To study the role of a novel MDR-related gene MGr1-Ag in the multidrug resistant (MDR) gastric cancer. The whole length MGr1-Ag gene was cloned, and eukaryotic vectors carrying the full length of MGr1-Ag cDNA and its antisense expression vector were constructed. The sense vector and anti-sense were then transfected into MGC803 cells and SGC7901/VCR cells respectively by lipofectamine. The result showed that a 1.0kb fragment was successfully amplified by RT-PCR and cloned into pUCm-T vector. DNA sequencing suggested that the fragment was the properly encoded MGr1-Ag gene. Recombinant eukaryotic plasmids harboring MGr1-Ag and its antisense expression vector were also obtained by subcloning the gene into pCDNA3.1/V5-His, which was confirmed by endonuclease digestion. As confirmed by Western blot, stable cell strains with up and down-regulated MGr1-Ag expression were constructed. Those cell strains provided the basis for further study on the MGr1-Ag in MDR of gastric cancer.

Objective To analyse the dynamic changes of cardiac diseases over the past 30 years. Method All hospitalized patients of internal medicine department over the past 30 years in the 2nd Xiangya Hospital of Central South University were analyzed in order to understand the dynamic changes of cardiac diseases. Results The results showed the proportion of the heart diseases to all medical diseases was gradually increased during the past three decades. The constitution ratio of the classification of the heart disease was also changing continuously and progressively, with coronary heart disease ranking the first and, in contrast, rheumatic heart disease the second since 1980s. Meanwhile, the number and the proportion of the patients with cardiomyopathy and arrhythmias of unknown causes were increased. Anemic heart disease and cardiovascular syphilis had been wiped out since 1980s and 1990s respectively. Conclusion Classification of cardiac diseases changes gradually and continually during the past 30 years.

ObjectiveTo investigate the role of productive factors in the development of primary biliary cholangitis (PBC). MethodsA total of 273 female patients with a definite diagnosis of PBC who visited Xijing Hospital from October 2013 to August 2015 were enrolled. The patients with autoimmune hepatitis, primary sclerosing cholangitis overlap syndrome, and hepatic encephalopathy and those with incomplete data after telephone follow-up were excluded, and 54 female PBC patients who had female relatives were finally enrolled (PBC group). The female relatives who were less than 10 years older or younger than the patients were collected, and those with severe systemic diseases and incomplete data after telephone follow-up were excluded; finally 88 relatives were enrolled (relative group). The questionnaire for female reproductive factors in PBC was used to survey all these enrolled patients and collect data. The t-test was used for comparison of continuous data between groups, the chi-square test was used for comparison of categorical data between groups, and a multivariate logistic regression analysis was used for the analysis of dose-response relationship. ResultsThe PBC group had a significantly higher number of births than the relative group (2.55±1.84 vs 1.84±0.95, t=2.708, P=0.009). Furthermore, there was a significant dose-response pattern between the number of births and the development of PBC (P=0.002). ConclusionThe number of births may be associated with the development of PBC in a dose-response manner. As for the female population susceptible to PBC, a reduction in the number of births may reduce the possibility of PBC.

OBJECTIVETo develop an oral DNA vaccine based on MG(7)-Ag mimotope of gastric cancer using attenuated Salmonella typhimurium and evaluate its efficacy and protective effect.

METHODSThe eukaryotic expression vector including the MG(7)-Ag mimotope and a Th epitope was constructed, and then transduced into an attenuated Salmonella typhimurium to get the oral DNA vaccine. C57BL/6 J mice were orally immunized with 1 x 10(8) cfu Salmonella transfectants, with Salmonella harboring empty plasmid, with phophate buffered saline (PBS) as control. At the 6th week, serum titer of MG(7) antibody was detected by ELISA. In the 8th week, a [(3)H]-thymidine incorporation assay was performed to test the proliferation of murine spleen cells to the stimulant of MG(7)-Ag mimicry peptide. At the same time, Ehrlich ascites carcinoma cells expressing MG(7)-Ag were used in tumor challenge assay to evaluate the protective effect of the immunization.

RESULTSThe oral DNA vaccine induced MG(7) antibody in mice, while in vivo unprimed proliferation assay of the spleenocytes showed no difference among the three groups. Two weeks after tumor challenge, 2 in 7 immunized mice were tumor free, while none in the control group was protected.

CONCLUSIONOral DNA vaccine based on the MG(7)-Ag momitope is immunogenic. It is able to induce specific immunity response against tumor in mice, and the vaccine is partially protective.

Administration, Oral ; Amino Acid Sequence ; Animals ; Antigens, Neoplasm ; blood ; genetics ; immunology ; Base Sequence ; Cancer Vaccines ; genetics ; immunology ; therapeutic use ; Epitopes ; genetics ; immunology ; Female ; Mice ; Mice, Inbred C57BL ; Molecular Mimicry ; genetics ; immunology ; Molecular Sequence Data ; Plasmids ; genetics ; Polymerase Chain Reaction ; Stomach Neoplasms ; drug therapy ; immunology ; Treatment Outcome ; Vaccines, DNA ; genetics ; immunology ; therapeutic use

Objective To estimate the diagnostic value of cytology, DNA-ICM(DNA-image cytometry),cytology combined with DNA-ICM for pancreatic malignancy,and to explore the cut-off value for DNA-ICM. Methods Patients with suspicious pancreatic malignancy were retrospectively identified. In total,145 EUS-FNA specimens acquired from 140 separate patients were examined by cytology and DNA-ICM. Diagnostic values among cytology, DNA-ICM and the combination of the techniques in detecting pancreatic malignancy were compared. Results Compared with cytology, DNA-ICM had a lower sensitivity (63.0% VS 82.4%)and accuracy(69.7% VS 85.5%). After combining the techniques, the diagnostic value for pancreatic malignancy significantly improved compared with that by cytology(0.941 VS 0.912, P=0.007 0)or DNA-ICM only(0.941 VS 0.815, P<0.000 1). By using the Youden index, the cut-off value for DNA-ICM to detect pancreatic malignancy was one cell with DI(DNA index)≥2.5. Notably,with this standard, the sensitivity and accuracy of DNA-ICM significantly increased to 72.3% and 77.2%, and those of the combined techniques increased to 91.6% and 93.1%, respectively. Conclusion Automated DNA-ICM is an objective and effective method for pancreatic malignancy. Although DNA-ICM has a lower diagnostic value than that of conventional cytology, an improved value was obtained after combining the techniques.

Objective:To investigate the relationship between common functional gastrointestinal diseases symptoms with psychological factors, diet and lifestyles by using the network analysis method which has achieved great success in the field of psychology in recent years.Method:A questionnaire survey was conducted in two military units using the cluster sampling method during July 2020, and a total of 1 805 subjects were included. Functional gastrointestinal disease symptoms were evaluated with the Gastrointestinal Symptom Rating Scale (GSRS). The state, trait anxiety scale and stress response scale were used to evaluate the mental and psychological state by self-evaluation. R was used to build the network and calculate statistical parameters.Results:1 486 of the 1 805 subjects (82.3%) had experienced functional gastrointestinal diseases symptoms within 2 weeks, but most of them were mild. Network analysis shows that there was a strong interaction between digestive system symptoms with different clinical manifestations (Spearman coefficient ranges 0.31-0.56). There was a clear relationship between functional gastrointestinal symptoms and mental and psychological factors (Spearman coefficient ranges 0.16-0.27), but there was no clear interaction with diet, age, education level, body mass index, etc. Functional gastrointestinal diseases symptoms were connected with mental and psychological factors through two nodes: stress and indigestion. The stability coefficient of node strength correlation was 0.75, indicating that the network was stable.Conclusions:The current study revealed the network structure and features of functional gastrointestinal diseases symptoms with mental and psychological factors. The key linking nodes provided potential interfering target for controlling functional gastrointestinal symptoms related to mental and psychological factors.

Objective: To examine the regulatory effect of bone marrow mesenchymal stem cells (BM-MSCs) on the polarization of bone marrow-derived macrophages, and to provide a theoretical support for the application of mesenchymal stem cells in the treatment of liver fibrosis. Methods: MSCs and macrophages were first isolated from the bone marrow of mice. Macrophages were polarized to M1 macrophages with lipopolysaccharide (LPS) and interferon-γ (IFN-γ), and to M2 macrophages with interleukin-4 (IL-4). The macrophages were then co-cultured with BM-MSCs in a Transwell for 24 h, and changes in the percentages of M1 and M2 macrophages were examined using flow cytometry. The mRNA levels of the M1 macrophage-associated cytokines, tumor necrosis factor-α (TNF-α) and interleukin-23a (IL-23a), and M2 macrophage-associated molecules, arginase-1 (Arg-1) and CD163, were measured by real-time quantitative PCR. The two samples were compared using the t test, and P < 0.05 was considered as statistically significant. Results: Flow cytometry showed that the percentage of M1 macrophages was significantly lower in the (macrophage + LPS + IFN-γ + BM-MSC) co-culture group than in the (macrophage + LPS + IFN-γ) group (62.5% ± 4.6% vs 86.6% ± 6.9%, t = 5.034, P = 0.0073). In addition, the relative mRNA expression of TNF-α and IL-23a was also significantly reduced in the co-culture group compared with those in the macrophage control group as measured by RT-qPCR (t = 11.57 and 10.57, respectively, P < 0.05). Compared with that in the macrophage control group, the percentage of M2 macrophages in the (macrophage+BM-MSC) co-culture group was significantly increased (89.5% ± 5.8% vs 70.1% ± 6.3%, t = 3.924, P = 0.0172), along with significantly elevated relative mRNA expression of Arg1 (14.35±1.05 vs 1.00±0.03, t = 21.96, P < 0.05) and CD163 (3.04 ± 0.27 vs 1.00 ± 0.03, t = 13.14, P < 0.05). Conclusion BM-MSCs can inhibit LPS + IFN-γ-induced polarization to M1 macrophages and promote polarization to M2 macrophages through the release of paracrine factors.

Objective To investigate the efficacy and safety of programmed death receptor-1 (PD-1) inhibitor combined with transarterial chemoembolization (TACE) and anti-angiogenic drug tyrosine kinase inhibitor (TKI) versus TACE combined with TKI in the treatment of advanced hepatocellular carcinoma (HCC) and related influencing factors for prognosis. Methods An analysis was performed for all patients who received TACE+TKI+PD-1 inhibitor and some patients who received TACE+TKI in The First Affiliated Hospital of Air Force Medical University from June 2018 to July 2021. Related clinical data were collected, and propensity score matching (PSM) was used to balance the baseline characteristics between groups. The chi-square test was used for comparison of categorical data between two groups; the Wilcoxon rank-sum test was used for comparison of the number of TACE procedures between two groups; the Kaplan-Meier method was used to analyze overall survival (OS), and univariate and multivariate Cox regression models were used to analyze the influencing factors for prognosis. Results A total of 181 patients with advanced HCC were screened out, among whom 50 patients were treated with TACE+TKI+PD-1 inhibitor; after PSM, 40 patients treated with TACE+TKI+PD-1 inhibitor were enrolled as observation group and 40 patients treated with TACE+TKI were enrolled as control group. At the end of follow-up, the median follow-up time was 28.6 (95% confidence interval [ CI ]: 22.1-35.1) months, and the median OS was 15.9 (95% CI : 7.5-24.2) months in the observation group and 11.2 (95% CI : 5.0-17.5) months in the control group. The Cox regression analysis showed that the application of PD-1 inhibitor (hazard ratio [ HR ]=0.42, 95% CI : 0.23-0.80, P =0.008), the number of TACE procedures ( HR =0.67, 95% CI : 0.46-0.99, P =0.043), Child-Pugh class ( HR =2.40, 95% CI : 1.15-5.00, P =0.019), and vascular invasion ( HR =3.42, 95% CI : 1.11-9.42, P =0.031) were independent influencing factors for prognosis. The incidence rate of grade > 2 adverse events was 40% for both the observation group and the control group, and there was no significant difference between the two groups ( P =0.818). Conclusion Compared with TACE+TKI, TACE+TKI+PD-1 inhibitor can significantly prolong the OS of patients in advanced HCC, with relatively controllable adverse events.