BACKGROUND/OBJECTIVES: 4-Hydroxy-2-nonenal (HNE) is a biomarker for oxidative stress to induce inflammation. Methionine is an essential sulfur-containing amino acid with antioxidative activity. On the other hand, the evidence on whether and how methionine can depress HNE-derived inflammation is lacking. In particular, the link between the regulation of the nuclear factor-κB (NF-κB) signaling pathway and methionine intake is unclear.This study examined the link between depression from HNE accumulation and the antiinflammatory function of L-methionine in rats.MATERIALS/METHODS: Male Wistar rats (3-week-old, weighing 70–80 g) were administered different levels of L-methionine orally at 215.0, 268.8, 322.5, and 430.0 mg/kg body weight for two weeks. The control group was fed commercial pellets. The hepatic HNE contents and the protein expression and mRNA levels of the inflammatory mediators were measured. The interleukin-10 (IL-10) and glutathione S-transferase (GST) levels were also estimated. RESULTS: Compared to the control group, hepatic HNE levels were reduced significantly in all groups fed L-methionine, which were attributed to the stimulation of GST by L-methionine. With decreasing HNE levels, L-methionine inhibited the activation of NF-κB by up-regulating inhibitory κBα and depressing phosphoinositide 3 kinase/protein kinase B. The mRNA levels of the inflammatory mediators (cyclooxygenase-2, interleukin-1β, interleukin-6, inducible nitric oxide synthase, tumor necrotic factor alpha) were decreased significantly by L-methionine. In contrast, the protein expression of these inflammatory mediators was effectively down regulated by L-methionine. The anti-inflammatory action of L-methionine was also reflected by the up-regulation of IL-10. CONCLUSIONS This study revealed a link between the inhibition of HNE accumulation and the depression of inflammation in growing rats, which was attributed to L-methionine availability. The anti-inflammatory mechanism exerted by L-methionine was to inhibit NF-κB activation and to up-regulate GST.

Objective:To establish a disease risk prediction model for the newborn screening system of inherited metabolic diseases by artificial intelligence technology.Methods:This was a retrospectively study. Newborn screening data ( n=5 907 547) from February 2010 to May 2019 from 31 hospitals in China and verified data ( n=3 028) from 34 hospitals of the same period were collected to establish the artificial intelligence model for the prediction of inherited metabolic diseases in neonates. The validity of the artificial intelligence disease risk prediction model was verified by 360 814 newborns ' screening data from January 2018 to September 2018 through a single-blind experiment. The effectiveness of the artificial intelligence disease risk prediction model was verified by comparing the detection rate of clinically confirmed cases, the positive rate of initial screening and the positive predictive value between the clinicians and the artificial intelligence prediction model of inherited metabolic diseases. Results:A total of 3 665 697 newborns ' screening data were collected including 3 019 cases ' positive data to establish the 16 artificial intelligence models for 32 inherited metabolic diseases. The single-blind experiment ( n=360 814) showed that 45 clinically diagnosed infants were detected by both artificial intelligence model and clinicians. A total of 2 684 cases were positive in tandem mass spectrometry screening and 1 694 cases were with high risk in artificial intelligence prediction model of inherited metabolic diseases, with the positive rates of tandem 0.74% (2 684/360 814)and 0.46% (1 694/360 814), respectively. Compared to clinicians, the positive rate of newborns was reduced by 36.89% (990/2 684) after the application of the artificial intelligence model, and the positive predictive values of clinicians and artificial intelligence prediction model of inherited metabolic diseases were 1.68% (45/2 684) and 2.66% (45/1 694) respectively. Conclusion:An accurate, fast, and the lower false positive rate auxiliary diagnosis system for neonatal inherited metabolic diseases by artificial intelligence technology has been established, which may have an important clinical value.

OBJECTIVE: To investigate the mechanism by which epigallocatechin gallate (EGCG) induces gene demethylation and promotes the apoptosis of acute myeloid leukemia KG-1 and THP-1 cell lines. METHODS: KG-1 and THP-1 cells treated with 25, 50, 75, 100 or 150 μg/mL EGCG for 48 h were examined for gene methylation using MSP and for cell proliferation using MTT assay. The changes in cell cycle and apoptosis of the two cell lines after treatment with EGCG for 48 h were detected using flow cytometry. The mRNA and protein expressions of DNMT1, CHD5, p19, p53 and p21 in the cells were detected using RT-quantitative PCR and Western blot. RESULTS: EGCG dose-dependently reversed hypermethylation of gene and reduced the cell viability in both KG-1 and THP-1 cells ( < 0.05). EGCG treatment caused obvious cell cycle arrest in G1 phase, significantly increased cell apoptosis, downregulated the expression of DNMT1 and upregulated the expressions of CHD5, p19, p53 and p21 in KG-1 and THP-1 cells ( < 0.05). CONCLUSIONS EGCG reduces hypermethylation of gene in KG-1 and THP-1 cells by downregulating DNMT1 to restore its expression, which results in upregulated expressions of p19, p53 and p21 and induces cell apoptosis.

BACKGROUND:A short-peptide small molecule hydrogel (SMH) developed in the previous study has more obvious advantages than other hydrogels to improve local microenvironment, carry bioactive substances and interfere with stem cell signal transduction pathways. OBJECTIVE:To explore the effect of SMHs on bone marrow mesenchymal stem cells (BMSCs) proliferation, apoptosis and differentiation into myocardial cells. METHODS: (1) Passage 9 rat BMSCs in vitro were divided into control group and experimental group, followed by routine culture and culture in SMHs, respectively. At 7 days of culture, cell proliferation and apoptosis were detected. Cells in the two groups were exposed to anaerobic environment for 12 hours, and expression levels of Bcl-2, Bax and Caspase-3 in BMSCs were detected. (2) Passage 9 BMSCs were divided into four groups and then cultured in 5-azacytidine, SMHs, SMHs+5-azacytidine, and L-DMEM (normal control), respectively. After 4 weeks of induction, expression of CTnT, desmin and Cx-43 proteins was detected and expression levels of early cardiac transcription factors, NKX2.5 and GATA-4, were also measured. RESULTS AND CONCLUSION: (1) Compared with the control group, better proliferation and lower apoptosis of BMSCs were found in the experimental group. Under anaerobic conditions, the number of survival cells was reduced in both groups, but less apoptosis or necrosis was found in the experimental group than the control group (P < 0.05). Moreover, the level of Bcl-2 was higher in the experimental group than the control group (P < 0.01), while the levels of Bax and Caspases-3 protiens were lower in the experimental group than the control group (P < 0.01). (2) NKx2.5 and GATA-4 mRNA expression was found in both 5-azacytidine and SMHs+5-azacytidine groups, and moreover, the mRNA levels of early cardiac transcription factors were significantly higher in the SMHs+5-azacytidine group than in the 5-azacytidine group (P < 0.05). In the normal control group, cTnT expressed negatively, and desmin and Cx-43 expressed weakly. The expression of cTnT, desmin and Cx-43 proteins was higher in the SMHs+5-azacytidine group than in the 5-azacytidine and SMHs groups, while there was no significant difference between the latter two groups. To conclude, SMHs as a culture medium is conducive to the proliferation of BMSCs, reduces cell apoptosis, and promotes myocardial differentiation of BMSCs.

Type II innate lymphoid cells ( ILC2s) are widely distributed in the blood , intestines, trachea, lung, spleen, liver, animal fat and skin, and involved in the innate immune responses .ILC2s have attracted much atten-tion for its important roles in the conversion of white adipose to beige adipose .Studies have shown that ILC2s are essential for the proliferation and differentiation of adipocyte precursor cells , and they also play a vital role in anti-parasitic infection and allergic inflammation .This review discusses the discovery , differentiation , development , distribution and function of ILC2s, and their relationships with the browning of white adipose tissue for providing valuable references on understanding the pathogenesis , prevention and treatment of obesity and fat metabolism disorders .

Interleukin (IL)-36, a member of the IL-1 family, plays an important role in mediating inflammatory diseases .IL-36 subfamily consists of IL-36α, IL-36β, IL-36γand IL-36 receptor antagonists (IL-36Ra).IL-36 receptor (IL-36R) and IL-1 receptor accessory protein (IL-1RAcP) are involved in the IL-36-mediated signal transduction .In human, mutations in the gene encoding IL-36Ra will lead to IL-36R signaling disorders , mainly manifested as diffuse pustular psoriasis .It suggests that IL-36 plays an important role in psoriasis.Recently, it has also been found that IL-36 is related to inflammation in other organs . These emerging roles of IL-36 provide new thoughts for the treatment of human inflammatory diseases .

Objective To explore the inhibition role of 1 ,25 dihydroxyvitamin D3 on laryngeal cancer Hep‐2 cell proliferation and its influence on mTOR signal pathway .Methods Hep‐2 cells were treated with different concentrations of 1 ,25 dihydroxyvitamin D3 (10-8 ,10-7 ,10-6mol/L) for 24 ,48 ,72 h respectively .The proliferation situation of Hep‐2 cells was detected by the MTT meth‐od and the inhibition rate was calculated .The effect of 1 ,25 dihydroxyvitamin D3 on Hep‐2 cell cycle distribution was analyzed by flow cytometry .The influence of 1 ,25 dihydroxyvitamin D3 on mTOR signaling pathway was detected by Western blot .Results Different concentrations of 1 ,25 dihydroxyvitamin D3 could inhibit the proliferation of Hep‐2 cells ,changed the cell cycle distribu‐tion and increased the proportion of Hep‐2 cells in G0/G1 phase .The expressions of TSC1 and TSC2 protein after 1 ,25 dihydroxyvi‐tamin D3 intervention were increased compared with the control group (P<0 .01) ,while the Rheb protein expression was signifi‐cantly decreased(P<0 .01):mTOR protein and phosphorylation level were significantly decreased compared with the control group (P<0 .01) ,the decrease of mTOR protein phosphorylation was especially obvious (P<0 .01);4EBP‐1 protein expression was in‐creased compared with the control group (P<0 .01) .Conclusion 1 ,25‐dihydroxyvitamin D3 alters the Hep‐2 cell cycle distribution , affects the protein expression of mTOR signaling pathway ,thus inhibits the cell proliferation .

Nuclear factor(NF)-κB is widely expressed in various types of tissue cells. Abnormal activation of NF-κB signaling pathway can lead to the initiation and progression of asthma,chronic ob?structive pulmonary disease(COPD)and other lung inflammatory diseases,but inhibition of its activity can effectively alleviate the occurrence and development of these diseases. In this paper ,we review biological characteristics of NF-κB,role of NF-κB signaling in asthma,COPD and other inflammatory lung diseases,and potential application of NF-κB inhibitors in clinical treatment of these inflammatory lung diseases. It will provide a valuable reference for further study on pathogenesis,prevention,and control of lung inflammatory diseases.

AIM:Tostudywhe ther theangiotens in-(1-7)[Ang-(1-7)]/Mas receptor axis protects cardio-myocytes against high glucose (HG)-induced injury by inhibiting nuclear factor-κB (NF-κB) pathway.METHODS:The cell viability was measured by CCK-8 assay.The intracellular levels of reactive oxygen species ( ROS) were detected by DCFH-DA staining .The number of apoptotic cells was tested by Hoechst 33258 nuclear staining .Mitochondrial membrane potential ( MMP) was examined by JC-1 staining.The levels of NF-κB p65 subunit and cleaved caspase-3 protein were de-termined by Western blotting.RESULTS: Treatment of H9c2 cardiac cells with 35 mmol/L glucose (HG) for 30, 60, 90, 120 and 150 min significantly enhanced the levels of phosphorated ( p) NF-κB p65, peaking at 60 min.Co-treatment of the cells with 1 μmol/L Ang-(1-7) and HG for 60 min attenuated the up-regulation of p-NF-κB p65 induced by HG. Co-treatment of the cells with Ang-(1-7) at concentrations of 0.1~30μmol/L and HG for 24 h inhibited HG-induced cy-totoxicity, evidenced by an increase in cell viability .On the other hand, 1 μmol/L Ang-(1-7) ameliorated HG-induced apoptosis, oxidative stress and mitochondrial damage , indicated by decreases in the number of apoptotic cells , cleaved caspase-3 level, ROS generation and MMP loss .However, the above cardioprotective effects of Ang-(1-7) were markedly blocked by A-779, an antagonist of Ang-(1-7) receptor (Mas receptor).Similarly, co-treatment of H9c2 cardiac cells with 100 μmol/L PDTC ( an inhibitor of NF-κB) and HG for 24 h also obviously reduced the above injuries induced by HG.CONCLUSION:Ang-(1-7)/Mas receptor axis prevents the cardiomyocytes from the HG-induced injury by inhibiting NF-κB pathway .

Objective To investigate the single nucleotide polymorphisms(SNPs) rs3733591(C>T) of SLC2A9 gene in Chinese Han population, and to explore the association of this gene polymorphisms with gout susceptibility, tophi, serum uric acid levels, other clinical and laboratory data and the levels of SLC2A9 mRNA of peripheral blood mononuclear cells(PBMCs). Methods ① A total of 297 primary gout arthritis patients(GA) and 211 normal controls(NC) were enrolled into this study. The clinical and laboratory data of patients were collected. The genotypes and alleles frequencies were measured by using TaqMan ?SNP Geno-typing Assays and the possible association between gene polymorphism of SLC2A9 and gout was investigated by Chi-square test. The odds ratios(OR) and 95% confidence intervals(95%CI) were calculated. ② The lev-els of SLC2A9 mRNA on PBMCs of 86 gout patients(46 patients in remission) and controls were measured by real-time quantitative polymerase chain reaction (RT-qPCR). The nonparametric test was used to analyze the expression in different groups. Results The frequencies of genotypes and alleles of rs3733591(C>T) in gout patients were different from controls(P<0.05). The frequency of TT genotype was significantly lower than that in controls (P<0.05) and the relative risk of this genotype to develop gout was 0.647 (95%CI: 0.452-0.925). Moreover, the frequency of T allele in cases was much lower than in controls (60.9% vs 69.2%, χ2=7.324, P=0.007, OR=0.695), but the frequency of C allele was much higher(39.1% vs 30.8%, χ2=1.440, P=0.007, OR=1.440). Interestingly, the levels of SLC2A9 mRNA on PBMCs in gout patients who carried TC genotype of rs3733591 was higher than those who carried TT genotype(P<0.05). There was no difference in the expression of SLC2A9 mRNA on PBMCs among different genotype carriers of rs3733591 in controls (P>0.05). However, there was no significant difference in the distribution of genotypes and alleles between 30 tophaceous gout patients and 190 non-tophaceous gout patients(P>0.05). Conclusion Results of present study suggest the rs3733591(C>T) polymorphism of the SLC2A9 gene might be associated with gout development, but not with tophaceous gout. The C allele predisposes to gout, and TT genotype and T allele might protect Chinese Han population from developing gout. The rs3733591(C>T) polymorphism probably affects the susceptibility to gout by influencing the f expression of SLC2A9 mRNA susceptibility.