ObjectiveThis study aims to identify cuproptosis-related gene biomarkers in Alzheimer's disease（AD） using bioinformatics and machine learning methods， validate them at the clinical specimen level， and predict potential traditional Chinese medicine（TCM）. MethodsDifferentially expressed genes in the AD group and normal group were obtained using the Gene Expression Omnibus （GEO） database， and intersections were taken with reported cuproptosis-related genes to obtain differentially expressed cuproptosis-related genes. Machine learning methods were applied to identify core differential genes of cuproptosis in AD. Peripheral blood's single nucleated cells from clinical AD patients were collected， and the relative gene expression was clinically verified by real-time polymerase chain reaction（Real-time PCR）. Potential TCM regulating cuproptosis for AD were predicted by COREMINE database. ResultsA total of 12 cuproptosis-related genes were obtained， mainly involved in pathways of tricarboxylic acid cycle， 2-oxocarboxylic acid metabolism， and carbon metabolism. Five core cuproptosis-related genes， dihydrolipoamide dehydrogenase （DLD）， glutaminase （GLS）， pyruvate dehydrogenase E1 subunit beta （PDHB）， full name nuclear factor （erythroid-derived 2）-related factor 2 （NFE2L2）， and dihydrolipoamide branched-chain transacylase E2 （DBT） were finally screened using four machine methods. Thirty cases each of normal and AD patients were collected clinically. Compared with those in the normal group， minimum mental state examination （MMSE） and Montreal cognitive assessment （MoCA） were significantly decreased in the AD group （P<0.01）， Homocysteine（Hcy）， interleukin（IL）-6， C-reactive protein（CRP） ， and β amyloid protein（Aβ） indexes were significantly increased （P<0.01）， and malondialdehyde（MDA） indexes were decreased （P<0.05）. Superoxide dismutase（SOD） levels were significantly decreased （P<0.01）. The mRNA relative expressions of NFE2L2 and DBT were up-regulated （P<0.05）， and those of DLD， GLS， and PDHB were significantly down-regulated （P<0.01）. The TCM regulating cuproptosis-related genes for the treatment of AD were mainly based on the four Qi such as warmth， calmness， and cold， and the five flavors including bitterness， sweetness， and pungency， and it was attributed to the meridians of the liver， spleen， stomach， and kidney， with the efficacy of tonifying Qi， activating blood， eliminating phlegm， and resoling dampness. ConclusionDLD， GLS， NFE2L2， PDHB， and DBT can be used as novel diagnostic molecular markers for AD cuproptosis-related genes， and the corresponding potential therapeutic TCM can provide new ideas for the treatment of AD by TCM.

OBJECTIVES: To explore the mechanism of Gandou Fumu Decoction (GDFMD) for improving Wilson's disease (WD) in tx-J mice. METHODS: With 6 syngeneic wild-type mice as the control group, 30 tx-J mice were randomized into WD model group, low-, medium- and high-dose GDFMD treatment groups, and Fer-1 treatment group. Saline (in control and model groups) and GDFMD (3.48, 6.96 or 13.92 g/kg) were administered by gavage, and Fer-1 was injected intraperitoneally once daily for 14 days. Oil red and HE staining were used to observe lipid deposition and pathological conditions in the liver tissue; ALT, AST, albumin, AKP levels were determined to assess liver function of the mice. Western blotting and RT-qPCR were used to detect hepatic protein and mRNA expressions of GPX4, ACSL4, ALOX15, FTH1, FLT, TFR1, FAS, SCD1, and ACOX1, and Fe²⁺, MDA, ROS, SOD, GSH and 4-HNE levels were analyzed to assess oxidative stress. RESULTS: The mouse models of WD showed obvious fatty degeneration in the liver tissue significantly increased serum levels of ALT, AST and AKP, decreased albumin level, increased Fe²⁺, MDA, ROS, 4-HNE levels, decreased SOD and GSH levels (P<0.05), lowered protein expressions of ACOX1, GPX4, FTH1, FLT, FAS, and SCD1, and increased protein contents of TFR1, ACSL4 and ALOX15 in the liver. Treatment with GDFMD and Fer-1 improved liver histopathology and liver function of the mouse models, decreased the levels of Fe²⁺, MDA and ROS, increased SOD and GSH levels, and reversed the changes in hepatic protein expressions. CONCLUSIONS GDFMD improves liver steatosis in mouse models of WD possibly by inhibiting hepatocyte ferroptosis through the GPX4/ACSL4/ALOX15 signaling pathway.
Animals ; Ferroptosis/drug effects* ; Mice ; Signal Transduction/drug effects* ; Drugs, Chinese Herbal/therapeutic use* ; Hepatolenticular Degeneration/drug therapy* ; Hepatocytes/metabolism* ; Phospholipid Hydroperoxide Glutathione Peroxidase ; Fatty Liver/metabolism* ; Arachidonate 15-Lipoxygenase/metabolism* ; Coenzyme A Ligases/metabolism* ; Liver/metabolism* ; Male

Objective To explore the mechanism of Gandou Fumu Decoction(GDFMD)for improving Wilson's disease(WD)in tx-J mice.Methods With 6 syngeneic wild-type mice as the control group,30 tx-J mice were randomized into WD model group,low-,medium-and high-dose GDFMD treatment groups,and Fer-1 treatment group.Saline(in control and model groups)and GDFMD(3.48,6.96 or 13.92 g/kg)were administered by gavage,and Fer-1 was injected intraperitoneally once daily for 14 days.Oil red and HE staining were used to observe lipid deposition and pathological conditions in the liver tissue;ALT,AST,albumin,AKP levels were determined to assess liver function of the mice.Western blotting and RT-qPCR were used to detect hepatic protein and mRNA expressions of GPX4,ACSL4,ALOX15,FTH1,FLT,TFR1,FAS,SCD1,and ACOX1,and Fe2+,MDA,ROS,SOD,GSH and 4-HNE levels were analyzed to assess oxidative stress.Results The mouse models of WD showed obvious fatty degeneration in the liver tissue significantly increased serum levels of ALT,AST and AKP,decreased albumin level,increased Fe2+,MDA,ROS,4-HNE levels,decreased SOD and GSH levels(P<0.05),lowered protein expressions of ACOX1,GPX4,FTH1,FLT,FAS,and SCD1,and increased protein contents of TFR1,ACSL4 and ALOX15 in the liver.Treatment with GDFMD and Fer-1 improved liver histopathology and liver function of the mouse models,decreased the levels of Fe2+,MDA and ROS,increased SOD and GSH levels,and reversed the changes in hepatic protein expressions.Conclusion GDFMD improves liver steatosis in mouse models of WD possibly by inhibiting hepatocyte ferroptosis through the GPX4/ACSL4/ALOX15 signaling pathway.

Objective To investigate the effects of Gandou Fumu Decoction on liver fibrosis,iron metabolism,and ferroptosis in patients with hepatolenticular degeneration(Wilson disease,WD).Methods Seventy-eight hospitalized patients with WD characterized by kidney and liver deficiency,with phlegm and blood stasis,from the Department of Neurology,the First Affiliated Hospital of Anhui University of Chinese Medicine,were randomly assigned to two groups using a random number table method.The control group(n=39)received sodium dimercaptosulfonate in combination with a low-copper and high-protein diet.The observation group(n=39)received the same treatment as the control group,with the addition of Gandou Fumu Decoction(one dose per day,taken twice daily,in the morning and evening).Both groups underwent six treatment cycles,each lasting eight days.Ultrasonographic parameters,including portal vein main trunk diameter(PVMD),portal vein velocity(PVV),shear wave velocity(SWV),liver stiffness measurement(LSM),serum liver fibrosis markers(hyaluronic acid[HA],laminin[LN],procollagen typeⅢN-terminal peptide[PⅢNP],collagen type Ⅳ[CⅣ],aspartate aminotransferase to platelet ratio index[APRI],fibrosis-4 index[FIB-4]),and iron metabolism indicators(serum iron[SI],ferritin[FT])were compared before and after treatment.The relationship between baseline iron metabolism markers and ultrasonographic parameters,as well as serum liver fibrosis markers,was analyzed.Clinical efficacy,traditional Chinese medicine(TCM)syndrome scores,and adverse reactions were also compared between the groups.Additionally,bioinformatics analysis was performed to identify potential targets of Gandou Fumu Decoction for WD treatment.Peripheral blood mononuclear cells were collected from a normal group of 20 healthy individuals,as well as from both the control and observation groups before and after treatment.Real time fluorogenic quantitative PCR was performed to validate the expression changes of these targets across the groups.Results Compared with pre-treatment values,no significant changes were observed in PVMD levels in either group after treatment.No significant change in PVV was observed in the control group,whereas a significant decrease was noted in the observation group(P＜0.01).SWV,LSM,HA,LN,PⅢNP,CⅣ,APRI,FIB-4,and FT levels were significantly reduced compared to pre-treatment levels(P＜0.05,P＜0.01),whereas SI remained unchanged.Compared with the control group,the observation group had no significant difference in PVMD but had significantly lower PVV,SWV,LSM,HA,LN,PⅢNP,CⅣ,APRI,FIB-4,and FT levels(P＜0.05,P＜0.01),whereas SI remained unchanged.The total effective rate of treatment in the observation group was significantly higher than that in the control group(P＜0.05).Both groups showed a significant reduction in TCM syndrome scores after treatment(P＜0.01),with a significantly greater reduction observed in the observation group(P＜0.01).No significant adverse reactions were reported during treatment.Before treatment,there was no significant correlation between the SI of both groups and PVMD,PVV,SWV,LSM,HA,LN,PⅢNP,CⅣ,APRI,and FIB-4.In the observation group,FT showed a positive correlation with SWV,LSM,LN,PⅢNP,CⅣ,APRI,and FIB-4(P＜0.05,P＜0.01),while in the control group,FT showed a positive correlation with HA,LN,PⅢNP,CⅣ,APRI,and FIB-4(P＜0.01).After treatment,in the control group,SI showed a positive correlation with APRI and FIB-4(P＜0.05,P＜0.01),but there was no significant correlation between SI in the observation group and FT in both groups with the above-mentioned indicators.Bioinformatics analysis identified four potential targets of Gandou Fumu Decoction for treating WD,namely heme oxygenase 1(HMOX1),peroxisome proliferator-activated receptor alpha(PPARα),small heat shock protein B1(HSPB1),and mitogen-activated protein kinase 3(MAPK3).Compared to the normal group,both the control and observation groups had significantly lower PPARαand HSPB1 expression and significantly higher HMOX1 and MAPK3 expression before treatment(P＜0.01).Compared to before treatment within the same group,both groups showed significantly increased PPARα and HSPB1 expression and significantly decreased HMOX1 and MAPK3 expression after treatment(P＜0.05,P＜0.01).After treatment,the observation group had significantly higher PPARα and HSPB1 expression and lower HMOX1 and MAPK3 expression than the control group(P＜0.05,P＜0.01).Conclusion Gandou Fumu Decoction demonstrates remarkable advantages in improving clinical efficacy,Chinese medicine syndrome scores,iron metabolism,liver fibrosis progression,ultrasound imaging parameters,and ferroptosis-related biomarkers expression in patients with WD,with a favorable safety profile.

Objective·To explore the clinical efficacy and safety of sticky bone[deproteinized porcine bone mineral granules combined with advanced platelet-rich fibrin(A-PRF)and liquid platelet-rich fibrin(L-PRF)]in the repair of unilateral alveolar cleft.Methods·Patients diagnosed with unilateral alveolar cleft who met the inclusion and exclusion criteria were recruited from December 1,2023 to August 31,2024.The patients were randomly divided into 2 groups.The experimental group received sticky bone grafts,and the control group received autologous iliac cancellous bone grafts for alveolar cleft repair.The primary efficacy index was the bone resorption rate at 6 months post-surgery,calculated by measuring the grafted bone volume immediately after surgery and at 6 months using Simplant Pro 17.01 software based on patients'computed tomography data collected before surgery,immediately after surgery,and 6 months post-surgery.Secondary efficacy indices included bone density at 6 months after surgery,the occurrence of postoperative complications in patients,and the scores on the oral health-related quality of life scale.Prism 10 software was used for data analysis,and t-test and Pearson correlation analysis methods were adopted.Results·Seventeen patients with unilateral alveolar cleft were included in the experimental group,and 15 in the control group.The remaining bone volume in the experimental group at 6 months after surgery was more than that in the control group.The bone resorption rate in the experimental group was 33.24%±17.16%,significantly lower than 66.31%±17.98%in the control group(P<0.001).One patient in the experimental group had vestibular mucosal dehiscence accompanied by bone powder discharge 2 weeks after surgery;3 patients in the control group had vestibular mucosal dehiscence accompanied by cancellous bone discharge 1 month after surgery,with one case of grafted bone necrosis 4 months after surgery.The bone density in the experimental group at 6 months after surgery was significantly higher than that in the control group(P<0.001),and the scores on the oral health-related quality of life scale were lower.Conclusion·Compared with iliac cancellous bone,when sticky bone is used in the treatment of patients with unilateral alveolar cleft,it shows a lower bone resorption rate and higher bone density,indicating better osteogenic ability.Fewer number of complications and lower scores on the oral health-related quality of life scale suggest greater safety,thereby supporting its strong potential for clinical promotion and application.

Objective To investigate the effects of Gandou Fumu Decoction on liver fibrosis,iron metabolism,and ferroptosis in patients with hepatolenticular degeneration(Wilson disease,WD).Methods Seventy-eight hospitalized patients with WD characterized by kidney and liver deficiency,with phlegm and blood stasis,from the Department of Neurology,the First Affiliated Hospital of Anhui University of Chinese Medicine,were randomly assigned to two groups using a random number table method.The control group(n=39)received sodium dimercaptosulfonate in combination with a low-copper and high-protein diet.The observation group(n=39)received the same treatment as the control group,with the addition of Gandou Fumu Decoction(one dose per day,taken twice daily,in the morning and evening).Both groups underwent six treatment cycles,each lasting eight days.Ultrasonographic parameters,including portal vein main trunk diameter(PVMD),portal vein velocity(PVV),shear wave velocity(SWV),liver stiffness measurement(LSM),serum liver fibrosis markers(hyaluronic acid[HA],laminin[LN],procollagen typeⅢN-terminal peptide[PⅢNP],collagen type Ⅳ[CⅣ],aspartate aminotransferase to platelet ratio index[APRI],fibrosis-4 index[FIB-4]),and iron metabolism indicators(serum iron[SI],ferritin[FT])were compared before and after treatment.The relationship between baseline iron metabolism markers and ultrasonographic parameters,as well as serum liver fibrosis markers,was analyzed.Clinical efficacy,traditional Chinese medicine(TCM)syndrome scores,and adverse reactions were also compared between the groups.Additionally,bioinformatics analysis was performed to identify potential targets of Gandou Fumu Decoction for WD treatment.Peripheral blood mononuclear cells were collected from a normal group of 20 healthy individuals,as well as from both the control and observation groups before and after treatment.Real time fluorogenic quantitative PCR was performed to validate the expression changes of these targets across the groups.Results Compared with pre-treatment values,no significant changes were observed in PVMD levels in either group after treatment.No significant change in PVV was observed in the control group,whereas a significant decrease was noted in the observation group(P＜0.01).SWV,LSM,HA,LN,PⅢNP,CⅣ,APRI,FIB-4,and FT levels were significantly reduced compared to pre-treatment levels(P＜0.05,P＜0.01),whereas SI remained unchanged.Compared with the control group,the observation group had no significant difference in PVMD but had significantly lower PVV,SWV,LSM,HA,LN,PⅢNP,CⅣ,APRI,FIB-4,and FT levels(P＜0.05,P＜0.01),whereas SI remained unchanged.The total effective rate of treatment in the observation group was significantly higher than that in the control group(P＜0.05).Both groups showed a significant reduction in TCM syndrome scores after treatment(P＜0.01),with a significantly greater reduction observed in the observation group(P＜0.01).No significant adverse reactions were reported during treatment.Before treatment,there was no significant correlation between the SI of both groups and PVMD,PVV,SWV,LSM,HA,LN,PⅢNP,CⅣ,APRI,and FIB-4.In the observation group,FT showed a positive correlation with SWV,LSM,LN,PⅢNP,CⅣ,APRI,and FIB-4(P＜0.05,P＜0.01),while in the control group,FT showed a positive correlation with HA,LN,PⅢNP,CⅣ,APRI,and FIB-4(P＜0.01).After treatment,in the control group,SI showed a positive correlation with APRI and FIB-4(P＜0.05,P＜0.01),but there was no significant correlation between SI in the observation group and FT in both groups with the above-mentioned indicators.Bioinformatics analysis identified four potential targets of Gandou Fumu Decoction for treating WD,namely heme oxygenase 1(HMOX1),peroxisome proliferator-activated receptor alpha(PPARα),small heat shock protein B1(HSPB1),and mitogen-activated protein kinase 3(MAPK3).Compared to the normal group,both the control and observation groups had significantly lower PPARαand HSPB1 expression and significantly higher HMOX1 and MAPK3 expression before treatment(P＜0.01).Compared to before treatment within the same group,both groups showed significantly increased PPARα and HSPB1 expression and significantly decreased HMOX1 and MAPK3 expression after treatment(P＜0.05,P＜0.01).After treatment,the observation group had significantly higher PPARα and HSPB1 expression and lower HMOX1 and MAPK3 expression than the control group(P＜0.05,P＜0.01).Conclusion Gandou Fumu Decoction demonstrates remarkable advantages in improving clinical efficacy,Chinese medicine syndrome scores,iron metabolism,liver fibrosis progression,ultrasound imaging parameters,and ferroptosis-related biomarkers expression in patients with WD,with a favorable safety profile.

Objective·To explore the clinical efficacy and safety of sticky bone[deproteinized porcine bone mineral granules combined with advanced platelet-rich fibrin(A-PRF)and liquid platelet-rich fibrin(L-PRF)]in the repair of unilateral alveolar cleft.Methods·Patients diagnosed with unilateral alveolar cleft who met the inclusion and exclusion criteria were recruited from December 1,2023 to August 31,2024.The patients were randomly divided into 2 groups.The experimental group received sticky bone grafts,and the control group received autologous iliac cancellous bone grafts for alveolar cleft repair.The primary efficacy index was the bone resorption rate at 6 months post-surgery,calculated by measuring the grafted bone volume immediately after surgery and at 6 months using Simplant Pro 17.01 software based on patients'computed tomography data collected before surgery,immediately after surgery,and 6 months post-surgery.Secondary efficacy indices included bone density at 6 months after surgery,the occurrence of postoperative complications in patients,and the scores on the oral health-related quality of life scale.Prism 10 software was used for data analysis,and t-test and Pearson correlation analysis methods were adopted.Results·Seventeen patients with unilateral alveolar cleft were included in the experimental group,and 15 in the control group.The remaining bone volume in the experimental group at 6 months after surgery was more than that in the control group.The bone resorption rate in the experimental group was 33.24%±17.16%,significantly lower than 66.31%±17.98%in the control group(P<0.001).One patient in the experimental group had vestibular mucosal dehiscence accompanied by bone powder discharge 2 weeks after surgery;3 patients in the control group had vestibular mucosal dehiscence accompanied by cancellous bone discharge 1 month after surgery,with one case of grafted bone necrosis 4 months after surgery.The bone density in the experimental group at 6 months after surgery was significantly higher than that in the control group(P<0.001),and the scores on the oral health-related quality of life scale were lower.Conclusion·Compared with iliac cancellous bone,when sticky bone is used in the treatment of patients with unilateral alveolar cleft,it shows a lower bone resorption rate and higher bone density,indicating better osteogenic ability.Fewer number of complications and lower scores on the oral health-related quality of life scale suggest greater safety,thereby supporting its strong potential for clinical promotion and application.

Objective:To investigate the alteration in iodine nutritional status and influence on thyroid function in the elderly aged≥65 years following water source modification in high iodine areas.Methods:Data from Yaoji Town, Xuzhou, Jiangsu(an area with high iodine due to water sources) of the national epidemiological survey on thyroid diseases, iodine nutrition, and diabetes(TIDE study) in 31 provinces and cities in China from 2015 to 2017 were utilized. Additionally, data from the screening, monitoring, and intervention on thyroid diseases(TOPS study) in the elderly(≥65 years) in Shunhe Town, Suqian, Jiangsu(an area with iodine levels exceeding the recommended amount), and Yaoji Town, Xuzhou from May to August 2021, are included. Each subject completed a questionnaire, physical examination, laboratory tests and thyroid ultrasound examinations. A total of 2 717 subjects aged≥65 years were included, including group 1, 258 subjects in TIDE study; Group 2, 1 313 subjects in TOPS Xuzhou area; Group 3, 1 146 subjects in TOPS Suqian area.Results:The urinary iodine concentration(UIC) in group 2 was significantly lower than that in group 1 [(235.16±67.09)μg/L vs (491.58±384.93)μg/L, P<0.001], but no significant difference compared with group 3 [(235.16±67.09) μg/L vs(231.62±66.11) μg/L, P>0.05]. The serum TSH level in group 2 was significantly lower than that in group 1 [(2.92±5.14)μIU/mL vs (4.15±9.19)μIU/mL, P<0.001]. Compared with group 2 and 3, the prevalence of subclinical hypothyroidism in the elderly in group 1 was the highest(22.48% vs 10.13% and 8.12%, P<0.001). TSH levels were linearly correlated with age in both excessive iodine and more than adequate iodine nutrition areas. TSH level was gradually increased with age. Conclusion:The alteration in TSH levels among the elderly is notably linked to both aging and iodine status. The prevalence of hypothyroidism in the elderly can be significantly reduced when the iodine nutrition status of the elderly returns to normal.

The treatment of craniomaxillofacial (CMF) defects and deformities is highly challenging due to the complex anatomical structures, significant individual variations and high demands for personalized reconstruction. In recent decades, the integration of cutting-edge technologies from fields such as computer science, mechanical engineering, and material science into medicine has greatly facilitated the transformation of CMF surgery from traditional, experience-based approaches to digitalized and intelligent methods. This review thoroughly analyzes the development and application areas of digital technology in CMF surgery including virtual surgical planning and facial prediction, three-dimensional printing, computer-assisted navigation and robotic surgery, virtual and mixed reality, as well as big data and artificial intelligence technologies. The aim is to offer comprehensive and in-depth reference information for the clinical practice in digital CMF surgery.

Objective:To retrieve, evaluate, and summarize the best evidence for the prevention and management of nipple pain or injury in breastfeeding puerperae, providing evidence-based basis for clinical practice of breastfeeding.Methods:According to the "6S" evidence pyramid model, clinical decisions, guidelines, expert consensus, evidence summaries, and systematic reviews on the prevention and management of nipple pain or injury in breastfeeding puerperae were systematically searched. The search period was from database establishment to March 10, 2023. The research team members independently evaluated the quality of the included article based on the corresponding quality evaluation standards, and combined professional judgment to extract and summarize evidence for the final included article.Results:A total of 15 articles were included, including two clinical decisions, 7 guidelines, and 6 systematic reviews. Finally, 19 best pieces of evidence were summarized, including four themes, namely, assessment of nipple pain or injury, prevention of nipple pain or injury, management of nipple pain or injury, and health education.Conclusions:Obstetrical nurses and midwives should provide standardized nursing for breastfeeding puerperae based on specific clinical scenarios, reduce the incidence of nipple pain or injury in breastfeeding puerperae, and promote puerperae to adhere to breastfeeding.