Objective To explore the regulatory mechanism of NFE2L2/KEAP1 in alveolar bone repair induced by periodontitis.Methods A rat periodontitis model was established and divided into four groups:Control group(n=6);Periodontitis model group(n=6);Periodontitis+lentivirus empty vector group(n=6);Periodontitis+NFE2L2 overexpression plasmid group(n=6).Histopathological changes in each group were observed using HE staining.TRAP staining was used to detect osteoclast positivity,while ELISA was employed to measure inflammatory cytokine levels in tissues.Immunofluorescence and qPCR were used to detect NFE2L2 expression,and western blot was used to assess the expression of osteogenic proteins ALPL2,RUNX2,and COL1.Primary periodontal ligament cells(hPDLCs)were cultured,and cells were transfected to overexpress NFE2L2 and KEAP1.The cells were divided into six groups:Normal group;Model group;pcDNA-NC group;pcDNA-NFE2L2 group;pc-NFE2L2+pcDNA-NC group;pc-NFE2L2+pcDNA-KEAP1 group.A cellular model was established,and the morphology of primary hPDLCs was observed under a microscope.Cell proliferation was assessed using CCK-8.Osteogenic mineralization was observed using alizarin red staining,and western blot was used to detect osteogenic proteins and autophagy markers.Cell migration was observed using a scratch assay.Results(1)After model induction,redness,swelling of the gums,extensive inflammatory infiltration,and alveolar bone resorption were observed,confirming successful model establishment.Partial tissue recovery occurred after NFE2L2 overexpression via lentivirus.(2)After model induction,osteoclast positivity increased,confirming successful model establishment.Overexpression of NFE2L2 reduced osteoclast positivity(P<0.001).(3)After model induction,levels of IL-1β,IL-10,and TNF-α were significantly higher than in the normal group(P<0.05),confirming successful model establishment.Transfection with NFE2L2 lentivirus reduced inflammatory cytokine levels(P<0.0001).After model induction,osteogenic protein expression decreased compared to the normal group,but overexpression of NFE2L2 increased osteogenic protein expression(P<0.05).(5)LPS treatment significantly reduced cell viability,while NFE2L2 overexpression enhanced it(P<0.0001).(6)LPS treatment reduced calcified nodules,while NFE2L2 overexpression increased them.Addition of pcDNA-KEAP1 reduced mineralized nodules.(7)LPS treatment decreased osteogenic protein expression,while NFE2L2 overexpression increased it.However,addition of pcDNA-KEAP1 reduced osteogenic protein expression(P<0.05).(8)LPS treatment reduced cell migration,whereas NFE2L2 overexpression enhanced it(P<0.0001).(9)Expression of autophagy markers decreased after LPS treatment,but increased after transfection with NFE2L2 plasmid.However,addition of pcDNA-KEAP1 reduced the expression of autophagy markers(P<0.05).Conclusion This study identified the regulatory role of NFE2L2/KEAP1 in periodontitis,providing a scientific basis for the treatment of periodontitis.

Objective To investigate the effects of artesunate (ART) on interstitial pneumonia and sialadenitis in MRL/lpr mice.Methods A total of 18 MRL/lpr mice were randomly allocated to a hydroxychloroquine sulfate (HCQ) group,a ART group and a control group.At the age of 18 weeks,the mice in the HCQ group and ART group were given HCQ 150 mg/kg daily and ART 50 mg/kg daily for 12 weeks,respectively.The histopathological changes of pneumonitis and submaxillaritis were assessed by hematoxylin and eosin staining.The levels of monocyte chemoattractant protein-1 (MCP-1) in the serum and urine were measured by the enzyme-linked immunosorbent assay.Results At the age of 30 weeks,the index of peribronchiolar lesion (1.62 ± 0.19,1.52 ± 0.30 vs.1.95 ± 0.34;all P＜0.05),the index of perivascular lesion (1.23 ± 0.18,1.28 ± 0.12 vs.1.57 ± 0.33;all P＜0.05),the alveolar lesions index (1.35 ± 0.16,1.05 ± 0.15 vs.1.72 ± 0.34;all P＜0.05) and the submaxillaritis index (1.48 ± 0.22,1.43 ± 0.15 vs.1.84 ± 0.34;all P＜0.05) in the HCQ group and the ART group were significantly decreased than those in the control group.The MCP-1 levels in the serum (1 103.02 ± 185.56 pg/ml,1 072.37 ± 242.43 pg/ml vs.1 490.67 ± 329.43 pg/ml;all P＜0.05) and urine (189.16 ± 70.85 pg/ml,198.79 ± 113.47 pg/ml vs.446.79 ± 192.31 pg/ml;all P＜0.05) in the HCQ group and the ART group were significantly lower than those in the control group.Conclusion ART can decrease the MCP-1 level,and ameliorate interstitial pneumonitis and sialadenitis in MRL/lpr mice.

Objective To investigate the prevalence status of overweight and obesity among the middle‐aged and elderly pop‐ulations in southwest area and their influence factors .Methods By adopting the stratified cluster sampling method ,the mode of the questionnaire investigation combined with the physical examination was adopted to collect the effective data in 7 205 permanent resi‐dents aged 40-79 years old in cities and countries of Chongqing and Chengdu ,including the contents of age ,waistline ,hipline ,dias‐tolic blood pressure ,heart rate ,postprandial blood sugar ,triglyceride ,exercise frequency ,uric acid ,etc .The prevalence rates of over‐weight and obesity among the middle‐aged and elderly populations in southwest area and their distribution characteristics in cities and countries ,and in male and female populations were described .The related influence factors of overweight and obesity were in‐vestigatedResults The prevalence rate of overweight was 35 .91% among the middle‐aged and elderly populations in southwest area (37 .02% for cities ,32 .92% for countries ,33 .84% for males and 37 .41% for females);the prevalence rate of obesity was 11 .40%(11 .13% for cities ,11 .40% for countries ,8 .94% for males and 12 .84% for females);among them ,the overweight rate in cities was higher than that in countries ,the overweight rate in females was higher than that in males .The obesity rate in females was higher than that in males(P<0 .01) ,the obesity rate had no difference between cities and countries(P>0 .05) .The multivariate re‐gression analysis showed that lack of exercise ,overweight or obesity family history were the risk factors of male overweight and o‐besity ,while the difference between cities and countries ,lack of exercise were the risk factors of female overweight and obesity .Con‐clusion The overweight and obesity rates among the middle‐aged and elderly populations in southwest area are high ,the body mass in nearly half of people exceeds the normal standard .Strengthening the overweight and obesity related health education in communi‐ties and restraining the serious trend of overweight in the population of this area have an important significance for the prevention and treatment of cardiovascular diseases .

Objective To explore the epidemiological status of abnormal glucose metabolism and its influential factors among middle and aged population with hypertension in Chengdu area. Methods In 2008, after adopting the methods of stratified cluster sampling, the authors investigated 4685 subjects of the middle and aged population between the age of 40-79 in Chengdu urban and rural area by checking blood pressure and oral glucose tolerance test (OGTY). Patients with previously known diabetes mellitus (DM) were only asked to perform fasting glucose and to carry out a questionnaire. Comparison of the prevalence rates of abnormal glucose metabolism in hypertensive and non-hypertensive subjects was carried out. The prevalence rates of isolated impaired glucose tolerance (I-IGT) and isolated postprandial hyperglycemia (IPH) among middle and aged subjects with hypertension were acquired and the influential factors of abnormal glucose metabolism among middle and aged subjects with hypertension were analyzed. Results The prevalence rate of abnormal glucose metabolism in the hypertensive subjects was obviously higher than that in the non-hypertensive subjects; without using OGTT, 72.9% of the pre-diubetic and 54. 4% of the new diagnosed DM patients would remain undiagnosed if fasting plasma glucose detection was used alone. Age, diabetic history of first degree relatives ,overweight or obesity were the risk factors for the development of abnormal glucose metabolism among middle and aged male subjects with hypertension in Chengdu area. Exercise training and high education level were the protective factors. Age, diabetic history of first degree relatives,abdominal obesity and hypertriglyceridemia were the risk factors for the development of abnormal glucose metabolism among middle and aged female subjects with hypertension in Chengdu area. Conclusions More than 50% of middle and aged subjects with hypertension in Chengdu area has accompanying abnormal glucose metabolism. OGTT easily discloses the abnormal status and should be a routine procedure in the diagnosis of pre-diabetes or DM in such population. Appropriate exercise, learning diabetes-related knowledge to take reasonable lifestyle, and intervention of metabolic factors such as overweight or obesity are advised. Abdominal obesity and hypertriglyceridemia play important roles in leading to abnormal glucose metabolism among middle and aged population with hypertension.

Objective To investigate the efficacy of umbilical cord mesenchymal stem cells (UC-MSCs) transplantation in the treatment of the MRL/lpr mice. Methods Twenty four 18-week-old MRL/lpr female mice were divided into 3 groups:group 1 (G1) were transplanted with 1×106 UC- MSCs through caudal vein, group 2 (G2) were transplanted with 1×106 UC- MSCs three times and group 3 (G3) were treated with 0.5 ml normal saline as controls. Enzyme linked immunosorbent assay (ELISA) was used to measure the levels of serum anti-dsDNA antibodies. Twenty-four hours proteinuria and body weight were assessed every two weeks. The histopathology changes of the kidneys and lungs were observed. Results ① At the 25th weeks, the 24 hours proteinuria in group G1 (2.3±1.9) mg and G2 (1.8±1.4) mg was decreased than that in the control group (3.8±2.1) mg (P<0.05), and at the 27th weeks, that of groups G1 (2.5±1.5) mg and G2 (1.9±1.2) mg was also significantly decreased than in the control group (5.4±2.4) mg (P<0.01); ② From the 24th week, the body weight of groups G1 and G2 increased significantly than that of the control group (P< 0.05). At week 29, serum creatinine decreased significantly in both groups G1 (7.2±3.2) μmol/L and G2 (6.2±2.8) μmol/L than in the control group (12.5±2.3 ) μmol/L (P<0.05); ③One week after transplantation, the levels of anti-dsDNA antibodies in group G1 (46±11)×102 U/ml and G2(49×43)×102 U/ml were bothsignificantly decreased than those of the control groups (99±42)×102 U/ml (P<0.05) and the difference between group G2 (36±15)×102 U/ml and the controls (68±32)×102 U/ml was statistically significant; ④The nephron crescent formation in group G1 (0.12±0.07) and G2 (0.08±0.02) was significantly lower that of the control group (0.20±0.06) (P<0.05) and that of group G2 was significantly less that of froup G1 (P<0.05); ⑤ The interstitial pneumonitis was singnificantly milder in group G1 than group G2. Conclusions UC- MSCs is very effective in treating MRL/lpr mice. It is safe and free of rejection reactions.

Objective To investigate the therapeutic effects and mechanisms of hydroxychloroquine (HCQ) in the MRL/lpr mice. Methods MRL/lpr mice were divided into HCQ, the artesunate (ART) and proteinuria was detected with Coomassi Brilliant blue method. Enzyme linked immunosorbent assay (ELISA) was used to measure the anti-doubM-stranded DNA (ds-DNA) antibody. Renal tissue sections were dyed By PAS methods. The percentage of CD4+ Foxp3+ T cells in the spleen and lymph nodes were detected by flow 2.0) mg groups were decreased than in the control group (4.8±3.2) mg (P<0.05). And it was also lower in the HCQ (2.8±1.1) mg and ART (2.4±1.9) mg group than in the control group (6.4±1.9) mg (P<0.01) at 30 in the control group (37.1±1.0) g (P<0.01), while serum creatinine decreased significantly (7.8±4.0) μmol/L than in the control group (12.5±2.3) μmol/L (P<0.05), and the serum anti ds-DNA antibodies levels (3047±renal damage in the HCQ group and in the ART group was Both significantly improved than that in the entages of CD4+ Foxp3+ T cells in spleen when compared with the control group (1.5±0.5)% (P<0.05). The mice in the HCQ group (0.68±0.33)% and in the ART group (0.97±0.28)% had higher percentages of CD4+ Foxp3+ T cells in lymph nodes as compared with control group (2.15±0.72)%(P<0.01). Conclusion HCQ is effective in treating MRL/lpr lupus mice. It can improve the pathologic lesions of lupus nephritis, reduce proteinuria and antibody production. Both HCQ and ART can up-regulate the percentage of CD4+ Foxp3+ T cells in spleen of MRL/lpr mice.

Objective To investigate the muhilineage differentiation potential of bone marrow-derived mesenchymM stem eels (MSCs) in patients with systemic lupus erythematosus (SLE).Methods Density gradient centrifugation and plastic adherence methods were used for isolation of marrow-derived MSCs.Then tIIeir differentiation potentiality to lipoblasts and osteoblasts waft tested.MSCs loading on hydroxyapatite were elnbedded in the nude mouse's subcutaneous tissues.Eight weeks later.osteogenesis was evaluated by HE staining.PPA Rγ2,LPL,Runx2/CBFA1,osteocalcin gene expression in MSCs after differentiation were examined by RT-PCR.Results The positive rates of lipoblasts stained by oil red O and optical density in SLE were decreased than in the control group[(35±7)% vs (80±5)%] (0.14±0.04 vs 0.27±0.04),and the positive rates of osteoblasts stained by Alizarin Red S in SLE were decreased than those in the control group [(35±4)% vs (45±4)%].Osteoblast differentiation in the SLE group was less than that of the contro]group.The mRNA expression of LPL (0.369±0.020 vs 0.481±0.038).Runx2/CBFA1 (0.371±0.000 vs 0.563±0.069).osteoealcin (0.819±0.023 vs 0.962±0.049) of MSCs after difierentiation in the SLE group was decreased than that of the control group.There was no significant difference in the expression of PPARγ2 mRNA between SLE and controI group (0.421±0.052 vs 0.441±0.012).Conelusion MSCs from SLE have abnormalities in osteogenie and adipogenic differentiation potential.

Objective To investigate the effects of tumor necrosis factor-alpha (TNF-α) on insulin sensitivity and glucose-lipid metabolism in TNF-α-induced IR mice. Methods Male C57BL/6J mice were given an intraperitoneal injection of TNF-α (H group,6μg/kg; M group,3μg/kg; L group,1μg/kg;twice daily) and saline (NC group) for 7 days. The plasma glucose and insulin were assayed during intravenous glucose tolerance test (IVGTT) and hyperinsulinemic-euglycemic clamp combined with 3-[3H] glucose as a tracer was carried out. Results After TNF-α treatment,fasting blood glucose (FBG),plasma insulin and free fatty acids (FFA) were significantly elevated in H group compared with NC,L and M groups (P<0.01 and P<0.05,respectively). There was a lower glucose tolerance in H group versus other three groups during IVGTT. The insulin release by glucose stimulation was higher in H group versus NC and L groups (P<0.01 and P<0.05). Basal glucose disappearance rate (GDR) and hepatic glucose production (HGP) were significantly increased in H group compared with NC group (P<0.01). During the steady-state of clamp,plasma insulin levels were significantly increased in H group versus NC group (341.7±17.7 vs 84.7±5.5mU/L,P<0.01). The suppressive effect of insulin on FFA was significantly blunted in H group compared with NC group (0.82±0.03 vs 0.43±0.07mmol/L,P<0.01). Steady-state glucose infusion rate (GIR) was significantly decreased in H group compared with NC group (39.1±2.3 vs 54.2±2.2 mg·kg-1·min-1,P<0.01). Although GDR was increased in both group,but it was still lower in H group than in control group(47.9±0.8 vs 53.9±2.0 mg.kg-1.min-1,P<0.01).As compared with baseline,HGP in the controls was almost completely suppressed during steady state of clamp,but in H group suppressed by approximately 41%. Conclusions High-dose TNF-α treatment induces the abnormality of glucose-lipid metabolism and the insulin resistance of hepatic and peripheral tissue in mice

Objective To investigate the in vivo or in vitro immune regulatory effects of allogeneic bone-marrow mesenchymal stem ceils (MSC) and human umbilical cord MSC on CD4+ Foxp3+ T cells in the peripheral blood of patients with systemic lupus erythematosus (SLE) and in the spleen of MRL/Ipr mice. Methods Human MSC were isolated and expanded from bone marrow cells of healthy donors and infused into five SLE patients. The percentages of CD4+ Foxp3+ T cells in peripheral blood were detected by flow cytometry. Human peripheral blood mononuclear cells (PBMC) were prepared by centrifugation on a Ficoll Hypaque density gradient. The MSC and PBMC from unrelated donors (MSC:PBMC =1:1,1:10,1:50) were added into 24-well plates. After 72h of co-culture, the percentages of CD4+ Foxp3+ T cells were detected by flow cytome- try. Twenty four 18-week-old MRL/Ipr female mice were divided into 3 groups and were injected with umbilical cord MSC (1×106 cells for one time, 1×106 cells for three times and 0.5 ml sodium chloride as control respectively). The percentages of CD4+ Foxp3+ T cells in spleen and lymphoid nodes were detected by flow cytometry. Results The percentages of blood CD4+ Foxp3+ T cells at one week (4.8± 1.6)% and at three months (6.0±2.6)% post MSC transplantation for patients with SLE were both higher than that before transplantation (2.1±1.2)% (n=5,P<0.05). The co-culture of normal bone marrow MSC with PBMC from SLE patients resulted in a statistically significant increase of CD+ Foxp3+ T cells percentage in PBMC on a dose dependent manner (P<0.05). The percentages of CD4+ Foxp3+ T cells of PBMC from SLE patients co-cultured with lupus MSC were lower than that of normal MSC (P<0.05). The cultured supematant of normal MSC also upregulated the percentages of CD4+ Foxp3+ T cells of PBMC from SLE patients (P<0.05). The MRL/lpr mice that had been injected umbilical cord MSC for one time and three times had higher percentages of CD4+ Foxp3+T cells in the spleen but lower in the lymphoid nodes as compared with controls (P<0.01), but without statistical significant difference. Conclusion Allogeneic or heterogeneie MSC transplantation upregulates the percentages of CD4+ Foxp3+ T cells in SLE patients or in MRL/Ipr mice. Upregnlation of Treg population may be one of the mechanisms of MSC transplantation that is effective for SLE treatment.

Objective To investigate the effects of tumor necrosis factor-alpha (TNF-?) on insulin sensitivity and glucose-lipid metabolism in TNF-?-induced IR mice. Methods Male C57BL/6J mice were given an intraperitoneal injection of TNF-? (H group,6?g/kg; M group,3?g/kg; L group,1?g/kg;twice daily) and saline (NC group) for 7 days. The plasma glucose and insulin were assayed during intravenous glucose tolerance test (IVGTT) and hyperinsulinemic-euglycemic clamp combined with 3-[3H] glucose as a tracer was carried out. Results After TNF-? treatment,fasting blood glucose (FBG),plasma insulin and free fatty acids (FFA) were significantly elevated in H group compared with NC,L and M groups (P