Objective: To explore the mechanism of triptolide in the treatment of rheumatoid arthritis (RA) and analyze its safety. Methods: A total of 60 rats were randomly divided into control group, model group, methotrexate (MTX) group and triptolide (TP) low, medium and high dose groups with 10 rats in each group. In addition to the control group, the rats in the other groups were established type II collagen-induced RA model. After the successful establishment of the model, rats in MTX group were given 0.4 mg/kg MTX by gavage from the 3rd week. Rats in TP high, middle, and low dose groups were given 0.1, 0.2, and 0.4 mg/kg TP by gavage. Rats in control group and model group were given equal volume distilled water once a day for 4 weeks. The paw swelling of rats in each group was compared. The percentage of CD4+CD25+and CD4+Foxp3+ Treg was detected by flow cytometry. The levels of IL-10, IL-17, TNF-α, VEGF, IFN-γ, TGF-β, ALT, and AST in the serum were detected. The pathological morphology of ankle joint was observed under microscope. The expression levels of IL-10, IL-17, TNF-alpha, VEGF, IFN-γ, and TGF-β were detected by immunohistochemistry. Results: Pathological sections showed that synovial cells were proliferated significantly in the ankle joint of rats in the model group, with infiltration of a large number of inflammatory cells such as monocytes and lymphocytes, new capillaries, thinning of bone trabeculae and serious erosion of cartilage surface. The degree of pathological changes in other groups was significantly less than that in model group. After treatment, the degree of joint swelling and arthritis index in MTX and TP groups were significantly decreased compared with those before treatment (P < 0.05). Compared with model group, CD4+Foxp3+Treg and CD4+CD25+Treg were increased in MTX group and TP all dose groups (P < 0.05). Compared with the model group, the serum levels of IL-10, TGF-β in MTX and TP all dose groups were increased, while the levels of IL-17, TNF-α, VEGF and IFN-γ were decreased (P < 0.05). Compared with model group, the expressions of IL-10 and TGF-β in ankle joint tissue of rats in MTX and TP all dose groups were increased significantly, while the expressions of IL-17, TNF-α, VEGF and IFN-γ were decreased significantly (P < 0.05). Compared with control group or model group, there was no significant difference in serum ALT and AST levels between MTX group and TP all dose groups (P > 0.05). There was no significant difference between MTX group and TP high dose group (P > 0.05). Conclusion: TP is effective in treating type II collagen-induced arthritis in rats, which can significantly improve joint swelling. Its mechanism is related to promoting the expression of IL-10 and TGF-β, increasing the proportion of Treg cells, inhibiting the expression of IL-17, TNF-α, VEGF and IFN-γ, and has no obvious hepatotoxicity.

OBJECTIVE: To investigate inhibitory effect of FLOT1 gene expression on invasion and apoptosis of gastric cancer cells. METHODS: The designed FLOT1 siRNA lentiviral vector (si-FLOT1 group) was transfected into human gastric cancer MKN-45 cells, at the same time, the negative control lentivirus vector (negative group) was transfected, and the blank group was set up. Western blotting was used to detect the expression of FLOT1, E-cadherin, α-SMA, cleaved caspase 3, Bcl-2 and Bax proteins. After cells were transfected for 24, 48, 72 and 96 h, cell viability was detected by CCK-8 assay. After cells were transfected for 48 h, transwell chamber, flow cytometry and DCFH-DA assay were used to detect the invasiveness, apoptosis rate and ROS content, respectively. RESULTS: FLOT1 siRNA lentiviral vector inhibited significantly the expression of FLOT1 in MKN-45 cells, which was significantly different from the blank group (P<0.05). Compared with si-FLOT1 group and the blank group, the cell vitality decreased, the invasion ability decreased, the apoptosis rate increased, the expression of E-cadherin, cleaved caspase 3 and Bax protein increased, the expression of α-SMA and Bcl-2 protein decreased, and the content of ROS increased, and the difference was statistically significant (P<0.05). CONCLUSION: Down regulation of FLOT1 gene expression can reduce the invasiveness of gastric cancer cells by inhibiting EMT, and promote apoptosis by regulating the expression of apoptosis related proteins and increasing the ROS content of cells.

Postoperative intra-abdominal adhesion is one of the most common complications in the postoperative period. Current remedies are very ineffective to prevent the pathological outcomes except steroid hormones. Rhynchophylline is deemed as a pharmacologically active component from traditional Oriental medicine Uncaria rhynchophylla (Miq.) Jacks. (Rubiaceae). This study was designed to investigate the preventative effect of rhynchophylline on the abdominal adhesions in rats. Rhynchophylline relieved the experimental abdominal adhesion and decreased the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the blood serum in a dose-dependent manner. The levels of transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF) were reduced significantly in the peritoneal fluid. The potential mechanism of the activity is related to inhibition of the TGF-β1/Smad signaling pathway.

OBJECTIVETo investigate the effects of mannan-binding lectin (MBL) on the maturation and cytokine secretion of human dendritic cells (DC) induced by Candida albicans (C. albicans).

METHODSThe plastic-adherent mononuclear cells were prepared from the blood of healthy adult volunteers. The human peripheral blood mononuclear cells-derived dendritic cells (MNC-DC) were induced by 5-day-culture in medium supplemented with rhGM-CSF and rhIL-4, and then cultured for 2 days in presence or absence of C. albicans at varying concentration of human MBL ranging from 1 to 20 mg/L. DC's shape and characters were observed under inverted microscopy, the expression of CD83 and CD86 on DC was analyzed by FACS. The levels of TNF-α and IL-6 were detected by ELISA. FACS also was used to investigate the interaction of MBL with immature DC(imDC) and C. albicans. Western blot was used to detect C. albicans-induced IκBα phosphorylation and p65/NF-κB translocation in DC.

RESULTSMBL at higher concentrations (10-20 mg/L) down-regulated the expression of CD83 and CD86 on the monocyte-derived dentritic cells(MoDC) induced by C. albicans, and inhibited the production of TNF-α and IL-6 induced by C. albicans. FACS showed that MBL could not only bind to C. albicans but also bind to imDCs in a Ca2+-dependent manner. Western blot showed that MBL could decrease the phosphorylation of IκBα and the nuclear translocation of p65/ NF-κB.

CONCLUSIONMBL may inhibit TNF-α and IL-6 production induced by C. albicans in DC through NF-κB signaling pathways, suggesting that MBL can play some roles in the regulation of C. albicans-induced immune response.

OBJECTIVE: To investigate the effect of sulforaphane (SFN) on G METHODS: KG1a and KG1cells were treated by different concentrations of SFN for 48 h. Flow cytometry (FCM) was used to analyze the phase distribution of cell cycle. High-throughput sequencing was used to detect the effect of SFN on the expression of cell cycle related genes in KG1a cells. The mRNA expression of P53, P21, CDC2 and CyclinB1 were detected by qPCR. The protein expression of P53, CDC2, P-CDC2 and CyclinB1 were detected by Western blot. RESULTS: Cells in the G CONCLUSION SFN induces leukemia cells to block in G
Cell Cycle ; Humans ; Isothiocyanates/pharmacology* ; Leukemia, Myeloid, Acute ; Mitosis ; Sulfoxides

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*

【Objective】 To investigate the value of urodynamic study (UDS) in guiding the treatment of lower urinary tract dysfunction (LUTD) in elderly patients with ischemic stroke (IS) during convalescence, in order to provide reference for clinical treatment. 【Methods】 A total of 50 LUTD patients with IS who were admitted to the First Affiliated Hospital of Xinxiang Medical University during Jan.2020 and Jan.2022 were selected.Oral tolterodine was administered to patients with detrusor overactivity (DO), clean intermittent catheterization (CIC) to those with no detrusor reflex and symptomatic increased residual urine, and oral administration of tamsulosin to those with functional obstruction of bladder outlet.The lower urinary tract symptoms (LUTS) relief rate, UDS parameters and quality of life (QoL) scores were compared before treatment and 3 months after treatment. 【Results】 The UDS examination results showed that 25 cases (50.0%) had simple DO, 9 cases (18.0%) had DO with impaired detrusor muscle contraction function, 5 cases (10.0%) had DO with bladder outlet functional obstruction, 4 cases (8.0%) had no detrusor reflex, and 7 cases (14.0%) had simple bladder outlet functional obstruction.After 3 months of treatment, the symptoms of LUTS, including frequent urination, urgent urination, incontinence, dysuria and urinary retention were significantly improved (P<0.05).The maximum urine flow rate and urine output were significantly increased, the residual urine volume was significantly reduced, QoL scores were significantly reduced, with significant differences (P<0.001). 【Conclusion】 UDS is significant in guiding the treatment of LUTD in elderly patients with IS during convalescence.

Central nervous system involvement in primary Sjögren's syndrome (pSS) is less common and usually presents as white matter lesions, neuromyelitis optica spectrum disorder (NMOSD), or transverse myelitis. NMOSD is an immune-mediated inflammatory demyelinating disease of the central nervous system with a high rate of relapse and significant disability. Studies have shown that patients with pSS combined with NMOSD have more severe symptoms and poorer prognosis. Here, we present a case of critical illness in pregnancy-associated NMOSD combined with Sjögren's syndrome. The patient was a 30-year-old pregnant woman with a history of Sjögren's syndrome who was diagnosed with NMOSD. She received combination therapy with steroids, intravenous immunoglobulin (IVIG), and hydroxychloroquine during pregnancy, resulting in partial resolution of numbness below the waist. However, due to irregular medication adherence outside the hospital setting, she developed weakness in her right lower limb accompanied by inability to move it, while her left lower limb still had some mobility but occasional numbness along with urinary and fecal incontinence. Ten days later, she was admitted to the emergency department where an emergency cesarean section was performed to deliver a healthy baby boy. However, her condition worsened postpartum as she developed high fever accompanied by bilateral lower limb paralysis and weakness along with loss of voluntary control over urination and defecation. The patient underwent ano-ther course of treatment consisting of steroids and IVIG; however there was limited improvement in symptoms observed after this intervention. Following administration of rituximab for the first time, the patient developed urinary tract infection which was successfully managed before continuing regular infusions. In later stages the patient could walk slightly with a limp and regained control over urination and defecation, allowing her to resume normal activities. This case suggests that combination therapy with steroids, IVIG, and hydroxychloroquine should be considered for the patients with pregnancy-associated NMOSD combined with Sjögren's syndrome. Rituximab can significantly improve symptoms such as postpartum paralysis in patients with NMOSD, however, there may be a risk of infection associated with its use.
Adult ; Female ; Humans ; Pregnancy ; Cesarean Section/adverse effects* ; Critical Illness ; Hydroxychloroquine/therapeutic use* ; Hypesthesia/complications* ; Immunoglobulins, Intravenous/therapeutic use* ; Inflammation/complications* ; Neuromyelitis Optica/diagnosis* ; Paralysis/complications* ; Pregnancy Complications/therapy* ; Rituximab/therapeutic use* ; Sjogren's Syndrome/complications* ; Steroids/therapeutic use* ; Vision Disorders

Objective: To investigate the effects of interferon inducible protein 16 (IFI16), a cytosolic DNA sensor, on the expression of human T-cell leukemia virus type 1 (HTLV-1) proteins and pro-inflammatory cytokines in adult HTLV-1-positive T cells. Methods: IFI16 expression in different HTLV-1-positive T cell lines was detected by immunoblot assay. Specific siRNA targeting the IFI16 gene was constructed and the gene silencing efficiency was detected by immunoblot assay. Expression of HTLV-1 Tax protein at mRNA and protein levels was respectively detected by real-time PCR and immunoblot assay after knocking down the expression of IFI16 in HTLV-1-positive T cells with siRNA. Expression of interferon (IFN)-α, IFN-γ, tumor necrosis factor (TNF)-α, Tax and Env were detected by real-time PCR. Results: Compared with the HTLV-1-negative T cell line Jurkat, IFI16 expression was enhanced in the HTLV-1-positive T cell lines MT2, MT4 and C8166. Tax expression was increased, while that of IFN-α, IFN-γ and TNF-α was decreased in MT2 and MT4 cells after silencing the expression of IFI16 with siRNA. Conclusions IFI16 expression was increased in HTLV-1-positive MT2 and MT4 cells. Meanwhile, IFI16 promoted the production of interferon and pro-inflammatory cytokines and inhibited the expression of HTLV-1 proteins.

OBJECTIVETo investigate the effects of arsenic trioxide (AsO) on K562 cell proliferation by regulating cell cycle protein D1 and cyclin-dependent kinase inhibitor p27kip1.

METHODSMTT was used to detect the effect of AsOon K562 cell proliferation, so as to screen out the appropriate drug concentration. Furthermore, the K562 cell apoptosis was observed by microscopy. The expression of CyclinD1 and p27kip1 in K562 cells treated with AsOwas analyzed by reverse transcription-polymerase chain reaction(RT-PCR), immunohistochemistry and Western blot.

RESULTSAsOcould inhibit the proliferation of K562 cells in a dose- and time- dependent manner (r= 0.967). And the apoptosis cell number in AsOgroup was significantly higher than that in the control group(P<0.05). AsOcould markedly inhibit the expression of CyclinD1 in K562 cells(P<0.05), but the expression of P27kip1 was not significantly changed after AsOtreatment.

CONCLUSIONSAsOcan induce K562 cell apoptosis and inhibit K562 cell proliferation by regulating the expression of CyclinD1.