face were synthesized,and their solid inclusion complexes with LG were successfully prepared by preparing their saturated solutions.The formation of the solid inclusion complexes was confirmed by scanning electron microscopy(SEM)and powder X-ray diffraction(PXRD),and their supramolecular binding behavior in solution was studied using multiple techniques.A 1∶1 inclusion stoichiometry of inclusion complexation was defined using Job plot by ultraviolet-visible(UV-vis)spectroscopy,and their binding stability constants(Ks)were determined as 2862.77,3494.70,6521.85 and 9599.48 L/mol using UV-vis spectroscopic titration,far more superior to that of nativeβ-CD(Ks=236.79 L/mol).This indicated that the amino side chains on CCDs could actively participate in the inclusion complexation through anion-cation interactions,significantly strengthening the host-guest binding between CCDs and LG.The inclusion modes were further elucidated based on proton and two-dimensional rotating-frame overhauser enhancement spectroscopy(2D-ROESY)nuclear magnetic resonance(NMR)experiments and molecular docking.Water solubility of LG was dramatically promoted up to 4.9 mg/mL,which was 70-fold higher than that of native LG.This study could draw inspiration for the binding and solubilization of phenols such as flavones by design of cationic macrocyclic molecules.

Current ambient mass spectrometry ionization often requires external auxiliary equipment such as high-voltage power supply,gas cylinder,and syringe pump.Moreover,the process of sample preparation is cumbersome,and the experimental operations are complex,which makes it difficult to adapt to real-time on-site detection.In this work,a novel method was proposed,in which direct sampling of raw samples,online extraction of interest analytes,and ionization of target molecules were integrated into a single unit.With the developed method,the in-situ extraction and nano-electrospray ionization for both liquid and solid raw samples were achieved.Also,a handheld ion source and its pose adjustment device were developed,and the position and angle parameters were subsequently optimized.The performance of the ionization device was tested using standard solutions of caffeine and reserpine.The limits of detection(LODs)were 0.08 μg/L and 0.14 μg/L,with relative standard deviations(RSDs)≤3.7% and≤5.6%,respectively,indicating that the device possessed high sensitivity and stability.Using this device,three different concentrations of reserpine standard solutions were continuously tested for five days.The intra-day RSDs were consistently≤4.7% and the inter-day RSDs were all≤10.3%,showing the good working stability of the device.Without any pretreatment,a rapid qualitative detection of medicinal components including astragaloside II and cycloastragenol in five traditional Chinese medicines was carried out,with RSDs≤8.0% and≤7.1%,respectively.Additionally,rapid qualitative detection of gallic acid,a medicinal component,in white peony roots,and hypaphorine as well as quercetin in cowherb seeds were carried out,with RSDs≤7.0%,≤6.4% and≤6.1%,respectively.These results demonstrated that the ionization technology and device exhibited good stability during qualitative detection of raw samples.

Objective:To compare the effect of empirical cefazolin and vancomycin on the adverse outcomes in peritoneal dialysis (PD)-associated peritonitis patients.Methods:This was a retrospective analysis of a single-centre prospective cohort. Clinical data of consecutive PD-related peritonitis episodes occurring for the first time in patients (≥18 years) between January 1, 2008 and December 31, 2021 were reviewed. Patients were classified into a cefazolin group or a vancomycin group according to the empirical antibiotic regimen. The primary endpoint was peritonitis-related death within 1 month of onset and the secondary endpoint was transfer to haemodialysis for peritonitis-related reasons within the same period. Differences in both endpoints between the two regimens were analysed in the overall population and in the Gram-positive peritonitis subgroup. Univariable and multivariable logistic regression models were used to estimate the risk of adverse outcomes associated with antibiotic choice. Propensity-score matching was performed to control for confounding bias.Results:A total of 516 eligible PD patients developed peritonitis during the study period were included, among whom 138 received empirical cefazolin and 322 received vancomycin. Baseline characteristics were significantly different between the cefazolin and vancomycin groups, including annual income >50 000 yuan ( χ2=17.854, P<0.001), cardiovascular disease history ( χ2=3.909, P=0.048), prior peritonitis history ( χ2=18.327, P<0.001), serum albumin ( t=2.430, P=0.013), triglycerides ( Z=-3.108, P=0.002), total cholesterol ( t=3.752, P<0.001), phosphate ( t=3.362, P=0.002) and sodium ( t=3.021, P=0.004). Neither peritonitis-related mortality nor transfer to haemodialysis differed between the cefazolin and vancomycin groups in the overall cohort or in the Gram-positive peritonitis subgroup (all P>0.05). Logistic regression analysis (univariable and multivariable) showed that empirical vancomycin was not associated with a higher risk of adverse outcome when compared with cefazolin in the overall cohort or in the Gram-positive peritonitis subgroup (all P>0.05). After propensity-score matching, results remained consistent (all P>0.05). Conclusion:Empirical cefazolin and vancomycin yield similar rates of short-term adverse outcomes in patients with PD-associated peritonitis, including those caused by Gram-positive organisms.

Objective:To review and analyze the results of cervical cancer screening for women aged 30-65 years in Tianjin from 2018—2023, and to explore the choice of cervical cancer screening strategies.Methods:The results of free cervical cancer screening in Tianjin from January 2018 to December 2023 were collected and divided into two periods: group A [2018—2020; screening programs included cervical cytology, Thinprep cytologic test (TCT), and human papillomavirus (HPV) DNA+TCT combined screening] and group B (2021—2023; the screening scheme of HPV E6/E7 mRNA primary screening and TCT diversion, referred to as “HPV E6/E7 mRNA primary screening”) according to the policy changes, and according to the primary screening method of cervical cancer, it was divided into four methods: cervical smear primary screening, TCT primary screening, HPV DNA+TCT combined primary screening and HPV E6/E7 mRNA primary screening. Cervical cancer and precancerous lesion detection rate, cervical cancer early diagnosis rate were analyzed.Results:(1) A total of 1 634 155 women of eligible age were screened, and the detection rate of cervical cancer and precancerous lesion was 216.3/100 000 (3 535/1 634 155), among which the detection rate of precancerous lesion was 200.3/100 000 (3 274/1 634 155) and cervical cancer was 16.0/100 000 (261/1 634 155). (2) The cervical cancer and precancerous lesion detection rate and cancer early diagnosis rate in group B were significantly higher than those in group A [480.3/100 000 (2 132/443 924) vs 117.9/100 000 (1 403/1 190 231), 97.56% (2 080/2 132) vs 92.37% (1 296/1 403); both P<0.001]. (3) The detection rates of cervical cancer and precancerous lesions in women who underwent initial screening with cervical cytology, TCT, HPV DNA+TCT combined screening, and HPV E6/E7 mRNA screening were 106.8/100 000 (10 980/1 028 227), 167.7/100 000 (190/113 319), 236.2/100 000(115/48 685) and 480.3/100 000 (2 132/443 924), respectively. HPV E6/E7 mRNA primary screening had the highest cervical cancer and precancerous lesion detection rate among the four screening methods ( χ2=2 017.59, P<0.001). Conclusion:After 2021, the cervical cancer screening method of HPV E6/E7 mRNA primary screening and TCT triage adopted in Tianjin is suitable for general screening of large populations and is an effective method for initial screening of cervical cancer.

Objective:To explore the differences in early clinical features between Kawasaki disease shock syndrome (KDSS) and septic shock (SS).Methods:A retrospective case-control study was conducted. Clinical data was collected from 64 children who were diagnosed with KDSS or SS and admitted to the Department of Critical Care Medicine of Capital Center for Children′s Health, Capital Medical University from January 2018 to February 2025. Mann-Whitney U test, χ2 test, or Fisher′s exact test were used to compare the differences in clinical features, treatment, and outcomes between children with KDSS and SS. Lasso regression was applied to screen predictive variables, and multivariable logistic regression analysis was performed to identify factors associated with KDSS. Receiver operating characteristic (ROC) curve was used to evaluate the predictive value of parameters for KDSS. Results:Among the 64 children (30 males and 34 females), the age was 3.6 (1.2, 6.5) years. There were 51 cases in the SS group and 13 cases in the KDSS group. Compared to children with SS, children with KDSS had a longer pre-shock fever duration, lower lactate levels and serum albumin levels, and higher soluble interleukin-2 receptor (sIL-2R) levels (all P<0.05). Additionally, they exhibited a higher incidence of coronary involvement, pericardial effusion, and ascites, a higher utilization rate of intravenous immunoglobulin, and a lower utilization rate of invasive mechanical ventilation (all P<0.05). There was no significant difference in in-hospital mortality between KDSS and SS ( P=0.574). Multivariate logistic regression analysis identified pre-shock fever duration and sIL-2R as independent factors associated with KDSS ( OR=1.52 and 1.54 per 1 000 U increase, 95% CI 1.12-2.05 and 1.06-2.24, respectively; both P<0.05). ROC curve analysis showed that the areas under the curve for pre-shock fever duration and sIL-2R in identifying KDSS were 0.83 (95% CI 0.73-0.94, P=0.001) and 0.70 (95% CI 0.53-0.87, P=0.042), respectively. The optimal cutoff values were 3.5 d and 3.8×10 6 U/L, with sensitivities of 0.91 and 0.82, and specificities of 0.71 and 0.62, respectively. Conclusions:Children with KDSS have higher incidences of coronary involvement, pericardial effusion, and ascites compared to those with SS. Pre-shock fever duration and sIL-2R may serve as potential early indicators for distinguishing KDSS from SS.