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 analyze white matter hyperintensities(WMH)and 11C-CFT PET/CT characteristics in patients with PD,and explore their relationship.Methods A total of 84 PD pa-tients admitted to our department between March 2021 and March 2025 were retrospectively en-rolled,and they were divided into young group(33 cases)and elderly group(aged＞65 years,51 cases)and also into non-WMH group(22 cases)and WMH group(62 cases)based on the severity of WMH.The degree of white matter lesions was scored,including periventricular white matter hyperintense(PVH)score and deep white matter hyperintense(DWMH)score.All patients un-derwent head MRI and 11C-CFT-PET/CT scanning.The T/N value and asymmetry index were calculated.Results The elderly PD patients exhibited significantly advanced age and higher H-Y stages,but lower T/N values of affected side of caudate nucleus,contralateral caudate nucleus,affected side of anterior putamen,contralateral anterior putamen and contralateral posterior puta-men,and posterior putamen asymmetry index than the young patients(P＜0.05,P＜0.01).Simi-lar results were observed in the WMH group when compared with the non-WMH group(P＜0.05,P＜0.01).The PVH score was negatively correlated with the T/N values of affected side of caudate nucleus(r=-0.282,P=0.009),contralateral caudate nucleus(r=-0.215,P=0.049),affected side of anterior putamen(r=-0.249,P=0.022),contralateral anterior putamen(r=-0.280,P=0.010)and contralateral posterior putamen(r=-0.285,P=0.009),and DWMH score was also negatively correlated with the T/N values in the affected side of caudate nucleus and affected side of anterior putamen(P＜0.05).Conclusion Both age and disease severity impact WMH and dopaminergic system in PD patients.A higher WMH load is associated with dopamin-ergic neuronal damage.

ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus， in order to obtain the optimal fermentation conditions and flora structure， and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2（ITS2） high-throughput sequencing， combined with fungal community diversity analysis and fungal community structure analysis， were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7， 14， 21 d（numbered Y1-Y3 for the former， and numbered F1-F3 for the latter）， respectively. At the same time， the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， combined with principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and compound retention time， parent ions， characteristic fragment ions and other information， the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method， while in the non-pressure-shelf natural fermentation method， there was a significant difference with the fermentation process， and at the genus level， the dominant genus of samples Y1， Y2， Y3 and F2 was Aspergillus， while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable， and the microbial community structure was more stable， which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis， including 70 flavonoids， 38 coumarins， 10 alkaloids， 34 organic acids and 3 other compounds. After fermentation， two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition， multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods， mainly including flavonoids， organic acids and coumarins. Comprehensively， the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable， the species richness was high and the overall content of differential compounds was high， which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation， the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds， and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus， as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.

ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus， in order to obtain the optimal fermentation conditions and flora structure， and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2（ITS2） high-throughput sequencing， combined with fungal community diversity analysis and fungal community structure analysis， were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7， 14， 21 d（numbered Y1-Y3 for the former， and numbered F1-F3 for the latter）， respectively. At the same time， the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， combined with principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and compound retention time， parent ions， characteristic fragment ions and other information， the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method， while in the non-pressure-shelf natural fermentation method， there was a significant difference with the fermentation process， and at the genus level， the dominant genus of samples Y1， Y2， Y3 and F2 was Aspergillus， while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable， and the microbial community structure was more stable， which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis， including 70 flavonoids， 38 coumarins， 10 alkaloids， 34 organic acids and 3 other compounds. After fermentation， two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition， multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods， mainly including flavonoids， organic acids and coumarins. Comprehensively， the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable， the species richness was high and the overall content of differential compounds was high， which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation， the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds， and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus， as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.

Objective:Explore the effect of modified position on correction of breech presentation and the ma-ternal and infant outcomes in primiparas at 30-34 weeks of gestation in the midwifery clinic.Methods:From Jan-uary to December2024,274 primiparas at30-34 weeks of gestation who visited the midwifery clinic of The Gener-al Hospital of Northern Theater Command were selected.They were randomly divided into two groups according to the coin-tossing method.Through the inclusion exclusion,and elimination criteria,the selected pregnant women were finally categorized into the knee-chest position group(control group,n=126)and the modified position group(observation group,n=140).The success rate of breech presentation correction,delivery mode,duration of labor,1-minute and 5-minute Apgar scores of vaginally delivered infants,umbilical artery blood gas,postpartum re-covery,and postpartum depression incidence were compared between the two groups.Results:The success rate of breech presentation correction in the observation group was significantly higher than that in the control group(77.9%vs 58.7%,P<0.05).After breech presentation correction,the vaginal delivery rate was higher in the ob-servation group than in the control group(80.7%vs 66.2%,P<0.05).Among women who achieved vaginal de-livery,the intrapartum complication rate was significantly lower in the observation group than in the control group(22.7%vs.46.9%,P<0.05).The duration of the first stage of labor(557.7±195.5 min vs.629.6±208.2 min)and the second stage of labor(67.7±11.4 min vs.79.6±10.3 min)was significantly shorter in the observation group than in the control group(P<0.05).The pH of umbilical artery blood gas of newborns in the observation group delivered vaginally was significantly higher than that in the control group(7.367±0.028 vs.7.322±0.027,P<0.05).There were no statistically significant differences in the1-minute and5-minute Apgar scores of vaginally delivered infants,base excess,and lactate between the two groups(P>0.05).The initial time of lactation(21.7±4.4 h vs.23.5±4.1 h),time to ambulation(8.2±1.2 d vs.9.2±1.4 d),time for lochia to subside(29.6±4.8 d vs.34.1±4.8 d),and postpartum hemorrhage(298.1±56.1 ml vs.362.9±42.6 ml)in the observation group were significantly lower than those in the control group(P<0.05).There were no statistically significant differ-ences in the depression scores(EPDS)at 1 week and 6 weeks postpartum between the two groups of women with vaginal delivery(P>0.05).Conclusions:The modified position in the midwifery clinic significantly improves the success rate of breech presentation correction for primiparas at 30-34 weeks of gestation,good maternal and infant outcomes,and could be widely applied in clinical practice.

Objective:Explore the effect of modified position on correction of breech presentation and the ma-ternal and infant outcomes in primiparas at 30-34 weeks of gestation in the midwifery clinic.Methods:From Jan-uary to December2024,274 primiparas at30-34 weeks of gestation who visited the midwifery clinic of The Gener-al Hospital of Northern Theater Command were selected.They were randomly divided into two groups according to the coin-tossing method.Through the inclusion exclusion,and elimination criteria,the selected pregnant women were finally categorized into the knee-chest position group(control group,n=126)and the modified position group(observation group,n=140).The success rate of breech presentation correction,delivery mode,duration of labor,1-minute and 5-minute Apgar scores of vaginally delivered infants,umbilical artery blood gas,postpartum re-covery,and postpartum depression incidence were compared between the two groups.Results:The success rate of breech presentation correction in the observation group was significantly higher than that in the control group(77.9%vs 58.7%,P<0.05).After breech presentation correction,the vaginal delivery rate was higher in the ob-servation group than in the control group(80.7%vs 66.2%,P<0.05).Among women who achieved vaginal de-livery,the intrapartum complication rate was significantly lower in the observation group than in the control group(22.7%vs.46.9%,P<0.05).The duration of the first stage of labor(557.7±195.5 min vs.629.6±208.2 min)and the second stage of labor(67.7±11.4 min vs.79.6±10.3 min)was significantly shorter in the observation group than in the control group(P<0.05).The pH of umbilical artery blood gas of newborns in the observation group delivered vaginally was significantly higher than that in the control group(7.367±0.028 vs.7.322±0.027,P<0.05).There were no statistically significant differences in the1-minute and5-minute Apgar scores of vaginally delivered infants,base excess,and lactate between the two groups(P>0.05).The initial time of lactation(21.7±4.4 h vs.23.5±4.1 h),time to ambulation(8.2±1.2 d vs.9.2±1.4 d),time for lochia to subside(29.6±4.8 d vs.34.1±4.8 d),and postpartum hemorrhage(298.1±56.1 ml vs.362.9±42.6 ml)in the observation group were significantly lower than those in the control group(P<0.05).There were no statistically significant differ-ences in the depression scores(EPDS)at 1 week and 6 weeks postpartum between the two groups of women with vaginal delivery(P>0.05).Conclusions:The modified position in the midwifery clinic significantly improves the success rate of breech presentation correction for primiparas at 30-34 weeks of gestation,good maternal and infant outcomes,and could be widely applied in clinical practice.

Objective To analyze white matter hyperintensities(WMH)and 11C-CFT PET/CT characteristics in patients with PD,and explore their relationship.Methods A total of 84 PD pa-tients admitted to our department between March 2021 and March 2025 were retrospectively en-rolled,and they were divided into young group(33 cases)and elderly group(aged＞65 years,51 cases)and also into non-WMH group(22 cases)and WMH group(62 cases)based on the severity of WMH.The degree of white matter lesions was scored,including periventricular white matter hyperintense(PVH)score and deep white matter hyperintense(DWMH)score.All patients un-derwent head MRI and 11C-CFT-PET/CT scanning.The T/N value and asymmetry index were calculated.Results The elderly PD patients exhibited significantly advanced age and higher H-Y stages,but lower T/N values of affected side of caudate nucleus,contralateral caudate nucleus,affected side of anterior putamen,contralateral anterior putamen and contralateral posterior puta-men,and posterior putamen asymmetry index than the young patients(P＜0.05,P＜0.01).Simi-lar results were observed in the WMH group when compared with the non-WMH group(P＜0.05,P＜0.01).The PVH score was negatively correlated with the T/N values of affected side of caudate nucleus(r=-0.282,P=0.009),contralateral caudate nucleus(r=-0.215,P=0.049),affected side of anterior putamen(r=-0.249,P=0.022),contralateral anterior putamen(r=-0.280,P=0.010)and contralateral posterior putamen(r=-0.285,P=0.009),and DWMH score was also negatively correlated with the T/N values in the affected side of caudate nucleus and affected side of anterior putamen(P＜0.05).Conclusion Both age and disease severity impact WMH and dopaminergic system in PD patients.A higher WMH load is associated with dopamin-ergic neuronal damage.

Objective To investigate the effect of whole-course empowerment health intervention on the psychological resilience and self-care ability of older patients with severe pneumonia.Methods A total of 210 older patients with severe pneumonia admitted to Sichuan Provincial People's Hospital between January 2020 and December 2023 were enrolled.The patients were sequentially numbered according to the order of admission.Then,they were assigned to a conventional care group(105 cases)and a whole-course empowerment group(105 cases)by a 1∶1 ratio using a random number table.The conventional care group received conventional clinical intervention,while the whole-course empowerment group received the whole-course empowerment health intervention regimen in addition to the intervention administered in the conventional care group.Psychological resilience was assessed with the Connor-Davidson Resilience Scale(CD-RISC),self-care ability with Exercise of Self-Care Agency Scale(ESCA),and quality of life with the World Health Organization Quality of Life Brief Version(WHOQOL-BREF).The evaluation results obtained before and after the interventions were compared.Results After intervention,the scores for all dimensions of CD-RISC in the whole-course empowerment group were higher than those in the conventional care group(t=10.326,P＜0.001;t=6.386,P＜0.001;t=9.581,P＜0.001).The scores for all dimensions of ESCA after intervention were higher in whole-course empowerment group than those in the conventional care group(t=1 1.599,P＜0.001;t=9.072,P＜0.001;t=4.353,P＜0.001;t=16.646,P＜0.001).After intervention,the scores for all dimensions of WHOQOL-BREF in the whole-course empowerment group were higher than those in the conventional care group(t=9.164,P＜0.001;t=13.809,P＜0.001;t=5.001,P＜0.001;t=6.695,P＜0.001).Conclusion Whole-course empowerment health intervention significantly enhances psychological resilience,self-care capacity,and quality of life in older patients with severe pneumonia.

Aim To design and synthesize a series of phenylacetamide derivatives with different substituted phenylacetic acid as raw materials,and to investigate the protective effects of the compound on the damage of pancreatic β cells induced by palmitate acid(PA).Methods Min6 cells were cultured and divided into B blank control group,PA treatment group and PA+compounds group.The viability of Min6 cells was de-tected by CCK-8.The protein expressions of TXNIP and NLRP3 were observed by Western blot.MDA con-tent and SOD activity were detected by MDA and SOD kit.The insulin secretion of Min6 islet cells was meas-ured with insulin ELISA kit.Results A total of 10 phenylacetamide derivatives were designed and synthe-sized.Their structures were confirmed by 1H NMR and ESI-MS.Pharmacological activity study showed that most of the compounds had protective effects on islet βcells,among which LY-6 and LY-8 had stronger pro-tective effects than PA model group,with the cell via-bility of 61.4％,and LY-6 had the highest cell activi-ty,reaching to 104.9％.Compared with PA group,the protein expression of TXNIP and NLRP3 decreased in LY-6 and LY-8 groups,MDA content decreased and SOD activity increased,and insulin secretion of Min6 cell increased.Conclusions LY-6 and LY-8 inhibit TXNIP expression and decrease the activation of NL-RP3 inflammasome,and decrease the production of MDA and increase SOD activity,and thus reducing is-let β cells apoptosis and increasing insulin secretion.Therefore,the compound LY-6 could serve as a poten-tial anti-diabetic new chemical entity.