The effects of all-trans-retinoic acid (ATRA) in low doses supplementation on concentrations of polar retinoid metabolites (PRM) and retinoids in the ethanol-fed rat liver, and on hepatocyte injury were investigated. The rat model of alcoholic liver disease (ALD) was induced by intragastric infusion of ethanol, and then the rats were administrated with ATRA in two different doses (150 microg/kg body weight and 1.5 mg/kg body weight) for 4 weeks. Concentrations of retinoids in rat liver and plasma were determined by using HPLC. Liver tissues pathologic changes were observed under the light microscopy and electron microscopy. The serum transaminases concentrations were measured. The results showed that the HPLC analysis of retinoids revealed that retinoids (vitamin A, RA, retinyl palmitate) concentrations in ethanol-fed rat liver and RA concentration in ethanol-fed rat plasma were markedly diminished (P<0.01) after ethanol feeding for 12 weeks. Furthermore, obvious peaks of PRM were formed in livers of ethanol-fed rats. ATRA 150 microg/kg supplementation in ethanol-fed rats for 4 weeks raised RA concentration in both liver and plasma, and also raised vitamin A concentration in liver to control levels, partially restored retinyl palmitate concentration (P<0.05) in liver. ATRA 1.5 mg/kg supplementation raised not only RA concentrations in liver and plasma but also retinyl palmitate concentrations in liver. However, the vitamin A concentration in liver of ATRA-supplemented rats (1.5 mg/kg) was higher than that of controls (P<0.05). The histologic observation of liver tissues indicated that ATRA treatment notably alleviated hepatocellular swelling, steatosis, the swelling of mitochondria and proliferation of smooth endoplasmic reticulum (SER). ATRA treatment greatly decreased levels of serum transaminases as compared with the only ethanol-fed group (P<0.05). It was concluded that low-dose ATRA treatment could restore retinoids concentrations and abolish the PRM formation in liver of ALD rats, and then ameliorate the injury of liver cells.

Objective:To investigate the effect of segmental Le FortⅠosteotomy and bilateral sagittal split ramus osteotomy ( BSSRO ) on the condyle position in skeletal class Ⅲ malocclusion patients . Methods:In this retrospective study , 19 patients with skeletal class Ⅲmalocclusion who met the inclu-sion criteria were enrolled .All the patients underwent the segmental Le FortⅠ osteotomy and BSSRO . Cone beam computed tomography ( CBCT) scans were performed in the following phases:T1:within one week before the surgeries;T2:within one week post-surgery;T3:three months post-surgery;T4:6 to 14 months post-surgery .The posterior spaces , anterior spaces and the superior spaces of the bilateral tem-poromandibular joints were measured according to the Kamelchuk method respectively .The fossa ratios of the condyle and the distribution of the condyle positions related to the glenoid fossa ( anterior , concentric and posterior position ) were calculated .The results were analyzed statistically .Results:The posterior space , the anterior space and the superior space of bilateral temporomandibular joints in T 2 phase [ right:(2.78 ±1.23) mm, (2.47 ±0.89) mm, (3.07 ±0.85) mm; left: (2.93 ±0.83) mm, (2.69 ± 1.14) mm, (3.44 ±1.16) mm] showed significantly larger spaces than those in T 1 phase [right:(1.81 ±0.95) mm, (1.65 ±0.55) mm, (2.13 ±0.52) mm;left:(2.12 ±1.05) mm, (1.79 ±0.59) mm, (2.15 ±0.93) mm],in T3 phase [right:(2.08 ±1.25) mm, (1.79 ±0.68) mm, (1.80 ±0.76) mm;left: (2.05 ±0.75) mm, (1.99 ±0.94) mm, (2.14 ±0.71) mm] and in T4 phase [right:(1.94 ±0.77) mm, (1.81 ±0.69) mm, (2.05 ±0.69) mm;left:(1.89 ±0.69) mm, (1.80 ±0.61) mm, (2.19 ±0.75) mm], P<0.05.No significant differences were observed among T 1,T3 and T4 pha-ses in the terms of the joint spaces of both sides ( P >0.05).The fossa ratio and the condyle position related to the glenoid fossa had no significant difference in all the four phases (P>0.05).The results suggested that the condyle moved downward in T 2 phase and changed to the original pre-surgery position in T3 phase, then keot stable in T4 phase.Conclusion:Segmental Le FortⅠ osteotomy and BSSRO caused significant and transient changes of the condyle position in skeletal class Ⅲmalocclusion patients . However , the condyle tended to move back to the original pre-surgery position and might keep stable .

Objective To explore the possible role of proline-rich tyrosine kinase (Pyk2) in the pathogenesis of systemic lupus erythematosus (SLE).Methods The expression of Pyk2 in the peripheral blood mononuclear cells (PBMCs) of 50 patients with SLE and 36 healthy controls were tested with RT-PCR assay.The activation of Pyk2 was inhibited using specific inhibitor Pyk2 (TyrA9).Semi-quantitative PCR methodwas used to detect the Blys expression of PBMCs.One-way ANOVA and Pearson's correlation analysis were used for statistical analysis.Results The Pyk2 expression level (28.31 ±0.91) of SLE patients was significantly higher than that in healthy controls (33.69±0.04),the difference was statistically significant (P＜0.05).The activation of Pyk2 was stimulated and the expression levels of Blys in the PBMCs of patients with SLE was elevated.By inhibiting the activation of Pyk2,the BLyS expression levels decreased significantly.Conclusion Pyk2 may be involved in the abnormal activation of lymphocytes which lead to the pathogenesis of SLE.Pyk2 expression is associated with SLE disease activity,disease aggravation,and the Pyk2 expression levels is also increased significantly.In addition,the expression level of Pyk2 is higher in patients with renal involvement than those patients with other organ involvement.

Objective By detecting the expression levels of Foxp3 in CD4+CD25+ regulatory T cells of peripheral blood mononuclear cells (PBMCs) in rheumatoid arthritis (RA),and the Foxp3 gene promoter region methylation to explore its role in the pathogenesis of RA.Methods Twenty-five RA patients and 10 healthy controls were selected,and the PBMCs were extracted by density gradient centrifugation.Foxp3 expression levels of CD4+CD25+ regulatory T cells were detected by flow cytometry.The real-time fluorescence quantitative PCR assay was used to detect the Foxp3 mRNA expression in PBMCs; and bisulfate processing gene sequencing was used to determinethe differences in Foxp3 gene promoter sequence methylation level of PBMCs.The comparison between groups was analyzed using one-way ANOVA; two sets of qualitative data were compared using Fisher's exact test.Results The expression levels of Foxp3 mRNA in the CD4+CD25+regulatory T cells of active RA patients (2.31±0.25) was significantly lower than inactive RA group (3.68±0.26) and healthy controls (5.67±0.34),the difference was statistically significant (P＜0.05).The Foxp3 mRNA expression level in inactive RA group was lower than that of the healthy controls (P＜0.05).Foxp3 promoter region-67,-74 sites of methylation level in PBMCs of RA patients (46％) was significantly higher than that of the healthy controls (6％).Conclusion Reduction in the number of CD4+CD25+ regulatory T cells may be involved in the pathogenesis of RA and Foxp3 gene promoter methylation levels plays a key role in this process.

Objective To assess the efficacy and safety of tacrolimus (TAC) combined with methotrexate (MTX) for the treatment of refractory rheumatoid arthritis (RA),and to compare it with cyclophosphamide (CTX) added to MTX for the treatment of refractory RA.Methods Thirty-six cases of refractory RA patients were divided into the observation group and the control group.TAC+MTX were used in the observation group,and CTX+MTX were used in the control group.We used repeated measures to analyze the variance and Fisher exact probability method to analyze the efficacy at 8 weeks and 24 weeks.Results The effective rate of the observation group in 8 weeks,24 weeks were 77.8％(14 cases) and 100％(18 cases) respectively,while those of the control group were 11.1％ (2 cases) and 44.4％(8 cases),it showed that both TAC+MTX and CTX+MTX in the treatment of refractory RA were effective,but the efficacy of TAC+MTX was better than CTX+MTX,the difference of C reactive protein (CRP) and disease activity score (DAS)28 was statistically significant (P＜0.05),and it could significantly improve the clinical symptoms and laboratory indexes.Conclusion TAC+MTX is effective and safe in treating refractory RA,and is worth of spreading.

Objective:To study the effects of MYC-induced long non-coding RNA (MINCR) targeting miR-584-3p on the proliferation, invasion and migration of rheumatoid arthritis synovial fibroblasts (RASFs).Methods:Synovial tissue samples were collected from 25 rheumatoid arthritis (RA) patients and 25 patients with joint trauma undergoing joint replacement surgery. Expression of MINCR and miR-584-3p in synovial tissue was detected using Real-time quantitative polymerase chain reaction (PCR). RASFs were separated for in- vitro culture, and RASFs were transfected with MINCR small interfering RNA (si-MINCR), miR-584-3p mimics, si-MINCR and miR-584-3p inhibitors (anti-miR-584-3p). Changes in cellular viability, clone formation, migration and invasion were detected by cell counting kit (CCK-8), plate cloning experiment, scratch healing test, Transwell test, respectively. Dual luciferase reporter gene assay was applied to evaluate the effect of miR-584-3p on the luciferase activity of MINCR. The independent t-test was used to analyze the differences between the two groups, and the one-way analysis of variance (ANOVA) and SNK- q test were used to analyze the differences between multiple groups. Results:MINCR was significantly up-regulated in RA synovial tissue compared to normal synovial tissue [(3.27±0.36) vs (1.00±0.08), t=30.777, P<0.01], whereas miR-584-3p was significantly down-regulated in RA synovial tissue [(0.43±0.05) vs (1.00±0.06), t=36.491, P<0.01]. After interference with MINCR expression, the activity [(0.52±0.04) vs (1.05±0.09), t=16.144, P<0.01] and clone formation number [(45±5) vs (99±9), t=15.960, P<0.01], scratch healing rate [(28±3)% vs (69±6)%, t=18.013, P<0.01] and invasion number [(53±5) vs (113±12), t=14.019, P<0.01] of RASFs were significantly decreased. After overexpression of miR-584-3p, the activity [(0.65±0.05) vs (1.02±0.09), t=26.063, P<0.01], clone formation number [(52±5) vs (98±8), t=14.619, P<0.01], scratch healing rate [(35±3)% vs (68±6)%, t=13.711, P<0.01] and invasion number [(62±5) vs (117±11), t=13.346, P<0.01] of RASFs were significantly decreased. miR-584-3p could specifically bind to MINCR and significantly inhibit the luciferase activity [(0.45±0.04) vs (0.97±0.06), t=21.633, P<0.01]. Compared with interference with MINCR, the activity [(0.92±0.07) vs (0.49±0.04), t=16.000, P<0.01], the clone formation number [(86±8) vs (45±5), t=14.008, P<0.01], scratch healing rate [(59±7)% vs (27±3)%, t=13.200, P<0.01], and invasions number [(99±9)% vs (54±5)%, t=13.414, P<0.01] of RASFs were significantly increased after miR-584-3p and MINCR were inhibited simultaneously. Conclusion:Interfering lncRNA MINCR can inhibit the proliferation, migration and invasion of RASFs by targeting and up-regulating miR-584-3p.

The effects of all-trans-retinoic acid (ATRA) in low doses supplementation on concentrations of polar retinoid metabolites (PRM) and retinoids in the ethanol-fed rat liver, and on hepatocyte injury were investigated. The rat model of alcoholic liver disease (ALD) was induced by intragastric infusion of ethanol, and then the rats were administrated with ATRA in two different doses (150 μg/kg body weight and 1.5 mg/kg body weight) for 4 weeks. Concentrations of retinoids in rat liver and plasma were determined by using HPLC. Liver tissues pathologic changes were observed under the light microscopy and electron microscopy. The serum transaminases concentrations were measured. The results showed that the HPLC analysis of retinoids revealed that retinoids (vitamin A,RA, retinyl palmitate) concentrations in ethanol-fed rat liver and RA concentration in ethanol-fed rat plasma were markedly diminished (P＜0.01) after ethanol feeding for 12 weeks. Furthermore, obvious peaks of PRM were formed in livers of ethanol-fed rats. ATRA 150 μg/kg supplementation in ethanol-fed rats for 4 weeks raised RA concentration in both liver and plasma, and also raised vitamin A concentration in liver to control levels, partially restored retinyl palmitate concentration (P＜0.05) in liver. ATRA 1.5 mg/kg supplementation raised not only RA concentrations in liver and plasma but also retinyl palmitate concentrations in liver. However, the vitamin A concentration in liver of ATRA-supplemented rats (1.5 mg/kg) was higher than that of controls (P＜0.05). The histologic observation of liver tissues indicated that ATRA treatment notably alleviated hepatocellular swelling,steatosis, the swelling of mitochondria and proliferation of smooth endoplasmic reticulum (SER).ATRA treatment greatly decreased levels of serum transaminases as compared with the only ethanol-fed group (P＜0.05). It was concluded that low-dose ATRA treatment could restore retinoids concentrations and abolish the PRM formation in liver of ALD rats, and then ameliorate the injury of liver cells.

Objective:In this study, the role of IL-17 in the pathogenesis of idiopathic myositis (IIM) was preliminarily investigated by detecting the expression of IL-17 in the muscle tissues of patients with idiopathic inflammatory myositis (IIM) and normal controls.Methods:Twenty-eight patients (20 in DM group with dermatomyositis and 8 in ASS group with anti-synthase syndrome) who were diagnosed with IIM after muscle biopsy and autoantibody detection in our hospital for the first time from October 2019 to August 2021 were included. Twelve cases with normal muscle tissue matched for age and sex were included as the control group. Western blot and immunohistochemical techniques were used to detect the expression level of IL-17 in muscle tissue, and enzyme-linked immunosorbent assay (ELISA) was used to detect serum IL-6. Mann-Whitney U rank sum test was used to compare the difference of IL-17 expression in muscle tissue between the two groups, and non-parametric test was used for comparison between multiple groups. Chi-square test and Spearman rank correlation analysis were used, and P<0.05 was considered statistically significant. Results:① The expression level of IL-17 in IIM muscle tissue[1.63(1.30, 2.05)pg/ml was higher than that in control group[1.00(0.96, 1.00)pg/ml, and the difference was statistically significant ( Z=-3.52, P<0.001). The difference be-tween DM[1.94(1.58, 2.14)pg/ml] and ASS[1.22(1.04,1.55)pg/ml was statistically significant ( Z=-3.20, P=0.001). ② Compared with healthy control group [4.08(3.01, 5.67)pg/ml, the expression of IL-6 in ⅡM serum[8.88(4.93, 13.64) was high ( Z=-3.01, P=0.003), which was positively correlated with the expression of IL-17 ( r=0.42, P=0.027). ③ The ex-pression of IL-17 in muscle tissue was higher in IIM associated with muscle weakness[1.91(1.56, 2.14) pg/ml vs 1.50(1.04, 2.00)pg/ml] ( Z=-1.38, P=0.020), dysphagia [2.06(1.99, 2.14)pg/ml vs 1.62(1.52, 2.04)pg/ml] ( Z=-2.74, P=0.010) and skin involvement[1.98(1.57, 2.14)pg/ml vs 1.04(0.86, 1.61)pg/ml] ( Z=-3.20, P<0.010), and the differences were statistically significant ( P<0.05). ④IL-17 was positively correlated with Myoact-total activity ( r=0.51, P=0.006), Myoact-muscle symptom ( r=0.45, P=0.016), erythrocyte sedimen tation ( r=0.48, P=0.020), and myoenzyme increase ( r=0.56, P=0.002). Conclusion:IL-17 and IL-6 are synergistically involved in the pathogenesis of IIM, suggesting that IL-17 is the therapeutic target of IIM.

Field experiments were conducted in Shangluo pharmaceutical base in Shaanxi province to study the effect of nitrogen, phosphorus and potassium in different fertilization levels on Platycodon grandiflorum soil microorganism and activities of soil enzyme, using three-factor D-saturation optimal design with random block design. The results showed that N0P2K2, N2P2K0, N3P1K3 and N3P3K1 increased the amount of bacteria in 0-20 cm of soil compared with N0P0K0 by 144.34%, 39.25%, 37.17%, 53.58%, respectively. The amount of bacteria in 2040 cm of soil of N3P1K3 increased by 163.77%, N0P0K3 increased the amount of soil actinomycetes significantly by 192.11%, while other treatments had no significant effect. N2P0K2 and N3P1K3 increased the amounts of fungus significantly in 0-20 cm of soil compared with N0P0K0, increased by 35.27% and 92.21%, respectively. N3P0K0 increased the amounts of fungus significantly in 20-40 cm of soil by 165.35%, while other treatments had no significant effect. All treatments decrease soil catalase activity significantly in 0-20 cm of soil except for N2P0K2, and while N2P2K0 and NPK increased catalase activity significantly in 2040 cm of soil. Fertilization regime increased invertase activity significantly in 2040 cm of soil, and decreased phosphatase activity inordinately in 0-20 cm of soil, while increased phosphatase activity in 2040 cm of soil other than N1P3K3. N3P0K0, N0P0K3, N2P0K2, N2P2K0 and NPK increased soil urease activity significantly in 0-20 cm of soil compared with N0P0K0 by 18.22%, 14.87%,17.84%, 27.88%, 24.54%, respectively. Fertilization regime increased soil urease activity significantly in 2040 cm of soil other than N0P2K2.
Bacteria ; enzymology ; growth & development ; isolation & purification ; metabolism ; Bacterial Proteins ; analysis ; metabolism ; Catalase ; analysis ; metabolism ; Fertilizers ; analysis ; Fungal Proteins ; analysis ; metabolism ; Fungi ; enzymology ; growth & development ; isolation & purification ; metabolism ; Nitrogen ; metabolism ; Phosphoric Monoester Hydrolases ; analysis ; metabolism ; Phosphorus ; metabolism ; Potassium ; metabolism ; Soil ; chemistry ; Soil Microbiology ; Urease ; analysis ; metabolism

Interferon-γ (IFN-γ), a key effector molecule in anti-tumor immune response, has been well documented to correlate with the intratumoral infiltration of immune cells. Of interest, however, a high level of IFN-γ has been reported in salivary adenoid cystic carcinoma (SACC), which is actually a type of immunologically cold cancer with few infiltrated immune cells. Investigating the functional significance of IFN-γ in SACC would help to explain such a paradoxical phenomenon. In the present study, we revealed that, compared to oral squamous cell carcinoma cells (a type of immunologically hot cancer), SACC cells were less sensitive to the growth-inhibition effect of IFN-γ. Moreover, the migration and invasion abilities of SACC cells were obviously enhanced upon IFN-γ treatment. In addition, our results revealed that exposure to IFN-γ significantly up-regulated the level of programmed death ligand 1 (PD-L1) on SACC cell-derived small extracellular vesicles (sEVs), which subsequently induced the apoptosis of CD8⁺ T cells through antagonizing PD-1. Importantly, it was also found that SACC patients with higher levels of plasma IFN-γ also had higher levels of circulating sEVs that carried PD-L1 on their surface. Our study unveils a mechanism that IFN-γ induces immunosuppression in SACC via sEV PD-L1, which would account for the scarce immune cell infiltration and insensitivity to immunotherapy.
B7-H1 Antigen/metabolism* ; CD8-Positive T-Lymphocytes/pathology* ; Carcinoma, Adenoid Cystic/pathology* ; Carcinoma, Squamous Cell/pathology* ; Cell Line, Tumor ; Humans ; Immunosuppression Therapy ; Interferon-gamma/pharmacology* ; Mouth Neoplasms/metabolism* ; Programmed Cell Death 1 Receptor/metabolism* ; Salivary Gland Neoplasms/pathology*