A growing body of evidence suggests that p300 histone acetyltransferase plays important roles in cancer cell differentiation and proliferation. Here, we employed structure-based hierarchical virtual screening method to identify novel lead compounds of p300 histone acetyltransferase. From a screening library containing approximate 100 000 diverse druglike compounds, 33 compounds were chosen for experimental testing and one compound, 4-acetyl-2-methyl-N-morpholino-3,4-dihydro-2H-benzo[b][1, 4]thiazine-7-sulfonamide (17), showed as micromolar inhibitor. Based on its predicted binding pose, we investigated its binding characteristics by designing two series of structural modifications. The obtained structure-activity relationship results are consistent with the predicted binding model. We expect that the identified novel p300 histone acetyltransferase inhibitors will serve as starting points for further development of more potent and specific histone acetyltransferase inhibitors.
Drug Design ; Enzyme Inhibitors ; chemical synthesis ; chemistry ; Molecular Structure ; Morpholines ; chemical synthesis ; chemistry ; Structure-Activity Relationship ; Sulfonamides ; chemical synthesis ; chemistry ; p300-CBP Transcription Factors ; antagonists & inhibitors ; chemistry

OBJECTIVETo understand antibody responses to and RNA sequences of Hantavirus in patients with hemorrhagic fever renal syndrome (HFRS) in Yantai areas and to demonstrate the type of the prevalent viruses caused HFRS.

METHODSSerum specimens collected at acute and convalescent stages from 90 patients with HFRS and IgM and IgG antibodies against Hantavirus were detected with ELISA, and cross plaque reduction neutralizing tests were performed to detect neutralizing antibody. Viral RNA was extracted from the patients? sera by using Trizol method and nested PCR was utilized to amplify the specific segments of the viral cDNA and the products of the PCR were TA cloned and then the nucleotide sequences were determined.

RESULTSThe IgM antibody was positive in 82.2% (88/107) of the patients while the IgG antibody was positive in 85.7% (66/77) of the patients. Both the serologic and sequence analyses demonstrated that the epidemic of HFRS in Yantai areas was caused by mixed types of Hantavirus. The prevalent strains of Hantavirus had higher homology with the strains isolated in Korea than with those isolated previously in China.

CONCLUSIONSThe serologic and sequencing analyses indicated that the epidemic of HFRS in Yantai areas was caused by mixed types of Hantavirus dominated by type SEO.

Antibodies, Viral ; blood ; Base Sequence ; China ; DNA, Viral ; analysis ; Disease Reservoirs ; Hantaan virus ; classification ; genetics ; immunology ; Hemorrhagic Fever with Renal Syndrome ; virology ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Molecular Sequence Data ; Sequence Analysis, DNA ; Serotyping

OBJECTIVETo develop quality of life questionnaire of Chinese medicine for postoperative patients with colorectal cancer (QLQ-CMPPCC), thus comprehensively and objectively evaluating the clinical efficacy of Chinese medicine and pharmacy in treating postoperative patients with colorectal cancer (CC).

METHODSThe theoretical structure model of the questionnaire was addressed in combined with basic theories of Chinese medicine according to the principle of WHO quality of life (QOL). The primary questionnaire was developed using methods of structuralization policy making after we extensively retrieve various universal and specific questionnaires for CC cancer patients at home and abroad. The 205 CC patients were tested by questionnaire. The items were screened using experts grading method, item selection analysis, dispersion trends of standard deviation, t-test, correlation coefficient method, factor analysis,and Cronbach's alpha.

RESULTSThe QLQ-CMPPCC was developed containing four domains of physical, psychological, independence, and social functions, involving 20 aspects and 54 items. Of them, non-fistula patients answered 43 items and fistula patients answered 46 items. One item covered the general QOL evaluation.

CONCLUSIONSQLQ-CMPPCC showed Chinese medical features. It comprehensively reflected the connotation of QOL for postoperative CC patients. It could be taken as a tool for evaluating Chinese medical efficacy for postoperative CC patients.

Colorectal Neoplasms ; surgery ; Humans ; Medicine, Chinese Traditional ; methods ; Postoperative Period ; Quality of Life ; Surveys and Questionnaires ; Treatment Outcome

OBJECTIVETo observe the association between job stress (effort-reward imbalance model)and blood lipids among university staff in Yunnan province.

METHODSA cross-sectional survey was conducted among 1244 university staff in Yunnan province. The job stress was measured by the validated Chinese self-reported Effort-Reward Imbalance Questionnaire (ERI). Blood lipids were measured in all participated staff members.

RESULTSAfter adjustment for relevant confounding factors, it was found that the risk of increased serum triglyceride was 3.5 folds higher in male staffs with high extrinsic effort compared those with low extrinsic effort (OR = 3.45, 95%CI: 1.32 - 9.04) while the risk of increased serum low density lipoprotein-cholesterol was 2.9 folds higher in male staffs with high overcommitment compared those with low overcommitment (OR = 2.86, 95%CI: 1.03 - 7.96). The risk of elevated serum triglyceride increased in proportion to increasing job stress: 3.5 folds increase in male staffs with moderate job stress (OR = 3.43, 95%CI: 1.24 - 9.53) and 4 folds increase in male staffs with high job stress (OR = 4.16, 95%CI: 1.42 - 12.17) compared those with low job stress. However, there was no significant association between job stress and lipid profile in female staffs.

CONCLUSIONOur results show that job stress (effort-reward imbalance) is positively associated with abnormal blood lipids in male university staffs.

Adult ; Cross-Sectional Studies ; Female ; Humans ; Lipids ; blood ; Male ; Middle Aged ; Stress, Psychological ; blood ; Surveys and Questionnaires ; Workload ; psychology ; statistics & numerical data

OBJECTIVETo study the effect of polyamine biosynthesis inhibition on growth characteristics of human lung carcinoma cells and its correlation with the expression of human lung carcinoma associated antigen ALT-04ag gene.

METHODSThe gene expression was detected by RT-PCR and immunocytochemical tests. The cell growth characteristics were studied by cell growth curves, morphological observation, FCM analysis and DNA electrophoresis.

RESULTSHuman lung squamous carcinoma cells L78 treated with 5 mmol/L alpha-difluoromethylornithine (DFMO) for 5 days showed significant growth inhibition and apoptosis induction. The mRNA and protein expressions of ALT-04ag gene in the cells were downregulated, while these changes resulted from DFMO treatment were prevented by provision of DFMO along with exogenous putrescine.

CONCLUSIONThe effect of polyamine biosynthesis inhibition induced by DFMO restrains the growth characteristics and promotes apoptosis of human lung carcinoma L78 cells, which is associated with down regulation of ALT-04ag gene expression.

Antigens, Neoplasm ; biosynthesis ; genetics ; Apoptosis ; Carcinoma, Squamous Cell ; pathology ; Cell Division ; Down-Regulation ; Eflornithine ; pharmacology ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; pathology ; Oncogene Proteins ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Cells, Cultured

Objective: To investigate the levels of Th9 cells and interleukin-9 (IL-9), and to assess the regulatory activity of Th9/IL-9 to anti-tumor immune response in patients with gastric cancer. Methods: Thirty-four patients with gastric cancer who received operation in the First Affiliated Hospital of Xinxiang Medical University between October 2018 and August 2019 were included. Twenty individuals who received physical examination in the same period were also enrolled. Peripheral blood was collected, and then plasma and peripheral blood mononuclear cells (PBMCs) were isolated. Tumor-infiltrating lymphocytes (TILs) and autologous gastric cancer cells were isolated from resected gastric cancer tissues. CD4(+) T cells, CD8(+) T cells, and CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells were purified from PBMCs and TILs. Plasma IL-9 level was measured by enzyme linked immunosorbent assay (ELISA). The percentage of CD3(+) CD4(+) IL-9(+) Th9 cells in PBMCs and TILSs was assessed by flow cytometry. The mRNA levels of IL-9 and transcriptional factors purine-rich nucleic acid binding protein 1 (PU.1) were semi-quantified by real-time quantitative polymerase chain reaction (RT-qPCR). PBMCs and TILs from gastric cancer patients were stimulated with recombinant human IL-9. Cellular proliferation was measured by cell counting kit-8. The phosphorylation levels of signal transducer and activator of transcription 3 (STAT3) and STAT6 were investigated by western blot. Cytokine production was measured by ELISA. Purified CD8(+) T cells from TILs of gastric cancer patients were stimulated with recombinant human IL-9. CD8(+) T cells and autologous gastric cancer cells were cocultured in direct contact and indirect contact manner. The percentage of target cell death was calculated by measuring the lactate dehydrogenase (LDH) level. These cretion of γ-Interferon (γ-IFN) and tumor necrosis factor-α (TNF-α) was measured by ELISA. CD4(+) CCR4(-)CCR6(-)CXCR3(-)cells, CD8(+) T cells, and autologous gastric cancer cells were directly cocultured, and anti-IL-9 neutralizing antibody was added. The target cell death was measured. Results: The percentages of CD3(+) CD4(+) IL-9(+) Th9 cells in PBMCs of control group and PBMCs of gastric cancer group were (1.21±0.25)% and (1.14±0.19)%, respectively. The difference was not statistically significant (P=0.280). The percentage of CD3(+) CD4(+) IL-9(+) Th9 cells in TILs of gastric cancer group was (2.30±0.55)%, which was higher than those in PBMCs of control group and PBMCs of gastric cancer group (P<0.001). The plasma IL-9 level in control group and gastric cancer group were (5.04±1.51) and (4.93±1.25) ng/ml. The difference was not statistically significant (P=0.787). The relative levels of IL-9 mRNA in PBMCs of control group and PBMCs of gastric cancer group were 1.33±0.39 and 1.36±0.27. The difference was not statistically significant (P=0.691). The relative level of IL-9 mRNA in TILs of gastric cancer group was 2.90±0.75, which was higher than those in PBMCs of control group (P<0.001) and PBMCs of gastric cancer group (P<0.001). The relative levels of PU.1 mRNA in PBMCs of control group and PBMCs of gastric cancer group were 1.21±0.12 and 1.20±0.11. The difference was not statistically significant (t=0.21, P=0.833). PU.1 mRNA relative level in TILs of gastric cancer group was 2.81±0.65, which was higher than those in PBMCs of control group (P<0.001) and PBMCs of gastric cancer group (P<0.001). Recombinant human IL-9 stimulation did not affect the proliferation of PBMCs and TILs of gastric cancer patients (P>0.05), but elevated the phosphorylation level of STAT6 and induced the secretions of γ-IFN, IL-17, and IL-22 by TILs (P<0.05). In direct contact culture system, IL-9 stimulation promoted tumor-infiltrating CD8(+) T cells-induced autologous gastric cancer cell death [(20.62±2.27)% vs. (16.08±2.61)%, P<0.01)]. In indirect contact culture system, IL-9 stimulation did not increase CD8(+) T cell-induced autologous gastric cancer cell death [(5.21±0.70)% vs. (5.31±1.22)%, P=0.998)]. However, the secretion levels of γ-IFN were elevated in response to IL-9 stimulation in both culture systems [direct contact culture system: (100.40±12.05) pg/ml vs. (76.45±8.56) pg/ml; indirect contact culture system: (78.00±9.98) pg/ml vs. (42.09±10.71) pg/ml; P<0.01]. The TNF-α secretion level did not significantly changed (P>0.05). In direct contact culture system, the percentage of target cells was (22.01±3.05) % and γ-IFN secretion level was (104.5±12.84) pg/ml in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells group, which was higher than (16.08±2.61)% and (76.45±8.56) pg/ml in CD8(+) T cells+ gastric cancer cells group (P<0.01). However, the percentage of target cells was (14.47±3.14)% and γ-IFN secretion level was (70.45±19.43) pg/ml in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells+ anti-IL-9 neutralizing antibody group, which were lower than those in CD4(+) CCR4(-)CCR6(-)CXCR3(-) cells+ CD8(+) T cells+ gastric cancer cells group (P<0.01). Conclusion: Tumor-infiltrating Th9 cells and the secreting IL-9 promote the activity of CD8(+) T cells in gastric cancer patients, and enhance anti-tumor immune response.
Humans ; CD8-Positive T-Lymphocytes ; Stomach Neoplasms/pathology* ; Tumor Necrosis Factor-alpha/metabolism* ; Lymphocytes, Tumor-Infiltrating/pathology* ; Interferon-gamma/metabolism* ; RNA, Messenger/metabolism* ; Antibodies, Neutralizing/metabolism*