Objective To explore the effect of vasoactive intestinal peptide ( VIP) on the prevention and treatment of experimental autoimmune encephalomyelitis(EAE) rats by regulating the levels of IFN-γand IL-17A in brain tissue. Methods Sixty healthy female Wistar rats were randomly divided into 4 groups:normal control group, EAE control group, VIP low-dose group and VIP high-dose group.Myelin basic protein ( MBP)+complete Freurd′s adjuvant ( CFA) was used to establish an EAE model.The low and high-dose VIP groups were intraperitoneally injected with VIP 4 nmol/kg(0.2 ml) and 16 nmol/kg (0.8 ml) respectively every other day,while normal control group and EAE group with 0.8 ml saline for ten consecutive days.The incubation period, progression and peak of neurological dysfunction score ( NDS) changes of rats were recorded.The pathological changes, the GFAP+astrocyte activation in the brain at the morbidity peak of rats and the cytokine levels of IFN-γand IL-17A in brain tissue were observed.Results The incubation period was extended, the progression and peak NDS were shortened in the two VIP groups.In normal control group, there was no inflammatory cell infiltration or active astrocytes in brain tissue.The degree of infiltration of inflammatory cells and the degree of astrocyte activation in the VIP control group were significantly lower than in the EAE group.The cytokine levels of IFN-γand IL-17A in brain tissue were reduced in VIP groups.Conclusion By lowering IFN-γand IL-17A content in brain tissue, the infiltration of inflammatory cells and astrocyte activation are inhibited.VIP plays an important role in prevention and control of EAE.

Objective To investigate the impacts of forkhead box M1(FOXM1)on the proliferation,invasion,and drug resistance of gastric cancer cells by regulating the circular RNA circ_NOTCH1.Methods Western blotting and real-time quantitative PCR were performed to determine the expression of FOXM1 protein and circ_NOTCH1,respectively,in the gastric cancer tissue,para-carcinoma tissue,human normal gastric mucosa epithelial cell line GES-1 and gastric cancer cell lines MGC-803,HGC-27,and BGC-823.BGC-823 cells were classified into the following groups:control,short hairpin RNA FOXM1(sh-FOXM1)and negative control(sh-NC),small interfering RNA circ_NOTCH1(si-circ_NOTCH1)and negative control(si-NC),and sh-FOXM1+circ_NOTCH1 overexpression plasmid(sh-FOXM1+pcDNA-circ_NOTCH1)and sh-FOXM1+negative control(sh-FOXM1+pcDNA).CCK-8 assay and clone formation assay were employed to measure the cell proliferation,and Transwell assay to measure cell invasion.After treatment with 1.0 mg/L adriamycin for 48 h,the cell resistance in each group was analyzed.Western blotting was employed to determine the expression levels of FOXM1,proliferating cell nuclear antigen(PCNA),Bax,multi-drug resistance-associated protein 1(MRP1),and multi-drug resistance gene 1(MDR1).RNA pull-down and RNA immunoprecipitation were employed to examine the binding of circ_NOTCH1 to FOXM1 protein.Results Compared with those in the para-carcinoma tissue,the expression levels of FOXM1 protein and circ_NOTCH1 in the gastric cancer tissue were up-regulated(all P<0.001).Compared with GES-1 cells,MGC-803,HGC-27,and BGC-823 cells showed up-regulated expression levels of FOXM1 protein and circ_NOTCH1(all P<0.001).Compared with the control group and sh-NC group,the sh-FOXM1 group with down-regulated expression of FOXM1 protein and circ_NOTCH1 showed decreased optical density value,clone formation rate,cell invasion number,and cell viability,down-regulated expression of PCNA,MRP1,and MDR1,and up-regulated expression of Bax protein in BGC-823 cells(all P<0.001).Compared with the control group and the si-NC group,the si-circ_NOTCH1 group with down-regulated expression of circ_NOTCH1 showed decreased optical density value,clone formation rate,cell invasion number,and cell viability,down-regulated expression of PCNA,MRP1,and MDR1,and up-regulated expression of Bax protein in BGC-823 cells(all P<0.001).Compared with sh-FOXM1 group and sh-FOXM1+pcDNA group,the sh-FOXM1+pcDNA-circ_NOTCH1 group with up-regulated expression of circ_NOTCH1 showed increased optical density value,clone formation rate,cell invasion number,and cell viability,up-regulated expression of PCNA,MRP1,and MDR1,and down-regulated expression of Bax protein(all P<0.001).FOXM1 protein was able to interact with circ_NOTCH1.Conclusion Interference with FOXM1 may inhibit the proliferation,invasion,and drug resistance of gastric cancer cells by silencing circ_NOTCH1 expression.
Humans ; bcl-2-Associated X Protein/metabolism* ; Carcinoma ; Cell Line, Tumor ; Cell Proliferation/genetics* ; Drug Resistance ; Forkhead Box Protein M1/metabolism* ; Gene Expression Regulation, Neoplastic ; MicroRNAs/genetics* ; Proliferating Cell Nuclear Antigen/metabolism* ; Receptor, Notch1/metabolism* ; RNA, Small Interfering/genetics* ; Stomach Neoplasms/genetics*

Objective　The association of hepatitis B virus (HBV) infection with the clinical features and prognosis of diffuse large B-cell lymphoma (DLBCL) remains unclear. This study aimed to investigate the relationship between HBV infection and the clinical characteristics and prognosis of DLBCL.　Methods　This retrospective included 199 cases of DLBCL initially treated in our Department of Oncology from January 2012 to December 2017, which fell into an HBV group (n = 92) and a non-HBV group (n = 107). We recorded the clinical features, liver function before and during the treatment, progression-free survival and overall survival of the patients. Based on the level of lactate dehydrogenase (LDH) and the presence of HBV infection, we again divided the patients into four groups: normal (non-HBV infection and normal LDH, n = 67), high LDH (without HBV infection, n = 40), HBV (HBV infection with normal LDH, n = 59), and HBV+high LDH (with both HBV infection and high LDH, n = 33), and compared the results of treatment among different groups.　Results　The incidence rate of liver damage was significantly higher in the HBV than in the non-HBV group before chemotherapy (P < 0.05) but showed no statistically significant difference between the two groups during chemotherapy (P > 0.05). The rate of therapeutic effectiveness was remarkably lower in the HBV+high LDH than in the normal, high LDH and HBV groups (30.3% vs 82.1%, 87.5% and 88.1%, P < 0.01). The progression-free survival was markedly longer in the non-HBV than in the HBV group ([63.9 ± 2.4] vs [45.7 ± 2.9] mo, P = 0.004).　Conclusion　HBV infection is involved in the development and progression of DLBCL, and may act synergetically with high LDH and exerts a negative effect on the therapeutic efficacy.

PD-1/PD-L1 plays a pivotal role in the inhibition of T lymphocytes and tumor immune escape. Immunological checkpoint inhibitors of PD-1/PD-L1 can restore inactivated T cells and enhance the ability of killing tumor cell. At present, PD-1/PD-L1 inhibitors have been used in a variety of tumor types. Although some patients prove significant effects, there are still some patients with primary or acquired resistance to PD-1/PD-L1 inhibitors. The mechanism of resistance is related to the loss of the first signal or costimulatory signal, the expression of PD-1/PD-L1,the expression of other immune checkpoints on the surface of T cells, the immunosuppressive tumor microenvironment and systemic immunity. And we will find ways to prolong the survival time of patients through the combination therapy. This article mainly reviews the resistance mechanism and combination therapy of PD-1/PD-L1.

AIM: To investigate the postoperative ocular surface changes in acute attack eye and contralateral eye with primary angle-closure glaucoma(PACG)and cataract.METHODS: A total of 40 patients with monocular acute PACG combined with cataract who admitted to Zhengzhou Central Hospital Affiliated to Zhengzhou University from January 2021 to January 2022 were selected. Trabeculectomy combined with phacoemulsification and intraocular lens implantation was carried out in the acute attack eyes, and phacoemulsification and intraocular lens implantation were carried out in the contralateral eyes. The ocular surface disease index(OSDI)questionnaire, noninvasive first tear film break-up time(NifBUT), noninvasive average tear film break-up time(NiaBUT)and tear meniscus height(TMH)were assessed preoperatively and 1, 3 and 6mo postoperatively.RESULTS: The OSDI scores of the included patients at 1 and 3mo postoperatively(14.72±3.07, 11.39±2.24)were significantly higher than those preoperatively(9.68±1.98; all P<0.0083), and there was no significant difference between 6mo postoperatively(10.18±1.84)and preoperatively. NifBUT of the acute attack eyes at 1 and 3mo postoperatively was significantly lower than that preoperatively, and NiaBUT of the acute attack eyes at 1, 3 and 6mo postoperatively was significantly lower than that preoperatively(all P<0.0083). The NifBUT and NiaBUT of the contralateral eyes at 1mo postoperatively were significantly lower than those preoperatively(all P<0.0083), and there was no significant difference between 3 and 6mo postoperatively and preoperatively. There was no significant difference in TMH of the attack eyes and the contralateral eyes postoperatively and preoperatively(P>0.05).CONCLUSION: The stability of tear film after surgery of PACG and cataract is decreased. The acute attack eye needs 6mo or even longer to recover, while the contralateral eye needs roughly 3mo.

Objective To explore the role and mechanism of microRNA-204(miR-204) carried by the exosomes of human umbilical cord-derived mesenchymal stem cells(hUC-MSC) in regulating the polarization of macrophages in a mouse model of myocardial ischemia-reperfusion(I/R) injury. Methods After the hUC-MSCs were isolated,cultured,and identified,their adipogenic and osteogenic differentiation capabilities were determined.The exosomes of hUC-MSCs were separated by ultracentrifugation,and the expression of CD81,CD63,tumor susceptibility gene 101(Tsg101),and calnexin in the exosomes was determined by Nanoparticle Tracking Analysis software,transmission electron microscopy,and Western blotting.Three groups(hUC-MSC,miR-204 mimic,and negative control) were designed for the determination of the expression of miR-204 in the cells and their exosomes by qRT-PCR.The C57BL/6J mice were randomly assigned into a sham operation group,an I/R group,a hUC-MSC exosomes group,a negative control group,and a miR-204 mimic group.Except the sham operation group,the I/R model was established by ligating the left anterior descending artery.The echocardiography system was employed to detect the heart function of mice.HE staining was employed to observe the pathological changes of mouse myocardium.ELISA was employed to determine the levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),arginase 1(Arg-1),and IL-10 in the myocardial tissue.After the macrophages of mouse myocardial tissue were isolated,flow cytometry was employed to determine the expression of CD11c and CD206,and ELISA to measure the levels of IL-1β,TNF-α,Arg-1,and IL-10 in the macrophages. Results hUC-MSCs had adipogenic and osteogenic differentiation capabilities,and the exosomes were successfully identified.Compared with the negative control group,the miR-204 mimic group showed up-regulated expression of miR-204 in hUC-MSCs and their exosomes(P<0.001,P<0.001).Compared with the sham operation group,the modeling of I/R increased the left ventricular end-diastolic diameter(LVEDD)(P<0.001),left ventricular end-systolic diameter(LVESD)(P<0.001),myocardial injury score(P<0.001),and the levels of IL-1β(P<0.001),TNF-α(P<0.001),and CD11c(P<0.001).Meanwhile,it lowered the left ventricular ejection fraction(LVEF)(P<0.001),left ventricular fractional shortening(LVFS)(P<0.001),Arg-1(P<0.001),IL-10(P<0.001),and CD206(P<0.001).Compared with those in the I/R group,the LVEDD(P<0.001),LVESD(P<0.001),myocardial injury score(P<0.001),and the levels of IL-1β(P<0.001),TNF-α(P=0.010),and CD11c(P<0.001) reduced,while LVEF(P<0.001),LVFS(P<0.001),and the levels of Arg-1(P<0.001),IL-10(P=0.028),and CD206(P=0.022) increased in the hUC-MSC exosomes group.Compared with those in the negative control group,the LVEDD(P<0.001),LVESD(P<0.001),myocardial injury score(P=0.001),and the levels of IL-1β(P=0.048),TNF-α(P<0.001),and CD11c(P=0.007) reduced,while the LVEF(P<0.001),LVFS(P<0.001),and the levels of Arg-1(P<0.001),IL-10(P=0.001),and CD206(P=0.001) increased in the miR-204 mimic group. Conclusion The hUC-MSC exosomes overexpressing miR-204 can inhibit the polarization of macrophages in the I/R mouse model to M1-type and promote the polarization to M2-type.
Animals ; Humans ; Mice ; Disease Models, Animal ; Exosomes/pathology* ; Interleukin-10/metabolism* ; Macrophages ; Mesenchymal Stem Cells ; Mice, Inbred C57BL ; MicroRNAs/genetics* ; Myocardial Reperfusion Injury ; Osteogenesis ; Stroke Volume ; Tumor Necrosis Factor-alpha/metabolism* ; Umbilical Cord/pathology* ; Ventricular Function, Left

Objective:To explore the mechanism of resveratrol （RSV） in the treatment of lung adenocarcinoma （LUAD） based on bioinformatics and molecular biology. Method:The targets of RSV were retrieved from DrugBank and then imported into STRING for constructing a protein-protein interaction （PPI） network.TCGA database was utilized to analyze the expression of target genes in tumor and normal tissues， followed by the prediction of their impacts on tumor occurrence and development and the screening of target genes using random forest and univariate Cox regression models.With the results of bioinformatics taken into consideration， the mechanism of RSV in inhibiting LUAD was further explored by molecular biology. Result:Ten Hub genes were screened out from the PPI network of RSV targets.Among them， solute carrier family 2 member 1 （SLC2A1）， arachidonate 5-lipoxygenase （ALOX5）， peroxisome proliferative activated receptor gamma （PPARG）， and arachidonate 15-lipoxygenase （ALOX15） differed significantly in their expression in tumor and normal tissues.As revealed by random forest and univariate COX regression analysis， SLC2A1 was of great significance to the survival and prognosis of patients with LUAD.The survival analysis through Kaplan-Meier （KM） plotter indicated that the SLC2A1 expression was closely related to the overall survival （OS）， first progression （FP）， and post-progression survival （PPS） of LUAD patients.The molecular biological experiments further proved that RSV inhibited the proliferation and migration of LUAD cells by reducing the expression of SLC2A1.As verified by immunohistochemical scoring， SLC2A1 protein expression in tumor tissue was significantly different from that in normal tissue. Conclusion:RSV inhibits the proliferation and migration of LUAD cells by reducing the expression of SLC2A1， which has far-reaching significance in the clinical treatment of LUAD.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Pneumonia ; diagnosis ; drug therapy ; epidemiology ; Retrospective Studies

OBJECTIVETo investigate the expression of aquaporin 3, 4, and 8 in the colonic mucosa of rat models with slow transit constipation (STC).

METHODSSTC rat model was established by giving the rats the compound solution of diphenoxylate. Real time polymerase chain reaction (RT-PCR) was used to measure the expression of aquaporin mRNA in colonic mucosa of STC rat models (study group,n=16) and normal rats (control group,n=16). Gray scale ratio of aquaporin to β-action (internal reference) was used for quantification.

RESULTSRT-PCR revealed that the mean gray scale ratios of aquaporin 3 in the proximal colon of the study group and control group were 0.344 and 0.602 (P<0.05), and were 0.419 and 0.509 in the distal colon (P>0.05), respectively. The mean gray scale ratios of aquaporin 4 in the proximal and the distal colon were 0.764 and 0.759 in the study group (P>0.05), and were 0.776 and 0.736 in the control group (P>0.05), respectively. However, there was no expression of aquaporin 8 in the proximal and the distal colon in either the study group or the control groups.

CONCLUSIONSExpression of aquaporin 3 in the proximal colon of STC rat models is down-regulated, which regulates water absorption. There are no significant changes in the expressions of aquaporin 4 and 8.

Animals ; Aquaporin 3 ; metabolism ; Aquaporin 4 ; metabolism ; Aquaporins ; metabolism ; Colon ; metabolism ; Constipation ; metabolism ; Disease Models, Animal ; Female ; Intestinal Mucosa ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective To investigate immunological mechanism underlying leflunomide ( LEF ) against experimental allergic encephalomyelitis ( EAE) by studying the effects of LEF on peripheral blood CD28/CTLA-4 costimulatory molecules expression in guinea pig with EAE.Methods 50 adult female guinea pigs were randomly divided into normal control group,low dose group,EAE control group,middle dose group,and high dose group, 10 guinea pigs in each group respectively.Low, medium and high dose group were treated with LEF 10 mg/kg· D, 20 mg/kg· D and 40 mg/kg· D orally, 1 times/day, to the termination of the experiment.The expression of CD28, CTLA-4 were detected by flow cytometry, incidence was recorded at the same time.Results Compared with EAE control group,high dose and middle dose group incubation period were prolonged, progress period were shorten and neurological dysfunction score decreased(P<0.05).Compared with normal control group, the expression of CD28 of EAE control group increased and the decreased expression of CTLA-4 (P<0.05).High, middle dose treatment group compared with EAE control group CD28 down regulated expression (P<0.05), but the expression of CTLA-4 increased(P<0.05).Among the treatment groups, of CD28 molecules with high dose group decreased significantly(P<0.05), the expression of CTLA-4 molecules with high dose treatment groups upregulated significantly (P<0.05).Correlation analysis showed that the EAE control group, the treatment group the CD28/CTLA-4 ratio in peripheral blood and nerve dysfunction score is proportional ( r=0.85, P=0.002; r=0.77, P=0.000).Conclusion LEF can reduce EAE guinea pig neurological symptoms, the better and the higher dose effect.Its mechanism may be through down-regulation of CD28 molecules in peripheral blood, upregulation of the expression of CTLA-4 molecule.so as to reduce the inflammatory response, relieve the clinical symptoms.