【Abstract】Objective This study was conducted to build experimental model of orthotopic liver transplantation in rat (ROLT) with the character of acute rejection; and to study the effect of cytotoxic T lymphocyte antigen 4 immunoglobulin G (CTLA4Ig) on prevention of rejection and the induction of immune tolerance of ROLT. Methods Build model of Wistar→SD ROLT(performed by the two cuff method) with character of acute rejection.Recipients were injected with CTLA4Ig 75 μg per ROLT into abdominal cavity after 2 days of operation. Contrast was made with no treatment group, the clinical characters, the liver function, the transplantated liver pathologic character and the concentrations of TNFα in serum were observed and measured on postoperative day 7. In treatment group, all above observation were done on postoperative month 4. Above all, determination of the effect of CTLA4Ig on preventing acute rejection and inducing tolerance in ROLT was observed.Results ①Recipients (no treatment group) died one by one within 6th～14th days; pathologic character of rejection in transplantation liver could be found; ② In treatment group, on postoperative day 7 and month 4, no clinical rejection character and no pathologic character of rejection in transplantation liver were found and serum concentration of cytokins related to TNFα found lower than that of contrast group(P＜0.05), and serum concentration of ALT、AST、TBIL、DBIL found lower too(P＜0.05); But serum concentration of TP and Alb was found higher than that of contrast group(P＜0.05). Conclusion ① CTLA4Ig treatment alone inhibits the rejection responce in ROLT. ② CTLA4Ig treatment can Prevent rejection and induce immune tolerance in ROLT model with characters of acute rejection; the serum concentration of cytokins related to TNFα can probably be used as evaluation standard of rejection in ROLT rejection.
ObjectiveTo study the contents of CD44 that shared exon variant 5 (CD44v5) in peripheral blood lymphocytes (PBL) of patients with gastric carcinoma and the expression of CD44v5 in tumor tissue and their clinical significance. MethodsThe contents of CD44v5 were determined by FlowCytometry in PBL of 31 patients with gastric carcinoma before surgery and 10 normal controls. Tissue expression of CD44v5 in 33 patients with gastric carcinoma was investigated by immunohistochemistry. ResultsThe contents of CD44v5 were significantly higher in PBL of patients with gastric carcinoma before surgery than those of controls (P＜0.01). Nodepositive gastric cancer patients showed significantly elevated contents of CD44v5 in PBL in comparison with nodenegative gastric cancer (P＜0.01）. Significant correlations were noted between the contents of CD44v5 in PBL of patients with gastric carcinoma before surgery and tumor size, depth of invasion, lymph node metastasis and different the Vnion International Centre Le Cancer (VICC) stages of tumor (P＜0.05). The expression of CD44v5 could be detected in 69.7% of tumor tissue,but was not detected in adjacent normal gastric mucosa. Significant correlations were noted between CD44v5 expression and depth of invasion,and lymph node metastasis.The presence of CD44v5 protein was correlated with the lymph node involvement rate. Conclusion CD44v5 in PBL or tumor tissue may be useful as a metastatic marker. It may be of important clinical value in the diagnosis of metastasis and judgement of development for the patients with gastric cancer.
Effect of radical operation on expression of interleukin-2(IL-2)mRNA and production of IL-2 were markedly reduced preoperatively and four weeks after operation,expression of IL-2 mRNA significantly enhanced,but it was still lower than that in the normal group.Production of IL-2 nearly reached normal level,When PBL was activated by phytohemagglutinin(PHA),expresseion of IL-2 mRNA and production of IL-2 were much higher than that in non-activated PBL.These results suggested that expression of IL-2 were much higher than that in non-activated PBL.These results suggested that expression of IL-2 mRNA and production of IL-2 are dificient in gastric cancer patients,and radical surgery will help them to recover and they can also be improved through activation with PHA.
ObjectiveTo investigate the predictive value of preoperative D-dimer, CA19-9, neutrophil-to-lymphocyte ratio (NLR) in the diagnosis of pancreatic cancer. MethodsThe clinicopathologic data of patients who underwent pancreaticoduodenectomy for pancreatic occupying lesions in the First and Fifth Affiliated Hospitals of Xinjiang Medical University from January 2016 to December 2019 were analyzed retrospectively. According to the pathological characteristics, the patients were divided into pancreatic cancer and pancreatic benign tumour. The gender, age, accompanying diseases, preoperative D-dimer, CA19-9, lymphocyte count, neutrophil count, NLR and so on were compared between them. The receiver operating characteristic (ROC) curves of preoperative D-dimer, CA19-9, and NLR for diagnosis of pancreatic cancer were draw. ResultsA total of 142 patients were collected, including 79 patients with pancreatic cancer and 63 patients with pancreatic benign tumour. Compared with the patients with pancreatic benign tumour, the age was older (P<0.05), the number of neutrophils was more (P<0.05), the number of lymphocytes was less (P<0.05), the preoperative levels of CA19-9 and D-dimer, and the preoperative NLR were higher (P<0.05) for the patients with pancreatic cancer. The multivariate analysis of logistic regression revealed that the preoperative higer CA19-9, D-dimer, and NLR increased the incidence of pancreatic cancer (P<0.05), then the optimal cut-off value of preoperative levels of CA19-9 and D-dimer, and preoperative NLR on diagnosis of pancreatic cancer was 108 U/mL, 306 μg/L and 3.2, respectively, the corresponding area under ROC curve was 0.900, 0.891, 0.768, respectively, and which of combination of preoperative CA19-9, D-dimer, and NLR was 0.931. For the pancreatic cancer patients with preoperative NLR, CA19-9, and D-dimer higher than the optimal cut-off value, the proportions of patients with pTNM stage Ⅲ and lymph node metastasis were higher than those for pancreatic cancer patients with below than optimal cut-off value (P<0.05). ConclusionFrom preliminary study results, preoperative CA19-9, D-dimer, and NLR values have certain values in diagnosis of pancreatic cancer, and diagnostic value of combined detection of 3 indexes is higher than single index.
ObjectiveTo compare the clinical recovery and immune response between laparoscopic-assisted and open D2 gastrectomy for advanced gastric cancer. MethodsThe clinical data of 53 patients with advanced gastric cancer from January 2012 to October 2013 were studied prospectively. According to random number table, patients were randomly divided into laparoscopic-assisted group(LA group, n=27) and open operation group(OO group, n=26). Operative time, blood loss, time to passage of flatus, time to resume soft diet, after bed time, postoperative hospital stay, and number of retrieved lymph nodes were compared respectively between the two groups. The changes in CD3, CD4+, CD8+, IgG, IgA, IgM, and CRP were examined respectively by using flow cytometry and immunoturbidimetric assays on the preoperative day 1, and on the postoperative day 1 and 7. ResultsThe operative time was longer significantly in LA group than that in OO group(P < 0.05). The mean blood loss, the first flatus time, after bed time, and postoperative hospital stay in the two groups were all different statistically(P < 0.05), and all were better in LA group. However, the mean number of retrieved lymph nodes and the time to resume soft diet were not significantly different in the two groups(P > 0.05). On the day 1 and 7 after operation, the CD3, CD4+, and CD8+ significantly decreased as compared with those preoperatively in two groups(P < 0.01, P < 0.05). On the day 1 after operation, the levels of IgG, IgA, and IgM significantly decreased as compared with those preoperatively in two groups(P < 0.05). Those immunoglobulin in LA group recovered to close to the level before surgery, but in OO group sustained lower level(P < 0.05). On the day 1 and 7 after operation, CRP level significantly increased as compared with those preoperatively in two groups(P < 0.01, P < 0.05). Those changes of above index were not significantly different between the LA group and OO group on the day 1 after operation(P > 0.05). All index recovered gradually in the two groups on the day 7 after operation and were better in LA group(P < 0.05, except IgA). ConclusionLaparoscopic radical gastrectomy for advanced gastric cancer resulted in a quicker clinical recovery and a lesser depression to the perioperative cellular and humoral immune function.
Objective To study effect of carcinoembryonic antigen (CEA) positive targeted lymphocytes on gastric cancer cells in vitro and in vivo. Methods The peripheral blood mononuclear cells (PBMCs) were isolated from the peripheral blood of healthy volunteers. The recombinant vector anti-CEA-scFv-CD3ζ-pcDNA3.0 was transfected into the PBMCs by lipofectamine 2000, by this means, the CEA special lymphocytes were obtained. Meanwhile, the PBMCs transfected with empty plasmid pcDNA3.0 were used as control (empty vector lymphocytes). The different lymphocytes and gastric cancer cells (CEA positive KATOⅢ gastric cancer cells and CEA negative BGC-823 gastric cancer cells) were co-cultured, then the ability to identify the gastric cancer cells and it’s effect on apoptosis of gastric cancer cells were observed at 24 h or 36 h later respectively. The CEA special lymphocytes and empty vector lymphocytes were injected by the tail vein of nude mice bearing gastric cancer cells, then it’s effect on the tumor was observed. Results ① The CEA special lymphocytes could strongly identify the KATOⅢ gastric cancer cells (identification rate was 72.3%), which could weakly identify the BGC-823 gastric cancer cells (identification rate was 7.8%). ② The apoptosis rate of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly higher than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (P=0.032), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (P=0.118). ③ The tumor volume of the co-culture of CEA special lymphocytes and KATOⅢ gastric cancer cells was significantly smaller than that of the co-culture of empty vector lymphocytes and KATOⅢ gastric cancer cells (F=5.010, P<0.01) or the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells (F=4.982, P<0.01), which had no significant difference between the co-culture of CEA special lymphocytes and BGC-823 gastric cancer cells and the co-culture of empty vector lymphocytes and BGC-823 gastric cancer cells (F=1.210, P>0.05). Conclusion CEA special lymphocytes can promote cell apoptosis and inhabit tumor reproduction of CEA positive gastric cancer cells in vitro and in vivo.
ObjectiveTo explore the combined application of neutrophil to lymphocyte ratio (NLR) and systemic immune inflammation index (SII) on the prognosis of hepatitis B-related hepatocellular carcinoma after resection.MethodsRetrospectively collected data of 180 patients with hepatitis B-related hepatocellular carcinoma who were hospitalized in the Department of Infectious Diseases and Hepatobiliary Surgery of the Affiliated Hospital of Southwest Medical University and received surgical treatment from January 2013 to December 2019, including general information, laboratory examination and abdominal CT or MRI results. NLR and SII values were measured at one week before operation, and their critical values of NLR and SII were determined by ROC curve analysis. Univariate and multivariate analysis were performed to determine the risk factors to predict the survival status of patients with hepatitis B-related hepatocellular carcinoma after hepatectomy.ResultsUnivariate analysis showed that AFP, platelets, TNM staging, portal vein tumor thrombus, tumor differentiation, NLR, SII, and NLR+SII combined score were significantly correlated with the prognosis of patients with hepatitis B-related hepatocellular carcinoma (P<0.05). Multivariate analysis showed that PLT [HR=1.791, 95%CI (1.124, 2.854), P=0.014], NLR [HR=4.289, 95%CI (2.571, 7.156), P<0.001], SII [HR=5.317, 95%CI (3.016, 9.374), P<0.001], and NLR+SII combined score [HR=7.901, 95%CI (4.124, 15.138), P<0.001] were independently correlated with the survival of patients with hepatitis B-related hepatocellular carcinoma.ConclusionsThe preoperative NLR+SII combined score can be used to evaluate the postoperative prognosis of patients with hepatitis B-related hepatocellular carcinoma. The higher the score, the lower the postoperative survival rate.
Objective To observe the therapeutic effect of mensenchymal stem cells (MSCs) for experimental autoimmune uveitis (EAU). Methods MSCs were obtained from Wistar rats and selected by plastic adherence. Lewis rats were divided into treatment group and control group, six rats in each group. EAU models were induced by immunization with an emulsion (0.2 ml) containing 30 mu;g interphotoreceptor retinoid-binding protein derived peptide R16 and complete Freundprime;s adjuvant. The clinical manifestations of two groups were observed. Nine to 11 day after modeling, 1 ml MSCs suspension, which contained 5times;106 MSCs, were injected into the rats in treatment group via tail vein, and the rats in control group were given equal volume of phosphate buffer solution. Fifteen day after modeling, the eyes were collected to test the proportion of interferon gamma;, interleukin-17 and Foxp3 positive cells by flow cytometry. The clinical scores were analyzed by mixed linear model and statistical analysis of variance of repeated measurement data. The results of flow cytometry were analyzed using independent-sample t test. Results Six days after immunization, mild dilatation and congestion of iris vascular was observed. Nine days after immunization, mild muddy anterior chamber, myosis and absent pupillary reaction to light were observed. Twelve days after immunization, muddy anterior chamber, occlusion of pupil and dimmed or disappeared red reflex were observed, and then inflammation was slowly reduced. From 11 to 15 days after immunization, the clinical score of treatment group was lower than that in control group, the difference was statistically significant (t=2.42, 2.21, 4.16, 5.24, 4.03; P＜0.05). The results of flow cytometry showed that MSCs treatment could decrease the proportion of CD4＋T cells, Th1 cells and Th17 cells, increase the proportion of Treg cells. Conclusion MSCs treatment can ameliorate EAU, up-regulate the expression of Treg cells and down-regulate the expression of CD4＋T cells, Th1 cells and Th17 cells.
Objective To explore the effect of cytotoxic T lymphocyte-associated antigen 4 (CTLA4-Ig) fusion protein on the function of orthotopic liver allograft. Methods Orthotopic liver allograft models of rhesus monkeys were established in this study. The survival time, liver function and morphologic changes of graft were observed, respectively. The levels of IL-2 and IL-10 were detected by ELISA. Apoptosis was monitored by TUNEL.Results The average survival of control group was 6.57 d, while the average survival of CTLA4-Ig group was 14.92 d, which was statistically prolonged (Plt;0.05). Serum ALT level was highly increased, and Alb level decreased obviously in control group. While the levels of ALT and Alb kept in normal in CTLA4-Ig group. After day 3-7, the expressions of IL-2 were highly expressed in control group, while the expressions of IL-10 in CTLA4-Ig group were higher than those of control group. The severity of rejection reaction after day 3 was weaker in CTLA4-Ig group than that of control group by histological assessment. The apoptosis index after day 3 in the liver cells was highly increased in control group as compared with the CTLA4-Ig group. Conclusions CTLA4-Ig fusion protein therapy can induce immunotolerance and prolong the survival of recipients. The increasing of cytokines IL-2 or the decreasing of cytokines IL-10 may be one of the laboratory indexes in monitoring immunotolerance of transplantation.
Objective To explore the predictive value of neutrophil-to-lymphocyte ratio (NLR) in peripheral blood for postoperative complications of elective endovascular repair for abdominal aortic aneurysm (AAA). Methods From August 2016 to November 2021, the clinical data of patients with AAA who received endovascular isolation repair for the first time in the Department of Vascular Surgery of Beijing Hospital were retrospectively analyzed, including the basic information of the patients, comorbid diseases, and the largest diameter of AAA, preoperative blood labotry test, postoperative complications, long-term survival rate and other indicators. The optimal NLR in peripheral blood was determined, and the differences in postoperative complications and long-term survival rates between the high NLR group and the low NLR group were analysed. Results A total of 120 patients with AAA underwent endovascular isolation for the first time were included in this study, including 105 males and 15 females. The age ranged from 52 to 94 years, with an average of (73.3 ± 8.26) years. The largest diameter of abdominal aortic aneurysm was 35 to 100 mm, with an average of (58.5 ± 12.48) mm. The best cut-off value of NLR for predicting postoperative complications of AAA was 2.45 by using Yoden index screening. Those with NLR ≥2.45 were in the high NLR group (n=66), and those with NLR <2.45 were in the low NLR group (n=54). There was no statistically significant difference between the two groups in the incidence of overall complications and the incidence of sub-complications (P>0.05). The results of logistic regression analysis suggested that NLR was an independent risk factor for complications after endovascular repair of AAA (P<0.05). The median survival time of patients in the high NLR group and the low NLR group was 31.47 months and 35.28 months, respectively, and there was no statistically significant difference between the two groups (P>0.05). Conclusion NLR can be used as a reference predictor of complications after elective endovascular repair of AAA, but more research results are still needed to confirm.