The aim of this study is to establish an gene editing method of Mesomycoplasma hyo-pneumoniae(Mhp)based on the homologous recombination principle.The restriction enzyme di-gestion and ligation method combined with gene synthesis were used to construct a shuttle plasmid to achieve replication in both Mhp and Escherichia coli(E.coli).The pGEM?-T vector was used as the skeleton.The oriC sequence of Mhp which can achieve the replication of the plasmid in Mhp was inserted into the vector.Sequences of the Spiroplasma promoter and puromycin resistance gene were then inserted into the above constructed plasmid to screen recombinant clones.The up-stream and downstream homologous arms of p97 were constructed to initiate homologous recombination.The recA gene of E.coli is inserted to improve the efficiency of homologous recom-bination.The obtained shuttle plasmid was then delivered into Mhp by electro-transformation or chemical transformation.A shuttle plasmid,pGEM?-Mhp-oriC-p 97,which can replicate in both Mhp and E.coli was constructed.With the transformation of this plasmid,the carried puromycin gene and recA gene can be expressed,the p97 gene can be edited.Finally,the genetically unstable p97 gene mutant was initially obtained.In this study,a tool for Mhp gene editing based on the principle of homologous recombination was established,which laid a foundation for the develop-ment of tools for studying the pathogenesis of Mhp.

Objective To investigate the effect of fumigation decoction on the expression of aggrecan(AGG)and type Ⅱ collagen(COL2A1)in an intervertebral disc degeneration(IDD)model,and to gain an in-depth understanding of the cellular and molecular mechanisms underlying traditional Chinese medicine fumigation therapy.Methods An IDD model was established using Sprague-Dawley rats by inducing disc injury via needle puncture.The rats were divided into three groups:normal control(CON),model(PUN),and fumigation(CHF).Only the CHF group received treatment with fumigation decoction for 4 weeks.Imaging analysis was conducted to assess the relative height of intervertebral discs,while Masson's trichrome staining was used to observe structural changes in the discs.The expression of NLRP3 inflammasome was detected by immunohistochemistry.Quantitative RT-PCR and Western blotting were employed to evaluate the expression levels of AGG and COL2A1 genes and proteins.Results A total of 12 rats were randomly divided into 3 groups,with 4 rats in each group.Compared with the CON group,disc height was significantly lost in the PUN group(P＜0.05),and disc height could be partially restored in the CHF group(P＜0.05).In the PUN group,the expression levels of AGG mRNA and COL2A1 mRNA and protein in the nucleus pulposus tissue were significantly lower than those in the CON group(P＜0.05).Compared with the PUN group,the CHF group showed significantly increased expression levels of AGG mRNA and COL2A1 mRNA and protein in the nucleus pulposus tissue(P＜0.05).Histological analysis revealed that,compared with the PUN group,the CHF group exhibited partial restoration of the gel-like properties of the nucleus pulposus and an increase in proteoglycan content.Additionally,NLRP3 expression in the nucleus pulposus tissue of the CHF group was markedly reduced.Conclusion Fumigation decoction treatment can enhance the expression of AGG and COL2A1 and decrease the expression of NLRP3 in IDD models,which may be one of its mechanisms for treating lumbar degenerative diseases.This research provides new insights into the treatment of IDD.

Objective To evaluate the therapeutic efficacy of endoscopic total thyroidectomy performed via a combined transoral and submental approach in patients with thyroid cancer Methods This study enrolled 227 patients diagnosed with papillary thyroid carcinoma(PTC)who underwent total thyroidectomy combined with unilateral central lymph node dissection at the Department of Thyroid and Hernia Surgery,the First Affiliated Hospital of Gannan Medical University,between October 2019 and March 2024.Of these patients,68 were treated using the fully endoscopic combined transoral and submental approach(endoscopic group),whereas 159 underwent the conventional open anterior cervical approach(open group).A retrospective analysis was performed to evaluate and compare the therapeutic outcomes between the two groups.Results Surgical procedures were successfully completed in both groups without complications,and no additional incisions were required in the endoscopic group.Compared with the open surgery group,the endoscopic group had a longer operative duration,reduced intraoperative blood loss,and greater total postoperative drainage volume.Furthermore,on the first postoperative day,patients in the endoscopic surgery group exhibited lower levels of serum calcium and intact parathyroid hormone(iPTH).The incidence of temporary parathyroid dysfunction was significantly higher in the endoscopic group(51.47%)than in the open surgery group(30.82%),and this difference was statistically significant(P<0.05).No significant differences were found between the two groups regarding preoperative serum iPTH levels,preoperative serum calcium levels,number of dissected lymph nodes,incidence of temporary recurrent laryngeal nerve injury,postoperative hemorrhage,incision infection,or submental sensory loss(P>0.05).Conclusions Compared with traditional open surgery,total thyroidectomy via TOETSMVA under full endoscopy demonstrates comparable efficacy and favorable long-term safety.However,it is associated with a higher risk of transient hypoparathyroidism in the short term,highlighting the necessity for enhanced intraoperative protection of the parathyroid glands.

Objective To evaluate the therapeutic efficacy of endoscopic total thyroidectomy performed via a combined transoral and submental approach in patients with thyroid cancer Methods This study enrolled 227 patients diagnosed with papillary thyroid carcinoma(PTC)who underwent total thyroidectomy combined with unilateral central lymph node dissection at the Department of Thyroid and Hernia Surgery,the First Affiliated Hospital of Gannan Medical University,between October 2019 and March 2024.Of these patients,68 were treated using the fully endoscopic combined transoral and submental approach(endoscopic group),whereas 159 underwent the conventional open anterior cervical approach(open group).A retrospective analysis was performed to evaluate and compare the therapeutic outcomes between the two groups.Results Surgical procedures were successfully completed in both groups without complications,and no additional incisions were required in the endoscopic group.Compared with the open surgery group,the endoscopic group had a longer operative duration,reduced intraoperative blood loss,and greater total postoperative drainage volume.Furthermore,on the first postoperative day,patients in the endoscopic surgery group exhibited lower levels of serum calcium and intact parathyroid hormone(iPTH).The incidence of temporary parathyroid dysfunction was significantly higher in the endoscopic group(51.47%)than in the open surgery group(30.82%),and this difference was statistically significant(P<0.05).No significant differences were found between the two groups regarding preoperative serum iPTH levels,preoperative serum calcium levels,number of dissected lymph nodes,incidence of temporary recurrent laryngeal nerve injury,postoperative hemorrhage,incision infection,or submental sensory loss(P>0.05).Conclusions Compared with traditional open surgery,total thyroidectomy via TOETSMVA under full endoscopy demonstrates comparable efficacy and favorable long-term safety.However,it is associated with a higher risk of transient hypoparathyroidism in the short term,highlighting the necessity for enhanced intraoperative protection of the parathyroid glands.

The aim of this study is to establish an gene editing method of Mesomycoplasma hyo-pneumoniae(Mhp)based on the homologous recombination principle.The restriction enzyme di-gestion and ligation method combined with gene synthesis were used to construct a shuttle plasmid to achieve replication in both Mhp and Escherichia coli(E.coli).The pGEM?-T vector was used as the skeleton.The oriC sequence of Mhp which can achieve the replication of the plasmid in Mhp was inserted into the vector.Sequences of the Spiroplasma promoter and puromycin resistance gene were then inserted into the above constructed plasmid to screen recombinant clones.The up-stream and downstream homologous arms of p97 were constructed to initiate homologous recombination.The recA gene of E.coli is inserted to improve the efficiency of homologous recom-bination.The obtained shuttle plasmid was then delivered into Mhp by electro-transformation or chemical transformation.A shuttle plasmid,pGEM?-Mhp-oriC-p 97,which can replicate in both Mhp and E.coli was constructed.With the transformation of this plasmid,the carried puromycin gene and recA gene can be expressed,the p97 gene can be edited.Finally,the genetically unstable p97 gene mutant was initially obtained.In this study,a tool for Mhp gene editing based on the principle of homologous recombination was established,which laid a foundation for the develop-ment of tools for studying the pathogenesis of Mhp.

Objective To investigate the effect of fumigation decoction on the expression of aggrecan(AGG)and type Ⅱ collagen(COL2A1)in an intervertebral disc degeneration(IDD)model,and to gain an in-depth understanding of the cellular and molecular mechanisms underlying traditional Chinese medicine fumigation therapy.Methods An IDD model was established using Sprague-Dawley rats by inducing disc injury via needle puncture.The rats were divided into three groups:normal control(CON),model(PUN),and fumigation(CHF).Only the CHF group received treatment with fumigation decoction for 4 weeks.Imaging analysis was conducted to assess the relative height of intervertebral discs,while Masson's trichrome staining was used to observe structural changes in the discs.The expression of NLRP3 inflammasome was detected by immunohistochemistry.Quantitative RT-PCR and Western blotting were employed to evaluate the expression levels of AGG and COL2A1 genes and proteins.Results A total of 12 rats were randomly divided into 3 groups,with 4 rats in each group.Compared with the CON group,disc height was significantly lost in the PUN group(P＜0.05),and disc height could be partially restored in the CHF group(P＜0.05).In the PUN group,the expression levels of AGG mRNA and COL2A1 mRNA and protein in the nucleus pulposus tissue were significantly lower than those in the CON group(P＜0.05).Compared with the PUN group,the CHF group showed significantly increased expression levels of AGG mRNA and COL2A1 mRNA and protein in the nucleus pulposus tissue(P＜0.05).Histological analysis revealed that,compared with the PUN group,the CHF group exhibited partial restoration of the gel-like properties of the nucleus pulposus and an increase in proteoglycan content.Additionally,NLRP3 expression in the nucleus pulposus tissue of the CHF group was markedly reduced.Conclusion Fumigation decoction treatment can enhance the expression of AGG and COL2A1 and decrease the expression of NLRP3 in IDD models,which may be one of its mechanisms for treating lumbar degenerative diseases.This research provides new insights into the treatment of IDD.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To explore and analyze the predictive value of EuroSCORE Ⅱ and SYNTAX Ⅱ scores for clinical outcomes in patients undergoing coronary artery bypass grafting (CABG) surgery.Methods:A total of 500 coronary artery disease (CAD) patients who underwent CABG in Shanxi Cardiovascular Hospital from April 2014 to July 2023 were selected as the study subjects, all patients were given EuroSCORE Ⅱand SYNTAX Ⅱ scores to evaluate the predictive value of EuroSCOREⅡfor perioperative mortality and SYNTAX Ⅱ for 4-year mortality. Univariate and multivariate Logistic analysis were employed to analyze the independent risk factors for perioperative and 4-year mortality.Results:There were 3 deaths during the perioperative period, with a mortality rate of 0.60%, the predicted mortality rate of EuroSCOREⅡwas 1.71%; there were 21 deaths at 4 years after surgery, with a mortality rate of 4.23% and the predicted mortality rate of SYNTAX Ⅱwas 9.02%. Logistic regression analysis showed that left ventricular ejection fraction (LVEF) was the only independent protective factor for perioperative mortality, and advanced age was the only independent risk factor for 4-year postoperative mortality in patients ( P<0.05). The analysis of the working characteristic curve of the subjects found that the area under the receiver operating characteristic curve ( ROC) of EuroSCORE Ⅱ for perioperative mortality was 0.782, and the area under ROC curve of SYNTAX Ⅱfor postoperative 4-year mortality was 0.743. Conclusion:Both EuroSCORE Ⅱand SYNTAX Ⅱhave certain predictive value for perioperative mortality and postoperative 4-year mortality in patients undergoing CABG, respectively, but the predicted mortality rate is relatively higher.

Objective To investigate the expression of cyclooxygenase-2(COX-2)and Ki-67 in rectal cancer tissues and their predictive value for the sensitivity to neoadjuvant chemoradiotherapy(NAC).Methods The clinical data of 87 patients receiving chemoradiotherapy before rectal cancer surgery at Nanjing Hospital Affiliated to Nanjing University of Chinese Medicine(the Second Hospital of Nanjing)from June 2021 to September 2022 were retrospectively analyzed.In addition,40 normal rectal tissue samples were selected from the Department of Pathology of Nanjing University of Chinese Medicine(the Second Hospital of Nanjing)as control.The expression levels of COX-2 and Ki-67 in tumor and adjacent tissues of patients with rectal cancer as well as in normal rectal tissues were detected by using the immunohistochemical method.The patients were divided into chemoradiotherapy-sensitive group(n=62)and chemoradiotherapy-resistant group(n=27)according to whether they were sensitive to chemoradiotherapy.The correlation between the expression levels of COX-2,Ki-67 in tumor tissues and adjacent tissues and the sensitivity to chemoradiotherapy was analyzed.The relative factors affecting the effect of chemoradiotherapy on rectal cancer patients were analyzed by using the logistic regression model.The receiver operating characteristic(ROC)curve was drawn,and the area under the curve(AUC)was used to evaluate the predictive value of COX-2 and Ki-67 expression levels in tumor tissues of rectal cancer patients for the sensitivity to NAC.Results Among the 87 patients with rectal cancer,60 patients were sensitive to chemoradiotherapy,with a sensitivity rate of 68.97％.The positive expression rates of COX-2 and Ki-67 in tumor tissues and adjacent tissues were significantly higher than those in normal rectal tissues(x2=53.187,7.131,53.047,14.613;P＜0.05).The positive expression rates of COX-2 and Ki-67 in tumor tissues were significantly higher than those in adjacent tissues(x2=72.572,67.616;P＜0.05).The positive expression rates of COX-2 and Ki-67 in tumor tissues of patients in the chemoradiotherapy-sensitive group were significantly lower than those in the chemoradiotherapy-resistant group(x2=3.965,6.264;P＜0.05).Logistic regression analysis showed that the positive expression of COX-2 and Ki-67 in tumor tissues were factors affecting the efficacy of NAC in rectal cancer patients(P＜0.05).ROC curve analysis results showed that the sensitivity of COX-2 and Ki-67 expression and their combination for predicting sensitivity of patients to NAC was 100.00％,100.00％,and 100.00％,respectively;while the specificity was 13.33％,20.00％,and 31.67％,respectively;and the AUC was 0.567,0.600,and 0.658,respectively.Conclusion The positive expression of COX-2 and Ki-67 in tumor tissues are factors affecting the efficacy of NAC in rectal cancer patients,and the combined detection of COX-2 and Ki-67 expression has a high predictive value for the sensitivity of NAC.

Objective To investigate the value of multiparametric CT features for predicting the risk classification of gastric stro-mal tumor(GST).Methods The clinical data from 139 patients with GST were retrospectively collected.According to the patho-logical risk results,the patients were divided into two groups:a low-risk GST group(including very low-and low-risk)with 75 patients and a high-risk GST group(including medium and high-risk)with 64 patients.The CT features between low-risk GST group and high-risk GST group were compared using chi-squared test or t-test.The risk factors of high-risk GST were identified by univariate analysis.The prediction models were built by multivariate logistic regression.The performance of models were evaluated by receiver oper-ating characteristic(ROC)curve.Results There were significant differences in the maximum tumor diameter,minimum tumor diameter,arterial phase enhancement degree,venous phase enhancement rate,arterial phase enhancement degree rate,venous phase enhance-ment degree rate,cystic,and necrosis between low-risk GST group and high-risk GST group,which were associated with the risk classification of GST.The area under the curve(AUC)of the quantitative features-based model that combined maximum tumor diam-eter,minimum tumor diameter,arterial phase enhancement degree,venous phase enhancement rate,arterial phase enhancement degree rate and venous phase enhancement degree rate,showed a significantly higher performance than the qualitative features-based model that incorporated cystic and necrosis(0.981 vs 0.850,P<0.001).Conclusion Maximum tumor diameter,minimum tumor diameter,arterial phase enhancement degree,venous phase enhancement rate,arterial phase enhancement degree rate,venous phase enhance-ment degree rate,as well as cystic and necrosis,are associated with the risk classification of GST and can predict the high-risk GST.