Objective To explore the clinical efficacy of the"S.L.O.T.S."(Separate,Loop,Observe,Tension,Sign)three-layer five-step method for preserving testicular artery in microsurgical subinguinal varicocelectomy.Methods Clinical data of 190 patients with varicocele in our hospital from March 2020 to September 2022 were retrospectively analyzed.A total of 97 cases underwent"S.L.O.T.S."three-layer five-step method with preservation of testicular artery in microsurgical subinguinal varicocelectomy(microscopic group),and 93 cases underwent laparoscopic spermatic vein bundle ligation(laparoscopic group).The semen quality and postoperative complications of the two groups were compared before and after surgery.Results The laparoscopic group had significantly shorter operation time[(32.2±7.3)min vs.(65.1±8.3)min,t=28.959,P=0.000]compared to the microscopic group.The proportion of postoperative secondary hydrocele[1.0％(1/97)vs.8.6％(8/93),x2=4.470,P=0.034].epididymitis[2.1％(2/97)vs.9.7％(9/93),x2=5.049,P=0.025].and total complications[3.1％(3/97)vs.21.5％(20/93),x2=15.128,P=0.000]in the microscopic group were significantly lower than those in the laparoscopic group.The indicators of semen quality such as total number of sperm[(53.3±5.8)× 106 vs.(50.4±4.9)× 106,t=3.676,P=0.000].sperm concentration[(19.6±2.2)× 106/ml vs.(18.1±2.4)×106/ml,t=4.418,P=0.000].rate of sperm anterior motility[(46.1±5.6)％vs.(41.5±4.1)％,t=6.476,P=0.000].total sperm motility[(55.7±4.9)％vs.(51.2±3.8)％,t=7.115,P=0.000].and percentage of normal sperm[(9.0±1.7)％vs.(7.6±1.6)％,t=5.550,P=0.000]were significantly better in the microscopic group than those in the laparoscopic group.Conclusion "S.L.O.T.S."three-layer five-step method has advantages of programmed surgical steps,and effective identification and preservation of testicular arteries and lymphatic vessels,which is beneficial for improving semen quality and reducing postoperative complications such as secondary hydrocele and epididymitis.

Immediate implant-supported fixed restoration in edentulous jaws demonstrates a success rate comparable to that of conventional implant restoration. However, this approach still presents a certain degree of technique sensitivity. In the field of immediate implant-supported fixed restoration in dentistry, a repeatable and stable jaw relation is the prerequisite for the design and fabrication of prostheses. It also reduces chairside denture placement and occlusal adjustment time and lowers the risk of occlusion-related complications. For patients with terminal dentition, the precise transfer of jaw relation following full-arch implantation serves as the fundamental basis for implant-supported occlusal reconstruction and functional restoration. This process is also a key research focus and challenge in the area of implant-supported occlusal rehabilitation. This review summarizes the procedures and methods for determining and transferring jaw relation in immediate implant-supported fixed restoration. It aims to serve as a basis for clinical decision making in implant-supported fixed restorations for terminal dentition patients.
Humans ; Dental Prosthesis, Implant-Supported ; Clinical Decision-Making ; Immediate Dental Implant Loading ; Jaw, Edentulous/surgery*

Objective:To successfully isolate and identify patient-derived ovarian cancer stem cells(OCSC),and to investigate the expression of monocarboxylate transporter 2(MCT2)in OCSC and its effects on the stem-ness and invasiveness of OCSC.Methods:Primary ovarian cancer cells were obtained,and OCSC models were established through screening and culturing.The screened OCSC were designated as the OCSC group,while the adherent primary cells were employed as control group 1.Two MCT2 inhibition constant Ki-specific inhibitors,namely[α-cyano-4-hydroxycinnamic acid(CHC)and ARC155858],were utilized for intervention,setting up the CHC inhibition group and the ARC155858 inhibition group.The group with only the solvent DMSO added was set respectively as CHC control group 2 and ARC 155858 control group 2.Flow cytometry and qRT-PCR were used for identification of OCSC markers.The expression levels and locations of MCT2 were detected by qRT-PCR and immunofluorescence staining.The effects of MCT2 inhibitors on the stemness and invasiveness of OCSC were verified using sphere formation assays and Matrigel invasion assays.Results:A primary OCSC cell bank consis-ting of 23 clinical models was established.qRT-PCR results showed that the expression level of MCT2 in OCSC group(5.78±3.55 and 122.89±19.90)was significantly higher than that in the control group 1(1.01±0.10 and 1.55±1.45),the difference is statistically significant(P＜0.05).Fluorescent quantitative analysis confirmed that MCT2 expression in OCSC group was higher than that in the control group 1(6.79±0.67 AU vs.2.49±1.78 AU,P=0.030).Treatment with MCT2 inhibitors in OCSC resulted in a significant reduction in sphere formation in the CHC inhibition group(6.00±2.17 spheres/1000 cells)compared to the CHC control group 2(14.62±3.07 spheres/1000 cells),the difference is statistically significant(P＜0.01),and in the ARC155858 inhibition group(11.00±4.42 spheres/1000 cells)compared to the ARC155858 control group 2(23.90±4.72 spheres/1000 cells),the difference is statistically significant(P＜0.01).In Matrigel invasion assays,the invasiveness in the in-hibitor group was significantly lower than in the control group(ARC155858:inhibitor group 10.20±5.98 cells vs.control group 67.20±28.96 cells,P＜0.05).Conclusions:This study observed that MCT2 expression significantly increased in OCSC.Inhibition of MCT2 significantly reduces the stemness and invasiveness of OCSC,providing potential research directions for targeted therapy of ovarian cancer.

Objective To analyze differentially expressed proteins(DEPs)in the hippocampal tissue of an amyloid-beta 1-42(Aβ1-42)-induced Alzheimer's disease(AD)rat model using Tandem mass tag(TMT)-based quantitative proteomics,followed by bioinformatic analysis to explore potential AD mechanisms.Methods Twelve male Sprague-Dawley(SD)rats were randomly assigned to a control group(n=6)and a model group(n=6).The AD model was established by bilateral hippocampal injection of Aβ1-42 in the model group,while the control group received an equivalent volume of saline.Cognitive function was assessed using the novel object recognition test,and hippocampal Aβ deposition was detected by immunofluorescence.DEPs were identified using TMT-based proteomics and subsequently analyzed via Gene Ontology(GO)annotation,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment,and protein-protein interaction(PPI)network analysis.Key DEPs were validated using parallel reaction monitoring(PRM)technology.Results The model group exhibited a significantly lower novel object recognition index(P<0.01)and significantly increased hippocampal Aβ deposition(P<0.01)compared to the control group.Proteomic analysis identified 183 DEPs(87 upregulated,96 downregulated).GO analysis revealed that DEPs were primarily enriched in processes such as amyloid-beta binding and ion transmembrane transport.KEGG analysis indicated significant enrichment in 42 pathways,including dopaminergic synapse,glutamatergic synapse,cholinergic synapse,and long-term potentiation.Ten core DEPs were identified from the PPI network,and PRM validation confirmed expression trends consistent with the TMT results.Conclusion Aβ1-42-induced AD involves the synergistic action of multiple targets,biological processes,and pathways.The activation of glutamatergic and dopaminergic synaptic signaling pathways,mediated by core DEPs(e.g.,Th、D1、VGLUT2、GluN2A、GluA1、GluA3、Shank1、DARPP-32、PKC-δ、PKC-α、PKA C-β、CaMKⅡα、PTK2B),likely represents a key molecular mechanism in this AD model,providing a basis for identifying potential therapeutic targets.

Objective To investigate the mechanism by which Yangmai Tongluo formula improves microcirculation disorders induced by high homocysteine(Hcy)levels via regulation of the acid sphingomyelinase(ASM)and endoplasmic reticulum(ER)stress pathways.Methods Fifty male C57BL/6J mice were divided randomly into a control group,Hcy model group,Yangmai Tongluo formula low-/high dose groups(5.3,10.4 g/kg,respectively),and a folic acid group(0.08 g/kg).Except for the control group,microcirculation disorders were induced in all mice using drinking water containing 1.8 g/L Hcy for 6 weeks.After modeling for 2 weeks,mice were administered the corresponding treatments by gavage for 4 weeks.Serum Hey concentrations and the blood perfusion volume of the lower extremity microvessels were measured.Protein expression levels of zonula occludens ZO-1,ZO-2,intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1),ASM,glucose-regulated protein 78(GRP78),and C/EBP homologous protein(CHOP)in the cardiac microvascular endothelium were analyzed using immunofluorescence.Results Serum Hcy levels were significantly increased in the Hcy model group compared with the control group(P＜0.05).Yangmai Tongluo formula did not significantly reduce Hey levels compared with the Hey model group,but blood perfusion in the lower extremities was significantly increased(P＜0.01)and expression levels of ZO-1 and ZO-2 in the cardiac microvascular endothelium were restored(P＜0.001)in the Yangmai Tongluo formula high dose group.It also inhibited the expression of ICAM-1,VCAM-1,ASM,GRP78,and CHOP(P＜0.05),with comparable effects to folic acid.Conclusions Yangmai Tongluo formula improves Hcy-induced microcirculation disorders and endothelial dysfunction by inhibiting ASM activity and alleviating ER stress,via a mechanism closely related to the regulation of endothelial inflammation and barrier stability.These result provide experimental evidence to support the use of traditional Chinese medicine to treat microvascular diseases.

Objective:To construct and validate a clinical model combining CYP11B2 gene polymorphisms with clinical parameters to predict complete postoperative hypertension remission in primary aldosteronism patients.Methods:The clinical data of a total of 116 patients with primary aldosteronism who underwent unilateral adrenalectomy from April 2018 to August 2024 were retrospectively included. There were 63 males and 53 females，with a body mass index（BMI）of（25.50 ± 2.03）kg/m 2. Genomic DNA was extracted from venous blood leukocytes before surgery，and polymerase chain reaction-restriction fragment length polymorphisms（PCR-RFLP）were used to detect CYP11B2（rs1799998）promoter region 344（C > T）base substitution. The follow-up duration was more than 6 months，with the following parameters recorded at the last follow-up：plasma aldosterone，renin，serum potassium，and sodium levels. Blood pressure progression and antihypertensive medication usage were also assessed. The postoperative outcome was determined according to the Primary Aldosteronism Surgical Outcome score（PASO）for primary aldosteronism，and the specific criteria were as follows. ① Clinical complete remission：the patient's blood pressure returned to normal（< 140/90 mmHg，1 mmHg = 0.133 kPa）and all antihypertensive drugs were discontinued；②Partial clinical remission：blood pressure returns to normal，and the number or dose of antihypertensive drugs is reduced compared with before；③Clinical non-remission：blood pressure does not drop and antihypertensive drugs do not change or increase compared with before surgery. Patients were divided into complete and incomplete remission groups. The chi-square test was used for univariate analysis，followed by binary logistic forward conditional regression for multivariate analysis，and a variety of machine learning algorithms such as random forest，logistic regression，support vector machine and gradient lifter were integrated，and the results of multivariate analysis were included to construct a postoperative blood pressure outcome model，and the predictive performance of the model was evaluated by using receiver operating characteristic（ROC）curve，calibration curve and clinical decision curve. Results:The PCR-RFLP detection results of 116 cases showed the genotype distribution of CYP11B2（344C > T）（rs1799998）as follows：CC type in 50 cases（43.1%），CT type in 46 cases（39.7%），and TT type in 20 cases（17.3%）. There were 74 cases in the complete remission group and 42 cases in the incomplete remission group，and the rate of complete remission with hypertension at the end of the operation was 63.8%. Univariate analysis showed that the the differences between complete remission group and incomplete remission group in body mass index［（24.27 ± 2.90）kg/m 2 vs.（26.98 ± 3.17）kg/m 2， P<0.001］，preoperative hypertension grade（grade 1/2/3：29/29/16 cases vs. 9/13/20 cases， P = 0.012），preoperative antihypertensive drugs（0/1/≥ 2：25/32/17 cases vs. 7/15/20 cases， P = 0.016），and CYP11B2（344C > T）（CC/TT + CT：39/35 cases vs. 11/31 cases， P = 0.006）were statistically significant. Multivariate analysis showed that the type of preoperative antihypertensive drugs［≥ 2： OR = 5.26（95% CI 1.12?24.61， P = 0.016；1： OR = 4.55（95% CI 1.23?22.47）， P = 0.025］was the strongest independent predictor，followed by CYP11B2（344C > T）［ OR = 4.02（95% CI 1.16?13.82）， P = 0.028］and BMI［ OR = 3.96（95% CI 2.26?6.92）， P < 0.001］. Comparing the receiver operating feature（ROC）curves of the four types of machine learning models，the best model was the support vector machine model with an area under the curve（AUC）of 0.88（95% CI 0.82?0.95），followed by the gradient elevator model of 0.83（95% CI 0.76?0.91），the logistic regression model of 0.78（95% CI 0.68?0.88），and the random forest model of 0.77（95% CI 0.68?0.86）. The optimal threshold of the Yoden index of the support vector machine model was 0.588，with a sensitivity of 78.5% and a specificity of 86.5%. The clinical decision curve and calibration curve show that the support vector machine model has a higher net benefit and acceptable stability and reliability. Conclusions:The support vector machine model incorporating CYP11B2 gene polymorphisms，BMI，and types of preoperative antihypertensive medications could effectively predict postoperative hypertension remission in primary aldosteronism patients，providing new evidence for personalized treatment strategies

Objective:To construct and validate a clinical model combining CYP11B2 gene polymorphisms with clinical parameters to predict complete postoperative hypertension remission in primary aldosteronism patients.Methods:The clinical data of a total of 116 patients with primary aldosteronism who underwent unilateral adrenalectomy from April 2018 to August 2024 were retrospectively included. There were 63 males and 53 females，with a body mass index（BMI）of（25.50 ± 2.03）kg/m 2. Genomic DNA was extracted from venous blood leukocytes before surgery，and polymerase chain reaction-restriction fragment length polymorphisms（PCR-RFLP）were used to detect CYP11B2（rs1799998）promoter region 344（C > T）base substitution. The follow-up duration was more than 6 months，with the following parameters recorded at the last follow-up：plasma aldosterone，renin，serum potassium，and sodium levels. Blood pressure progression and antihypertensive medication usage were also assessed. The postoperative outcome was determined according to the Primary Aldosteronism Surgical Outcome score（PASO）for primary aldosteronism，and the specific criteria were as follows. ① Clinical complete remission：the patient's blood pressure returned to normal（< 140/90 mmHg，1 mmHg = 0.133 kPa）and all antihypertensive drugs were discontinued；②Partial clinical remission：blood pressure returns to normal，and the number or dose of antihypertensive drugs is reduced compared with before；③Clinical non-remission：blood pressure does not drop and antihypertensive drugs do not change or increase compared with before surgery. Patients were divided into complete and incomplete remission groups. The chi-square test was used for univariate analysis，followed by binary logistic forward conditional regression for multivariate analysis，and a variety of machine learning algorithms such as random forest，logistic regression，support vector machine and gradient lifter were integrated，and the results of multivariate analysis were included to construct a postoperative blood pressure outcome model，and the predictive performance of the model was evaluated by using receiver operating characteristic（ROC）curve，calibration curve and clinical decision curve. Results:The PCR-RFLP detection results of 116 cases showed the genotype distribution of CYP11B2（344C > T）（rs1799998）as follows：CC type in 50 cases（43.1%），CT type in 46 cases（39.7%），and TT type in 20 cases（17.3%）. There were 74 cases in the complete remission group and 42 cases in the incomplete remission group，and the rate of complete remission with hypertension at the end of the operation was 63.8%. Univariate analysis showed that the the differences between complete remission group and incomplete remission group in body mass index［（24.27 ± 2.90）kg/m 2 vs.（26.98 ± 3.17）kg/m 2， P<0.001］，preoperative hypertension grade（grade 1/2/3：29/29/16 cases vs. 9/13/20 cases， P = 0.012），preoperative antihypertensive drugs（0/1/≥ 2：25/32/17 cases vs. 7/15/20 cases， P = 0.016），and CYP11B2（344C > T）（CC/TT + CT：39/35 cases vs. 11/31 cases， P = 0.006）were statistically significant. Multivariate analysis showed that the type of preoperative antihypertensive drugs［≥ 2： OR = 5.26（95% CI 1.12?24.61， P = 0.016；1： OR = 4.55（95% CI 1.23?22.47）， P = 0.025］was the strongest independent predictor，followed by CYP11B2（344C > T）［ OR = 4.02（95% CI 1.16?13.82）， P = 0.028］and BMI［ OR = 3.96（95% CI 2.26?6.92）， P < 0.001］. Comparing the receiver operating feature（ROC）curves of the four types of machine learning models，the best model was the support vector machine model with an area under the curve（AUC）of 0.88（95% CI 0.82?0.95），followed by the gradient elevator model of 0.83（95% CI 0.76?0.91），the logistic regression model of 0.78（95% CI 0.68?0.88），and the random forest model of 0.77（95% CI 0.68?0.86）. The optimal threshold of the Yoden index of the support vector machine model was 0.588，with a sensitivity of 78.5% and a specificity of 86.5%. The clinical decision curve and calibration curve show that the support vector machine model has a higher net benefit and acceptable stability and reliability. Conclusions:The support vector machine model incorporating CYP11B2 gene polymorphisms，BMI，and types of preoperative antihypertensive medications could effectively predict postoperative hypertension remission in primary aldosteronism patients，providing new evidence for personalized treatment strategies

Objective To analyze differentially expressed proteins(DEPs)in the hippocampal tissue of an amyloid-beta 1-42(Aβ1-42)-induced Alzheimer's disease(AD)rat model using Tandem mass tag(TMT)-based quantitative proteomics,followed by bioinformatic analysis to explore potential AD mechanisms.Methods Twelve male Sprague-Dawley(SD)rats were randomly assigned to a control group(n=6)and a model group(n=6).The AD model was established by bilateral hippocampal injection of Aβ1-42 in the model group,while the control group received an equivalent volume of saline.Cognitive function was assessed using the novel object recognition test,and hippocampal Aβ deposition was detected by immunofluorescence.DEPs were identified using TMT-based proteomics and subsequently analyzed via Gene Ontology(GO)annotation,Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment,and protein-protein interaction(PPI)network analysis.Key DEPs were validated using parallel reaction monitoring(PRM)technology.Results The model group exhibited a significantly lower novel object recognition index(P<0.01)and significantly increased hippocampal Aβ deposition(P<0.01)compared to the control group.Proteomic analysis identified 183 DEPs(87 upregulated,96 downregulated).GO analysis revealed that DEPs were primarily enriched in processes such as amyloid-beta binding and ion transmembrane transport.KEGG analysis indicated significant enrichment in 42 pathways,including dopaminergic synapse,glutamatergic synapse,cholinergic synapse,and long-term potentiation.Ten core DEPs were identified from the PPI network,and PRM validation confirmed expression trends consistent with the TMT results.Conclusion Aβ1-42-induced AD involves the synergistic action of multiple targets,biological processes,and pathways.The activation of glutamatergic and dopaminergic synaptic signaling pathways,mediated by core DEPs(e.g.,Th、D1、VGLUT2、GluN2A、GluA1、GluA3、Shank1、DARPP-32、PKC-δ、PKC-α、PKA C-β、CaMKⅡα、PTK2B),likely represents a key molecular mechanism in this AD model,providing a basis for identifying potential therapeutic targets.

Objective:To successfully isolate and identify patient-derived ovarian cancer stem cells(OCSC),and to investigate the expression of monocarboxylate transporter 2(MCT2)in OCSC and its effects on the stem-ness and invasiveness of OCSC.Methods:Primary ovarian cancer cells were obtained,and OCSC models were established through screening and culturing.The screened OCSC were designated as the OCSC group,while the adherent primary cells were employed as control group 1.Two MCT2 inhibition constant Ki-specific inhibitors,namely[α-cyano-4-hydroxycinnamic acid(CHC)and ARC155858],were utilized for intervention,setting up the CHC inhibition group and the ARC155858 inhibition group.The group with only the solvent DMSO added was set respectively as CHC control group 2 and ARC 155858 control group 2.Flow cytometry and qRT-PCR were used for identification of OCSC markers.The expression levels and locations of MCT2 were detected by qRT-PCR and immunofluorescence staining.The effects of MCT2 inhibitors on the stemness and invasiveness of OCSC were verified using sphere formation assays and Matrigel invasion assays.Results:A primary OCSC cell bank consis-ting of 23 clinical models was established.qRT-PCR results showed that the expression level of MCT2 in OCSC group(5.78±3.55 and 122.89±19.90)was significantly higher than that in the control group 1(1.01±0.10 and 1.55±1.45),the difference is statistically significant(P＜0.05).Fluorescent quantitative analysis confirmed that MCT2 expression in OCSC group was higher than that in the control group 1(6.79±0.67 AU vs.2.49±1.78 AU,P=0.030).Treatment with MCT2 inhibitors in OCSC resulted in a significant reduction in sphere formation in the CHC inhibition group(6.00±2.17 spheres/1000 cells)compared to the CHC control group 2(14.62±3.07 spheres/1000 cells),the difference is statistically significant(P＜0.01),and in the ARC155858 inhibition group(11.00±4.42 spheres/1000 cells)compared to the ARC155858 control group 2(23.90±4.72 spheres/1000 cells),the difference is statistically significant(P＜0.01).In Matrigel invasion assays,the invasiveness in the in-hibitor group was significantly lower than in the control group(ARC155858:inhibitor group 10.20±5.98 cells vs.control group 67.20±28.96 cells,P＜0.05).Conclusions:This study observed that MCT2 expression significantly increased in OCSC.Inhibition of MCT2 significantly reduces the stemness and invasiveness of OCSC,providing potential research directions for targeted therapy of ovarian cancer.

Objective To investigate the mechanism by which Yangmai Tongluo formula improves microcirculation disorders induced by high homocysteine(Hcy)levels via regulation of the acid sphingomyelinase(ASM)and endoplasmic reticulum(ER)stress pathways.Methods Fifty male C57BL/6J mice were divided randomly into a control group,Hcy model group,Yangmai Tongluo formula low-/high dose groups(5.3,10.4 g/kg,respectively),and a folic acid group(0.08 g/kg).Except for the control group,microcirculation disorders were induced in all mice using drinking water containing 1.8 g/L Hcy for 6 weeks.After modeling for 2 weeks,mice were administered the corresponding treatments by gavage for 4 weeks.Serum Hey concentrations and the blood perfusion volume of the lower extremity microvessels were measured.Protein expression levels of zonula occludens ZO-1,ZO-2,intercellular adhesion molecule-1(ICAM-1),vascular cell adhesion molecule-1(VCAM-1),ASM,glucose-regulated protein 78(GRP78),and C/EBP homologous protein(CHOP)in the cardiac microvascular endothelium were analyzed using immunofluorescence.Results Serum Hcy levels were significantly increased in the Hcy model group compared with the control group(P＜0.05).Yangmai Tongluo formula did not significantly reduce Hey levels compared with the Hey model group,but blood perfusion in the lower extremities was significantly increased(P＜0.01)and expression levels of ZO-1 and ZO-2 in the cardiac microvascular endothelium were restored(P＜0.001)in the Yangmai Tongluo formula high dose group.It also inhibited the expression of ICAM-1,VCAM-1,ASM,GRP78,and CHOP(P＜0.05),with comparable effects to folic acid.Conclusions Yangmai Tongluo formula improves Hcy-induced microcirculation disorders and endothelial dysfunction by inhibiting ASM activity and alleviating ER stress,via a mechanism closely related to the regulation of endothelial inflammation and barrier stability.These result provide experimental evidence to support the use of traditional Chinese medicine to treat microvascular diseases.