In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.

In this study,the carboxylated magnetic beads were coupled with bivalent nanobodies a-gainst porcine epidemic diarrhea virus(PEDV)M protein to construct immunomagnetic beads(IM-NBs-Ⅱ),The capture and enrichment function of IMNBs-Ⅱ was verified by using PEDV propaga-ted in Vero cells.A one-step RT-qPCR detection method for PEDV was established by combining the characteristics of IMNBs-Ⅱ with the detection advantages of reverse transcription fluorescence quantitative polymerase chain reaction(RT-qPCR).Specific analysis found that this method has no cross reactivity with swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,porcine circovirus,indicating that it has good specificity.Sensitivity analysis re-vealed that the detection sensitivity of the RT-qPCR based on IMNBs-Ⅱ was increased 10 times compared to traditional RT-qPCR methods.Detection of the clinical samples confirm that the RT qPCR method based on IMNBs-Ⅱ is suitable for rapid and accurate detection of clinical feces and tissue samples.The method established in this study effectively avoids contamination issues during nucleic acid extraction,simplifies experimental procedures,and saves detection time,which pro-vides a method for efficient detection of PEDV.

Objective:To explore the safety key points of total pancreaticoduodenectomy in the era of vascular resection technology and the important factors affecting rapid postoperative recovery.Methods:The clinical data of 52 patients with pancreatic cancer who underwent total pancreaticoduodenectomy in Beijing Chaoyang Hospital Affiliated to Capital Medical University from November 2014 to September 2022 were retrospectively analyzed, including 34 males and 18 females, aged (62±9). The intraoperative situation, incidence of postoperative complication, postoperative blood glucose control and postoperative survival rate were analyzed.Results:All operations of the 52 patients were successfully completed, including 48 patients underwent total pancreaticoduodenectomy combined with portal vein resection and allograft vascular grafts via artery approach. The portal vein occlusion time was (20±5) min. The incidence of postoperative complications was 28.8% (15/52), including 2 cases of abnormal gastric empty, 2 cases of diarrhea, 2 cases of chylous fistula, 4 cases of abdominal infection, 1 case of gastrointestinal fistula, 3 cases of gastrointestinal bleeding, and 1 case of pulmonary infection. Subcutaneous short-acting insulin injection was used to control blood glucose in the early stage after surgery, short-acting insulin combined with long-acting insulin was used for subcutaneous injection before sleep for diet recovery. All patients did not experience uncontrolled hyperglycemia. The median survival time of 52 patients was 13 months, and the longest follow-up time was 38 months. There were 37 patients died of tumor recurrence, 4 patients died of cardiovascular and cerebrovascular accidents, and 1 patient died of pulmonary infection in the 42 died patients.Conclusions:Total pancreaticoduodenectomy via artery approach can improve the R 0 resection rate in pancreatic cancer patients with vascular invasion, the rate of postoperative complication and mortality has no significant increase. The postoperative blood sugar control is satisfactory and the quality of life is guaranteed.

Purpose/Significance To study the extraction method of adverse drug event(ADE)data from medical texts,and to pro-vide support for clinical drug risk management and scientific decision-making.Method/Process Based on pre-trained model,by com-bining the correlation between the two subtasks of entity recognition and relation extraction,a entity relation joint extraction method for ADE monitoring is designed.Result/Conclusion Experiments on the published ADE extraction dataset show that the proposed method is superior to existing methods and can effectively extract ADE information and its relation from medical texts,providing a powerful means for the discovery and monitoring of ADE.

Objective:To evaluate superior mesenteric artery preferential approach in the borderline resectable pancreatic head cancer.Methods:The clinical and follow-up data of 90 patients with borderline resectable pancreatic head cancer who underwent radical pancreatoduodenectomy at Beijing Chaoyang Hospital,Capital Medical University from Jan 2015 to Dec 2021 were analyzed.Results:After exploring the superior mesenteric artery in the lower colon area to confirm the vascular invasion meet the resection criteria, the blood supply is cut off first, then the tumors were resected en bloc, with the invaded vessels resected and reconstructed or replaced. All 90 patients successfully completed the operation without perioperative death. Pathology established pancreatic ductal adenocarcinoma. The 1-year, 2-year, and 3-year disease-free survival rates of patients in the arterial priority approach group were 68.2%, 60.4%, and 54.3%, while the 1-year, 2-year, and 3-year disease-free survival rates of patients by conventional approach were 58.4%, 26.4%, and 11.7% ( P=0.001). Conclusion:The superior mesenteric artery preferential approach in the inferior colon region can prolong the survival time of patients after surgery, and reduce the recurrence.

Rejection and adverse reactions caused by long-term use of immunosuppressants severely affect the survival rate and quality of life of organ transplant recipients. Immune tolerance induction plays a key role in improving the survival rate and quality of life of organ transplant recipients. In recent years, tremendous progress has been achieved in adoptive re-transfusion of regulatory cells. In this article, research progress in regulatory T cell (Treg), myeloid-derived suppressor cell (MDSC) and regulatory B cell (Breg) in animal experiment and clinical application was reviewed, and the main clinical problems of adoptive re-transfusion of regulatory cells, the application of chimeric antigen receptor Treg and the concept of cell therapy in immune evaluation were summarized, aiming to deepen the understanding of regulatory cell therapy, promote the application of regulatory cells in immune tolerance of organ transplantation, and improve clinical efficacy of organ transplantation and the quality of life of recipients.

Objective To investigate the guiding role of quantitative evaluation system of immune status in the individualized management of immunosuppressants for the recipients after liver transplantation. Methods Clinical data of 239 liver transplant recipients were retrospectively analyzed. MingDao Immune Cell Analysis (MICA) was established. All recipients were divided into two groups according to the adjustment regimens of immunosuppressants. The immunosuppressant regimen was adjusted according to MingDao Immune System Score (MISS) in the MISS group (n=84), and the medication plan was empirically adjusted during the same period in the control group (n=155). According to the time of postoperative detection (t), the recipients in the MISS group were divided into subgroup A (t ≤ 28 d, n=78), subgroup B (28 d < t ≤ 6 months, n=68), subgroup C (6 months < t ≤ 12 months, n=18), subgroup D (12 months < t ≤ 24 months, n=18) and subgroup E (t > 24 months, n=19). In the MISS group, postoperative MISS scores of recipients in subgroups A-E were analyzed. The incidence of acute rejection and opportunistic infection and the overall survival rate were statistically compared between the MISS and control groups. Results The MISS scores in subgroups A-E were -7.0 (-13.2, -2.0), -2.0 (-5.8, 1.8), -0.5 (-7.3, 2.8), -2.0 (-4.5, 3.3) and -3.0 (-6.0, 1.0), respectively. The immune status of the recipients was gradually improved over postoperative time, and the difference between two groups was statistically significant (P < 0.05). In the MISS group, 15% (13/84) of the recipients developed acute rejection, and 27% (42/155) in the control group, and the difference was statistically significant (P < 0.05). In the MISS group, the MISS score of the recipients with acute rejection was 0 (-2.5, 3.5), and -5.0 (-12.0, -1.0) for their counterparts without acute rejection, and the difference was statistically significant (P < 0.05). In the MISS group, 2% (2/84) of the recipients presented with postoperative opportunistic infection, and 9% (14/155) in the control group, and the difference was statistically significant (P < 0.05). In the MISS group, the 1- and 3-year overall survival rates were 86.9% and 79.8%, and 83.2% and 76.8% in the control group, and no significant difference was observed between two groups (P > 0.05). Conclusions MICA and MISS score may reflect the immune status of liver transplant recipients, and guide the individualized management of administration of immunosuppressants after liver transplantation.

Objective To evaluate the changes and significance of lymphocyte subsets in the recipients with acute rejection after liver transplantation. Methods The recipients presenting with acute rejection after liver transplantation were assigned into the rejection group (n=17), and their counterparts with stable liver function were allocated into the control group (n=17) according to the ratio of 1∶1 by propensity score matching method. The incidence of acute rejection after liver transplantation was analyzed, and the concentration of tacrolimus in the recipients was compared between two groups. The absolute value and proportion of lymphocyte subsets in peripheral blood were compared between two groups. The diagnostic value of lymphocyte subsets for acute rejection after liver transplantation was assessed by the receiver operating characteristic (ROC) curve. The absolute value and proportion of lymphocyte subsets in the rejection group were compared before and after treatment. Results Among 17 recipients in the rejection group, 4 cases developed acute rejection within postoperative 28 d, and 13 cases had acute rejection within postoperative 29-180 d. No significant difference was noted in the tacrolimus concentration between two groups (P=0.295). Compared with the control group, the proportions of peripheral blood T cells, CD4⁺T cells, B cells and natural killer (NK) T cells were significantly increased in the rejection group (all P < 0.05). The elevated proportion of NKT cells in the early stage after liver transplantation was an independent risk factor for acute rejection following liver transplantation[odds ratio (OR) 1.774, 95% confidence interval (CI) 1.059-2.971, P=0.029]. ROC curve analysis showed that the area under curve (AUC) of CD4⁺T cells, B cells and NKT cells was 0.76, 0.73 and 0.77, respectively. The AUC of combined use of CD4⁺T cells, B cells and NKT cells was 0.89, with a cut-off value of 0.69, sensitivity of 0.706 and specificity of 0.941. After corresponding treatment, all recipients were gradually recovered, and liver functions were eventually restored to normal in the rejection group. After treatment, the proportion of T cells, CD4⁺T cells, CD8⁺T cells and NK cells was significantly decreased (all P < 0.05). Conclusions The elevated proportion of NKT cells indicates an increased risk of acute rejection after liver transplantation. Combined use of CD4⁺T cells, B cells and NKT cells may deliver early detection and diagnosis of acute rejection after liver transplantation.

Objective:To investigate the clinical efficacy of superior mesenteric artery priority approach in infracolic compartment in the surgical treatment of resectable pancreatic head cancer.Methods:The clinical data of 79 patients with resectable pancreatic head cancer who underwent radical pancreaticoduodenectomy (PD) in the Department of Hepatobiliary Surgery of Beijing Chaoyang Hospital Affiliated to Capital Medical University from January 2016 to December 2019 were analyzed retrospectively. According to the surgical approach, they were divided into arterial priority approach group (arterial priority group, 48 cases) and conventional approach group (conventional approach group, 31 cases). The arterial priority group was to first explore the superior mesenteric artery in the subcolone, then cut off the tumor blood supply after ascertaining that the tumor can be removed, and finally remove the tumor and anastomose it routinely. The conventional approach group underwent traditional radical PD. The operation time, intraoperative bleeding, intraoperative blood transfusion, lymph node metastasis, number of lymph node dissections, resection criteria, tumor length diameter, degree of tumor differentiation, occurrence of postoperative complications (pancreatic fistula, impaired gastric emptying, abdominal bleeding, biliary fistula and diarrhea), postoperative hospital stay and postoperative chemotherapy were observed. The patients were followed up by the way of outpatient and/or telephone. The follow-up time was up to March 2021. The survival rate was calculated by Kaplan Meier method.Results:Compared with the conventional approach group, the amount of intraoperative bleeding in the arterial priority group was significantly reduced (400 ml vs 600 ml, P=0.005), the intraoperative blood transfusion rate was significantly reduced (39.6% vs 64.5%, P=0.030), the number of lymph node dissections was significantly increased (19 vs 13, P=0.024), the R 0 resection rate was significantly increased (83.3% vs 58.1%, P=0.013), but the incidence of diarrhea was significantly increased (39.6% vs 16.1%, P=0.027). The differences were statistically significant. The median survival time of arterial priority group and conventional approach group was both 19 months. The overall survival rates of 1, 2 and 3 years were 72.0%, 39.5%, 28.5% and 64.1%, 33.7% and 15.4% respectively. The median tumor free survival time was 15 months and 12 months. There was no significant difference. However, the 1, 2 and 3-year tumor free survival rate of patients in arterial priority group was significantly improved (61.2%, 39.5% and 25.9% vs 46.0%, 21.3% and 7.1%, P=0.042) , and the difference was statistically significant. Conclusions:Superior mesenteric artery priority approach in infracolic compartment can improve R 0 resection rate, prolong postoperative survival time, reduce recurrence and improve prognosis.

Objective:To explore the risk factors of Pneumocystis carinii pneumonia (PCP) after orthotopic liver transplantation (OLT), and optimize the treatment strategy. Methods:From May 2015 to March 2019, patients undergoing OLT and suffering from postoperative PCP were selected into PCP group ( n=8). Using the propensity score matching method, controls without postoperative PCP were selected from concurrent OLT patients at a ratio of 1: 4 ( n=32). Clinical data were collected and counted for analyzing the risk factors of PCP post-OLT. Results:During this period, 385 cases of OLT were performed. The incidence of PCP was 2.1% (8/385). PCP group were all males with an average age of (52.63±12.99)(27-69) years. PCP has an average onset time of (19.88±13.22)(9-50) weeks post-OLT. There were benign liver disease ( n=2) and malignant liver tumor ( n=6). All operative approaches were modified camel OLT. Univariate analysis revealed significant differences in rejection, peripheral blood lymphocyte count and percentage of peripheral blood lymphocyte after OLT ( P<0.05) and no significant differences existed in other factors ( P>0.05). Logistic regression analysis indicated that a lower count of peripheral blood lymphocyte post-OLT was an independent risk factor for postoperative PCP. Conclusions:A lower count of peripheral blood lymphocyte post-OLT elevates the risk of PCP. For high-risk patients, prophylaxis with TMP-SMX (trimethoprim-sulfamethoxazole) may effectively lower the incidence of PCP post-OLT.