Objective:To investigate the effect of thunder-fire moxibustion combined with pulsed electromagnetic fields(PEMFs)on bone metabolism and motor function in individuals with primary osteoporosis.Method:Ninety patients with primary osteoporosis were randomly assigned to three groups using the envelope method:PEMFs group(n=30),thunder-fire moxibustion group(n=30),and PEMFs+thunder-fire moxibustion group(n=30).The PEMFs treatment frequency was 16Hz with an intensity of 20mT,while thunder-fire moxi-bustion was applied at bilateral Ganshu(BL18)and Shenshu(BL23).Each treatment session lasted 30 min-utes,conducted three times a week for three consecutive months.The timed up-and-go(TUG)test,serum typeⅠprocollagen amino-terminal peptide(PINP)and type Ⅰ collagen carboxy-terminal peptide(CTX-Ⅰ)were as-sessed before treatment,after 3-month treatment,and at 6-month follow-up.Bone mineral density(BMD)was measured at 6-month follow-up.Result:①Inter-group comparison:after 3-month treatment,the PEMFs+Thunder-Fire Moxibustion group showed a significant reduction in TUG time and serum CTX-Ⅰ levels compared to the PEMFs group(P＜0.05),and a more significant reduction in serum PINP levels(P＜0.01).At the 6-month follow-up,the TUG time in PEMFs+thunder-fire moxibustion group was significantly lower than PEMFs group(P＜0.01).Compared to the thunder-fire moxibustion group,the PEMFs+thunder-fire moxibustion group serum level of CTX-Ⅰ of decreased significantly after 3-month treatment(P＜0.05).②Intra-group comparison:After three months of treatment com-pared to baseline,the PEMFs group showed a significant reduction in TUG time and serum CTX-Ⅰ levels(P＜0.05),while both the Thunder-Fire Moxibustion group and the PEMFs+Thunder-Fire Moxibustion group showed significant reductions in TUG time and serum PINP and CTX-Ⅰ levels(P＜0.01).Comparison between 6-month follow-up and before treatment,the TUG time of PEMFs group decreased significantly(P＜0.05).The TUG time and the serum level of PINP in PEMFs+thunder-fire moxibustion group significantly decreased(P＜0.01).There were no significant differences observed among any of the three groups regarding total BMD measure-ments at waist and hip regions after 6-month follow-up and before treatment(P＞0.05).Conclusion:The combination of Thunder-fire moxibustion and PEMFs can enhance motor function and bone metabolism indexes in patients with primary osteoporosis,BMD shows a trend of improvement.

Objective:To investigate the effect of thunder-fire moxibustion combined with pulsed electromagnetic fields(PEMFs)on bone metabolism and motor function in individuals with primary osteoporosis.Method:Ninety patients with primary osteoporosis were randomly assigned to three groups using the envelope method:PEMFs group(n=30),thunder-fire moxibustion group(n=30),and PEMFs+thunder-fire moxibustion group(n=30).The PEMFs treatment frequency was 16Hz with an intensity of 20mT,while thunder-fire moxi-bustion was applied at bilateral Ganshu(BL18)and Shenshu(BL23).Each treatment session lasted 30 min-utes,conducted three times a week for three consecutive months.The timed up-and-go(TUG)test,serum typeⅠprocollagen amino-terminal peptide(PINP)and type Ⅰ collagen carboxy-terminal peptide(CTX-Ⅰ)were as-sessed before treatment,after 3-month treatment,and at 6-month follow-up.Bone mineral density(BMD)was measured at 6-month follow-up.Result:①Inter-group comparison:after 3-month treatment,the PEMFs+Thunder-Fire Moxibustion group showed a significant reduction in TUG time and serum CTX-Ⅰ levels compared to the PEMFs group(P＜0.05),and a more significant reduction in serum PINP levels(P＜0.01).At the 6-month follow-up,the TUG time in PEMFs+thunder-fire moxibustion group was significantly lower than PEMFs group(P＜0.01).Compared to the thunder-fire moxibustion group,the PEMFs+thunder-fire moxibustion group serum level of CTX-Ⅰ of decreased significantly after 3-month treatment(P＜0.05).②Intra-group comparison:After three months of treatment com-pared to baseline,the PEMFs group showed a significant reduction in TUG time and serum CTX-Ⅰ levels(P＜0.05),while both the Thunder-Fire Moxibustion group and the PEMFs+Thunder-Fire Moxibustion group showed significant reductions in TUG time and serum PINP and CTX-Ⅰ levels(P＜0.01).Comparison between 6-month follow-up and before treatment,the TUG time of PEMFs group decreased significantly(P＜0.05).The TUG time and the serum level of PINP in PEMFs+thunder-fire moxibustion group significantly decreased(P＜0.01).There were no significant differences observed among any of the three groups regarding total BMD measure-ments at waist and hip regions after 6-month follow-up and before treatment(P＞0.05).Conclusion:The combination of Thunder-fire moxibustion and PEMFs can enhance motor function and bone metabolism indexes in patients with primary osteoporosis,BMD shows a trend of improvement.

Objective:To investigate correlation between neutrophil extracellular traps(NETs) formation and T cell subsets in mice with experimental autoimmune thyroiditis(EAT) and the impact of active vitamin D intervention.Methods:Six-week-old female BALB/c mice were randomly divided into Control group, EAT group and 1, 25 dihydroxy vitamin D 3[1, 25(OH) 2D 3] treatment group(VitD group; n=6/group). HE staining was used to observe thyroid pathology. Plasma thyroglobulin antibody(TGAb), thyroid peroxidase antibody(TPOAb), and 1, 25(OH) 2D 3 were measured by ELISA. Peripheral NETs formation, Th1, Th2, and Th17 cell ratio from spleen were measured by flow cytometry. Correlation between NETs formation rate and Th1, Th2, and Th17 cell ratio was analyzed. Results:Compared with Control group, mice in EAT group had significantly increased thyroid inflammation scores, thyroiditis morbidity, TPOAb, TGAb levels, NETs formation rate, Th2(CD4 + IL-4 + or CD4 + IL-13 + )and Th17 cell proportions( P were <0.001, 0.002, 0.007, <0.001, <0.001, 0.003, 0.001, and 0.002, respectively), and significant decreased 1, 25(OH) 2D 3, Th1 cell proportions, Th1/Th2(CD4 + IL-4 + ), Th1/Th2(CD4 + IL-13 + ), and Th1/Th17 ratios( P were 0.010, 0.018, 0.010, 0.005, and 0.007, respectively). Compared with the EAT group, the VitD group had lower thyroid inflammation scores, TPOAb, TGAb levels, NETs formation rate, Th2(CD4 + IL-4 + or CD4 + IL-13 + ) and Th17 cell proportions( P were 0.044, 0.007, <0.001, 0.001, 0.014, 0.008, and 0.001, respectively), and significant higher Th1 cell ratio, Th1/Th2(CD4 + IL-13 + ) and Th1/Th17 ratio( P were 0.011, 0.009, and 0.003, respectively). The Th1/Th2(CD4 + IL-4 + ) was not significantly increased in VitD group compared with EAT group( P=0.174). NETs formation rate was positively correlated with Th2(CD4 + IL-4 + or CD4 + IL-13 + ) and Th17 cell proportion( r were 0.65, 0.59, and 0.61; and P were 0.004, 0.010, and 0.007, respectively), but not with Th1 cell proportion( r=-0.47, P=0.051). Conclusion:EAT mice were more prone to NETs formation. Active vitamin D may relieve immune imbalance with increased Th2 and Th17 cell ratio and decreased Th1 cell ratio by reducing the formation of NETs in EAT mice. Vitamin D played the protective role in thyroid by reducing thyroid pathological damage and thyroid autoantibody levels, and relived overall lymphocyte imbalance.

OBJECTIVETo observe the effect of crocetin on autophagy in rat hepatocytes exposed to lipopolysaccharide (LPS) and D-galactosamine (D-gal) and explore the mechanism.

METHODSCultured rat hepatocytes were exposed to LPS (1 mg/L) and Dgal (60 mg/L) to induce cell injury and treated with crocetin, 3MA, or crocetin+3MA. Twelve hours after the treatments, the cells were examined for levels of ALT, AST and LDH in the supernatant using ELISA. LC3 fluorescence in the cells following immunofluorescence staining was observed using fluorescence microscopy. Autophagosomes in the cells were observed by transmission electron microscopy, and the cellular expressions of LC3, p62 and SIRT1 were detected using Western blotting.

RESULTSThe levels of ALT, AST and LDH in the hepatocytes were elevated after LPS- and D-gal-induced injury, reached the highest levels after 3MA treatment, but were decreased significantly by crocetin treatment. LC3 fluorescence increased obviously in the injured hepatoctyes, and the increment was the most obvious in crocetin-treated cells; LC3 fluorescence was decreased significantly after 3MA treatment. Cell injury induced obvious increase in autophagy in the hepatocytes, and the number of autophagosomes increased significantly after crocetin treatment but was reduced significantly after 3MA treatment. The cell injury caused an obvious up-regulation of LC3 and SIRT1 expression and down-regulated p62 expression. LC3 and SIRT1 expression levels were the highest and the expression of p62 was the lowest in cells with crocetin treatment. 3MA treatment significantly reduced the expression of LC3 and SIRT1 and increased the expression of p62 in the injured cells.

CONCLUSIONSAutophagy is increased in injured rat hepatocytes, and crocetin can promote autophagy in the injured cells to reduce further cell injury.

Objective To study multiple-drug resistance expression of CNE1 and CNE2 cell lines induced by radiation and its reversal by cyclosporine A and interferon. Methods Drug-resistant CNE1 and CNE2 cell lines were induced by radiation, and expression of P-gp, MRP, TOPⅡ and GST-? on protein and mRNA level were analyzed by flow cytometry and reverse transcription-polymerase chain reaction methods before and after radiation. At the same time, effect of cyclosporine A and interferon on multiple-drug resistance expression induced by radiation was studied. Results The results showed that expression of P-gp, MRP and GST-? proteins on CNE1 and CNE2 cell lines were up-regulated by radiation, and cyclosporine A and interferon could reverse the up-regulation of P-gp and MRP proteins induced by radiation. Conclusion Radiation can induce multiple-drug resistance in CNE1 and CNE2 cell lines, and it can be reversed by cyclosporine A and interferon.

AIM To study effects of antioxidant NADH on damage of irradiated normal cell lines. METHODS L02 liver cells were cultured in RPMI 1640, exposed to X ray irradiation, and continued to culture in the presence or absence of NADH for 24 h. The cellular viability was determined by routine MTT method. Using fluorescence probe and confocal microscope, the level of cellular H 2O 2 was detected. Positive rate of bcl 2 and Bax protein expression in the L02 cells were analyzed by flow cytometry. RESULT NADH can not only antagonized growth inhibition of X ray irradiated L02 cells, decreased cellular H 2O 2 production, but also significantly increased positive rate of L02 cells expressed bcl 2 protein and decrease positive rate of L02 cells expressed Bax proteins ( P