Objective To explore the correlation of MET status in patients with advanced prostatic acinar adenocarcinoma with the clinical pathological parameters and prognosis. Methods The specimen from 135 patients with advanced prostatic acinar adenocarcinoma was included. The expression of c-MET protein was detected via immunohistochemistry, and MET gene amplification was assessed by fluorescence in situ hybridization. The relationships of c-MET expression and gene amplification with clinicopathological features and prognosis were analyzed. Results The positive expression rate of c-MET was 52.60% (71/135). Compared with the c-MET expression in adjacent tissues, that in tumor tissues showed lower heterogeneous expression. Among the cases, 1.71% (2/117) exhibited MET gene polyploidy, but no gene amplification was detected. Positive c-MET expression was significantly correlated with high Gleason scores and grade groups (P=0.0073). However, no significant relationship was found between c-MET expression and progression-free survival or post-treatment t-PSA levels. Conclusion c-MET protein is expressed at a relatively low level in advanced prostatic acinar adenocarcinoma, suggesting that c-MET may not be a viable target for ADC drug development in prostatic acinar adenocarcinoma.

Peanut, a major economic and oil crop known for the high protein and oil content, is extensively cultivated in China. Peanut plants have the ability to form nodules with rhizobia, where the nitrogenase converts atmospheric nitrogen into ammonia nitrogen that can be utilized by the plants. Analysis of nodule fixation is of positive significance for avoiding overapplication of chemical fertilizer and developing sustainable agriculture. In this study, AhNIGT1.2, a member of the NIGT family predominantly expressed in peanut nodules, was identified by bioinformatics analysis. Subsequent spatiotemporal expression analysis revealed that AhNIGT1.2 was highly expressed in nodules and showed significant responses to high nitrogen, low nitrogen, high phosphorus, low phosphorus, and rhizobia treatments. Histochemical staining indicated that the gene was primarily expressed in developing nodules and at the connection region between mature nodules and peanut roots. The fusion protein AhNIGT1.2-GFP was located in the nucleus of tobacco epidermal cells. The AhNIGT1.2-OE significantly increased the number of peanut nodules, while AhNIGT1.2-RNAi reduced the number of nodules, which suggested a positive regulatory role of AhNIGT1.2 in peanut nodulation. The AhNIGT1.2-OE in roots down-regulated the expression levels of NRT1.2, NRT2.4, NLP1, and NLP7, which indicated that AhNIGT1.2 influenced peanut nodulation by modulating nitrate transport and the expression of NLP genes. The transcriptome analysis of AhNIGT1.2-OE and control roots revealed that overexpressing AhNIGT1.2 significantly enriched the differentially expressed genes associated with nitrate response, nodulation factor pathway, enzymes for triterpene biosynthesis, and carotenoid biosynthesis. These findings suggest that AhNIGT1.2 play a key role in peanut nodulation by regulating nitrate transport and response and other related pathways. This study gives insights into the molecular mechanisms of nitrogen and phosphorus in regulating legume nodulation and nitrogen fixation, and sheds light on the development of legume crops that can efficiently fix nitrogen in high nitrogen environments.
Arachis/physiology* ; Nitrates/metabolism* ; Plant Proteins/physiology* ; Transcription Factors/metabolism* ; Plant Root Nodulation/physiology* ; Gene Expression Regulation, Plant ; Root Nodules, Plant/metabolism* ; Nitrogen Fixation

Objective:To explore the anal function and postoperative complications of 2-stage Turnbull-Cutait pull-through coloanal anastomosis (TCA) for low rectal cancer.Methods:Patients undergoing radical rectal cancer resection from Feb 2023 to Nov 2024 in the First Hospital of Jilin University were divided into the TCA surgery group and the low anterior resection combined with prophylactic stoma (LAR) surgery group.Results:Among the 102 patients, there were 50 cases in the TCA group and 52 cases in the LAR group. In the single-arm analysis of the TCA group, the overall complication rate was 44%. The incidence rates of severe LARS at 1 month, 3 months, and 6 months after surgery were 97%, 77%, and 64% respectively. There was no significant difference in the complication rate within 30 days after surgery between the two groups,(44% vs. 38%, χ2=0.135, P>0.05). There was no significant difference in the incidence rate of severe LARS between the TCA group and the LAR group (77% vs. 69%, χ2=0.202, P>0.05), and there was not significant difference in the incidence rate of severe LARS between the two groups at the 6th month after surgery,(64% vs. 48%, χ2=1.132, P>0.05). Conclusion:In patients who underwent TCA surgery, the LARS symptoms gradually decreased over time. Compared with patients undergoing low anterior resection and stoma reversal, there were no significant differences in complications within 30 days after surgery and LARS symptoms within half a year.

Objective:To screen out the disinfection procedure and disinfectant suitable for the actual production of specific pathogen free(SPF)chicken embryo eggs,so as to ensure the disinfection effect of specific pathogen free(SPF)chicken embryo eggs in vaccine production.Methods:This study compares the microbial counting methods of soaking,swabbing with cotton swabs and pouring after thin-film filtration for SPF chicken embryo eggs in a GMP production workshop,and selects the most suitable method for SPF chicken embryo egg microbial counting.Experi-mental groups A,B,and C use self-prepared concentrations of 1∶50 sporicidal agent dilution solution,1∶128 alka-line phenol salt dilution solution,and ready-to-use compound quaternary ammonium disinfectant(sterile),respec-tively,and follow the actual disinfection procedures in the workshop to disinfect and sample the surfaces of SPF chicken embryo eggs entering different cleanliness grades,while using sterile water instead of disinfectant as the control group.The average bactericidal rate is calculated by recording the number of colonies and monitoring the viability of chicken embryo cells using microbial culture,and the disinfection effect of the three disinfectants on SPF chicken embryo eggs is evaluated.Results:The comparison of the results from the three methods shows that the method of sampling SPF chicken embryo eggs by immersion and then counting the microbial colonies through membrane filtration is superior to the other two methods.The final cleaning rate of the control group,which used sterilized injection water to clean the SPF chicken embryo eggs,was 91.67％to 96.97％,while the final steriliza-tion rate of the experimental group,which used the above three disinfectants to disinfect the SPF chicken embryo eggs,was 100.00％.By comparing the cell counts of the experimental group and the control group,it was found that the live cell density of the control group was(6.03-6.25)× 105 cells·mL-1,and that of the experimental groups A-C was(6.08-6.17)× 105 cells·mL-1,(5.99-6.25)× 105 cells·mL-1,and(5.87-6.21)× 105 cells·mL-1 respectively;the cell viability of the control group was 90.33％to 91.35％,and that of the experi-mental groups A-C was 88.25％to 92.12％,89.45％to 93.59％,and 88.02％to 92.89％respectively.Through statistical analysis,it was found that the P values of all experimental groups compared with the control group were greater than 0.05,indicating no statistically significant difference.Conclusion:By comparing the dis-infection effects,cell density and cell viability of the three disinfectants and comprehensively considering factors such as cost and risk of the three disinfectants,1∶50 sporicide dilution,1∶128 alkaline phenolate dilution and ready-to-use compound quaternary ammonium salt disinfectant(sterile)can all be used for the daily surface disin-fection of SPF chicken embryo eggs in the production workshop.The selection of an appropriate disinfectant should be based on specific application scenarios and requirements.

Objective:To investigate affinity of monoclonal antibody(MAb)of different cardiac troponinⅠ(cTnⅠ)epi-topes to cTnⅠ-troponin C(cTnⅠ-TnC)antigen,and to screen novel cTnⅠ-specific antibodies for clinical detection kit development.Methods:Based on technology platform of microarray chemiluminescence immunoassay,cTnⅠ-TnC antigen spot sample hard matrix chip was used,mouse derived cTnⅠ MAb was used as antibody to be detected,HRP-sheep-anti-mouse-IgG was used as antibody to detect,and experimental conditions were optimized for anti-origin solution,antigen spot sample concentration and antibody to be detected concentration.Affinity of cTnⅠ MAb with different antigenic epitopes to cTnⅠ-TnC was detected by competition method.Graded concentrations of cTnⅠ-TnC were added to experimental group,and diluents of equal volume were added to control group.Reaction signals of different cTnⅠ MAb were detected and affinity constants were calculated.Results:Antigen spot sample liquid was P105,antigen spot sample concentration was 0.1 mg/ml,and antibody concentration to be detected was 80 ng/ml were optimal experi-mental conditions.20c6cc and 7B9cc MAb containing TnC antibodies had the best affinity,and 20c6cc MAb reached 3.18×106 L/mol.cTnⅠ MAb located at C-end of peptide chain in a.a.r 130～145,169～178 and 190～196 regions showed good affinity,among which MAb 625(a.a.r 169～178)was 6.37×105 L/mol,showing the strongest affinity among cTnⅠ MAb.Conclusion:MAb has good affinity with cTnⅠ-TnC antigen at C-end of cTnⅠ peptide chain a.a.r 130～145,169～178,190～196 and TnC region.Designing a new type of cTnⅠ complement antibody targeting this region can be a new way to solve negative interference of autoantibody and peptide proteolysis.

To achieve efficient and rapid detection of duck astrovirus type 2(DAstV-2),RPA prim-ers and crRNA were designed and synthesized based on the conserved sequence of the ORF2 gene of DAstV-2.A detection method for DAstV-2 was constructed,integrating RPA nucleic acid ampli-fication,LwCas13a cleavage,and colloidal gold lateral flow dipstick visualization.The specificity,sensitivity,and concordance of this detection method were evaluated.The experimental results showed that the detection limit for the DAstV-2 recombinant plasmid standard was 1.2×101 cop-ies/μL,which is superior to the conventional RT-PCR method.The method can specifically detect DAstV-2 pathogenic nucleic acids without cross-reactivity with DAstV-1,DAstV-3,DAstV-4,duck plague virus(DEV),and duck tembusu virus(DTMUV).When testing liver tissue samples from ducks suspected of being infected with DAstV-2,the results obtained using this method were com-pletely consistent with those from real-time quantitative PCR,with a 100％concordance rate.How-ever,this method is simpler and faster to perform.The research indicates that the established RPA-CRISPR/Cas13a-LFD detection system has high sensitivity,strong specificity,and high accuracy,capable of completing rapid visual detection of DAstV-2 nucleic acids within 1 h at a constant tem-perature of 37 ℃,providing a new technical platform for the rapid diagnosis of DAstV-2.

Objective:To screen out the disinfection procedure and disinfectant suitable for the actual production of specific pathogen free(SPF)chicken embryo eggs,so as to ensure the disinfection effect of specific pathogen free(SPF)chicken embryo eggs in vaccine production.Methods:This study compares the microbial counting methods of soaking,swabbing with cotton swabs and pouring after thin-film filtration for SPF chicken embryo eggs in a GMP production workshop,and selects the most suitable method for SPF chicken embryo egg microbial counting.Experi-mental groups A,B,and C use self-prepared concentrations of 1∶50 sporicidal agent dilution solution,1∶128 alka-line phenol salt dilution solution,and ready-to-use compound quaternary ammonium disinfectant(sterile),respec-tively,and follow the actual disinfection procedures in the workshop to disinfect and sample the surfaces of SPF chicken embryo eggs entering different cleanliness grades,while using sterile water instead of disinfectant as the control group.The average bactericidal rate is calculated by recording the number of colonies and monitoring the viability of chicken embryo cells using microbial culture,and the disinfection effect of the three disinfectants on SPF chicken embryo eggs is evaluated.Results:The comparison of the results from the three methods shows that the method of sampling SPF chicken embryo eggs by immersion and then counting the microbial colonies through membrane filtration is superior to the other two methods.The final cleaning rate of the control group,which used sterilized injection water to clean the SPF chicken embryo eggs,was 91.67％to 96.97％,while the final steriliza-tion rate of the experimental group,which used the above three disinfectants to disinfect the SPF chicken embryo eggs,was 100.00％.By comparing the cell counts of the experimental group and the control group,it was found that the live cell density of the control group was(6.03-6.25)× 105 cells·mL-1,and that of the experimental groups A-C was(6.08-6.17)× 105 cells·mL-1,(5.99-6.25)× 105 cells·mL-1,and(5.87-6.21)× 105 cells·mL-1 respectively;the cell viability of the control group was 90.33％to 91.35％,and that of the experi-mental groups A-C was 88.25％to 92.12％,89.45％to 93.59％,and 88.02％to 92.89％respectively.Through statistical analysis,it was found that the P values of all experimental groups compared with the control group were greater than 0.05,indicating no statistically significant difference.Conclusion:By comparing the dis-infection effects,cell density and cell viability of the three disinfectants and comprehensively considering factors such as cost and risk of the three disinfectants,1∶50 sporicide dilution,1∶128 alkaline phenolate dilution and ready-to-use compound quaternary ammonium salt disinfectant(sterile)can all be used for the daily surface disin-fection of SPF chicken embryo eggs in the production workshop.The selection of an appropriate disinfectant should be based on specific application scenarios and requirements.

To achieve efficient and rapid detection of duck astrovirus type 2(DAstV-2),RPA prim-ers and crRNA were designed and synthesized based on the conserved sequence of the ORF2 gene of DAstV-2.A detection method for DAstV-2 was constructed,integrating RPA nucleic acid ampli-fication,LwCas13a cleavage,and colloidal gold lateral flow dipstick visualization.The specificity,sensitivity,and concordance of this detection method were evaluated.The experimental results showed that the detection limit for the DAstV-2 recombinant plasmid standard was 1.2×101 cop-ies/μL,which is superior to the conventional RT-PCR method.The method can specifically detect DAstV-2 pathogenic nucleic acids without cross-reactivity with DAstV-1,DAstV-3,DAstV-4,duck plague virus(DEV),and duck tembusu virus(DTMUV).When testing liver tissue samples from ducks suspected of being infected with DAstV-2,the results obtained using this method were com-pletely consistent with those from real-time quantitative PCR,with a 100％concordance rate.How-ever,this method is simpler and faster to perform.The research indicates that the established RPA-CRISPR/Cas13a-LFD detection system has high sensitivity,strong specificity,and high accuracy,capable of completing rapid visual detection of DAstV-2 nucleic acids within 1 h at a constant tem-perature of 37 ℃,providing a new technical platform for the rapid diagnosis of DAstV-2.

Objective:To investigate affinity of monoclonal antibody(MAb)of different cardiac troponinⅠ(cTnⅠ)epi-topes to cTnⅠ-troponin C(cTnⅠ-TnC)antigen,and to screen novel cTnⅠ-specific antibodies for clinical detection kit development.Methods:Based on technology platform of microarray chemiluminescence immunoassay,cTnⅠ-TnC antigen spot sample hard matrix chip was used,mouse derived cTnⅠ MAb was used as antibody to be detected,HRP-sheep-anti-mouse-IgG was used as antibody to detect,and experimental conditions were optimized for anti-origin solution,antigen spot sample concentration and antibody to be detected concentration.Affinity of cTnⅠ MAb with different antigenic epitopes to cTnⅠ-TnC was detected by competition method.Graded concentrations of cTnⅠ-TnC were added to experimental group,and diluents of equal volume were added to control group.Reaction signals of different cTnⅠ MAb were detected and affinity constants were calculated.Results:Antigen spot sample liquid was P105,antigen spot sample concentration was 0.1 mg/ml,and antibody concentration to be detected was 80 ng/ml were optimal experi-mental conditions.20c6cc and 7B9cc MAb containing TnC antibodies had the best affinity,and 20c6cc MAb reached 3.18×106 L/mol.cTnⅠ MAb located at C-end of peptide chain in a.a.r 130～145,169～178 and 190～196 regions showed good affinity,among which MAb 625(a.a.r 169～178)was 6.37×105 L/mol,showing the strongest affinity among cTnⅠ MAb.Conclusion:MAb has good affinity with cTnⅠ-TnC antigen at C-end of cTnⅠ peptide chain a.a.r 130～145,169～178,190～196 and TnC region.Designing a new type of cTnⅠ complement antibody targeting this region can be a new way to solve negative interference of autoantibody and peptide proteolysis.

Objective:To explore the anal function and postoperative complications of 2-stage Turnbull-Cutait pull-through coloanal anastomosis (TCA) for low rectal cancer.Methods:Patients undergoing radical rectal cancer resection from Feb 2023 to Nov 2024 in the First Hospital of Jilin University were divided into the TCA surgery group and the low anterior resection combined with prophylactic stoma (LAR) surgery group.Results:Among the 102 patients, there were 50 cases in the TCA group and 52 cases in the LAR group. In the single-arm analysis of the TCA group, the overall complication rate was 44%. The incidence rates of severe LARS at 1 month, 3 months, and 6 months after surgery were 97%, 77%, and 64% respectively. There was no significant difference in the complication rate within 30 days after surgery between the two groups,(44% vs. 38%, χ2=0.135, P>0.05). There was no significant difference in the incidence rate of severe LARS between the TCA group and the LAR group (77% vs. 69%, χ2=0.202, P>0.05), and there was not significant difference in the incidence rate of severe LARS between the two groups at the 6th month after surgery,(64% vs. 48%, χ2=1.132, P>0.05). Conclusion:In patients who underwent TCA surgery, the LARS symptoms gradually decreased over time. Compared with patients undergoing low anterior resection and stoma reversal, there were no significant differences in complications within 30 days after surgery and LARS symptoms within half a year.