Progress of ruxolitinib in the treatment of hemophagocytic lymphohistiocytosis / 国际儿科学杂志

Hemophagocytic lymphohistiocytosis (HLH) is a rare immune-mediated disorder characterized by hyperactivation of antigen-presenting cells and T cells, massive secretion of inflammatory cytokines, and impaired function of natural killer cells and CD8 + T cells.Ruxolitinib is a Januse kinase(JAK)inhibitor that reduces cytokine release and retards the inflammatory response by competitive binding to the JAK catalytic site, to achieve the goal of curing HLH.In recent years, ruxolitinib has been gradually applied in the treatment of HLH, and its effectiveness has also been verified.However, studies have also found that there are efficacy differences in the treatment of HLH caused by different etiologies.This article reviews the mechanism of ruxolitinib in the treatment of HLH and the differences in the efficacy of ruxolitinib in the treatment of HLH of different etiologies.

Targeting the JAK-STAT signaling pathway in the treatment of psoriasis / 中华皮肤科杂志

The Janus kinase (JAK) -signal transducer and activator of transcription (STAT) signaling pathway is closely related to the occurrence of psoriasis. Various cytokines, including interleukin (IL) -23, IL-22, interferon (IFN) -γ, etc., can promote some key pathologic processes (such as the proliferation and abnormal differentiation of keratinocytes, and infiltration of inflammatory cells) via the JAK-STAT pathway in psoriasis, which suggests that targeting JAK-STAT pathway is a new strategy for the treatment of psoriasis. In recent years, small-molecule JAK inhibitors have shown good efficacy and safety in the treatment of psoriasis, and drugs targeting STAT pathway have been under development, which provide more treatment options for psoriasis. This review summarizes progress in drugs targeting the JAK-STAT signaling pathway in the treatment of psoriasis.

In vitro study of dendritic cell derived exosomes transporting hepatitis D antigen induced specific cytotoxic T lymphocyte response / 中华传染病杂志

Objective:To explore the effect and mechanism of dendritic cell derived exosomes (Dexs) loading ubiquitinated (Ub) hepatitis D antigen (HDAg) on activating specific cytotoxic T lymphocytes (CTL).Methods:Ub-S-HDAg-Dexs were co-cultured with dendritic cells (DC) which were from the femora of C57BL/6 mice for 48 h, then flow cytometry was used to detect the maturity of DC (CD86, CD80 and major histocompatibility complex (MHC) Ⅱ). The spleen-derived T lymphocytes from C57BL/6 mice were added in vitro to activate DC and co-cultivated for 72 h. The T cells were divided into Ub-S-HDAg-Dexs group (add 50 μg/mL Ub-S-HDAg-Dexs), Blank-Dexs group (add 50 μg/mL DC derived exosomes without plasmid transfection), Con-Dexs group (add 50 μg/mL DC derived exosomes transfected by cantrol plasmid), PBS group (add 50 μL/mL phosphate buffered saline), and Ub-S-HDAg-Dexs+ AG490 group (add 50 μg/mL Ub-S-HDAg-Dexs, DC and T lymphocytes stimulated by exosomes, and 50 μmol/L AG490 was also added to the cell mix). Flow cytometry was used to detect CD8 + T cells secreting interferon-gamma, non-radioactive lactate dehydrogenase release test to detect the killing activity of specific CTL. Real-time quantitative polymerase chain reaction (PCR) and Western blotting were used to detect the mRNA and protein expressions of JAK kinase (JAK) 2, GATA-binding protein 3 (GATA3), T-bet, signal transduction and activator of transcription (STAT) 1 and STAT4. Independent sample t test were used for statistical analysis. Results:The positive rates of the surface molecules CD80, CD86, MHCⅡof DC stimulated by Ub-S-HDAg-Dexs were 83.850%±0.219%、68.910%±0.134%、84.320%±0.445%, respectively.In the Ub-S-HDAg-Dexs group, the rate of CD8 + T cells secreting interferon-gamma was 6.420%±0.028%, which was higher than those of other groups, including PBS group, Blank-Dexs group, Con-Dexs group and Ub-S-HDAg-Dexs+ AG490 group ( t=90.78, 30.32, 63.06 and 85.42, respectively, all P<0.001). The cytotoxicity of T cells in the Ub-S-HDAg-Dexs group was 82.4%±3.9%, which was higher than those of other groups ( t=17.28, 9.74, 3.95 and 15.89, respectively, all P<0.050). The relative mRNA expressions of JAK2, T-bet, STAT1, STAT4 in Ub-S-HDAg-Dexs group were higher than those in other groups, including Con-Dexs group ( t=10.74, 32.34, 13.00 and 16.28, respectively, all P<0.001), Blank-Dexs group ( t=15.05, 21.51, 6.46 and 13.12, respectively, all P<0.050), PBS group ( t=21.83, 41.42, 7.30 and 17.50, respectively, all P<0.050), Ub-S-HDAg-Dexs+ AG490 group ( t=35.75, 20.69, 17.02 and 17.07, respectively, all P<0.001), and the differences were all statistically significant. The protein expressions of T-bet, STAT1, STAT4 in Ub-S-HDAg-Dexs group increased compared with those in PBS group ( t=346.70, 57.54 and 55.81, respectively, all P<0.001), with statistical significance. In the presence of AG490, the protein expressions of T-bet, STAT1 and STAT4 decreased compared with those in Ub-S-HDAg-Dexs group, and the differences were statistically significant ( t=355.40, 52.79 and 126.10, respectively, all P<0.001). Conclusions:Ubiquitinated HDAg transported by exosomes could effectively promote DC maturation, induce T lymphocyte differentiation, and generate specific CTL responses, which provides a new idea for the treatment of hepatitis D.

Monomorphic epithelial intestinal T-cell lymphoma: a clinicopathological characteristic analysis / 白血病·淋巴瘤

Objective:To investigate the histopathological morphology, immunophenotype, molecular pathological features, clinical prognosis and treatment of monomorphic epithelial intestinal T-cell lymphoma (MEITL).Methods:The clinicopathological data of 5 patients with MEITL in Sichuan Jinyu Medical Laboratory Center Co., Ltd from March 2019 to February 2021 were retrospectively analyzed, and literatures were reviewed. All cases were tested by using histopathology, immunohistochemistry, in situ hybridization of Epstein-Barr virus (EBV) and T-cell clonability assessment, and 1 case had second-generation sequencing (NGS) test. Clinical follow-up was performed in 2 patients.Results:All 5 MEITL cases were middle-aged and old men. The histopathology showed that intestinal wall was diffuse with tumor cells infiltrating, and the cells were obviously epitheliophilic. All the tumor cells CD3, CD8, CD56, GrB were positively expressed, and expressions of other T-cell markers were different, among which 1 case had CD30 positive and 1 case had CD20 positive. All 5 cases were negative for EBV by in situ hybridization. Monoclonal rearrangement of T-cell receptor gene was detected in all 5 cases. Mutations of BCOR, JAK3, STAT5B and ATM were detected in 1 case by using NGS. Among 2 cases followed-up, 1 patient relapsed 7 months after he had the initial onset and underwent the first operation, and then he had another operation. This patient finally died of extensive metastasis in the lung, liver and abdominal cavity as well as ascites 13 months later; another patient died 1 month after emergency surgery for perforation.Conclusions:MEITL is a rare primary T-cell lymphoma of the digestive tract. The oncogenic event in the pathogenesis of MEITL mainly involves mutations in the tumor suppressor gene SETD2 and mutations in one or more genes of the JAK/STAT pathway. Currently, there is no standard treatment for MEITL. Most treatment options include surgical resection and anthracycline-based chemotherapy.

IL-27 in combination with IL-15 regulates anti-tumor effect of NK92 cells by phosphorylating the STATs pathway / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of IL-27 in combination with IL-15 on the anti-tumor effects of NK92 cells and the possible molecular and signaling mechanisms. Methods: NK92 cells with high IL-15 expression (IL-15-NK92 cells) were cultured in different mass concentrations of IL-27 (0, 10, 20, 30 and 60 ng/ml) for 24 h. The effects of IL-27 on IL-15 secretion, migration and proliferation of IL-15-NK92 cells were detected by ELISA, Transwell and CCK-8 assay, respectively. Flow cytometry was used to detect the expression levels of IL-15-NK92 cell surface receptors NKG2D, NKp30 and NKp46, as well as the secretion levels of perforin and granzyme B. LDH method was used to detect the cytotoxic effect of IL-15-NK92 cells on hematologic tumor cells and solid tumor cells, and WB was used to detect the expressions and phosphorylation level of STATs pathway-related proteins. Results: IL-27 at the concentration of 30 ng/ml promoted IL-15-NK92 cells secreting IL-15 (P<0.01), significantly enhanced the cell migration (P<0.05) but inhibited the proliferation of IL-15-NK92 cells (P<0.05). 30 ng/ml IL-27 could significantly promote the expressions of NKG2D, NKp30 and NKp46 on surface of IL-15-NK 92 cells, as well as elevate the secretion of perforin (all P<0.05), but didn’t affect the secretion of granzyme B (P>0.05); moreover, it also significantly enhanced the cytotoxicity of IL-15-NK92 cells against hematologic malignancies and solid tumor cells (P<0.05 or P<0.01), and up-regulated the phosphorylation levels of STAT1, STAT3 and STAT5 (all P<0.01). Conclusion: IL-27 can enhance the cytotoxicity of IL-15-NK92 cells against hematologic tumor cells and solid tumor cells, which might be related with its upregulation of phosphorylation level of STAT1, STAT3 and STAT5 in JAK-STAT pathway and multiple activating receptors in IL-15-NK92 cells.

Research progress on the promoting effect of interleukin-11 on tumors / 上海交通大学学报（医学版）

According to the lastest statistics, the overall morbidity and mortality of cancer in China still show an upward trend compared with historical data. An in-depth understanding of the molecular mechanisms of tumorigenesis and development is important to formulate future treatment strategies. Interleukin 11 (IL-11) is a member of cytokines that traditionally promote megakaryocyte maturation and regulate immune activity. In recent years, the promoting effect of IL-11 on tumor has been gradually discovered. This review mainly expounds that IL-11 is regu-lated by transforming growth factor-β/drosophila mothers against decapentaplegic protein (TGF-β/Smad) pathway, and may play a role in tumor-igenesis, drug resistance, metastasis and tumor microenvironment through signal transduction pathways such as Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway and phosphatidylinositol 3 kinase/protein kinase B (PI3K/AKT) pathway, and explores the application prospect of interfering IL-11 signal transduction in tumor therapy.

Therapeutic effect of methotrexate combined with electroacupuncture in rheumatoid arthritis rats / 中国组织工程研究

BACKGROUND: Currently, methotrexate for rheumatoid arthritis has been explored; however, there are still some problems such as unsatisfactory treatment effects. OBJECTIVE: To investigate the therapeutic effect and mechanism of methotrexate combined with electroacupuncture in rheumatoid arthritis rats. METHODS: A total of 100 Wistar rats were selected, 80 of which were injected type II collagenase to induce rheumatoid arthritis models. After successful modeling, the model rats were randomly divided into model group, methotrexate group, electroacupuncture group, and methotrexate+electroacupuncture group. The remaining 20 normal animals untreated were used as the normal control group. The arthritis index was evaluated at 1, 6, and 12 weeks of treatment. Hematological changes in the ankle were observed with hematoxylin-eosin staining at 12 weeks after treatment. RT-PCR and western blot were used to detect changes in tumor necrosis factor α, JAK3, and STAT3 expression levels. Flow cytometry was used to detect the apoptotic rate of chondrocytes. RESULTS AND CONCLUSION: Compared with the model group, methotrexate and electroacupuncture alone or in combination could significantly reduce the arthritis index of rheumatoid arthritis rats at 1, 6, and 12 weeks after administration (P < 0.05), and the reduction tendency was more significant in the methotrexate+electroacupuncture group. There was no synovial tissue lesion and joint capsule hyperplasia in the normal control group. In the model group, synovial tissue was infiltrated by extensive inflammatory cells; vasodilation and vascular hyperplasia in the synovium triggered the formation of vascular pannus; and intrathecal ligament nerve injury and proliferation of synovial and fibrous tissues were observed. Histological changes in the synovium were less after treatment with methotrexate and electroacupuncture alone than in combination. Treatment with methotrexate plus electroacupuncture obviously decreased inflammatory cell infiltration and synovial hyperplasia. Compared with the model group, the levels of tumor necrosis factor-α, JAK3 and STAT3 mRNA and protein were significantly reduced in the methotrexate group, electroacupuncture group, and methotrexate+electroacupuncture group (P < 0.05), and the reduction was more significant in the methotrexate+electroacupuncture group (P < 0.05). The apoptotic rate of chondrocytes was highest in the model group, followed by the methotrexate group, electroacupuncture group, and methotrexate+electroacupuncture group (P < 0.05). However, the apoptotic rate of chondrocytes in the methotrexate+electroacupuncture group was still higher than that in the normal control group (P < 0.05). To conclude, the combination of methotrexate and electroacupuncture for rheumatoid arthritis can inhibit the activity of tumor necrosis factor and JAK-STAT pathways, reduce the apoptosis of chondrocytes, and protect the articular cartilage.

microRNA-146a Promotes Growth of Acute Leukemia Cells by Downregulating Ciliary Neurotrophic Factor Receptor and Activating JAK2/STAT3 Signaling

PURPOSE: Acute leukemia (AL) is classified as acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). This study aimed to investigate the effect of miR-146a on childhood AL and its underlying molecular mechanisms. MATERIALS AND METHODS: Bone marrow samples were obtained from 39 AL children and 10 non-cancer controls. The expressions of miR-146a and ciliary neurotrophic factor receptor (CNTFR) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) in ALL and AML pediatric patients, as well as ALL (Jurkat) and AML (HL-60) cells. Correlations between miR-146a and clinical indicators were explored. A targeting relationship between miR-146a and CNTFR was detected by dual luciferase reporter gene assay. Cell proliferation, apoptosis, migration, and invasion of Jurkat and HL-60 cells were measured by MTT assay, flow cytometry, and transwell assay, respectively. LIF expression was detected by qRT-PCR in Jurkat and HL-60 cells. The expression of p-JAK2, JAK2, p-STAT3, and STAT3 in HL-60 cells was measured by Western blot. RESULTS: miR-146a was increased in ALL and AML pediatric patients, while CNTFR was decreased. miR-146a expression was associated with immunophenotype, karyotype, fusion gene, and SIL-TAL1. CNTFR was a target gene of miR-146a. miR-146a could promote cell proliferation, migration, and invasion, as well as inhibit cell apoptosis in Jurkat and HL-60 cells by downregulating CNTFR. Meanwhile, miR-146a inhibited the expression of LIF and activated JAK2/STAT3 pathway by downregulating CNTFR. CONCLUSION: miR-146a could promote the proliferation, migration, and invasion and inhibit the apoptosis of AL Jurkat and HL-60 cells by downregulating CNTFR and activating the JAK2/STAT3 pathway.

Magnolol exerts anti-asthmatic effects by regulating Janus kinase-signal transduction and activation of transcription and Notch signaling pathways and modulating Th1/Th2/Th17 cytokines in ovalbumin-sensitized asthmatic mice

Allergic asthma, is a common chronic inflammatory disease of the airway presenting with airway hyperresponsiveness and airway remodelling. T helper cells-derived cytokines are critically associated with asthma pathogenesis. Janus kinase-signal transduction and activation of transcription (JAK/STAT) signaling is found to be involved in asthma. Magnolol is a plant-derived bioactive compound with several pharmacological effects. The study aimed to assess the effects of magnolol in ovalbumin (OVA)-induced asthmatic model. BALB/c mice were sensitized and challenged with OVA. Magnolol (12.5, 25, or 50 mg/kg body weight) was administered to separate groups of animals. Dexamethasone was used as the positive control. Cellular infiltration into the bronchoalveolar lavage fluid (BALF) were reduced on magnolol treatment. The levels of Th2 and Th17 cytokines were reduced with noticeably raised levels of interferon gamma. Lung function was improved effectively along with restoration of bronchial tissue architecture. OVA-specific immunoglobulin E levels in serum and BALF were decreased by magnolol. Magnolol reduced Th17 cell population and effectively modulated the JAK-STAT and Notch 1 signaling. The results suggest the promising use of magnolol in therapy for allergic asthma.

Regulation of cytochrome P450 1A1 on M2 macrophage polarization / 中华危重病急救医学

Objective@#To investigate the potential effects of cytochrome P450 1A1 (CYP1A1) in regulating macrophages polarize to M2 type and explore the molecular mechanism.@*Methods@#All trials were completely randomized. ① Experiment 1: 6-8 weeks old healthy male C57BL/6J mice were collected, and primary peritoneal cells were extracted, then the cells were divided into phosphate buffered saline (PBS) group and interleukin-4 (IL-4) group. The cells in the IL-4 group were stimulated with 10 mg/L IL-4 (M2 macrophage inducer); and those in the PBS group were given with an equal amount of PBS. The mRNA expressions of intracellular M2 type polarized marker molecules including arginase-1 (Arg-1) and chitinase 3 like protein 1 (YM1) at 2, 4, 6 hours after IL-4 challenge were determined by quantitative reverse transcription-polymerase chain reaction (RT-qPCR). The phosphorylation of tyrosine protein kinase 1/signaling transcriptional and transduced activator 6 (JAK1/STAT6) signaling pathway and protein expressions of CYP1A1 and Arg-1 at 6, 12, 24 hours after IL-4 challenge were determined by Western Blot. ② Experiment 2: RAW264.7 cells with high expression CYP1A1 (CYP1A1/RAW) and their negative control cells (NC/RAW) were cultured in vitro. The cells in logarithmic growth phase were collected, and then they were divided into PBS control group and IL-4 group. The treatment method was the same as experiment 1. The phosphorylations of intracellular JAK1/STAT6 and phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathways in different cells at 1 hour and 2 hours after IL-4 challenge were determined by Western Blot. The mRNA and protein expressions of Arg-1 in different cells at 2 hours and 4 hours after IL-4 challenge were determined by RT-qPCR and Western Blot, respectively.@*Results@#① Experiment 1: after IL-4 challenge for 2 hours, the mRNA expressions of Arg-1 and YM1 in the primary peritoneal macrophages of mice were significantly increased as compared with the PBS control group [Arg-1 mRNA (2-ΔΔCt): 1.75±0.82 vs. 1.00±0.21; YM1 mRNA (2-ΔΔCt): 2.58±0.53 vs. 1.00±0.20, both P < 0.05] which indicated that IL-4 induced macrophage to M2 type polarization successfully. Meanwhile, the mRNA expression of CYP1A1 in polarized mouse peritoneal primary macrophages was also elevated as compared with the PBS control group, and peaked at 4 hours [CYP1A1 mRNA (2-ΔΔCt): 2.25±0.69 vs. 1.00±0.17, P < 0.01]. The results indicated that CYP1A1 expression was enhanced when macrophages polarized to M2 type. Compared with the PBS control group, the bands of phosphorylated JAK1 (p-JAK1), phosphorylated STAT6 (p-STAT6), Arg-1 and CYP1A1 were enhanced in primary peritoneal macrophages of mice in the IL-4 group, and reached the peak value at 12 hours then gradually decreased. This result indicated that the phosphorylation of JAK1/STAT6 pathway was enhanced in M2 macrophages with high expression of CYP1A1, and the pathway was activated. ② Experiment 2: after IL-4 challenge for 2 hours, the expression of Arg-1 mRNA in CYP1A1/RAW cells was significantly higher than that in NC/RAW cells (2-ΔΔCt: 3.02±0.60 vs. 1.47±0.43, P < 0.05), and the protein band signal was also stronger, and both peaked at 4 hours. This indicated that CYP1A1 could promote the polarization of macrophages to M2. After IL-4 challenge for 2 hours, the expression of p-JAK1 and p-JAK3 protein bands in both cells were significantly enhanced as compared with the PBS control group, but the enhancement of p-STAT6 band in CYP1A1/RAW cells was stronger than that of NC/RAW cells, indicating that CYP1A1 promoted macrophage polarization by promoting phosphorylation of the JAK1/STAT6 pathway. In the meantime, the protein band of Akt, a downstream protein of the PI3K/Akt pathway, in CYP1A1/RAW cells was significantly lower than that of NC/RAW cells, indicating that CYP1A1 did not promote macrophage polarization through this pathway.@*Conclusions@#CYP1A1 promotes the polarization of macrophage to M2 type. The mechanism is related to promoting the phosphorylation of JAK1/STAT6 pathway.

Regulation of cytochrome P450 1A1 on M2 macrophage polarization / 中华危重病急救医学

To investigate the potential effects of cytochrome P450 1A1 (CYP1A1) in regulating macrophages polarize to M2 type and explore the molecular mechanism. Methods All trials were completely randomized. ① Experiment 1: 6-8 weeks old healthy male C57BL/6J mice were collected, and primary peritoneal cells were extracted, then the cells were divided into phosphate buffered saline (PBS) group and interleukin-4 (IL-4) group. The cells in the IL-4 group were stimulated with 10 mg/L IL-4 (M2 macrophage inducer); and those in the PBS group were given with an equal amount of PBS. The mRNA expressions of intracellular M2 type polarized marker molecules including arginase-1 (Arg-1) and chitinase 3 like protein 1 (YM1) at 2, 4, 6 hours after IL-4 challenge were determined by quantitative reverse transcription-polymerase chain reaction (RT-qPCR). The phosphorylation of tyrosine protein kinase 1/signaling transcriptional and transduced activator 6 (JAK1/STAT6) signaling pathway and protein expressions of CYP1A1 and Arg-1 at 6, 12, 24 hours after IL-4 challenge were determined by Western Blot. ② Experiment 2: RAW264.7 cells with high expression CYP1A1 (CYP1A1/RAW) and their negative control cells (NC/RAW) were cultured in vitro. The cells in logarithmic growth phase were collected, and then they were divided into PBS control group and IL-4 group. The treatment method was the same as experiment 1. The phosphorylations of intracellular JAK1/STAT6 and phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) pathways in different cells at 1 hour and 2 hours after IL-4 challenge were determined by Western Blot. The mRNA and protein expressions of Arg-1 in different cells at 2 hours and 4 hours after IL-4 challenge were determined by RT-qPCR and Western Blot, respectively. Results ① Experiment 1: after IL-4 challenge for 2 hours, the mRNA expressions of Arg-1 and YM1 in the primary peritoneal macrophages of mice were significantly increased as compared with the PBS control group [Arg-1 mRNA (2-ΔΔCt): 1.75±0.82 vs. 1.00±0.21; YM1 mRNA (2-ΔΔCt): 2.58±0.53 vs. 1.00±0.20, both P < 0.05] which indicated that IL-4 induced macrophage to M2 type polarization successfully. Meanwhile, the mRNA expression of CYP1A1 in polarized mouse peritoneal primary macrophages was also elevated as compared with the PBS control group, and peaked at 4 hours [CYP1A1 mRNA (2-ΔΔCt): 2.25±0.69 vs. 1.00±0.17, P < 0.01]. The results indicated that CYP1A1 expression was enhanced when macrophages polarized to M2 type. Compared with the PBS control group, the bands of phosphorylated JAK1 (p-JAK1), phosphorylated STAT6 (p-STAT6), Arg-1 and CYP1A1 were enhanced in primary peritoneal macrophages of mice in the IL-4 group, and reached the peak value at 12 hours then gradually decreased. This result indicated that the phosphorylation of JAK1/STAT6 pathway was enhanced in M2 macrophages with high expression of CYP1A1, and the pathway was activated. ② Experiment 2: after IL-4 challenge for 2 hours, the expression of Arg-1 mRNA in CYP1A1/RAW cells was significantly higher than that in NC/RAW cells (2-ΔΔCt:3.02±0.60 vs. 1.47±0.43, P < 0.05), and the protein band signal was also stronger, and both peaked at 4 hours. This indicated that CYP1A1 could promote the polarization of macrophages to M2. After IL-4 challenge for 2 hours, the expression of p-JAK1 and p-JAK3 protein bands in both cells were significantly enhanced as compared with the PBS control group, but the enhancement of p-STAT6 band in CYP1A1/RAW cells was stronger than that of NC/RAW cells, indicating that CYP1A1 promoted macrophage polarization by promoting phosphorylation of the JAK1/STAT6 pathway. In the meantime, the protein band of Akt, a downstream protein of the PI3K/Akt pathway, in CYP1A1/RAW cells was significantly lower than that of NC/RAW cells, indicating that CYP1A1 did not promote macrophage polarization through this pathway. Conclusions CYP1A1 promotes the polarization of macrophage to M2 type. The mechanism is related to promoting the phosphorylation of JAK1/STAT6 pathway.

The JAK-STAT pathway and hemophagocytic syndrome / 国际儿科学杂志

Hemophagocytic syndrome (HPS),also known as hemophagocytic lymphohistiocytosis (HLH),is a syndrome with multiple clinical manifestations and severe impairment of organ function caused by abnormal activation of lymphocytes and histiocytes leading to hypercytokinemia.The pathogenesis of this disease is very complicated.So far,it can not be fully clarified.Studies have shown that activation of JAK-STAT signaling pathway can activate inflammatory factor-related gene transcription,which can cause inflammatory factor cascade reaction,and ultimately lead to the occurrence of hemophagocytic syndrome.In recent years,a series of targeted drugs for JAK-STAT pathway have been developed.A JAK1/JAK2 inhibitor,ruxolitinib,has been found to be useful in the treatment of HLH.This article reviews the mechanism of action of ruxolitinib in HLH and its clinical research results.

Study on the effects of target-silencing CXCR3 expression on malignant proliferation of hepatocellular carcinoma / 中华肝脏病杂志

Objective@#To explicit, the expression of chemokine receptor 3 in HCC tissues and its relationship with overall survival of patients, and to explore the effect of targeted silencing CXCR3 gene on proliferation of hepatocellular carcinoma cells and its mechanism of action.@*Methods@#The expression of CXCR3 in 60 cases of hepatocellular carcinoma and its adjacent tissues were detected by immunohistochemistry. The clinicopathological correlations between the expression levels of CXCR3 in hepatoma tissues of liver cancer patients were analyzed and univariate Kaplan-Meier survival analysis was performed in combination with follow-up data. Huh7 hepatoma cells were infected with lentivirus LV-CXCR3-shRNA. The effects of CXCR3 deletion on proliferation of hepatoma cells were determined by CCK-8 assay and tumor-bearing nude mice experiment.@*Results@#CXCR3 was highly expressed in HCC tissues, and the overall survival rate (OS) of patients with high CXCR3 expression was significantly lower than that of patients with low expression. After the CXCR3 gene was successfully silenced in Huh7 hepatocellular carcinoma cells, the proliferation ability of Huh7 cells was significantly inhibited in vitro, and the tumor growth rate of nude mice was slowed down, and the activity of JAK-STAT pathway in Huh7 cells was decreased, and the levels of c-MYC and Bcl-xl protein were decreased. In addition, deletion of CXCR3 can effectively inhibit IL-6-mediated JAK-STAT pathway activation.@*Conclusion@#CXCR3 high expression indicated that the survival rate was poor, and the target silencing of CXCR3 gene could inhibit the proliferation of hepatocellular carcinoma cells and maybe related to inhibition of JAK-STAT pathway activity. CXCR3 may be a potential target for the treatment of hepatocellular carcinoma.

Balance of ruxolitinib to anti-GVHD and GVL effect after allo-HSCT / 中华器官移植杂志

Objective To observe the anti-relapse and anti-graft versus host disease (GVHD) effects and side effects of ruxolitinib on patients who have relapsed leukemia after allo-hematopoietic stem cell transplantation (HSCT).Methods The clinical data of four patients sufferring from relapsed leukemia were collected and analyzed retrospectively.Three cases had a positive gene and 1 case had a extramedullary recurrence.All of them had serious GVHD involving multiparts,as the result of attenuating immunosuppressant aggressively.One case had central nervous system leukemia before allo-HSCT.Those patients were treated with ruxolitinib,according to the degree of GVHD,the treatment strategy and curative effect of GVHD,and the residual condition of original leukemia.Then,the degree of GVHD,the residual condition of original leukemia and the side effects of ruxolitinib were revaluated once a month after taking ruxolitinib.Results One case achieved completer remission (CR) and there partial remission (PR) in consideration of GVHD.Up to date,2 cases had no relapse in any level and 2 cases replased according to any of the results related to bone marrow aspiration.Conclusion Ruxolitinib is effective in patients with GVHD after allo-HSCT and doesn't influence GVL effect or increase the risk of relapse at the same time.Ruxolitinib doesn't have obvious side effects when treating GVHD.

Effect of Yiqigubiao Pill Regulate Inflammatory Cytokines Through JAK/STAT Pathway in COPD Rats Model / 世界科学技术-中医药现代化

The research observed the effect of Yiqigubiao Pill regulating inflammatory reaction mRNA expression in JAK/STAT pathway in COPD rats model and explored protective effect of Yiqigubiao pills in the treatment or auxiliary possible mechanism in COPD. 60 SFP Wistar rats were divided randomly into control group (CN), COPD model group (M) and Yiqigubiao Pill group (Yqgb). The rats in group M and group YQGB were establish by using the method of LPS and smoking as COPD, rats in group M and group CN were given normal saline intragastric administration, and rats in the YQGB group were given water solution intragastric administration at the corresponding dose, twice a day. After using drugs for 12 weeks, we used ELISA method to detect changes inIL-17a, IL-23, IFN-γ and RORγt expression in peripheral blood, RT-PCR method to detect changes in the expression of JAK1, JAK3, STAT1 and STAT3 mRNA in lung tissues. Compared with the group CN, the infiltration of inflammatory cells stained by HE in the experimental group was reduced, and the diameter of the bronchial lumen was broadened (P ＜ 0.01). Both of ELISA and RT-PCR results showed that, compared with group CN, the levels of IL-17a, IL-23 and RORγt, the levels of JAK1, JAK3, STAT1, and STAT3 mRNA were increased in group M. Compared with group M, the mRNA levels of above indicators in group Yqgb were decreased, and the difference was statistically significant (P ＜ 0.01). Correlation analysis revealed that JAK1, JAK3, STAT1 and STAT3 were associated with IL-17a, IL-23, and RORγt had statistical significance (P ＜ 0.05). Therefore, Yiqgubiao Pill can inhibit the inflammatory response in COPD rats model, and its mechanism may be associated with inhibition of JAK/STAT pathway.

Impact of Catgut Embedding Intervention on JAK 2-STAT 3 Signaling in Gastric Mucosa of Chronic Atrophic Gastritis Rats / 针刺研究

OBJECTIVE: To observe the impact of catgut embedding on histopathological changes and expression of phosphorylated-Janus kinase-2 (p-JAK 2), phosphorylated-signal transducer and activator of transcription 3 (p-STAT 3), Cyclin D 1, Bcl-2, and suppressor of cytokine signaling (SOCS) of the gastric mucosa in chronic atrophic gastritis (CAG) rats, so as to explore its mechanisms underlying improving CAG. METHODS: Forty-one SD rats were randomly divided into normal (n＝12), model (n＝9), natural recovery (n＝10) and catgut embedding (n＝10) groups. The CAG model was established by free drinking of N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) solution (100µg/mL) and irregular diet for 24 weeks. Catgut embedding was applied at bilateral "Zusanli" (ST 36), "Pishu" (BL 20) and "Zhongwan" (CV 12), once every 10 days, 6 times altogether. The histopathological changes of the gastric mucosa were observed by naked eye and light microscope after H．E. staining. Wes-tern blot was used to detect the expression of p-JAK 2, p-STAT 3, CyclinD 1, Bcl-2 and SOCS 3 proteins in the gastric antrum tissue. RESULTS: After modeling, the regional gastric tissue showed pale thinner gastric wall, and poorer elasticity with fewer plica, congestion of blood vessels, reduction of regional glands with necrosis and disordered arrangement as well as evident hyperplasia of the connective tissue, which was evidently milder in the catgut embedding group. Compared with the normal group, the expression levels of p-JAK 2, p-STAT 3, CyclinD 1, Bcl-2, SOCS 3 proteins were significantly up-regulated in both model group and natural recovery group (P0.05). After catgut embedding intervention, the expression levels of p-JAK 2, p-STAT 3, Cyclin D 1, Bcl-2 proteins were considerably down-regulated (P<0.01), and that of SOCS 3 protein was notably further increased relevant to the natural recovery group (P<0.01)．. CONCLUSION: Catgut embedding may improve pathological changes of gastric mucosa in CAG rats, which possibly associated with its effects in up-regulating gastric SOCS 3 protein expression and in inhibiting the activation of JAK 2-STAT 3 signaling pathway to reduce the expression of CyclinD 1 and Bcl-2 proteins.

Effect of JAK2/STAT1 on acute kidney injury induced by liver cold ischemia reperfusion in rats / 中华器官移植杂志

Objective To investigate the effect of Janus kinase2/signal transducer and activator of transcription 1 (JAK2/STAT1) signaling pathway on acute kidney injury induced by liver cold ischemia reperfusion (IR) in rats.Methods Thirty healthy male Sprague-Dawley rats,weighing 220-250 g,were assigned randomly to 3 groups (n =10/group):sham operation group (Sham group);liver cold ischemia reperfusion model group (I/R group);JAK2 kinase inhibitor AG490 group (AG490 group) (AG490 at dose of 10 mg/kg was intraperitoneally injected 30 min before establishment of the model).Other groups were given the equal volume of normal saline at the same time points.Then the rats were sacrificed at 6 h after reperfusion (at 6 h after the end of operation in Sham group).The renal function and oxidative stress level were observed.The pathological changes of the renal tissues and nephritic cell apoptosis were analyzed,and the expression of p-JAK2,pSTAT1,Bcl-2 and Bax was detected by Western blotting.Results As compared with Sham operation group,renal histological lesion and renal dysfunction were aggravated,level of oxidative stress and apoptosis rate were increased in I/R group,the Bcl-2/Bax ratio was decreased and the expression of pJAK2 and p-STAT1 was up-regulated.As compared with I/R group,AG490 dramatically attenuated histological lesions and oxidative stress,restored the renal function,and reduced the number of apoptotic tubular epithelial cells.AG490 significantly increased the Bcl-2/Bax ratio,and inhibited the expression of p-JAK2 and p-STAT1.Conclusion Blockage of JAK2/STAT1 signaling pathway can alleviate acute kidney injury after liver cold ischemia reperfusion probable through inhibiting the oxidative stress and apoptosis.

Anti-inflammatory mechanism of low dose methotrexate and its application in spinal cord injury / 中国药理学通报

Methotrexate (MTX)has dual effects of anti-inflam-matory and immune suppression,and its pharmacological mecha-nism is complex,diverse and synergistic.This paper summari-zes the main anti-inflammatory mechanism of low-dose MTX,in-cluding inhibition of JAK/STAT pathway,inhibition of inflam-matory reaction and immune response,increasing the accumula-tion of adenosine and the function of intracellular metabolites (methotrexate polyglutamate).In addition,low-dose MTX can inhibit oxidation by decreasing the level of lipid peroxidation, suppress the inflammatory response to secondary spinal cord in-jury,reduce spinal cord ischemia reperfusion injury and neuro-pathic pain,thus playing a neuroprotective role by a series of pharmacological mechanism.The anti-inflammatory mechanism of low-dose MTX and its application in spinal cord injury were reviewed,to guide the further research on the anti-inflammatory effect of MTX,and provide a theoretical basis for new drugs for clinical treatment of spinal cord injury.

JAK/STAT Pathway Modulates on Porphyromonas gingivalis Lipopolysaccharide- and Nicotine-Induced Inflammation in Osteoblasts / 치위생과학회지

Bacterial infection and smoking are an important risk factors involved in the development and progression of periodontitis. However, the signaling mechanism underlying the host immune response is not fully understood in periodontal lesions. In this study, we determined the expression of janus kinase (JAK)/signal transducer and activator of transcription (STAT) on Porphyromonas gingivalis lipopolysaccharide (LPS)- and nicotine-induced cytotoxicity and the production of inflammatory mediators, using osteoblasts. The cells were cultured with 5 mM nicotine in the presence of 1 µg/ml LPS. Cell viability was determined using MTT assay. The role of JAK on inflammatory mediator expression and production, and the regulatory mechanisms involved were assessed via enzyme-linked immunosorbent assay, reverse transcription-polymerase chain reaction, and Western blot analysis. LPS- and nicotine synergistically induced the production of cyclooxgenase-2 (COX-2) and prostaglandin E₂ (PGE₂) and increased the protein expression of JAK/STAT. Treatment with an JAK inhibitor blocked the production of COX-2 and PGE₂ as well as the expression of pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin-1β (IL-1β), and IL-6 in LPS- and nicotine-stimulated osteoblasts. These results suggest that JAK/STAT is closely related to the LPS- and nicotine-induced inflammatory effects and is likely to regulate the immune response in periodontal disease associated with dental plaque and smoking.

Exogenous hydrogen sulfide attenuates high glucose-induced injury by in-hibiting JAK/STAT pathway in human umbilical vein endothelial cells / 中国病理生理杂志

AIM: To explore whether exogenous hydrogen sulfide (H2S) depresses high glucose (HG)-in-duced injury by modulating the Janus kinase/signal transducer and activator of transcription ( JAK/STAT) pathway in hu-man umbilical vein endothelial cells (HUVECs).METHODS:The protein levels of JAK2, STAT3 and cleaved caspase-3 were determined by Western blot.The cell viability was measured by CCK-8 assay.Mitochondrial membrane potential ( MMP) was detected by rhodamine 123 staining followed by photofluorography.The intracellular level of reactive oxygen species (ROS) was analyzed by DCFH-DA staining followed by photofluorography.The activity of superoxide dismutase (SOD) was also measured.RESULTS:Pretreatment of the HUVECs with 400 μmol/L NaHS (a donor of H2S) for 30 min prior to exposure to 40 mmol/L glucose ( HG) markedly attenuated HG-induced upregulation of the phosphorylation of JAK2 and STAT3.Pretreatment with 400μmol/L NaHS for 30 min or with 20μmol/L AG490 (inhibitor of the JAK/STAT pathway) for 30 min attenuated the injury of HUVECs induced by HG, as indicated by the increases in cell viability and SOD activity, and decreases in the protein level of cleaved caspase-3, ROS generation and dissipation of MMP.CONCLU-SION:Exogenous H2 S protects HUVECs against HG-induced injury by inhibiting JAK/STAT pathway.