Objective To investigate the effect of diosgenin on the proliferation and apoptosis of breast cancer cells and its potential molecular mechanism. Methods The breast cancer cell line MCF-7 was treated with low, medium, and high doses of diosgenin, and cell proliferation was detected through the MMT method. Flow cytometry was used to detect cell apoptosis. Nuclear-cytoplasmic-protein separation method was applied to detect the subcellular localization of death associated protein (DAXX). qRT-PCR and Western blot were used to detect the expressions of DAXX and c-Jun N-terminal kinase pathway (JNK)-related proteins. Results Diosgenin considerably inhibited the proliferation of MCF-7 cells and promoted cell apoptosis in a concentration-dependent manner. Diosgenin can promote the movement of DAXX from nucleus into the cytoplasm. Diosgenin upregulated the expression of cell surface death receptor (Fas), increased the phosphorylation levels of JNK and mitogen activated protein kinase (p38), and activated the JNK/p38 signaling pathway with concentration dependence. Conclusion Diosgenin inhibits the proliferation and promotes the apoptosis of the breast cancer cell line MCF-7, whose mechanism may be related to the regulation of DAXX subcellular localization and the activation of JNK/p38 signaling pathway.

Protein tyrosine phosphatase nonreceptor type 2 (PTPN2) is a promising target for sensitizing solid tumors to immune checkpoint blockades. However, the highly polar active sites of PTPN2 hinder drug discovery efforts. Leveraging small interfering RNA (siRNA) technology, we developed a novel glutathione-responsive nano-platform HPssPT (HA/PEIss@siPtpn2) to silence PTPN2 and enhance immunotherapy efficacy in hepatocellular carcinoma (HCC). HPssPT showed potent transfection and favorable safety profiles. PTPN2 deficiency induced by HPssPT amplified the interferon γ signaling in HCC cells by increasing the phosphorylation of Janus-activated kinase 1 and signal transducer and activator of transcription 1, resulting in enhanced antigen presentation and T cell activation. The nano-platform was also able to promote the M1-like polarization of macrophages in vitro. The unique tropism of HPssPT towards tumor-associated macrophages, facilitated by hyaluronic acid coating and CD44 receptor targeting, allowed for simultaneous reprogramming of both tumor cells and tumor-associated macrophages, thereby synergistically reshaping tumor microenvironment to an immunostimulatory state. In HCC, colorectal cancer, and melanoma animal models, HPssPT monotherapy provoked robust antitumor immunity, thereby sensitizing tumors to PD-1 blockade, which provided new inspiration for siRNA-based drug discovery and tumor immunotherapy.

Objective:To grasp the distribution of fine antigenic epitope profiles of nucleoprotein (NP) and glycoprotein (GP) fragments of Crimean-Congo hemorrhagic fever virus (CCHFV) and to clarify the value of dominant antigenic epitopes in laboratory testing of Crimean-Congo hemorrhagic fever (CCHF).Methods:In a minimal synthetic short peptide consisting of 8 amino acids was segmentally expressed by CCHFV YL04057 strain using a modified bio-peptide synthesis method from 2014 to 2021 in the laboratory of Xinjiang University, College of Life Sciences. Using CCHFV polyclonal antibody or monoclonal antibody 14B7 (IgM) or CCHFV-positive sheep serum as antibodies, the minimal antigenic epitopes (BCEs) with antigenic activity on NP and GP fragments were identified by immunoblotting, and the obtained BCEs with sequence polymorphism were spatially clustered with CCHFV from different regions using the neighbor-joining method to determine the combination mode of BCEs with geographical correlation of regional distribution, to explore its application in establishing serological diagnosis. A prokaryotic expression plasmid (pET-32a), an E. coli expression plasmid (pGEX-KG) and a prokaryotic expression plasmid with an incomplete glutathione (GST188) tag (pXXGST-ST-1) were used to construct and express six dominant antigenic epitopes of different peptide lengths on NP fragments, and an indirect Enzyme-linked immunosorbent assay (ELISA) was established. CCHF sheep serum identified by immunofluorescence assay (IFA) was used as a control, and the specificity, sensitivity and overall compliance of the recombinant proteins with different peptide lengths of antigenic epitopes with IFA assay results were statistically analyzed. Results:CCHFV, NP and GP fragments had a total of 30 antigenically active BCEs, among which the core intermediate fragment NP2 (aa 170 th-305 th), which had a concentration of antigenic epitopes in the NP fragment, has 6 BCEs, and the NP1 (aa 1 st-200 th) and NP3 (aa 286 th-482 nd) at both ends have 9 BCEs; the Gc (aa 1 st-558 th) and Gn (aa 533 th-708 th) fragments of the GP fragment have 14 BCEs and a long antigenic peptide (AP) containing 15 amino acids, and the amino acid sequence homology of the NP fragment BCEs was 97.1% and that of the GP fragment BCEs was 89.1%. There was a significant difference ( P=0.0281, P<0.05). Among the 9 BCEs with sequence polymorphism in the GP fragment, 6 combined BCEs from GnEc1, GnE2, GnE4, GcE3, GcE6 and GcAP-4 (Ap) could cluster 15 CCHFV strains from different regions of the world into 5 geographical taxa, AsiaⅠ, AsiaⅡ, AficaⅠ, AficaⅡ and Europe. The constructs expressing PET-32a-NP (full length), PGEX-KG-NP2 (aa 170 th-305 th), pGEX-KG-NP2-1 (aa 235 th-275 th), PGEX-KG-NP2-1-1 (aa 237 th-256 th), pXXGST-1-NP2-1-2 (aa 250 th-265 th) and PGEX KG-NP2-1-3 (aa 260 th-276 th), six recombinant proteins CCHFV NP rabbit polyclonal antiserum (pAb) Western Blotting reaction positive, 33 sheep sera tested by IFA XHF as a reference, the sensitivity of the assay established by indirect ELISA using the recombinant proteins constructed from two fragments of NP2 and NP2-1 as antigens. The sensitivity, specificity and overall compliance were the best, with 73.4% (11/15) and 66.7% (10/15) for sensitivity, 100% (18/18) and 94.4% (17/18) for specificity, and 87.9% (29/33) and 81.8% (27/33) for overall compliance. Conclusion:CCHFV NP and GP are distributed with a high number of BCEs with antigenic immunoreactivity, among which the dominant antigenic epitopes are of high value in the laboratory serological diagnosis of CCHF.

Objective:To investigate the predictive value of the National Institutes of Health Stroke Scale (NIHSS) score at 24 h after endovascular treatment on the outcomes in patients with acute basilar artery occlusion (ABAO).Methods:Consecutive patients with ABAO received endovascular treatment at the Department of Neurology, Shengli Oilfield Central Hospital from January 2019 to December 2020 were retrospectively included. According to the modified Rankin Scale scores at 90 days after onset, the patients were divided into a good outcome group (0-3) and a poor outcome group (4-6), as well as a survival group and a death group. The demographic and clinical data between the groups were compared respectively. Multivariate logistic regression analysis was use to identify independent influencing factors for clinical outcomes and mortality. The predictive value of postprocedural 24 h NIHSS score on the outcomes was evaluated using the receiver operating characteristic (ROC) curves. Results:A total of 35 patients with ABAO were included. Their age was 62 years (interquartile range, 56-66 years), and 28 patients were males (80%); 19 (54.3%) had a good outcome, 16 (45.7%) had a poor outcome, and 7 (20.0%) died. Univariate analysis showed that there were statistically significant differences in hypertension, low-density lipoprotein cholesterol, fasting blood glucose, collateral circulation grading, vascular recanalization, and postprocedural 24 h NIHSS scores between the good outcome group and the poor outcome group (all P<0.05). Multivariate logistic regression analysis showed that the postprocedural 24 h NIHSS score was independently correlated with the poor outcome (odds ratio 1.131, 95% confidence interval 1.017-1.258; P=0.023). Multivariate analysis did not find the independent influencing factors for death. ROC curve analysis showed that the area under the curve of the postprocedural 24 h NIHSS score for predicting poor outcome was 0.814 (95% confidence interval 0.668-0.960; P=0.011). The optimal cutoff value was 19 points, and the corresponding sensitivity and specificity were 85.7% and 71.4% respectively. Conclusions:In patients with ABAO receiving endovascular treatment, the postprocedural 24 h NIHSS score has good predictive value for poor outcomes at 90 d after procedure.

Abstract: To investigate the effect and mechanism of pituitary adenylate cyclase activated peptide recombinant 13 peptide(PACAP13) on thymus at cell, organ and individual levels. Methods: Thymic epithelial cells(TEC) and bone marrow mesenchymal stem cells(MSC) were culturedin vitro, lymphocyte group, lymphocyte+TEC group, lymphocyte+MSC group and lymphocyte+TEC+MSC group were set up to detect the proliferation of lymphocytes in each group treated by PACAP13 peptide, and contents of cytokines interleukin(IL)-1, IL-2, IL-6 and transforming growth factor(TGF) in culture medium were detected. Dexamethasone(Dex) was added to induce cell apoptosis, and Annexin V-FITC/PI flow cytometry double staining was used to detect the effect of PACAP13 peptide on cell apoptosis. Thymus group, thymus+ TEC group, thymus+ MSC group, thymus+ TEC+ MSC group were set up, and PACAP13 peptide was added, the content of signal joint T cell receptor rearrangement excision cricles(sjTREC) in thymus was detected by real-time quantitative PCR(qRCR). The pristane-induced lupus model mice were randomly divided into model group and PACAP13 group. Model group was injected with 0.9% saline, and PACAP13 group was injected with PACAP13 peptide, thymus and spleen sjTREC levels were detected by qPCR, serum lupus-related autoantibodies were detected by ELISA and thymus-related mRNA expression levels were detected by reverse transcription quantitative PCR(qRT-RCR). Results: Compared with the lymphocyte group, the proliferation rate of thymus lymphocytes in the lymphocyte+TEC group, lymphocyte+ MSC group, lymphocyte+ TEC+ MSC group increased, and the apoptosis rate decreased(P<0.05), compared with the control group, the proliferation rate of lymphocytes in the PACAP13 group increased, the rate of apoptosis decreased, and the cytokines IL-1, IL-2, IL-6 and TGF in the cell culture medium increased(P<0.05). Compared with the thymus group, thymus+ TEC group, thymus+ MSC group, thymus+ TEC+ MSC group increased the content of thymus sjTREC, compared with the control group, PACAP13 peptide group thymus sjTREC content increased. Compared with the model group, the content of sjTREC in the thymus and spleen of lupus mice in the PACAP13 group increased(P<0.05), thymus mRNA expression level of AIRE, Fezf2, Foxp3 and TGF-β increased(P<0.01), and CXCL13 mRNA expression level decreased(P<0.01), the autoantibodies of anti-dsDNA antibody, anti-RNP/sm antibody and ANA decreased in serum(P<0.05). Conclusion Compared with the lymphocyte group, the proliferation rate of thymus lymphocytes in the lymphocyte+TEC group, lymphocyte+ MSC group, lymphocyte+ TEC+ MSC group increased, and the apoptosis rate decreased(P<0.05), compared with the control group, the proliferation rate of lymphocytes in the PACAP13 group increased, the rate of apoptosis decreased, and the cytokines IL-1, IL-2, IL-6 and TGF in the cell culture medium increased(P<0.05). Compared with the thymus group, thymus+ TEC group, thymus+ MSC group, thymus+ TEC+ MSC group increased the content of thymus sjTREC, compared with the control group, PACAP13 peptide group thymus sjTREC content increased. Compared with the model group, the content of sjTREC in the thymus and spleen of lupus mice in the PACAP13 group increased(P<0.05), thymus mRNA expression level of AIRE, Fezf2, Foxp3 and TGF-β increased(P<0.01), and CXCL13 mRNA expression level decreased(P<0.01), the autoantibodies of anti-dsDNA antibody, anti-RNP/sm antibody and ANA decreased in serum(P<0.05).

Objective To observe the preventing effect of optimal programs (improving turn over postures and elongating the turn over intervals)in preventing pressure sore occurrence in patients with acute stroke. Methods Acute stage stroke patients (time from on-set less than or equal to one week)were selected and divided randomly into 4 groups, one group adopted 2.0 hours turn over intervals as the routine group, others adopted 2.5 hours, 3.0 hours, 3.5 hours turn over intervals as the study groups. The incidence of pressure sores and complications were recorded. Results None of the groups (2.0 h, 2.5 h,3.0 h, 3.5 h)suffered from pressure sores. The incidence of complications including malnutrition, pulmonary infections and constipations was not increased in the study groups either. Conclusions Optimal turn over nursing programs including elongating turn over intervals to 2.5h, 3.0h, 3.5h and adopting 30 degree lateral posture can effectively prevent occurrence of pressure sores in acute stroke patients.

OBJECTIVETo compare the ettect and side-ettect of fluoxetine and combination of fluoxetine and Chinese or Tibetan medicine in treating senile depression in plateau district. Methods Ninety patients with diagnosis of senile depression conformed to CCMD-3 standard, in plateau district of 2260 - 3200 m altitude were randomly divided into three groups and treated with fluoxetine (group A), fluoxetine plus Sanpu Xinnao Xin granule (group B) and fluoxetine plus Xiaoyao pill (group C), respectively, 30 cases in each group. Therapeutic effects were evaluated with Hamilton' s depressive scale (HAMD) and treatment emergent symptom scale (TESS) after 6 weeks treatment.

RESULTSThere was no significant difference in the therapeutic effects between the three groups. The adverse reaction in Group B and C was less than that in Group A (P<0.01). Conclusion Sanpu Xinnao Xin granule and Xiaoyao pill can raise the tolerance of patients with senile depression in plateau area against the adverse reaction of fluoxetine.

Aged ; Altitude ; Antidepressive Agents, Second-Generation ; therapeutic use ; Depressive Disorder ; drug therapy ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Fluoxetine ; therapeutic use ; Humans ; Middle Aged ; Phytotherapy ; Serotonin Uptake Inhibitors ; therapeutic use

This study investigated the feasibility of using an hammerhead ribozyme against C II TA, a major regulator of MHC II antigens, to repress the expression of MHC II molecules on Hela cells. A hammerhead ribozyme (Rz464) specific to 463-465 GUC triplet of C II TA and its target gene were transcribed, then mixed up and incubated in vitro. The cleavage products were analyzed by PAGE and silver-staining. Rz464 was then inserted into the pIRES2-EGFP vector (pRz464). Stable transfectants of Hela with pRz464 were tested for class II MHC induction by recombinant human interferon-gamma (IFN-gamma). mRNA of C II TA was measured by RT-PCR. Our results showed that Rz464 could exclusively cleave C II TA RNA. When induced with IFN-gamma, the expression of HLA-DR, -DP, -DQ on pRz464+ Hela was induced, and the mRNA content of C II TA decreased too. It is concluded that Rz464 could inhibit C II TA and thus the family of genes was regulated by C II TA:MHC II molecules. These results provided insight into the future application of Rz464 as a new nucleic acid drug against auto-immune diseases.
Autoimmune Diseases ; therapy ; Base Sequence ; Genetic Therapy ; HLA-DP Antigens ; metabolism ; HLA-DQ Antigens ; metabolism ; HLA-DR Antigens ; metabolism ; HeLa Cells ; Humans ; Interferon-gamma ; pharmacology ; Molecular Sequence Data ; Nuclear Proteins ; immunology ; RNA, Catalytic ; genetics ; metabolism ; pharmacology ; physiology ; Recombinant Proteins ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; antagonists & inhibitors ; genetics ; immunology ; Transfection

Objective To investigate the feasibility of using anti-sense RNA against classⅡmajor histocompatibility complex(MHCⅡ)transactivator(CⅡTA),which might regulate MHCⅡexpression,to suppress the relative immune response.Methods Stable transfectants of dermal fibroblasts with pDarⅡ(pDarⅡ-D)were tested for the expression of classic MHCⅡ(HLA-DR,-DP,-DQ)antigens induced with recombinant human interferon-gamma(IFN-?).mRNA abundance of CⅡTA,and classic MHCⅡwas mea-sured by RT-PCR.IL-2mRNA expressed in T cells,stimulated by transfected dermal fibroblasts,was de-termined by mixed lymphocyte reaction.Results When induced with IFN-?,the expression of HLA-DR and-DP antigens on pDarⅡ-D was reduced by95.63%and87.89%,respectively.Meanwhile,the mRNA contents of CⅡTA and classic MHCⅡwere decreased significantly(P

Objective To analyze the efficacy and debase the toxicity of radiotherapy combined with composite Kushen injection in treatment of esophageal carcinoma patients. Methods A total of 126 cases of esophageal cancer were treated by radiotherapy were analysed. All patients were treated with the continuative hyperfractionated radiotherapy. The total dose was 66 ~ 74 Gy/5 ~ 7 wk, 2.2 ~ 2.4 Gy/d of esophageal carcinoma patients. Two groups were given respectively delivered with the same radioactive dosages and methods.But the synthetic group was treated by radiotherapy with composite Kushen injection. composite Kushen injection was given at the dosage of 20 ml with 500 ml of saline infusion for 20 consecutive days. Results X-ray grades of esophageal cancer after radiotherapy. The grade I of X-ray manifestation after radiotherapy for patients in synthetic group was superior to that patient in the radiotherapy alone group. (60.4 % vs 41 %; ?2 =4.57, P