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BACKGROUND: Previous studies found that treatment with awn needles can improve the inflammatory response after spinal cord injury, reduce the expression of high-mobility group protein 1 (HMGB-1) and pro-inflammatory factors, and inhibit the activity of nuclear transcription factors and the occurrence of neuronal apoptosis. OBJECTIVE: To analyze the relationship between the mechanism of inflammatory response and the HMGB-1/Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) signaling pathway after spinal cord injury. METHODS: The T9-10 spinal cord injury model was made according to the internationally recognized modified Allen’s modeling method. The blood samples of the spinal cord and tail artery were obtained for Basso, Beattie and Bresnahan scoring at 6, 24 hours, 3 and 7 days after modeling. The HMGB-1 content in the spinal cord and tail artery serum was measured by ELISA to determine the highest point of HMGB-1 in the spinal cord and the change rule of HMGB-1 content in serum and spinal cord. Taking the highest HMGB-1 content as the research point and using glycyrrhizic acid as the HMGB-1 inhibitor, the rats were divided into four groups: model group, glycyrrhizic acid group (intragastric administration of glycyrrhizic acid, 200 mg/kg), blank group and sham operation group. The highest HMGB-1 expression was determined by ELISA. The spinal cord was obtained at the highest point of HMGB1 expression. The correlation between HMGB-1/TLR4/NF-κB signaling pathway and inflammation after spinal cord injury was explored by immunoblotting and RT-PCR detection, and the pathological changes of spinal cord in rats were observed. Approval for this study was obtained by the Animal Experimental Ethics Committee of Zhejiang Chinese Medical University. RESULTS AND CONCLUSION: The expression of HMGB-1 in the spinal cord and blood at 3 days after spinal cord injury was significantly higher than that at 6 hours, 24 hours, and 7 days (P < 0.05). At 3 days after spinal cord injury, the expressions of HMGB-1, TLR4, NF-κB, interleukin-6, and tumor necrosis factor-α in the glycyrrhizic acid group were lower than those in the model group, but higher than those in the blank group and sham operation group (P < 0.05). The expressions of HMGB-1, TLR4 and NF-κB at gene and protein levels in the glycyrrhizic acid group were lower than those in the model group, but higher than those in the blank group and the sham operation group (P < 0.05). To conclude, after spinal cord injury, HMGB-1 in blood and spinal cord increased significantly and reached the highest value at 3 days. After inhibiting HMGB-1, it was found that the HMGB-1/TLR4/NF-κB pathway is one of the important pathways to induce inflammation after spinal cord injury.
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OBJECTIVE@#To choose the disease-causing gene in a Chinese pedigree with ankylosing spondylitis (AS) by whole-exome sequencing (WES), and provide theory basis for mechanism of disease.@*METHODS@#Clinical data of AS pedigree were collected, including 2 males, the age were 48 and 18 years old, the course of disease were 23 and 4 years. Whole blood genomic DNA of AS was extracted to perform whole exome sequencing, the results were compared with human databases, common variations which had been reported were wiped out, then non synonymous single nucleotide variants(SNVs) from the family members were combined, and candidate genes was selected initially.@*RESULTS@#Totally 80 G data was obtained from AS family with high quality.By comparing results between patient and normal subject, and filtering with number of biological database, the result showed heterozygous mutation of JAK2 gene 12 exon c.1709 A>G (p.Tyr570Cys) may be the potential disease-causing gene. The variant c.1151T>C of MUC3A gene may be one of the causes of intestinal symptoms in the family members.@*CONCLUSION@#It is feasible to find t candidate gene mutations of AS by Exon sequencing. The mutation c.1709 A>G in gene JAK2 identified by whole exome sequencing might be the pathogenic mutation in this AS pedigree.
Subject(s)
Humans , Male , Exome , Mucin-3 , Mutation , Pedigree , Spondylitis, Ankylosing , Exome SequencingABSTRACT
Objective: To construct bone morphogenetic protein 2 (BMP-2) gelatin/chitosan hydrogel sustained-release system, co-implant with induced pluripotent stem cells (iPS) derived mesenchymal stem cells (MSCs) to hydroxyapatite (HA)/zirconium dioxide (ZrO 2) bio porous ceramic foam, co-culture in vitro, and to explore the effect of sustained-release system on osteogenic differentiation of iPS-MSCs. Methods: BMP-2 gelatin/chitosan hydrogel microspheres were prepared by water-in-oil solution. Drug encapsulation efficiency, drug loading, and in vitro sustained release rate of the microspheres were tested. HA/ZrO 2 bio porous ceramic foam composite iPS-MSCs and BMP-2 gelatin/chitosan hydrogel sustained release system co-culture system was established as experimental group, and cell scaffold complex without BMP-2 composite gelatin/chitosan hydrogel sustained release system as control group. After 3, 7, 10, and 14 days of co-culture in the two groups, ALP secretion of cells was detected; gene expression levels of core binding factor alpha 1 (Cbfa1), collagen type Ⅰ, and Osterix (OSX) were detected by RT-PCR; the expression of collagen type Ⅰ was observed by immunohistochemical staining at 14 days of culture; and cell creep and adhesion were observed by scanning electron microscopy. Results: BMP-2 gelatin/chitosan hydrogel sustained-release system had better drug encapsulation efficiency and drug loading, and could prolong the activity time of BMP-2. The secretion of ALP and the relative expression of Cbfa1, collagen type Ⅰ, and OSX genes in the experimental group were significantly higher than those in the control group at different time points in the in vitro co-culture system ( P<0.05). Immunohistochemical staining showed that the amount of fluorescence in the experimental group was significantly more than that in the control group, i.e. the expression level of collagen type Ⅰ was higher than that in the control group. The cells could be more evenly distributed on the materials, and the cell morphology was good. Scanning electron microscopy showed that the sustained-release system could adhere to cells well. Conclusion: iPS-MSCs have the ability of osteogenic differentiation, which is significantly enhanced by BMP-2 gelatin/chitosan hydrogel sustained-release system. The combination of iPS-MSCs and sustained-release system can adhere to the materials well, and the cell activity is better.
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OBJECTIVE@#To explore the clinical effects of acupuncture with elongated needle on urinary retention after spinal cord injury.@*METHODS@#A total of 66 patients with urinary retention after spinal cord injury were randomized into an elongated needle group (34 cases) and a filiform needle group (32 cases). In both of the groups, the routine orthopedic treatment and the bladder function training were adopted. Additionally, in the elongated needle group, the acupuncture at Zhibian (BL 54) and Shuidao (ST 28) was used with the elongated needle. In the filiform needle group, the acupuncture with filiform needle was used at Zhibian (BL 54) and Shuidao (ST 28). In both of the groups, acupuncture treatment was provided once every two days, for 2 months consecutively. Separately, before treatment and after 1, 2 month treatment, the residual urine volume, urodynamic parameters (bladder capacity, peak urinary flow rate and bladder pressure) and safety indicators were observed in the patients of the two groups. The clinical therapeutic effects were compared between the two groups.@*RESULTS@#As compared with the results before treatment, the residual urine volume was reduced obviously (all <0.01), bladder capacity, peak urinary flow rate and bladder pressure were improved obviously (all <0.01) after 1, 2 month treatment in the two groups. After 1, 2 month treatment, the residual urine volume in the elongated needle group was lower than the filiform needle group (both <0.01). The improvements in bladder capacity, peak urinary flow rate and bladder pressure in the elongated needle group were superior to the filiform needle (all <0.01). The total effective rate was 94.1% (32/34) in the elongated needle group, higher than 71.9% (23/32) in the filiform needle group (<0.01). During treatment, there was no damage of kidney function, no fainting or broken needle happened in the two groups.@*CONCLUSION@#Acupuncture with elongated needle achieves the significant effects on urinary retention after spinal cord injury. This therapy is better than acupuncture with filiform needle and safe in operation.