5-Aza-2-deoxycytidine alleviates the progression of primary biliary cholangitis by suppressing the FoxP3 methylation and promoting the Treg/Th17 balance.

Primary biliary cholangitis (PBC) is a common autoimmune liver disease manifested by the infiltration of CD4+ T cells, and the subsequent targeted injury of biliary epithelial cells (BECs). As important components of CD4 subsets, the Treg/Th17 axis maintains an immunological balance between self-tolerance and inflammation in the liver microenvironment. However, the role and regulatory mechanism of the Treg/Th17 axis in PBC remain unclear. In this study, we examined the Treg/Th17 axis in PBC patients and found that the Treg/Th17 axis was imbalanced in PBC at both the transcriptional and cellular levels, with Treg being a weak candidate, which correlates with the PBC progression. This imbalanced Treg/Th17 axis was likely to be affected by the FoxP3 hypermethylation, which was related to the increase of DNA methyltransferase. Furthermore, the effect of 5-Aza-2-deoxycytidine (DAC)-mediated FoxP3 demethylation on PBC mice was investigated. We verified that DAC significantly suppressed the FoxP3 methylation and rebuilt the Treg/Th17 balance, resulting in the alleviation of liver lesions and inflammation. Taken together, our data indicate that DAC plays a positive role in alleviating the progression of PBC through the inhibition of DNA methylation of FoxP3 to rebuild the balanced Treg/Th17 axis. DAC could be considered as a potential candidate for the development of new anti-inflammation strategies in the treatment of PBC.

Expression of EZH2 and P53 and their correlation in ovarian cancer tissues.

Previous researches have demonstrated that EZH2 expression is increased in many solid tumors and is closely related to the worse progression, transcriptional silence, distal metastasis, and differential inhibition of tumors. P53 can regulate many cells signaling pathways and play an important role in cell cycle, cell apoptosis, and cell senescence. However, there are few reports on the expression of EZH2 and p53 in ovarian cancer and their correlation with the ovarian cancer. The purpose is to elucidate the expression of EZH2 and p53 in ovarian cancer and to study the relationship of EZH2 and p53 with the clinical parameters of ovarian cancer. In this study, both mRNA and protein level of EZH2 in ovarian cancer group was significantly higher than that in borderline, benign, and normal group; while the mRNA and protein level of p53 was significantly lower than that in borderline, benign, and normal group. The expression of EZH2 protein was mainly located in the cytoplasm and nucleus, while mutated p53 protein was mainly located in the nucleus. Furthermore, the expression of EZH2 is closely related to the FIGO stage and histological grade of ovarian cancer. EZH2 and P53 are closely related to the occurrence of ovarian cancer. We speculate that EZH2 may promote the development of ovarian cancer by inhibiting the expression of p53, suggesting that p53 may be the target gene of EZH2.

[Effects of interleukin 27 and its receptor on TGFßinduced murine pulmonary fibroblast proliferation and transformation].

OBJECTIVE: To investigate the effects of interleukin-27 (IL-27) and its receptor (WSX-1) on the proliferation, transformation and collagen synthesis of the mouse lung fibroblasts. METHODS: Cultured mouse lung fibroblasts were treated with TGF-ß1, recombinant murine IL-27, a IL-27 receptor (IL-27R) overexpression vector IL-27R/pCDNA3.1, IL-27 and IL-27R, or all the 3 combined. MTT assay was used to assess the proliferation of the cells, and RT-PCR and Western blotting were employed to examine the mRNA and protein expressions of a-smooth muscle actin (α-SMA) and types I and III collagen; immunofluorescence assay was used to test the expression and location of α-SMA. RESULTS: TGF-ß1 promoted the cell proliferation and obviously enhanced α-SMA expression and types I and III collagen synthesis in the fibroblasts. Both IL-27 and IL-27R significantly inhibited the proliferation of the pulmonary fibroblasts and obviously decreased their α-SMA expression and types I and III collagen synthesis, but when combined,they produced no obvious inhibitory effect on TGF-1-induced proliferation and transformation of pulmonary fibroblasts. CONCLUSION: Both IL-27 and IL-27R alone can suppress the proliferation, transformation, and collagen synthesis of mouse pulmonary fibroblasts, but their combined treatment produces no such inhibitory effect because of the neutralization of exogenous IL-27 by IL-27R to result in the failure of activating the cell signaling pathways.

Therapeutic effect of intravenous bone marrow-derived mesenchymal stem cell transplantation on early-stage LPS-induced acute lung injury in mice.

OBJECTIVE: To investigate the effect of intravenous bone marrow-derived mesenchymal stem cell (MSC) transplantation for early intervention of lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. METHODS: Thirty-six mice were randomized into control group, PBS-treated ALI group, and MSC-treated ALI group. In the latter two groups, mouse models of ALI were established by intranasal instillation of LPS, and 1 h later, the 4th passage of MSCs isolated from the bone marrow of mice or PBS were administered via the tail vein. The histological findings, lung wet/dry (W/D) weight ratio, neutrophil count and protein and cytokine contents in the bronchoalveolar lavage fluid (BALF), and myeloperoxidase (MPO) level in the lung tissue were analyzed at 24 h after MSC administration. Engraftment of MSCs in the recipient lung was determined by fluorescent PKH26 staining and flow cytometry. RESULTS: Compared with the control group, PBS-treated ALI group showed significantly higher protein levels, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and neutrophil count in the BALF and MPO content in the lung tissue, with also severe damage of lung histology. MSCs administration significantly reduced the lung W/D weight ratio, the levels of protein, TNF-α, IL-6 and neutrophil count in the BALF and MPO content in the lung tissue, and obviously lessened the lung injury 24 h after the transplantation. MSC administration also significantly increased the level of IL-10 in the BALF. CONCLUSION: Intravenous MSC transplantation can effectively improve the lung histology, attenuate the inflammatory response, reduce pulmonary edema in the early stage of LPS-induced ALI in mice, and such effects are independent of MSC engraftment in the lungs.

[Study on the association between the HLA-DRB1 alleles and type 2 diabetes in Yi nationality of Yunnan].

OBJECTIVE: To investigate the association between the polymorphism of HLA-DRB1 alleles and type 2 diabetes mellitus in Yi nationality of Yunnan. METHODS: Polymerase chain reaction-sequence specific primers (PCR-SSP) genotyping method was conducted in 79 patients with type 2 diabetes mellitus and 47 ethnically matched controls in Yi Nationality Autonomous Prefecture, Chuxiong. RESULTS: HLA-DR7 and DR11 allele frequencies in type 2 diabetic mellitus patients were significantly higher than those in non-diabetic control subjects respectively(P is 0.009, RR is 8.329;P is 0.029, RR is 7.734). CONCLUSION: DR7 and DR11 alleles are probably susceptible genes of type 2 diabetes mellitus in Yunnan Yi nationality.

[Association between the polymorphism of HLA-DQA1 alleles and type 2 diabetes in Yi nationality of Yunnan].

OBJECTIVE: To investigate the association between the polymorphism of HLA-DQA1 alleles and type 2 diabetes in Yi nationality of Yunnan. METHODS: Polymerase chain reaction with sequence-specific primers (PCR-SSP) genotyping method was conducted in 58 ethnic Yi patients with type 2 diabetes mellitus and 82 ethnically matched controls from Chuxiong of Yunnan. Then a study was made on the association between the polymorphism of HLA-DQA1 alleles and type 2 diabetes mellitus. RESULTS: The frequency of HLA-DQA1*0301 allele in the patients with type 2 diabetes mellitus was significantly higher than that in the healthy controls (P=0.002, RR=3.097), and the frequency of HLA-DQA1*0601 in the patients was significantly lower (P=0.025, RR=0.429). CONCLUSION: In Yi nationality of Yunnan, HLA-DQA1*0301 allele may be a susceptible gene and the HLA-DQA1*0601 allele may protect individuals from the risk of diabetes mellitus.

[Study on the association between the polymorphism of HLA-DQA1 alleles and type 2 diabetes in Yunnan Han nationality].

OBJECTIVE: To investigate the association between the polymorphism of HLA-DQA1 alleles and type 2 diabetes mellitus in Yunnan Hans. METHODS: Polymerase chain reaction-sequence specific primers(PCR-SSP) genotyping method was conducted in 108 Han patients with type 2 diabetes and 56 ethnically matched controls from the same area of Yunnan Province. RESULTS: HLA-DQA1*0301(RR=3.092, P<0.01) and DQA1*0501 (RR=3.257, P<0.05) allelic frequencies in type 2 diabetic patients were significantly higher than those in non-diabetic control subjects respectively. HLA-DQA1*0401 (RR=0.371, P<0.01) allelic frequencies in patients were significantly decreased, compared with controls; HLA-DQA1*0302 (RR=3.356, P<0.01) allelic frequencies in patients with type 2 diabetic nephropathy were significantly increased. CONCLUSION: HLA-DQA1*0301 and DQA1*0501 are susceptible genes of type 2 diabetes in Yunnan Han nationality; in reverse, HLA-DQA1*0401 is a resistant gene. HLA-DQA1*0302 is a susceptible gene of type 2 diabetic nephropathy.