Objective To discuss the application of therapeutic hemapheresis in clinical therapy and to observe the ad-verse reactions.Methods 126 times of therapeutic hemapheresis were performed in 121 patients by Nangel blood cell sepa-rator.Consumables and apheresis procedures were selected according to different treatment needs of patients.The clinical therapeutic efficacy laboratory indicators and adverse reactions during the treatment process were analyzed,and correspond-ing measures were taken.Results Among 100 patients who underwent platelet rich plasma(PRP)treatment,the success rate of collection was 98％.The average PRP platelet level achieved(981.22±3.55)×109/L,and the average concentration multiple of PRP achieved(4.01±0.52)times.All patients obtain symptomatic relief after treatment with PRP.The level of hemoglobin and red blood cell count of 6 polycythemia vera patients and the level of platelet of 10 essential thrombocythemia patients all decreased after therapeutic hemapheresis combined with hydroxycarbamide,with significant difference(P＜0.05).Five patients with preoperative autologous blood donation have been through the surgery successfully.Out of the 121 patients,10(8.26％)experienced mild adverse reaction(8.26％),15(12.40％)showed bruising and bleeding at puncture site,and 1(0.83％)had pseudoaneurysm.Conclusion Therapeutic hemapheresis has the advantages of good curative effect,high success rate and low incidence of adverse reactions,and can be applied in PRP collection,removal of blood pathological components and preoperative autologous blood donation.

Objective:To explore the expression level and clinicopathological characteristics of 14-3-3θ protein in distal cholangiocarcinoma tissues, and further analyze the long-term prognosis of patients with different expression levels.Methods:A retrospective cohort study was conducted to collect and analyze the clinical data of 135 patients with distal cholangiocarcinoma who underwent surgical resection at the Department of Hepatobiliary Surgery, Beijing Chaoyang Hospital Affiliated to Capital Medical University from January 2015 to December 2021, including 86 males and 49 females.(65.1±10.1) years old. Immunohistochemistry was used to detect the expression level of 14-3-3θ protein in cholangiocarcinoma tissue. The postoperative pathological sections of patients were evaluated based on the immune response score(IRS). The optimal cutoff value was determined through the receiver operating characteristic(ROC) curve was 3.5. Currently, the area under the curve was 0.741, the sensitivity was 73.5%, and the specificity was 71.3%. The patients were divided into two groups: IRS<4 was the 14-3-3θ low expression group( n=81), IRS≥4 was the 14-3-3θ high expression group( n=54). After surgery, the patient′s survival status was followed up through a combination of outpatient review and telephone follow-up. Normally distributed measurement data were expressed as mean ± standard deviation( ± s), and comparisons between groups were made using the t test; non-normally distributed measurement data were expressed as M( Q1, Q3), and comparisons between groups were made using the rank sum test. The chi-square test was used to compare enumeration data between groups. Results:The preoperative CA19-9 and lymph node metastasis in the 14-3-3θ low expression group were 44.3(20.8, 132.2) U/mL and 28 cases respectively. The preoperative CA19-9 and lymph node metastasis in the 14-3-3θ high expression group were 82.3(43.4, 396.9) U/mL and 32 cases respectively. The difference between the two groups was statistically significant( P<0.05). All patients had regular postoperative telephone calls or return to the hospital for follow-up. The median postoperative survival time of the 14-3-3θ low-expression group was 36 months. The 1-, 2-, and 3-year survival rates after surgery were 88.9%, 66.5%, and 66.5%, respectively. 49.4%, the median survival time after surgery in the 14-3-3θ high expression group was 13 months, and the 1, 2, and 3-year survival rates after surgery were 53.7%, 23.3%, and 13.3% respectively. The difference between the two groups was statistically significant. significance( P<0.01). Cox proportional hazard model performed multivariate analysis, CA19-9>37 U/mL ( RR=1.970, 95% CI: 1.186-3.272, P=0.009), lymph node metastasis( RR=1.681, 95% CI: 1.035-2.729, P=0.036) and 14-3-3θ staining intensity≥4 ( RR=2.438, 95% CI: 1.546-3.845, P<0.001) have worse long-term prognosis. Conclusions:The expression level of 14-3-3θ protein is related to CA19-9 and lymph node metastasis in distal cholangiocarcinoma. A high expression of 14-3-3θ protein indicates poor long-term prognosis in patients with extrahepatic cholangiocarcinoma.

Objective : Cas9-RNP biomimetic nanoparticles cas9-RNP@ MMs were prepared by encapsulating the Cas9 Ribonucleoprotein complex (RNP) using mouse macrophage membranes，with the aim of utilizing this biomimetic nanoparticle to deliver the Cas9-RNP complex for gene editing ，and further study the endocytosis of Cas9- RNP@ MMs and its gene editing effect in mouse macrophage RAW264. 7 in vitro ，providing evidence for the development oflow-toxicity biomimetic nanoparticle carriers that inhibit NLRP3 therapeutic targets． Methods : The purified mouse macrophage membrane was mixed with the prepared cas9-RNP mixture，and after ultrasound，the CAS9- RNP@ MMS was obtained by liposome extrusion instrument ; The particle size of Cas9-RNP@ MMswas measured by nanoparticle tracking analysis，and the particle morphology of Cas9-RNP@ MMs was observed under transmission electron microscope．Laser confocal Fluorescence microscope imaging was used to analyze the endocytosis Cas9-RNP @ MMs．The Biocompatibility of Cas9-RNP@ MMs was measured by MTT assay．The expression of NLRP3 was detected by qPCR and Western blot to verify the knockdown effect of Cas9-RNP@ MMs on NLRP3 gene. Results: The average particle diameter of Cas9-RNP@ MMs prepared from macrophages was about 216 nm．Under laser confocal fluorescence microscope，the Cas9-RNP@ MMs could be successfully endocysed by Raw246. 7 cell．MTT assay indicated that the Cas9-RNP@ MMs-treated mouse macrophage RAW246. 7 had good biocompatibility．qPCR and Western blot showed that two NLRP3-specific guide RNA were mediated by Cas9-RNP@ MMs，with good effect of knockdown NLRP3 gene expression． Conclusion Nano-scale vesicles Cas9-RNP@ MMs loaded with Cas9-RNP complexes were successfully prepared by biomimetic nanoparticles． Cas9-RNP@ MMs have good biocompatibility and can be efficiently endocytosed by RAW246. 7 cells．Cas9-RNP@ MMs containing NLRP3-specific sgRNA can specifically knock down NLRP3 gene expression.

Borderline resectable pancreatic cancer is a special subtype between resectable and unresectable pancreatic cancer. Although the tumor is technically suitable for resection, there is increased risk of positive margin after surgery. At present, there is no optimal diagnostical criteria and treatment options for borderline resectable pancreatic cancer. With the popularization of the concept of multidisciplinary diagnosis and treatment, neoadjuvant therapy has been widely used in borderline resectable pancreatic cancer, and received good outcomes in some centers. However, for patients with borderline resectable pancreatic cancer who are not sensitive to radiotherapy and chemotherapy, long time of neoadjuvant therapy may delay the best time for surgery. This article summarized the definition, classification criteria and the latest diagnosis and treatment progress of borderline resectable pancreatic cancer, and discussed the comprehensive treatment mode suitable for this kind of patients combined with the clinical experience of our center.

Objective:To analyse the correlation between the lymph nodes, the number of positive lymph nodes, the positive rate of lymph nodes and the long-term prognosis of patients with distal cholangiocarcinoma.Methods:The clinical data and follow-up results of 104 patients with distal cholangiocarcinoma admitted to Beijing Chaoyang Hospital Affiliated to Capital Medical University from January 2015 to December 2021 were retrospectively analyzed using a retrospective cohort study. All patients underwent pancreaticoduodenectomy. According to the nature of the patients′ lymph nodes, the number of positive lymph nodes, and the positive rate of lymph nodes, the X-tile software was used to analyze data respectively, and the critical value of SUVmax were determined, and the patients were divided into low-risk group and high-risk group by critical values. Based on this critical value, the patients were divided into low-risk groups and high-risk groups. There were 37 patients in the lymph node positive group and 67 patients in the negative group. In the lymph node positive group, according to the number of positive lymph nodes (the cut-off value=2), they were divided into the low-risk group( n=14), the high-risk group( n=23); grouped according to the lymph node positive rate (the cut off=0.13), and divided into the low-risk group( n=15), and the high-risk group( n=22). After grouping according to different lymph node parameters, the preoperative general data such as age, gender, laboratory examination, etc., the correlation between perioperative complications and long-term prognosis were analyzed. The measurement data conforming to the normal distribution was expressed as mean ± standard deviation( ± s), and the t test was used for comparison between groups; the measurement data that was not normally distributed was expressed as M ( Q1, Q3), and the comparison between groups was performed by the rank sum test. The enumeration data were expressed by the number of cases, and the comparison between groups was performed using the chi-square test. If the total number of cases was less than 50, and Fisher′s exact test was used. The optimal cut-off values for different lymph node parameters were determined using the x-tile 3.6.1 software. Survival curves were calculated and drawn using the Kaplan-Meier method, and the survival rates were compared using the Log-rank test. Results:All patients successfully underwent the operation, 5 patients(4.8%) died during the perioperative period. The median postoperative survival time of lymph node-positive patients and lymph node-negative patients was 17 months and 36 months, respectively, and the overall survival rates at 1, 3, and 5 years were 64.9%, 23.9%, 23.9%, and 81.5%, 49.8%, 41.7%( P=0.003). Among the patients grouped according to the number of positive lymph nodes, the median postoperative survival time of patients in the low-risk group and the high-risk group was 21 months and 17 months, and the 1-, 3-, and 5-year overall survival rates of the two groups were were 77.1%, 42.9%, 42.9% and 58.2%, 13.8%, 13.8%( P=0.284), respectively. Among the patients grouped according to the positive rate of lymph nodes, the median postoperative survival time of the patients in the low-risk group and the high-risk group was 30 months and 15 months, and the 1-, 3-, and 5-year overall survival rates of the two groups were 85.6%, 42.7%, 42.7% and 51.3%, 10.3%, 0.3%( P=0.020), respectively. Conclusions:Radical pancreaticoduodenectomy is the standard procedure for patients with distal cholangiocarcinoma. The prognosis of patients with positive lymph nodes is worse. For patients with positive lymph nodes, the positive rate of lymph nodes is a better predictor of the long-term prognosis compared with the number of positive lymph nodes.

Objective:This research aimed to study the relationship between preoperative CA19-9/GGT ratio and postoperative long-term survival in patients with distal cholangiocarcinoma.Methods:The clinical data of 121 patients with distal cholangiocarcinoma who underwent radical pancreaticoduodenectomy (PD) at the Department of Hepatobiliary Surgery, Beijing Chaoyang Hospital Affiliated to Capital Medical University from January 2013 to December 2019 were retrospectively analyzed. The ROC curve was drawn based on the preoperative CA19-9/GGT ratio and postoperative 1-year survival. Using the best cut-off value of CA19-9/GGT ratio, the 121 patients were divided into two groups: the low ratio group (CA19-9/GGT≤0.12, n=53) and the high ratio group (CA19-9/GGT>0.12, n=68). The clinical data of the two groups were compared, and the risk factors of long-term survival were analyzed. Results:There were 72 male and 49 female patients, aged (64.9±9.2) years. When compared with the high ratio group, the low ratio group had significantly less requirement for preoperative jaundice reduction, lower CA19-9, higher GGT, better tumor differentiation, and more patients without lymph node metastasis (all P<0.05). The median follow-up time was 26 months. The 1-, 3- and 5-year survival rates of the low vs. high ratio groups were 89.4% vs. 64.7%, 64.4% vs. 14.1%, 48.7% vs. 14.1%, respectively (all P<0.001). Multivariate analysis showed that CA19-9/GGT ratio>0.12 ( RR=2.802, 95% CI: 1.494-5.256), poor differentiation ( RR=1.855, 95% CI: 1.106-3.111) and lymph node metastasis ( RR=1.891, 95% CI: 1.129-3.169) were independent risk factors for long-term survival ( P<0.05). Conclusion:The ratio of CA19-9/GGT could be used as an index to predict long-term survival of patients with distal cholangiocarcinoma after PD. The smaller the ratio, the better was the long-term prognosis.

Organ transplantation is the first effective treatment for end-stage organ failure. However, early acute rejection after organ transplantation is often an important factor leading to transplantation failure. In order to reduce the incidence of rejection after organ transplantation, it is often necessary to use potent immunosuppressants to induce the immune system of recipients before or during transplantation. As a common method of covering immunosuppressive therapy in the early stage of organ transplantation, antibody immunoinducers can significantly reduce the occurrence of early acute rejection after organ transplantation, and delay or reduce the application of calcineurin inhibitor drugs, which is conducive to protecting renal function, promoting graft function recovery and long-term survival of recipients. This article reviews the application of antibody inducers in solid organ transplantation.

Objective:To investigate the infiltration of B10 cells in the anastomotic tissue after choledochojejunostomy by establishing a rat model of choledochojejunostomy.Methods:24 male SPF SD rats aged 6 weeks and weighing 180-200 g were randomly divided into control group, 1-week group, 2-week group and 4-week group, with 6 samples in each group. The rats were killed at different time after operation to obtain the anastomotic tissue and peripheral blood. The changes of anastomotic diameter were measured. The changes of inflammatory factors in peripheral blood were determined by ELISA. Flow cytometry was used to detect the proportion of B10 cells in peripheral blood and tissues around the anastomotic site. The formation of anastomotic scar was evaluated by histopathological section. IL-10 and TGF-β1 gene expression were detected by qPCR. The measurement data with normal distribution were expressed as ( Mean± SD). Analysis of variance was used to compare the measurement data of multiple groups. T test was used to compare the measurement data between the two groups. Results:The diameter of anastomotic stoma gradually narrowed with time after choledochojejunostomy, which was(2.7± 0.3) mm at the 4st week after operation; However, liver function and inflammation index reached the peak at the 2st week after operation, and then gradually returned to normal level; The proportion of B10 cells in peripheral blood did not change significantly in each period after operation, but the infiltration of B10 cells in anastomotic tissue increased significantly, which was significantly higher than that in the control group at the first week after operation [(16.6±4.0)% vs (1.1±0.3)%, P<0.05], and it was still higher than that of the control group at the 4st week after operation [(7.5±1.3)% vs (1.1±0.3)%, P<0.05]; The pathological staining of anastomotic tissue showed that with the passage of time, the infiltration of inflammatory cells in the bile duct wall increased, the proliferation of collagen fibers, the thickening of bile duct wall, and then led to scar formation; Expression of IL-10 and TGF-1β in anastomotic tissue after operation. At the 4st week after operation, IL-10 gene expression was still higher than that of the control group [(1.4±0.6) vs (0.5±0.2), P<0.05], the gene expression of TGF-1β was increased continuously and was higher than that of the control group [(3.9±0.9) vs (0.3±0.2), P<0.05]. Conclusion:The content of B10 cells in the anastomotic tissue of rats after choledochojejunostomy is significantly increased, and the expression of IL-10 gene is high, which may play a role in regulating local scar formation.

Objective:To analyze the risk factors of lymph node metastasis of distal cholangiocarcinoma.Methods:We retrospectively analyzed the clinical data of 123 patients of distal cholangiocarcinoma in Beijing Chaoyang Hospital between Jan 2011 and Dec 2019. All patients underwent pancreatoduodenectomy. The prognosis was evaluated according to the follow-up results. The survival rate was compared by log-rank test. Logistic regression was used to analyze the risk factors for lymph node metastasis.Results:One hundred and twenty-three patients successfully underwent the operations, 6 patients died during the perioperative time. The overall 1-year, 3-year and 5-year survival rates were 75.2%, 41.9% and 31.5%. The median survival time was 44.7 months. Fifty-one patients had lymph node metastasis, and the rate of lymph node metastasis was 41.5%. The median survival time was 55.5 months and 27.5 months for patients without and with lymph node metastasis, respectively. Correspondingly, the survival rates of 1-year, 3-year, 5-year were 83.0%, 50.7%, 42.5% vs. 63.5%, 19.0%, 19.0% ( P=0.000). Multivariate analysis showed that preoperative CA19-9 ( RR=7.064, 95% CI: 2.489-20.051) and portal venous system invasion ( RR=4.610, 95% CI: 1.252-16.972) were independent risk factors for lymph node metastasis. Conclusions:Lymph node metastasis is an important factor affecting the long-term survival of patients with distal cholangiocarcinoma. Preoperative CA19-9 level and portal venous system invasion are independent risk factors for lymph node metastasis.

Objective:To evaluate the prognosis and related risk factors of distal cholangiocarcinoma after surgical treatment.Methods:The clinical data of 123 patients of distal cholangiocarcinoma in Beijing Chaoyang Hospital between January 2011 and December 2019 were retrospectively analyzed, which including 72 males and 51 females, the average age was (64.9±9.2) years (range from 29 to 84 years). All patients underwent pancreatoduodenectomy. The observation measures contains: (1) Perioperative outcomes; (2) Follow-up outcomes; (3) Risk factors for long-term survival of distal cholangiocarcinoma. Follow-up was carried out to understand the long-term survival of patients, and follow-up method contains the outpatient reexamination and telephone. The deadline of follow-up date was March 2020. The normal distribution data were expressed by ( Mean± SD), and the non-normal distribution data were expressed by M ( P25, P75). Count data were expressed by cases and percentage. Kaplan-Meier method was used to calculate and draw the survival curve. Log-rank test was used to compare the survival rate. Cox proportional risk model was used in multivariate factor analysis. Results:(1) Perioperative outcomes: In our research, all of 123 patients were successfully completed the operations, and 6 patients dead during the perioperative. (2) Follow-up outcomes: The incidence of postoperative complications was 27.6%(34/123). One hundred and twenty-one patients were followed up, the follow-up rate was 98.4%, and the median follow-up time was 41.0 months.The overall 1-year, 2-year, 3-year and 5-year survival rates were 71.8%, 50.5%, 35.5% and 30.2%. And the median survival time was 42.7 months. (3) Risk factors for long-term survival of distal cholangiocarcinoma: Multivariate analysis showed that preoperative CA19-9 ( RR=1.470, 95% CI: 1.028-2.101), portal venous system invasion ( RR=2.020, 95% CI: 1.012-4.035) and tumor differentiation ( RR=1.735, 95% CI: 1.195-2.520) were independent risk factors for the prognosis. Conclusions:Radical pancreatoduodenectomy is the best treatment for distal cholangiocarcinoma. Preoperative CA19-9 level, portal venous system invasion and tumor differentiation are independent risk factors for the prognosis.