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Objective: To explore the clinical efficacy of spine subtle adjusting manipulation for postpartum low back pain (PLBP). Methods: A total of 76 patients with PLBP were randomized into a control group and a treatment group, with 38 cases in each group. The control group was treated with core muscle strengthening exercises, and the treatment group was treated with spine subtle adjusting manipulation. After 3 weeks of treatment, the clinical efficacy was observed, and the visual analog scale (VAS) score and Oswestry disability index (ODI), and the changes of lumbar Cobb angle and pelvic rotation were compared between the two groups. Results: The total effective rate of the treatment group was 92.1％, and that of the control group was 78.9％. The difference between the two groups was statistically significant (P<0.05). After treatment, the VAS score and ODI in both groups decreased, and the intra-group differences were all statistically significant (P<0.05). There were no intra-group statistical differences in the lumbar Cobb angle or pelvic rotation in the two groups (P>0.05). After treatment, there were no statistical differences in the lumbar Cobb angle or pelvic rotation between the two groups (P>0.05); the VAS score and ODI in the treatment group were significantly lower than those in the control group (P<0.05). Conclusion: Spine subtle adjusting manipulation can effectively relieve the pain for patients with PLBP, and improve their daily activity function.
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Interleukin-12 (IL-12) is a pro-inflammatory cytokine derived from antigen-presenting cells. IL-12 has a variety of effector mechanisms such as promoting T cells proliferation and cytotoxicity, inducing T cells and NK cells to secrete interferon-γ (IFN-γ), etc. IL-12 shows significant anti-tumor effect in preclinical stage and is quickly used in clinic, but its systemic administration has serious dose-limiting toxicity. At present, most of the research is about the topical use of IL-12 to reduce systemic toxicity, change tumor microenvironment and activate immune system. This article reviews the research progress and clinical trials of IL-12 in the treatment of solid tumors in recent years.
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Objective@#To investigate the effects of Paraquat on neural stem cell proliferation in vitro and explore the its mechanism based on DNA methylation pathway.@*Methods@#Nestin, β-tubulin III, and glial fibrillary acidic protein (GFAP) were detected by indirect immunofluorescence assay to evaluate self renewal and differentiation potentia of ReNcell CX human neural stem. The cells were treated with terminal concentrations of 0, 5, 25, 50, and 100μmol/L PQ for 24 hours, and the cells were induced by 50 μmol/L PQ for different time (6, 12, 24, 48 h). Cell viability was determined by MTT assay. The proliferation of neural stem cells was evaluated using Sox2/Brdu and Nestin/Brdu double immunofluorescence staining. The global DNA methylation level was assayed by MethyflashTM methylated DNA Quantification kit. The expression levels of Dnmts mRNA and protein were analyzed by quantitative reverse transcription polymerase chain reaction(qRT-PCR) and Western blot, respectively.@*Results@#Immunofluorescence showed that nestin was primarily expressed in proliferative neural stem cell and peotein biomarkers (β-tubulin III, GFAP) for neuron and astrocyte were expressed in differentiated cells. MTT assay showed PQ induced cell survival rate decrease in a time and dose dependent manner. Double immunfluorescence staining of cells showed colocalization of Sox2 and Brdu. The percentage of Brdu/Sox2 positive cells was significantly lower in the PQ-exposed (25, 50, 100μmol/L PQ treatment) groups compared to control (P<0.05); Meanwhile, The percentage of Brdu/Nestin positive cells was also significantly lower in the PQ-exposed(50,100μmol/L PQ treatment) groups compared to control (P<0.05). The results of global DNA methylation revealed a significant decrease in PQ-exposed groups (P<0.05). Western blot showed that compared with control group, the protein and mRNA levels of Dnmt1, Dnmt3a in PQ-exposed group were significantly decreased (P<0.05), but there was a significant increase in expression level of Dnmt3b in 50, 100 μmol/L PQ-treated group(P<0.05).@*Conclusion@#Paraquat could inhibite the proliferation of human neural stem cells through reducing the level of DNA methylation reaction by suppressing the protein expression and transcription of DNA methylated transferase(Dnmts).
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Objective@#To observe the effect of low concentration paraquat (PQ) on activation and phenotypic M1/M2 polarization of mouse microglia cells (BV2) .@*Methods@#BV2 cells were used as model, and cultured in vitro were exposed to paraquat at designed concentrations of 0, 0.015, 0.03, 0.06, 0.12, 0.24, 0.48 μmol/L and 0.05 μmol/L 1-methyl-4-phenylpyridinium (MPP+) for 24 h, and cell viability was determined by CCK8 assay. After induced by 0, 0.015, 0.03, 0.06, 0.12 μmol/L PQ and 0.05 μmol/L MPP+ for 24 h, the contents of tumor necrosis factor-α (TNF-α) , interleukin-6 (IL-6) and IL-1β in cell culture supernatant were determined by enzyme-linked inmunosorbent assay (ELISA) . Cell migration ability was determined by transwell. Immunofluorescence (IF) and flow cytometry were used to determine the phagocytic capacity of cells. Designed concentrations of 0, 0.03, 0.06, 0.12 μmol/L PQ and 0.05 μmol/L MPP+ for 24 h, the protein expressions of M1 markers of BV2 (TNF-α, IL-6, IL-1β, Nitric oxide synthase-iNOS, CD86) and M2 markers of BV2 (Arginase type-1 Arg-1 and Mannose recepteor-CD206) were determined by Western Blot after PQ expourse (0, 0.03, 0.06, 0.12 μmol/L) and 0.05 μmol/L MPP+ induction.@*Results@#Compared with 0 μmol/L PQ group, proliferation activity of BV2 cells was significantly increased by 0.03~0.12 μmol/L PQ while inhibited by 0.48 μmol/L PQ (P<0.05) . The cell proliferation activity of cells treated with 0.03 μmol/L PQ was significantly increased in 24 hours (P<0.05) . ELISA showed that TNF-α, IL-6 and IL-1β contents in the cell supernatant of the PQ group were significantly higher than those of 0 μmol/L PQ group, especially in 0.03 and 0.06 μmol/L PQ exposed group (P<0.05) . The results of IF and flow cytometry showed that phagocytic capacity of 0.015, 0.03 and 0.06 μmol/L PQ group was significantly enhanced compared with 0 μmol/L PQ group (P<0.05) . Transwell showed that the cell invasion ability of 0.03, 0.06, 0.12 μmol/L PQ was significantly higher than that of 0 μmol/L PQ group (P<0.05) . Western blot showed that compared with 0 μmol/L PQ group, the expression levels of M1 markers TNF-α, IL-6, IL-1β, iNOS and CD86 were significantly increased in 0.03 and 0.06 μmol/L PQ exposed group, while the expression levels of M2 markers Arg-1 and CD206 protein were decreased in 0.06 and 0.12 μmol/L PQ exposed group (P<0.05) .@*Conclusion@#Low concentration PQ can abnormally activate BV2 cell, making the transformation of BV2 cell into pro-inflammatory M1 type and inhibiting its transformation into anti-inflammatory M2 type.
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Objective@#To investigate the regulation of AMPK-mTOR signal transduction pathway in paraquat-induced autophagy of pheochromocytoma cells (PC12) .@*Methods@#The PC12 cell were treated with terminal concentrations of 0, 25, 50, 100, 200, 300 and 400 μmol/L PQ for 24 hours, and the cells were induced by 300 μmol/L PQ for different time (6, 12, 24, 48 h) . MTT was used to detect the relative survival rate of cells, and the dose/time-effect relationship was determined respectively. The cells were treated with PQ at concentrations of 0, 100, 200 and 300 μmol/L PQ for 24 hours, the lactate dehydrogenase (LDH) activity in the culture supernatant was detected by spectrophotometry. The expression and distribution of autophagic lysosomes were observed by MDC staining. The intracellular reactive oxygen species (ROS) was detected by dichlorofluorescein diacetate (DCFH-DA) . The expression of microtubule-related protein 1 light chain 3 (LC3) was measured by immunofluorescence. The protein level of LC3Ⅱ, p62, Beclin1 and p-AMPK, p-mTOR were detected by Western blot.@*Results@#Compared with the control group, the cell survival rate of the 100, 200, 300, 400 μmol/L PQ group decreased significantly, and showed a dose-dependent pattern (P<0.05) . The survival rate of cells treated with 300 μmol/L PQ decreased significantly with the prolongation of exposure time (12, 24, 48 h) (P<0.05) . Compared with the control group, the activity of LDH in 100, 200, 300 μmol/L PQ-treated group were significantly higher while The fluorescence intensity of ROS was significantly increased (P<0.05) . MDC staining showed the density of autophagic lysosomes and fluorescence intensity in PQ-treated group significantly decreased (P<0.05) . Immunofluorescence results showed the LC3 fluorescence intensity of PQ-treated group decreased which was consistent with MDC staining results. Western blot showed that compared with the control group, the expression levels of autophagy related proteins LC3Ⅱand Beclin1 in PQ-treated group were significantly lower, while the expression level of p62 protein was higher (P<0.05) . p-AMPK protein level decreased and p-mTOR protein expression increased in 200 and 300μmol/L PQ-treatd groups, with statistically significant difference (P<0.05) .@*Conclusion@#AMPK-mTOR signaling pathway played a regulatory role in PQ-induced decreased autophagy of PC12 cell.
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Objective@#To investigate the roles of p38 mitogen-activated protein kinases (p38 MAPK) , extracellular regulated protein kinases (ERK) and c-Jun N-tenninal kinases (JNK) of MAPK signaling pathway in Paraquat-induced epithelial to mesenchymal transition (EMT) of type II alveolarepithelial cells.@*Methods@#RLE-6NT cells were incubated with different concentrations of PQ (0, 25, 50, 100μmol/L) for 6, 12 and 24 h. Cell morphology alteration was observed under phase-contrast microscopy. Cell viability was determined using an MTT assay. Cell migration ability was detected using scratch wound assay. Protein expression of P-p38 MAP, P-Erk1/2, P-JNK, E-cad, ZO-1, Vimentin and а-SMA were detected by western blot. The level of genes related to fibrosis (COL-I, COL-III, FN and FSP-1) were analyzed via quantitative real-time RT-PCR.@*Results@#Cell morphology started to undergo EMT changes with a phenotype characteristic of mesenchymal cells, including an elongated shape and a lack of tight cell-cell adhesions induced by 100μmol/L PQ treatment in a time-dependent manner. MTT showed that cell viability decreased with increasing PQ concentration (50、100、200、300 μmol/L PQ treatment for 24 h) and increasing treatment time (200 μmol/L PQ treatment for 6, 12, 24, 36, 48 h) . Compared to control group, the expressions of the epithelial phenotype marker E-cad and ZO-1 significantly decreased with PQ treatment (50, 100μmol/L) in a time-dependent manner (P<0.05) . Additionally, the level of the mesenchymal marker (a-SMA, vimentin) dramatically increased with PQ treatment in the same concentration-and time-dependent manner (P<0.05) . Cell migration ability was markedly increased after 24 h of 100 μmol/L PQ treatment compared to control (P<0.05) . The phosphorylated forms of p38 MAPK, Erk1/2, and JNK were increased at 24 h after stimulation with PQ (P<0.05) . This PQ induced (100 μmol/L) phosphorylation was markedly attenuated in the presence of the p38 MAPK, ERK and JNK inhibitors (SB-203580, SP-600125 and PD98059) respectively. Furthermore, RT-PCR showed that PQ significantly induced the upregulation expression of COL I and III mRNA, Fn, and FSP-1 mRNA (P<0.05) .@*Conclusion@#PQ-induced pulmonary fibrosis occurs via EMT, which is mediated by the MAPK pathway.