To verify whether chaperones can promote the soluble expression of PlpE in Escherichia coli and whether the expressed protein is active,prokaryotic expression and Western blot detection were performed.The results showed that:The PlpE prokaryotic expression vector pET-32a(+)-plpE was expressed as inclusion body,and the expression form was not changed by changing the concentration of inducer,induction time and temperature.The companion proteins pG-KJE8,pGro7,pKJE7 and pG-Tf2 were co-expressed with pET-32a(+)-plpE in Eschierichia coli expres-sion system,respectively.When the final concentration of IPTG of 0.5 mmol/L,L-arabinose of 0.5 g/L or tetracycline of 5.0 μg/L were added as inducers and induced at 37 ℃ for 8 h,the results showed that the molecular companion pGro7 could change the expression of rp-PlpE from inclu-sion body to soluble expression.pG-KJE8,pKJE7 and pG-Tf2 had no effect on the expression of rp-PlpE.The soluble rp-PlpE can react specifically with the positive serum of Mannheimia haemolyti-ca.Therefore,the study showed that the co-expression of the chaperone protein pGro7 can make the rp-PlpE protein express in a soluble form,and the purified protein exhibits reactogenicity.These findings lay the foundation for the establishment of a subunit vaccine and serological diagno-sis methods for Mannheimia haemolytica.

To verify whether chaperones can promote the soluble expression of PlpE in Escherichia coli and whether the expressed protein is active,prokaryotic expression and Western blot detection were performed.The results showed that:The PlpE prokaryotic expression vector pET-32a(+)-plpE was expressed as inclusion body,and the expression form was not changed by changing the concentration of inducer,induction time and temperature.The companion proteins pG-KJE8,pGro7,pKJE7 and pG-Tf2 were co-expressed with pET-32a(+)-plpE in Eschierichia coli expres-sion system,respectively.When the final concentration of IPTG of 0.5 mmol/L,L-arabinose of 0.5 g/L or tetracycline of 5.0 μg/L were added as inducers and induced at 37 ℃ for 8 h,the results showed that the molecular companion pGro7 could change the expression of rp-PlpE from inclu-sion body to soluble expression.pG-KJE8,pKJE7 and pG-Tf2 had no effect on the expression of rp-PlpE.The soluble rp-PlpE can react specifically with the positive serum of Mannheimia haemolyti-ca.Therefore,the study showed that the co-expression of the chaperone protein pGro7 can make the rp-PlpE protein express in a soluble form,and the purified protein exhibits reactogenicity.These findings lay the foundation for the establishment of a subunit vaccine and serological diagno-sis methods for Mannheimia haemolytica.

Complete mesocolic excision in radical colorectal cancer surgery enhances both surgical quality and the accuracy of pathological staging. In the context of sigmoid colon cancer, the optimal extent of lymphadenectomy and the appropriate level of vascular ligation remain controversial. High ligation of the inferior mesenteric artery may facilitate more thorough lymph node dissection and allow for tension-free anastomosis. However, it requires a comprehensive assessment of postoperative complication risks and the preservation of organ function. Para-aortic lymph node dissection has shown potential survival benefits in patients with oligometastatic disease, yet its application should be individualized. Moreover, intraoperative navigation technologies, such as indocyanine green fluorescence imaging, can assist in accurately delineating the dissection field and support the feasibility of personalized surgical strategies. This review synthesizes current evidence and leading domestic and international clinical guidelines to systematically examine the latest developments in lymphadenectomy strategies for sigmoid colon cancer, focusing on mesenteric anatomy, D3 dissection, complete mesocolic excision, vascular ligation levels, para-aortic lymph node dissection, and fluorescence-guided imaging techniques.

Objective:To explore the effectiveness of shared decision-making intervention with nurses as "decision coaches" in perioperative decision management of bladder cancer patients.Methods:From March 2023 to February 2024, 152 bladder cancer patients in the Department of Urology Surgery of Xinxiang Central Hospital for surgical treatment were selected by convenience sampling. The patients were divided into control group and observation group of 76 cases each using the randomized numerical table method. Control group implemented conventional bladder cancer surgical decision management, and observation group implemented a shared decision-making intervention with nurses as "decision coaches" on this basis, and the intervention period was one week before surgery. Decisional Conflict Scale (DCS), Preparation Decision Making (PrepDM), Patients Satisfaction with Participation in Medical Decision-making Scale, and Decision Regret Scale (DRS) scores were compared between the two groups of patients before and after intervention.Results:Finally 74 patients in control group and 75 patients in observation group completed the study. After the intervention, the DCS scores of observation group were lower than those of control group and before the intervention, and the differences were statistically significant (all P<0.05). After the intervention, the PrepDM scores of observation group were higher than those of control group and before the intervention, and the differences were statistically significant (all P<0.05). One week after surgery, observation group had higher decision-making participation satisfaction scores and lower DRS scores than control group, and the differences were statistically significant (all P<0.05) . Conclusions:Preoperative shared decision-making interventions with nurses as "decision coaches" in bladder cancer patients can effectively reduce preoperative decision conflicts, increase preoperative decision readiness and participation satisfaction with decision-making, and decrease patients' postoperative decision regret.

Objective:To explore the effectiveness of shared decision-making intervention with nurses as "decision coaches" in perioperative decision management of bladder cancer patients.Methods:From March 2023 to February 2024, 152 bladder cancer patients in the Department of Urology Surgery of Xinxiang Central Hospital for surgical treatment were selected by convenience sampling. The patients were divided into control group and observation group of 76 cases each using the randomized numerical table method. Control group implemented conventional bladder cancer surgical decision management, and observation group implemented a shared decision-making intervention with nurses as "decision coaches" on this basis, and the intervention period was one week before surgery. Decisional Conflict Scale (DCS), Preparation Decision Making (PrepDM), Patients Satisfaction with Participation in Medical Decision-making Scale, and Decision Regret Scale (DRS) scores were compared between the two groups of patients before and after intervention.Results:Finally 74 patients in control group and 75 patients in observation group completed the study. After the intervention, the DCS scores of observation group were lower than those of control group and before the intervention, and the differences were statistically significant (all P<0.05). After the intervention, the PrepDM scores of observation group were higher than those of control group and before the intervention, and the differences were statistically significant (all P<0.05). One week after surgery, observation group had higher decision-making participation satisfaction scores and lower DRS scores than control group, and the differences were statistically significant (all P<0.05) . Conclusions:Preoperative shared decision-making interventions with nurses as "decision coaches" in bladder cancer patients can effectively reduce preoperative decision conflicts, increase preoperative decision readiness and participation satisfaction with decision-making, and decrease patients' postoperative decision regret.

Temporomandibular joint (TMJ) disc displacement is one of the most significant subtypes of temporomandibular joint disorders, but its etiology and mechanism are poorly understood. In this study, we elucidated the mechanisms by which destruction of inflamed collagen fibrils induces alterations in the mechanical properties and positioning of the TMJ disc. By constructing a rat model of TMJ arthritis, we observed anteriorly dislocated TMJ discs with aggravated deformity in vivo from five weeks to six months after a local injection of Freund's complete adjuvant. By mimicking inflammatory conditions with interleukin-1 beta in vitro, we observed enhanced expression of collagen-synthesis markers in primary TMJ disc cells cultured in a conventional two-dimensional environment. In contrast, three-dimensional (3D)-cultivated disc cell sheets demonstrated the disordered assembly of inflamed collagen fibrils, inappropriate arrangement, and decreased Young's modulus. Mechanistically, inflammation-related activation of the nuclear factor kappa-B (NF-κB) pathway occurs during the progression of TMJ arthritis. NF-κB inhibition reduced the collagen fibril destruction in the inflamed disc cell sheets in vitro, and early NF-κB blockade alleviated collagen degeneration and dislocation of the TMJ discs in vivo. Therefore, the NF-κB pathway participates in the collagen remodeling in inflamed TMJ discs, offering a potential therapeutic target for disc displacement.
Animals ; NF-kappa B/metabolism* ; Temporomandibular Joint Disorders/pathology* ; Temporomandibular Joint Disc/metabolism* ; Rats ; Rats, Sprague-Dawley ; Disease Models, Animal ; Male ; Collagen/metabolism* ; Cells, Cultured ; Joint Dislocations/pathology* ; Interleukin-1beta ; Arthritis, Experimental

Objective:To analyze the short-term efficacy and safety of robot-assisted laparoscopic partial nephrectomy（RAPN）for non-metastatic pathological stage T 3a renal cell carcinoma. Methods:The clinical and pathological data of 45 patients with pathologically confirmed non-metastatic T 3a renal cell carcinoma who underwent RAPN at Sun Yat-sen University Cancer Center between January 2016 and December 2023 were retrospectively reviewed. There were 30 males and 15 females. The average age of the cohort was（54.3±10.7）years，and the average clinical tumor diameter was（4.9±1.8）cm. Of all the patients，35（77.8%）were asymptomatic，7（15.6%）presented with hematuria，and 3（6.7%）presented with lumbar pain. Preoperative imaging assessed 34 patients（75.6%）as having clinical stage T 3a，all suspected of involving the collecting system or perirenal fat invasion；the remaining 11 patients（24.4%）were assessed as having stage T 1-2 disease. The median R.E.N.A.L. nephrectomy score was 8.0（7.0，10.0）. A history of hypertension，diabetes，or chronic kidney disease was present in 18 patients（40.0%）. The primary endpoint was progression-free survival，and the secondary endpoints included postoperative complications and short-term renal function outcomes. Survival curve was estimated using the Kaplan-Meier method，and renal function comparisons were made using the paired t-test. Results:The RAPN was performed through a transabdominal approach in 32 patients（71.1%），with a median estimated blood loss of 150.0（50.0，300.0）ml. Seven（15.6%）patients required intraoperative blood transfusion. The median length of postoperative hospital stay was 4.0（4.0，6.0）days. Postoperative complications occurred in 6 patients（13.3%），including 5（11.1%）with mild complications and 1（2.2%）with a severe complication. Renal function returned to baseline in 24 of 39 evaluable patients（61.5%），while 3 patients（7.7%）developed surgery-related chronic kidney disease 3 to 12 months postoperatively，but none required dialysis. The median follow-up time was 31.8（22.7，50.9）months，12（26.7%）patients received programmed cell death protein 1 inhibitor adjuvant therapy postoperatively. During follow-up，3 patients experienced tumor recurrence，the 3-year progression-free survival rate of the entire cohort was 95.4%.Conclusions:For some carefully selected patients with T 3a renal cell carcinoma，RAPN performed by experienced surgeons is a feasible and safe option，providing excellent short-term oncological outcomes，complication control，and renal function recovery. The long-term efficacy remains to be seen.

Complete mesocolic excision in radical colorectal cancer surgery enhances both surgical quality and the accuracy of pathological staging. In the context of sigmoid colon cancer, the optimal extent of lymphadenectomy and the appropriate level of vascular ligation remain controversial. High ligation of the inferior mesenteric artery may facilitate more thorough lymph node dissection and allow for tension-free anastomosis. However, it requires a comprehensive assessment of postoperative complication risks and the preservation of organ function. Para-aortic lymph node dissection has shown potential survival benefits in patients with oligometastatic disease, yet its application should be individualized. Moreover, intraoperative navigation technologies, such as indocyanine green fluorescence imaging, can assist in accurately delineating the dissection field and support the feasibility of personalized surgical strategies. This review synthesizes current evidence and leading domestic and international clinical guidelines to systematically examine the latest developments in lymphadenectomy strategies for sigmoid colon cancer, focusing on mesenteric anatomy, D3 dissection, complete mesocolic excision, vascular ligation levels, para-aortic lymph node dissection, and fluorescence-guided imaging techniques.

Objective:To analyze the short-term efficacy and safety of robot-assisted laparoscopic partial nephrectomy（RAPN）for non-metastatic pathological stage T 3a renal cell carcinoma. Methods:The clinical and pathological data of 45 patients with pathologically confirmed non-metastatic T 3a renal cell carcinoma who underwent RAPN at Sun Yat-sen University Cancer Center between January 2016 and December 2023 were retrospectively reviewed. There were 30 males and 15 females. The average age of the cohort was（54.3±10.7）years，and the average clinical tumor diameter was（4.9±1.8）cm. Of all the patients，35（77.8%）were asymptomatic，7（15.6%）presented with hematuria，and 3（6.7%）presented with lumbar pain. Preoperative imaging assessed 34 patients（75.6%）as having clinical stage T 3a，all suspected of involving the collecting system or perirenal fat invasion；the remaining 11 patients（24.4%）were assessed as having stage T 1-2 disease. The median R.E.N.A.L. nephrectomy score was 8.0（7.0，10.0）. A history of hypertension，diabetes，or chronic kidney disease was present in 18 patients（40.0%）. The primary endpoint was progression-free survival，and the secondary endpoints included postoperative complications and short-term renal function outcomes. Survival curve was estimated using the Kaplan-Meier method，and renal function comparisons were made using the paired t-test. Results:The RAPN was performed through a transabdominal approach in 32 patients（71.1%），with a median estimated blood loss of 150.0（50.0，300.0）ml. Seven（15.6%）patients required intraoperative blood transfusion. The median length of postoperative hospital stay was 4.0（4.0，6.0）days. Postoperative complications occurred in 6 patients（13.3%），including 5（11.1%）with mild complications and 1（2.2%）with a severe complication. Renal function returned to baseline in 24 of 39 evaluable patients（61.5%），while 3 patients（7.7%）developed surgery-related chronic kidney disease 3 to 12 months postoperatively，but none required dialysis. The median follow-up time was 31.8（22.7，50.9）months，12（26.7%）patients received programmed cell death protein 1 inhibitor adjuvant therapy postoperatively. During follow-up，3 patients experienced tumor recurrence，the 3-year progression-free survival rate of the entire cohort was 95.4%.Conclusions:For some carefully selected patients with T 3a renal cell carcinoma，RAPN performed by experienced surgeons is a feasible and safe option，providing excellent short-term oncological outcomes，complication control，and renal function recovery. The long-term efficacy remains to be seen.

OBJECTIVE To investigate the role and mechanism of microRNA-125b (miR-125b) in downregulating ion channel-related protein expression in a cardiac hypertrophy model.METHODS① In vivo:Lentiviral vectors for miR-125b overexpression and knockdown were constructed,and male C57BL/6 mice were divided into the following groups:sham group (thoracotomy without virus injection),LV-miR-125b group (mmu-miR-125b mimic),LV-miR-125b-inhibitor group (mmu-miR-125b-inhibitor),and negative control group (LV-NC).The mice were raised under normal conditions for 4 weeks.The ultrastructural changes in myocardium tissue sections of LV-miR-125b mice were observed using trans-mission electron microscopy.The cardiac hypertrophy model in mice was established using thoracic aortic constriction (TAC).Echocardiography was performed to measure ejection fraction (EF) and frac-tional shortening (FS),and the ratio of heart weight to body weight (HW/BW),ratio of heart weight to tibia length (HW/TL),as well as the expression level of the myocardial hypertrophy marker β-myosin heavy chain (β-MHC) were calculated to evaluate the success of the TAC-induced hypertrophy model.Subse-quently,C57BL/6 mice were divided into four groups:Sham group,TAC model group,LV-miR-125b-inhibitor+TAC group,and LV-NC+TAC group.Protein expression levels of cardiac sodium channel (Nav1.5) and calcium channel (Cav1.2) were detected using Western blotting.RT-qPCR was performed to assess the levels of miR-125b and mRNA expression of myocardial hypertrophy markers,including atrial natriuretic peptide (ANP),brain natriuretic peptide (BNP),and β-MHC.② In vitro:Primary cultured neonatal Kunming mouse cardiomyocytes were divided into four groups:cell control group (no treatment),miR-125b overexpression group,miR-125b-inhibitor group,and negative control group (NC).RT-qPCR was used to detect the levels of miR-125b,ANP,BNP,and β-MHC.Western blotting and immunofluorescence were performed to assess the expression levels of Nav1.5 and Cav1.2 in the cardiomyocytes.Luciferase reporter gene assay was used to evaluate the direct effect of miR-125b on the target proteins Nav1.5 and Cav1.2.RESULTS ① In vivo:Compared to the Sham group,the TAC model mice showed significantly increased the ratio of heart weight to body weight (HW/BW),the ratio of heart weight to tibia length (HW/TL),and expression levels of the myocardial hypertrophy marker β-MHC (P＜0.05),indicating the successful establishment of the TAC model.Furthermore,miR-125b expression was significantly elevated in the TAC model group (P＜0.01).In the LV-miR-125b group,compared to the LV-NC group,the expression levels of myocardial hypertrophy markers ANP,BNP,and β-MHC were significantly increased (P＜0.01),while the ejection fraction (EF) and fractional short-ening (FS) values of the mice were significantly reduced (P＜0.01).Additionally,Additionally,myocardium ultrastructure of LV-miR-125b group was damaged.Compared to the LV-NC+TAC group,the LV-miR-125b-inhibitor+TAC group showed a significant increase in ejection fraction (EF) and fractional shortening (FS) values (P＜0.05).Additionally,the levels of Nav1.5 and Cav1.2 in myocardium tissue were signifi-cantly elevated in the LV-miR-125b-inhibitor+TAC group compared to the LV-NC+TAC group (P＜0.05).② In vitro:Compared to the NC group,the miR-125b overexpression group showed a significant increase in miR-125b expression (P＜0.01),as well as elevated levels of ANP,BNP,and β-MHC (P＜0.01).However,miR-125b-inhibitor significantly reversed the increases in ANP,BNP,and β-MHC (P＜0.01).Western blotting and immunofluorescence results showed that,compared to the NC group,the miR-125b mimic group exhibited significantly decreased levels of Nav1.5 and Cav1.2 (P＜0.01),while miR-125b-inhibitor led to an increase in the levels of both Nav1.5 and Cav1.2.Luciferase assay results demon-strated that miR-125b directly binds to the ion channel proteins Nav1.5 and Cav1.2,encoded by the SCN5A and CACNA1C genes.CONCLUSION miR-125b promotes the development of cardiac hyper-trophy by inhibiting the voltage-gated ion channel proteins Nav1.5 and Cav1.2 Inhibition of miR-125b expression improves cardiac hypertrophy.