The crosstalk between macrophages and bone marrow mesen chymal stem cells （BMSCs） plays a vital role in regulating tendon-bone healing. Based on the traditional Chinese medicine （TCM） theory that the kidney governs the bones and generates marrow， yin-nourishing and kidney-tonifying Chinese medicines can promote the osteogenic differentiation of BMSCs and the M1-to-M2 polarization of macrophages， thereby facilitating tendon-bone healing. A variety of related medicinal monomers （such as diosgenin and oleanolic acid）， extracts and effective fractions （such as artificial tiger bone powder and total flavonoids of Drynaria fortunei ）， as well as compound prescriptions （such as Liuwei dihuang pill， Xujin jiegu decoction and Duhuo jisheng decoction）， can modulate the crosstalk between macrophages and BMSCs by regulating the Wnt/β-catenin， nuclear factor kappa-B， phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin and transforming growth factor-β/Smad signaling pathways. They further exert anti-inflammatory， antioxidant and osteogenic effects to effectively accelerate tendon-bone healing. However， current evidence was limited to animal experiments exhibiting such as limitations insufficent alignment between animal models and clinical injury types， research focusing mostly on single signaling pathways， and unclear core active ingredients. Future studies shall optimize animal models and carry out large-sample randomized controlled trials， clarify the interactive networks of multiple signaling pathways， and isolate the core active substances of yin-nourishing and kidney-tonifying Chinese medicines， so as to advance the precise application of traditional Chinese medicines.

The eIF3i gene was amplified from lamb testicular(LT)cells by PCR and cloned into pCold vector to construct the pCold-eIF3i plasmid.Plasmid PCR,double enzyme digestion and se-quencing were used to verify the results.The recombinant eIF3i protein was induced under the op-timized expression conditions.The expression and reactogenicity of the target protein were detected by SDS-PAGE and Western blot.New Zealand white rabbits were immunized with purified recom-binant eIF3i protein combined with Freund's complete and incomplete adjuvants for three times.Se-rum samples were collected after immunization.Indirect ELISA was used to detect antiserum titer,and Western blot was used to analyze antibody specificity.Indirect immunofluorescence assay(IFA)was used to detect the application effect of antibodies.The results showed that the size of LT eIF3i gene was 978 bp.The optimal expression conditions for the eIF3i recombinant protein were as follows:IPTG concentration of 0.2 mmol/L,temperature of 37 ℃,and induction time of 8 h.The recombinant eIF3i protein was expressed as an inclusion body with a size of about 36 kDa.The titer of polyclonal antibody against eIF3i protein was 1∶51 200.Western blot and IFA showed that the prepared polyclonal antibody against eIF3i protein had good reactivity and specificity.In conclusion,we successfully prepared rabbit anti-eif3i polyclonal antibody and confirmed that it could specifically recognize endogenous eIF3i protein,which laid a foundation for further study on the biological function of eIF3i protein.

This study aims to establish a convenient,new and visual detection method for the field diagnosis of Pasteurella multocida(Pm).With reference to the Pm kmt1 gene conserved sequence published in GenBank,PCR amplification primers were designed,the amplified kmt1 gene was cloned into pMD19-T vector,and the recombinant plasmid standard pMD19-T-kmt 1 was estab-lished and identified by PCR and sequencing.Using pMD1 9-T-kmt 1 plasmid as template and kmt1 gene as target gene,basic primers were designed and synthesized.According to the requirements of LFD,a probe(Pm-P)was designed,and the RPA-LFD method for Pm detection was established by optimizing the reaction conditions.Specificity and sensitivity tests were carried out,and 64 clini-cal samples were tested by the method.The results showed that the established Pm RPA-LFD method could be amplified at 37 ℃ for 15 min.Escherichia coli(E.coli),Salmonella enteriditis(SE),Riemerella anatipestifer(RA),Staphylococcus,goose parvovirus(GPV),duck plague virus(DPV),Muscovy duck parvovirus(MDPV)DNA was extracted as the template,and plasmid standard pMD19-T-kmt 1 was used as the positive control.All the positive controls were negative,indicating that the method had good specificity.The plasmid standard pMD1 9-T-kmt 1 was diluted with a 10-fold ratio,and the plasmid standard with a concentration of 107-100 copies/μL was used as the template.The sensitivity was 1.50×101 copies/μ,,which was 100 times higher than that of PCR.A total of 64 clinical samples with suspected RA were subjected to testing using PCR,RPA and LAMP-LFD,with a 100％compliance rate for all three detection tests.The results show that the established RPA-LFD method has the characteristics of strong specificity,high sensitivity,fast speed and visualization,and can be applied to the field detection of Pm.

Objective:To investigate the role of Ym2 in olfactory epithelium homeostasis and olfactory behavior.Methods:Differential expression of Ym2 in the olfactory epithelium of young and aged mice was analyzed using single-cell RNA sequencing. Ym2 and related genes in the aged olfactory epithelium were identified, and their biological function was analyzed by GO enrichment analysis. The expression of Ym2 and Ym2-related genes in the young and aged olfactory epithelium was detected by immunostaining and RNAscope in situ hybridization. Buried food pullet test was used to assess the impact of Ym2 knockout on olfactory function, while immunostaining was used to evalute the effect of Ym2 knockout of on the homeostasis of olfactory epithelium. Statistical analysis was performed using GraphPad Prism 8.0.1 software.Results:Single-cell RNA sequencing revealed that Ym2 was highly expressed in multiple cell types, including horizontal basal cells and respiratory ciliated cells of the olfactory epithelium in aged mice compared to young animals. In situ hybridization and immunostaining data showed that Ym2 mRNA level were higher in the aged olfactory epithelium than that in the young tissue ( t=4.50, P<0.001). At the protein level, Ym2 expression was higher in horizontal basal cells ( t=3.03, P<0.05) and supporting cells ( t=7.76, P<0.001) of the aged epithelium compared to the young tissue. Additionally, the mRNA levels of the two Ym2-related genes, Alox15 and Cxcl5, were also higher in the aged olfactory epithelium ( t=2.72 and 2.68, respectively, both P<0.05). Behavioral testing showed that Ym2-/- mice took significantly longer to find buried food pellets compared to wild-type (WT) mice ( t=2.35, P<0.05). Histological analysis revealed a significant reduction in the number of olfactory sensory neurons and supporting cells in the olfactory epithelium of Ym2-/- mice compared to WT mice ( t=5.86 and 3.69, respectively, both P<0.01). Conclusions:Ym2 plays a critical role in smell perception. Ym2 knockout leads to reduction in the number of olfactory epithelial cell and impairs olfactory behavior of food-searching in young mice.

The eIF3i gene was amplified from lamb testicular(LT)cells by PCR and cloned into pCold vector to construct the pCold-eIF3i plasmid.Plasmid PCR,double enzyme digestion and se-quencing were used to verify the results.The recombinant eIF3i protein was induced under the op-timized expression conditions.The expression and reactogenicity of the target protein were detected by SDS-PAGE and Western blot.New Zealand white rabbits were immunized with purified recom-binant eIF3i protein combined with Freund's complete and incomplete adjuvants for three times.Se-rum samples were collected after immunization.Indirect ELISA was used to detect antiserum titer,and Western blot was used to analyze antibody specificity.Indirect immunofluorescence assay(IFA)was used to detect the application effect of antibodies.The results showed that the size of LT eIF3i gene was 978 bp.The optimal expression conditions for the eIF3i recombinant protein were as follows:IPTG concentration of 0.2 mmol/L,temperature of 37 ℃,and induction time of 8 h.The recombinant eIF3i protein was expressed as an inclusion body with a size of about 36 kDa.The titer of polyclonal antibody against eIF3i protein was 1∶51 200.Western blot and IFA showed that the prepared polyclonal antibody against eIF3i protein had good reactivity and specificity.In conclusion,we successfully prepared rabbit anti-eif3i polyclonal antibody and confirmed that it could specifically recognize endogenous eIF3i protein,which laid a foundation for further study on the biological function of eIF3i protein.

Objective:To investigate the value of habitat radiomic features based on gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA)-enhanced MRI in establishing a predictive model for cytokeratin 19 (CK19) expression in hepatocellular carcinoma (HCC) and to evaluate its role in prognostic risk stratification.Methods:This multicenter case-control study retrospectively enrolled 489 patients with pathologically confirmed HCC who underwent Gd-EOB-DTPA-enhanced MRI between June 2016 and June 2024. Among them, 346 patients from the First Affiliated Hospital of Soochow University were divided into a training cohort ( n=245) and an internal test cohort ( n=101) via stratified sampling at a 7∶3 ratio. And 143 patients from Nantong Third Hospital Affiliated to Nantong University served as an external validation cohort. The training cohort included 53 CK19-positive and 192 CK19-negative patients. The internal test cohort included 21 CK19-positive and 80 CK19-negative patients. The external validation cohort included 30 CK19-positive and 113 CK19-negative patients. Univariate logistic regression analysis was performed to identify potential factors associated with CK19 expression, and a clinical-radiologic model was constructed. The k-means clustering algorithm was applied to segment target HCC lesions into 3 subregions. Radiomic features were extracted and selected from these habitat subregions. Habitat radiomics models were constructed for the arterial phase (AP), portal venous phase, hepatobiliary phase (HBP), and combined phases (CP). Multivariate logistic regression analysis identified independent clinical and radiologic predictors of CK19 expression, and the optimal habitat model score was integrated to build a clinical-radiologic-habitat combined model. The area under the receiver operating characteristic curve (AUC) was used to evaluate model predictive performance. Recurrence-free survival (RFS) was analyzed using the Kaplan-Meier method and the differences in survival curves were compared with the log-rank test. Results:Univariate logistic regression analysis revealed that alpha-fetoprotein (AFP) ( OR=2.629, 95% CI 1.412-4.896, P=0.002), AP enhancement ( OR=3.636, 95% CI 1.642-8.052, P=0.001), AP peritumoral enhancement ( OR=2.219, 95% CI 1.084-4.542, P=0.029), and HBP peritumoral hypointensity ( OR=2.010, 95% CI 1.004-4.021, P=0.049) were potential factors associated with CK19 expression, which were incorporated into the clinical-radiologic model. In the internal and external validation cohorts, the AUC of the clinical-radiologic model was 0.690 (95% CI 0.590-0.778) and 0.650 (95% CI 0.565-0.727), respectively. The habitat radiomics model based on CP images demonstrated the highest performance. It achieved AUC of 0.729 (95% CI 0.622-0.836) and 0.725 (95% CI 0.607-0.842) in the internal and external validation cohorts, respectively. Multivariate analysis identified AFP ( OR=2.494, 95% CI 1.163-5.348, P=0.019), AP enhancement ( OR=5.230, 95% CI 1.868-14.643, P=0.002) and habitat radiomics model score ( OR=4.105, 95% CI 2.643-6.368, P<0.001) as independent predictors of CK19 positivity. Based on these factors, a combined clinical-radiologic-habitat combined model was established. The clinical-radiologic-habitat combined model achieved AUCs of 0.767 (95% CI 0.671-0.846) and 0.730 (95% CI 0.649-0.801) in the internal and external validation cohorts, respectively. Significant differences in RFS were observed between the CK19-positive group (25.1 month) and CK19-negative group (51.0 month) as predicted by the clinical-radiologic-habitat model ( χ2=4.17, P=0.041). Conclusion:The clinical-radiologic-habitat combined model based on Gd-EOB-DTPA-enhanced MRI habitat radiomics demonstrates good predictive performance for CK19 expression in HCC and offers valuable prognostic stratification for clinical practice.

Objective:To analyze the influences of coronary angiography(CAG)of transradial artery on the success rate,radiation dose and safety in elderly patients with coronary heart disease(CHD).Methods:This study conducted a prospective study.A total of 150 elderly CHD patients who admitted to Heilongjiang Rad Cross Sengong General Hospital from November 2022 to November 2023 were selected,and they were randomly divided into three groups using a random number table,which included the femoral artery group(n=50),radial artery group(n=50),and ulnar artery group(n=50).The femoral artery group underwent CAG by using the femoral artery approach,and the radial artery group underwent CAG by using the radial artery approach,and the ulnar artery group underwent CAG by using the ulnar artery approach.The CAG success rate and CAG indicators[dosage of contrast agent,X-ray exposure time,time of conducting puncture,corrected TIMI frame count(CTFC)]were compared among the three groups.The radiation-related parameters[cumulative dose(CD),dose-area product(DAP)]also were compared among three groups.The patients'comfort degrees of three groups were assessed by adopting General Comfort Questionnaire(GCQ)at three time points(pre-CAG,during CAG,at the 12th hour post-CAG).In addition,the complications of three groups were recorded.Results:The numbers of success angiography were respectively 47 cases,48 cases and 46 cases in femoral artery group,radial artery group and ulnar artery group,and the success rates of them were respectively 94.00%(47/50),96.00%(48/50)and 92.00%(46/50),without statistically significant differences among three groups(P>0.05).There were not significant differences in the dosage of contrast agent,X-ray exposure time,time of conducting puncture,CTFC,CD and DAP among three groups(P>0.05).In the GCQ scores,the scores pre-CAG of all three groups were highest,followed by those at the 12th hour post-CAG,and then,those during CAG were lowest,which appeared a trend of rise after decline.The total incidence of complication was lowest(6.25%)in the radial artery group,followed by the ulnar artery group(10.87%),and that(21.28%)of the femoral artery group was the highest,but there was not statistically significant difference in that among three groups(P>0.05).Conclusion:The CAG success rates of the radial artery,femoral artery and ulnar artery are similar,and there is not significant difference in radiation dose.However,compared with CAG of femoral artery and ulnar artery,the CAG of radial artery has higher safety,and higher comfort degree at the same time.

With the continuous advancement of internet technology and the improvement of internet literacy among the general population, the concept of online-offline integration in internet hospitals has gradually gained acceptance and has been applied and developed both domestically and internationally. In thoracic surgery, the applicability of this model lies in enhancing efficiency and delivering comprehensive, diversified, and personalized medical services to address complex and severe conditions. However, challenges such as hardware limitations and diagnostic/treatment risks persist during the implementation of internet hospitals. Through future in-depth and localized research, the online-offline integration of internet hospitals is expected to undergo further development and refinement. This progress will facilitate its integration into clinical practice in thoracic surgery, ultimately providing patients with improved medical care services.

Objective:To investigate the feasibility and advantages of 30°reverse trendelenburg combined with semi-prone position in single-port thoracoscopic surgery for lung cancer.Methods:This study is a prospective, randomized controlled trial. Between October 2022 and December 2023, patients with lung cancer who met the inclusion and exclusion criteria were prospectively enrolled at the Department of Thoracic Surgery, the First Affiliated Hospital of Zhengzhou University,and randomized using a simple randomization design to generate a random number table. Patients were allocated in a 1∶1 ratio to either the modified positioning group (30°reverse trendelenburg combined with semi-prone position) or the conventional positioning group (lateral decubitus position). Comparative analysis was performed on feasibility metrics including operative time, postoperative pain scores, surgeon and assistant fatigue levels, number of lymph nodes dissected, duration of postoperative chest tube drainage, and postoperative hospital stay. Inter-group comparisons were conducted using independent sample t-tests, χ2 tests or Kruskal-Wallis. Results:A total of 211 patients were included.The modified positioning group comprised 105 patients (58 males, 47 females) with an age of (56.2±10.3）years (range: 23 to 74 years). The conventional positioning group included 106 patients (55 males, 51 females) with an age of (57.8±11.7）years (range: 21 to 76 years). All patients underwent successful single-port thoracoscopic lung cancer surgery without severe perioperative complications. No statistically significant differences were observed between groups in operative time ((98±34) minutes vs. (114±36) minutes, t=-1.642, P=0.221), number of lymph nodes dissected (13.4±2.5 vs. 12.8±3.2, t=0.702, P=0.763), postoperative chest tube duration ((4.3±1.5) days vs. (5.1±1.8) days, t=-1.348, P=0.247), or postoperative hospital stay ((5.0±1.2) days vs. (5.6±1.5) days, t=-0.831, P=0.682). The modified positioning group demonstrated significantly lower postoperative day 1 pain scores on the visual analog scale (3.1±2.0 vs. 4.6±1.9, t=-4.321, P=0.027) and postoperative surgeon-reported fatigue scores (primary surgeon: 13.6±2.5 vs. 18.5±3.3, t=-6.963, P=0.002; assistant surgeon: 12.1±2.0 vs. 17.1±3.1, t=-6.133, P=0.003) compared with the conventional group, with all differences reaching statistical significance. Conclusions:The application of 30°reverse trendelenburg combined with semi-prone position during single-port thoracoscopic lung cancer surgery is both safe and feasible. Compared with the conventional lateral decubitus position, this modified positioning demonstrates significant advantages in reducing postoperative pain and alleviating surgical team fatigue (including both primary and assistant surgeons).

This study aims to establish a convenient,new and visual detection method for the field diagnosis of Pasteurella multocida(Pm).With reference to the Pm kmt1 gene conserved sequence published in GenBank,PCR amplification primers were designed,the amplified kmt1 gene was cloned into pMD19-T vector,and the recombinant plasmid standard pMD19-T-kmt 1 was estab-lished and identified by PCR and sequencing.Using pMD1 9-T-kmt 1 plasmid as template and kmt1 gene as target gene,basic primers were designed and synthesized.According to the requirements of LFD,a probe(Pm-P)was designed,and the RPA-LFD method for Pm detection was established by optimizing the reaction conditions.Specificity and sensitivity tests were carried out,and 64 clini-cal samples were tested by the method.The results showed that the established Pm RPA-LFD method could be amplified at 37 ℃ for 15 min.Escherichia coli(E.coli),Salmonella enteriditis(SE),Riemerella anatipestifer(RA),Staphylococcus,goose parvovirus(GPV),duck plague virus(DPV),Muscovy duck parvovirus(MDPV)DNA was extracted as the template,and plasmid standard pMD19-T-kmt 1 was used as the positive control.All the positive controls were negative,indicating that the method had good specificity.The plasmid standard pMD1 9-T-kmt 1 was diluted with a 10-fold ratio,and the plasmid standard with a concentration of 107-100 copies/μL was used as the template.The sensitivity was 1.50×101 copies/μ,,which was 100 times higher than that of PCR.A total of 64 clinical samples with suspected RA were subjected to testing using PCR,RPA and LAMP-LFD,with a 100％compliance rate for all three detection tests.The results show that the established RPA-LFD method has the characteristics of strong specificity,high sensitivity,fast speed and visualization,and can be applied to the field detection of Pm.