Diagnostic value of serum TIM-3 in patients with liver cancer / 国际肿瘤学杂志

Objective:To analyze the changes of T cell immunoglobulin- and mucin-domain-containing molecule-3 (TIM-3) in serum of patients with liver cancer and its diagnostic value.Methods:From March 2021 to May 2021, 37 patients with viral hepatitis type B (hepatitis B group) , 44 patients with liver cirrhosis (liver cirrhosis group) and 27 patients with liver cancer (liver cancer group) were selected in the Second Affiliated Hospital of Air Force Medical University, and 35 healthy subjects who underwent physical examination during the same period were selected as the healthy control group. The serum alpha fetoprotein (AFP) , liver function indexes and TIM-3 levels were detected, and the differences among groups were analyzed. The correlations between TIM-3 and AFP and liver function indexes were analyzed by Spearman correlation. Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of TIM-3 in liver cancer.Results:There was a statistically significant difference in AFP among the hepatitis B group, liver cirrhosis group and liver cancer group ( χ2=11.75, P=0.003) . There were statistically significant differences in total bilirubin ( χ2=22.85, P<0.001) , direct bilirubin ( χ2=25.90, P<0.001) , indirect bilirubin ( χ2=19.92, P<0.001) , alanine aminotransferase ( χ2=36.64, P<0.001) , aspertate aminotransferase ( χ2=26.26, P<0.001) , aspertate aminotransferase/alanine aminotransferase ( χ2=34.67, P<0.001) and total bile acid ( χ2=13.10, P<0.001) among the hepatitis B group, liver cirrhosis group and liver cancer group. The serum levels of TIM-3 in the healthy control group, hepatitis B group, liver cirrhosis group and liver cancer group were 11.1 (4.2, 14.4) ng/ml, 12.7 (4.3, 23.9) ng/ml, 11.4 (3.4, 17.0) ng/ml and 15.7 (10.5, 21.2) ng/ml, with a statistically significant difference ( χ2=11.85, P=0.008) . There were statistically significant differences between the liver cancer group and healthy control group and liver cirrhosis group (both P<0.05) . Spearman correlation analysis showed that TIM-3 had no correlation with AFP in the four groups ( r=0.05, P=0.791; r=0.18, P=0.497; r=0.03, P=0.883; r=0.24, P=0.396) . There were correlations between serum TIM-3 and total protein in the healthy control group ( r=0.36, P=0.036) , serum TIM-3 and globulin in the hepatitis B group ( r=0.35, P=0.034) , and serum TIM-3 and total bile acid in the liver cancer group ( r=0.46, P=0.017) . ROC curve analysis showed that the sensitivity of serum TIM-3 for the diagnosis of liver cancer was 48.10%, and the specificity was 91.43%, when taking healthy subjects as the control group. The sensitivity of serum TIM-3 for the diagnosis of liver cancer was 96.30%, and the specificity was 41.77%, when taking healthy subjects and liver cirrhosis patients as the control group. The sensitivity of serum TIM-3 for the diagnosis of liver cancer was 96.30%, and the specificity was 40.52%, when taking healthy subjects, hepatitis B patients and liver cirrhosis patients as the control group. Conclusion:The serum level of TIM-3 in patients with liver cancer is significantly increased, which has certain diagnostic value for liver cancer, and can be used as a diagnostic marker and potential therapeutic target for liver cancer patients.

Evidence-based visualization analysis of literature of digestive endoscopic minimally-invasive resection in the past decade / 中华消化内镜杂志

Objective:To review publications in the field of digestive endoscopic minimally-invasive resection in the past 10 years in and outside China.Methods:Literature of digestive endoscopic minimally-invasive resection in the Web of Science and CNKI databases from January 1, 2011 to July 17, 2021 was retrieved. VOSviewer 1.6.11 was used for clustering and time series analysis of countries, institutions, authors and keywords, and drawing evidence-based visualization maps, so as to analyze the cooperation among countries, academic institutions and researchers, to compare the differences in research topics between Chinese and English databases, and to predict the future research hot spots and directions.Results:A total of 22 834 English articles and 4 636 Chinese articles were included. Over the past 10 years, the number of Chinese and English publications has been growing steadily, and most of them were published in professional journals. The publications were mainly from China, Japan, South Korea and the United States, where all exceeded 2 000. The National Cancer Center of Japan had 497 publications, ranking the first among all institutions. The cooperation between academic institutions showed obvious regional characteristics, and the inter-institutional and interpersonal cooperation needed improvement. In terms of keyword clustering, there was no significant difference between Chinese and English publications, but there were two additional clusters in Chinese publications, endoscopic nursing and submucosal tumor. Overlays analysis of key words showed that endoscopic surgery, tunneling technique, and submucosal tumor could be hot spots and future directions.Conclusion:Digestive endoscopic minimally-invasive resection has experienced a vigorous development in the past 10 years with a growing number of Chinese and English publications. China is playing an increasingly important role on the international stage. The advanced nature of research focus in Chinese publications is comparable to that in English publications, yet also showing Chinese characteristics. In the future, more efforts should be taken to strengthen regional cooperation and focus on research hot spots.

Analysis of risk factors of early pulmonary complications after orthotopic liver transplantation: a report of 313 cases / 中华器官移植杂志

Objective:To explore the effects of donor, basic recipient characteristics and intraoperative findings before orthotopic liver transplantation(OLT)on postoperative pulmonary complications and the influence of postoperative pulmonary complications on postoperative outcomes and long-term survival of patients.Methods:From January 2016 to December 2019, clinical data were retrospectively reviewed for 313 OLT recipients at First Affiliated Hospital, Nanjing Medical University.According to clinical manifestations, laboratory parameters and imaging findings within 7 days post-operation, they were divided into two groups of with and without postoperative pulmonary complications.Univariate and multivariate Logistic regression analyses were performed for general parameters of donors/recipients and perioperative variables of recipients in two groups.And Kaplan-Meier survival analysis was conducted for postoperative survival of two groups.Results:Multiariate Logistic regression analysis indicated that age, concurrent chronic diseases, low preoperative platelet count, intraoperative massive bleeding(>1 500 ml)and severe hypotension after perfusion were all independent risk factors for postoperative pulmonary complications.Compared with those without postoperative pulmonary complications, intensive care unit(ICU)stay, postoperative mechanical ventilation duration and hospital stay were significantly prolonged in patients with postoperative pulmonary complications.And rate of re-intubation, incidence of acute kidney injury and hospital mortality spiked markedly( P<0.05). Among OLT patients with benign liver diseases, overall survival rate of patients without postoperative pulmonary complications was significantly better than that of those with postoperative pulmonary complications( P<0.05). Conclusions:Age, concurrent chronic diseases, low preoperative platelet count, massive intraoperative bleeding(>1 500 ml)and severe hypotension after perfusion boost the risks of early postoperative pulmonary complications and adversely affect the outcomes of OLT recipients.

Effect of donor age factor on intraoperative and early postoperative complications in patients undergoing orthotopic liver transplantation / 中华麻醉学杂志

Objective:To evaluate the effect of donor age factor on intraoperative and early postoperative complications in the patients undergoing orthotopic liver transplantation (OLT).Methods:The clinical data of patients underwent OLT from January 2016 to December 2019 were retrospectively collected.The patients were divided into elderly donor group (age ≥65 yr) and non-elderly donor group (age<65 yr) according to the donor age.The preoperative and intraoperative clinical data of the two groups were matched by 1∶2 propensity score matching.The primary outcome measures were intraoperative postreperfusion syndrome and acute kidney injury and postoperative pulmonary complications within 7 days after surgery.The secondary outcome measures were time of intensive care unit stay, assisted mechanical ventilation time, postoperative length of hospital stay, rate of reintubation, and 6-month and 1-year survival and recurrence-free survival after surgery.Results:After propensity score matching was performed, 66 patients were enrolled, including 22 in elderly donor group and 44 in non-elderly donor group.There was no significant difference in the incidence of intraoperative postreperfusion syndrome, acute kidney injury and postoperative pulmonary complications within 7 days after operation, time of intensive care unit stay, assisted mechanical ventilation time, postoperative length of stay, 6-month and 1-year survival and recurrence-free survival rate between elderly donor group and non-elderly donor group ( P>0.05). Conclusion:Age factors do not exert effect on the incidence of intraoperative and early postoperative complications in the patients undergoing OLT.

Immunogenicity of the truncated NDV F protein surface-displayed on Lactobacillus casei / 生物工程学报

To evaluate immune efficacy of the recombinant Lactobacillus casei, we constructed pLA-Newcastle disease virus (NDV)-F/L. casei and obtained the expression products. PCR amplified the NDV F gene carrying part of the major epitopes. The target gene was inserted to the shuttle plasmid pLA, and then transformed into Escherichia coli BL21 (DE3) in order to screen positive recombinant plasmid. The positive recombinant plasmid was transformed into L. casei by electroporation to construct pLA-NDV-F/L. casei. The positive strains were identified by PCR. The reactivity of the recombinant bacteria was identified by Western blotting and the protein expression was detected by indirect immunofluorescence, flow cytometry and laser confocal microscopy. The 14-day-old chickens in each group were vaccinated by oral plus nose drops. The pLA-NDV-F/L. casei twice immunization group and three times immunization group, the commercial vaccine group, the pLA/L. casei group, the unchallenge PBS and the challenge PBS group were established. IgG in serum and sIgA in the lavage fluid of intestinal, nasal and lung were detected by ELISA. The protection rate of chickens was evaluated. The results showed that 94.10% of the recombinant bacteria expressed the F protein. The recombinant protein was highly expressed on the surface of L. casei with a protein size of 62 kDa, which specifically bound to anti-NDV serum. The levels of anti-F IgG and sIgA antibodies in each test group were significantly higher than those in the control groups. The duration of antibody in the pLA-NDV-F/L. casei three-time immunization group lasted 28 days longer than that in the twice immunized group, and there was no significant difference between antibody peak values. The attack protection rates in each group of immunized pLA-NDV-F/L. casei three times, twice, attenuated vaccine, pLA/L. casei and PBS were 80%, 80%, 90%, 0% and 0%, respectively. Therefore, the antigenic protein of NDV F was successfully expressed by L. casei expression system, which has of reactogenicity and immunogenicity, and could induce protective immune responses in chickens.

Effects of peroral endoscopic myotomy on achalasia in geriatric patients / 中华消化内镜杂志

Objective To evaluate the safety, feasibility, perioperative and long-term efficacy of peroral endoscopic myotomy ( POEM) for achalasia ( AC) in geriatric patients. Methods Data of 41 patients aged over 65 diagnosed with achalasia and treated with POEM in Zhongshan Hospital from August 2010 to December 2014 were retrospectively studied. Perioperative complications, preoperative and postoperative Eckardt score and pressure of the lower esophageal sphincter, esophageal reflux and clinical failure were analyzed. Results All 41 patients underwent POEM successfully, with median operation time of 42 min. Median hospitalization was 3 days. Major perioperative adverse events occurred in 4 cases ( 9. 75％) , whose hospitalization was longer than 5 days for perioperative adverse events. During median follow-up period of 40 months ( interquartile range 24-57 months) , median Eckardt score decreased from 8 to 1( P<0. 001 ) and pressure of the lower esophageal sphincter decreased from 23. 85 mmHg ( 1 mmHg=0. 133 kPa) to 9. 05 mmHg (P=0. 005). Clinical reflux occurred in 12 cases (29. 27％) and the 5-year success rate of POEM was 87. 80％ (36/41). Conclusion POEM is a safe and reliable treatment for geriatric AC patients with confirmed short-term and long-term effectiveness.

Prokaryotic expression and purification of anti-AEG-1 single-chain variable antibody / 国际检验医学杂志

Objective To construct anti-astrocyte elevated gene-1(AEG-1)single-chain variable antibody (V23)prokaryotic ex-pression vector,and to conduct the protein purification and immunological activity detection.Methods The Primer5 software was applied to design the primers aiming at the gene sequence of the antibody anti-AEG-1 single-chain variable region for constructing the prokaryotic expression plasmid of PRsetC/V23.After the enzyme digestion by the restriction enzyme Pst1 and correct DNA se-quencing,the prokaryotic expression plasmid was led to E.coli BL21 ,the prokaryotic expression engineering strain containing the V23 gene was constructed.After the induction with IPTG,the interest protein was purified by the magnetic beads with the HIS tag,and the content of interest protein was determined by the SDS-PAGE electrophoresis.Western blot and ELISA were adopted to detect the immune activity of the nti-AEG-1 single-chain variable region antibody.Results For the constructed prokaryotic expres-sion plasmid PRsetC/V23,the single enzyme digestion and sequencing analysis displayed that the constructed V23 gene was com-pletely consistent to the designing sequences.After IPTG induction,SDS-PAGE electrophoresis showed an apparent protein band at 31×103 ,the Western blot detection showed a specific AEG-1 response band at 80 ×103 ,the ELISA test showed the positive re-sults.Conclusion The PRsetC/V23 prokaryotic expression plasmid and the V23 prokaryotic expression engineering strain are suc-cessfully constructed,this engineering strain can express anti-AEG-1 single-chain variable region antibody protein,and the protein has good immune activity.

Genetic polymorphism of the phospholipase C epsilon 1 gene and risk of gastric cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The pathogenesis of gastric cancer (GC) involves environmental and genetic factors. Recently, two genome-wide association studies found that phospholipase C epsilon 1 (PLCE1) polymorphisms might be related to GC risk, and several studies further validated this finding. However, these studies yielded inconsistent results.</p><p><b>METHODS</b>A comprehensive database search was performed to identify eligible studies. Odds ratios with 95% confidence intervals were calculated to assess the strength of the association between PLCE1 rs2274223, rs753724, and rs11187842 and risk of GC. Subgroup analyses, publication bias, and sensitivity analyses were also conducted.</p><p><b>RESULTS</b>Eleven studies (12 cohorts) were included in the meta-analysis. Based on 13 676 cases and 23 569 controls, a significant association between PLCE1 rs2274223 and GC risk was detected under various genotypic models. In the subgroup analyses, the association was significant for cardia GC, but weak for non-cardia GC. The association under the heterozygote model was detected for PLCE1 rs753724 and rs11187842 based on three studies involving 2768 cases and 3890 controls.</p><p><b>CONCLUSIONS</b>Our findings demonstrate that the presence of the G allele at rs2274223 of the PLCE1 gene may contribute to susceptibility to GC, especially cardia GC. PLCE1 rs753724 and rs11187842 are associated with GC risk under the heterozygote model. Further well-designed large studies are warranted to validate these findings.</p>

Syphilis antibody enzyme-linked immunosorbent method grey area test results analysis / 国际检验医学杂志

Objective ELISA syphilis enzyme analysis(TP-ELISA) results for the gray zone two step after the replacement .To explore the suitability of syphilis detection procedures ,reduce misdiagnosis .Methods Randomly choose 50 288 cases of serum screening for syphilis enzyme linked immunosorbent assay ;in grey area of the sample ,then rapid plasma reagin test (RPR) and Treponema pallidum particle agglutination test(TPPA) for validation .According to disease and age groups ,statistics of anti TP an-tibody gray area ratio .Results 50 288 serum samples ,after TP-ELISA method to screen grey area in the early results of 61 cases . The highest age of grey area rate in different ages was more than 61 years old .Conclusion TP-ELISA detection still exist certain false positive ,set reasonable grey area ,the gray zone within the sample confirmation test has important clinical significance .

Recent progress in understanding the effects of autophagy in hepatocellular carcinoma / 中华肝胆外科杂志

This review discusses the relationship between autophagy and hepatocellular carcinoma (HCC).It will focus on the definition and mechanism of autophagy,the effects of autophagy on carcinogenesis,the reaction that chemotherapy and radiotherapy have on autophagy,developments of novel therapeutic methods aimed at autophagy,and will emphasize the bipolar role of autophagy in HCC.

The mechanism study of oxymatrine combined with radiotherapy on HO8910 cells of human ovarian cancer / 国际中医中药杂志

Objective To study the effect of oxymatrine (OMT) combined with radiotherapy on the growth of HO8910 cells of human ovarian cancer in vitro.Methods All human ovarian cancer samples were divided into four groups:control group,treated with culture solution; OMT group,treated with 4 mg/ml OMT; radiotherapy group,treated by 4 Gy radiotherapy; and combination group,firstly treated by 4Gy radiotherapy,and then cultured with 4 mg/ml OMT.Observed the changes of cell morphology by invert microscope at 10,24,48,and 72 hours respectively.Flow cytometry was adopted to detect cell apoptosis.Results ① invert microscope observation showed that compared with the control group,HO8910 cells demonstrated apoptosis of diminution in volume,thickening in cytoplasm,and gathering in nucleus in all the other three groups at 24 and 48 hours.② Flow cytometry showed that apoptosis rate of HO8910 cells and cell population in G1 phase increased in the combination group,which were significantly higher than the other three groups (P＜0.05).Conclusion Both OMT (4 mg/ml)and radiation (4Gy)can induce cell apoptosis,while the combination of them showed better therapeutic results.

Genetic construction of HLA-A2 restricted multi-epitopes gene of hepatitis C virus,expression and purification in E.coli for antigenic analysis / 中国免疫学杂志

Objective:To construct the recombinant prokaryotic plasmid to express HCV HLA-A2 restricted multi-CTL epitopes and to purify the fused protein for antigenic analysis.Methods:The human ubiquitin gene and multi-CTL epitopes gene was synthesized respectively,and digested by restrict enzyme before being cloned into pRSET-A.Then it was transformed into E.coli DH5α and the positive recombinant plasmid named pRSET-Ub-Mep was sequenced.Target protein was distinctly expressed after transformed into E.coli BL21 and induced with IPTG.Thus the protein was scanned and purified on Ni~(2+)-NTA column as well as Western blot performed after solubility analysis.Results:The recombinant plasmid pRSET-Ub-Mep was successfully constructed and it could efficiently express the target gene.Protein production was mainly in inclusion body and could be purified through Ni~(2+)-NTA column.The purified protein kept the antigen activity.Conclusion:The gene encoding for HCV HLA-A2-restricted multi-CTL epitopes is efficiently expressed and the target protein is purified,which establishes a foundation of further research to evaluate the cellular immune response induced by the target gene.