The application of quantitative MRI in the diagnosis of early intervertebral disc degeneration / 实用放射学杂志

Objective To evaluate the potential clinical value of T2 mapping and mDixon Quant in the diagnosis of early interver-tebral disc degeneration.Methods A total of 79 volunteers who underwent lumbar MRI examination were enrolled.All subjects were examined for 3.0T MR with T2WI,T2 mapping,and mDixon Quant while recording the condition of low back pain.The differ-ences between T2 mapping(map)value and fat fraction(FF)values of the vertebral(V)and nucleus pulposus(NP)within the Pfir-rmann Ⅰ and Pfirrmann Ⅱ intervertebral disc(grade Ⅰ 76,grade Ⅱ 87)were statistically analyzed.Receiver operating characteristic(ROC)curve analyses were performed for meaningful parameters.Results V-FF showed a mild positive correlation with degenera-tive intervertebral disc lesions,and NP-FF and NP-map values showed a mild negative correlation with lesions.There were statistically significant differences between the two groups in V-FF(P＜0.001),NP-FF(P=0.005),and NP-map(P＜0.001).Some measure-ments had statistically significant differences when different intervertebral disc segments were compared.Conclusion V-FF,NP-FF,and NP-map are associated with intervertebral disc degeneration.T2 mapping and mDixon Quant are potentially valuable as diagnostic tools to quantitatively assess early intervertebral disc degeneration and help diagnose.

Correlation between the expression of miRNA-142-3p,mixed lineage kinase 3 and interleukin-1beta in nucleus pulposus and the degree of lumbar intervertebral disc degeneration / 中国组织工程研究

BACKGROUND:MicroRNA(miRNA)levels are closely related to cell apoptosis and proliferation,extracellular matrix metabolism and inflammatory response in intervertebral disc cells.However,the specific role of miR-142-3p in lumbar intervertebral disc degeneration remains unclear. OBJECTIVE:To investigate the correlation between the expression of miRNA-142-3p,mixed lineage kinase 3 and interleukin-1β in nucleus pulposus tissue and degree of human lumbar intervertebral disc degeneration. METHODS:A total of 82 patients with lumbar intervertebral disc degenerative diseases in Suzhou Ninth People's Hospital from January 2020 to March 2022 were collected as the study subjects,all of whom underwent MRI examination before operation.According to the Videman classification,the patients were divided into mild degeneration group(n=36),moderate degeneration group(n=26)and severe degeneration group(n=20).Eighty-two specimens of the nucleus pulposus were obtained.The mRNA expression of miRNA-142-3p as well as the mRNA and protein expression of mixed lineage kinase 3,interleukin-1β,type I collagen,type II collagen in nucleus pulposus tissue were detected by qPCR and western blot assay.The correlation between the degree of human lumbar intervertebral disc degeneration and the expression levels of miRNA-142-3p,mixed lineage kinase 3,and interleukin-1β was also assessed using the Spearman correlation coefficient method.Thirty adult Sprague-Dawley rats were divided into sham-operated group(executed after puncturing skin and muscle only),mild degeneration group(executed 1 week after puncturing Co7/8 segments)and severe degeneration group(executed 2 weeks after puncturing Co7/8 segments),with 10 rats in each group.After that,we detected the protein expression of mixed lineage kinase 3 and interleukin-1β as well as the gene expression of miRNA-142-3p,mixed lineage kinase 3 and interleukin-1β in the nucleus pulposus tissue. RESULTS AND CONCLUSION:In human nucleus pulposus tissue,the miRNA-142-3p expression ranked from high to low as follows:mild degeneration group>moderate degeneration group>severe degeneration group(P<0.05);the gene and protein expression of mixed lineage kinase 3 and interleukin-1β from low to high was as follows:mild degeneration group<moderate degeneration group<severe degeneration group(P<0.05);the gene and protein expression of type I collagen from low to high was as follows:mild degeneration group<moderate degeneration group<severe degeneration group(P<0.05),and the gene and protein expression of type I collagen from high to low was as follows:mild degeneration group>moderate degeneration group>severe degeneration group(P<0.05).Spearman correlation analysis showed that the degree of disc degeneration was negatively correlated with miRNA-142-3p expression(P<0.05)and positively correlated with mixed lineage kinase 3 and interleukin-1β expression(P<0.05).In rat nucleus pulposus tissue,compared with the sham-operated group,the expression of mixed lineage kinase 3 and interleukin-1β gene and protein was elevated in the mild degeneration group(P<0.05)while miRNA-142-3p expression was decreased(P<0.05);compared with the mild degeneration group,the expression of mixed lineage kinase 3 and interleukin-1β gene and protein was increased in the severe degeneration group(P<0.05)while miRNA-142-3p expression was decreased(P<0.05).To conclude,the degree of human lumbar intervertebral disc degeneration is negatively correlated with miRNA-142-3p expression and positively correlated with mixed lineage kinase 3 and interleukin-1β expression in nucleus pulposus tissue.

Expression and clinical significance of lncRNA-Xist in lumbar disc degeneration nucleus pulposus tissue / 国际生物医学工程杂志

Objective:To explore the expression and clinical significance of long non-coding RNA (lncRNA) Xist in the nucleus pulposus tissue of lumbar intervertebral disc degeneration (LIDD).Methods:The Trizol extraction method was used to extract total RNA from LIDD nucleus pulposus tissue cells and in vitro cultured nucleus pulposus cells. The relative expression level of lncRNA Xist in LIDD nucleus pulposus tissue was verified through real-time fluorescence quantitative PCR. In vitro, a knockdown expression model was constructed by transfecting si-lncRNA Xist into a nucleus pulposus tissue cell line. The knockdown efficiency was identified using real-time fluorescence quantitative PCR. Cell proliferation behavior was detected using EdU staining and flow cytometry. Western Blot and flow cytometry were used to detect cell apoptosis. Results:Compared with control group, lncRNA Xist expression in LIDD nucleus pulposus tissues was increased ( P < 0.01). Compared with normal group, the number of EdU positive cells and the number of S phase cells in TNF-α group were decreased (all P < 0.01), cell apoptosis were increased ( P < 0.01), Bax protein expression was increased ( P < 0.01), and Bcl-2 protein expression was decreased ( P < 0.05). Compared with TNF-α group, the number of EdU positive cells and the number of S phase cells in TNF-α + siRNA-lncRNA Xist group were increased (all P < 0.05), cell apoptosis were decreased ( P < 0.05), Bax protein expression was decreased, and Bcl-2 protein expression was increased. Conclusions:lncRNA Xist is highly expressed in nucleus pulposus tissues of LIDD patients, inhibits the proliferation of nucleus pulposus cells, and promotes apoptosis, which suggests that lncRNA Xist may be used as a therapeutic target and biomarker of LIDD.

Research progress in mechanism of Chinese herbal compounds and monomers in delaying lumbar intervertebral disc degeneration / 中国中药杂志

Traditional Chinese medicine has unique advantages in the treatment of degenerative bone and joint diseases, and its widely used in clinical practice. In recent years, many scholars have conducted a large number of basic studies on the delay of intervertebral disc degeneration by herbal compound and monomeric components from different perspectives. In order to further elucidate its mechanism of action, this paper summarizes the in vivo and in vitro experimental studies conducted at the level of both herbal compound and single components, respectively, in order to provide references for the basic research on the treatment of lumbar intervertebral disc degeneration by Chinese medicine. A summary shows that commonly used herbal compound prescriptions include both classical prescriptions such as Duhuo Jisheng Decoction, as well as clinical experience prescriptions such as Yiqi Huoxue Recipe. Angelicae Sinensis Radix, Chuanxiong Rhizoma, Rehmanniae Radix Praeparata, Achyranthis Bidentatae Radix, and Eucommiae Cortex were used most frequently. Tonic for deficiency and blood stasis activators were used most frequently. The most utilized monomeric components include icariin, ginsenoside Re, salvianolic acid B and aucubin. The main molecular mechanisms by which herbal compound and monomeric components delay of lumbar intervertebral disc degeneration include improving the intervertebral disc microenvironment, promoting the synthesis of aggregated proteoglycans and type Ⅱ collagen in the intervertebral disc, reducing the degradation of the extracellular matrix, and inhibiting apoptosis in the nucleus pulposus cells, etc. The main signaling pathways involved include Wnt/β-catenin signaling pathway, MAPK-related signaling pathway, mTOR signaling pathway, Fas/FasL signaling pathway, PI3 K/Akt signaling pathway, NF-κB signaling pathway, JAK/STAT signaling pathway, and hedgehog signaling pathway, etc.

Effect of yaobishu on protein expression of aquaporin 1 and aquaporin 3 in nucleus pulposus of the degenerated lumbar intervertebral disc in a rabbit / 中国组织工程研究

BACKGROUND: The aging and lesions of the intervertebral disc are closely related to the lack of nutritional blood supply to the disc. Aquaporin plays an important role in the nutritional supply to the intervertebral discs, but the specific mechanism has not been fully defined. OBJECTIVE: To study the mechanism of Yaobishu on degenerated intervertebral disc in rabbits based on the changes of aquaporin (AQP) 1 and AQP3 protein expression. METHODS: Thirty-six New Zealand white rabbits were randomly divided into model group, low-dose Yaobishu group and high-dose Yaobishu group. Animal models of lumbar intervertebral disc prolapse were prepared through an injection of normal saline into L4/5 and L5/6 segments. The model group was intragastrically given normal saline 5 mL/kg per day, the low-dose group was intragastrically given Yaobishu 5 mL/kg per day, and the high-dose group was intragastrically given Yaobishu 10 mL/kg per day, twice a day, for 21 days. After 6 weeks of treatment, the intervertebral discs were taken for anatomical and histological observation using hematoxylin-eosin staining. Expression of AQP1 and AQP3 in the nucleus pulposus at protein and mRNA levels was quantified by RT-PCR and western blot assay. RESULTS AND CONCLUSION: In all the three groups, the annulus fibrosus was destroyed, abnormal cartilage tissue appeared, and the nucleus pulposus was reduced in number. Severest degeneration of the intervertebral disc was found in the model group, followed by the low-dose Yaobishu and high-dose Yaobishu groups in turn. The expression of AQP1 and AQP3 mRNA and protein in the high-dose Yaobishu group and low-dose Yaobishu group increased significantly after 6 weeks of treatment (P 0.05). There were significant differences in the expression of AQP1 and AQP3 mRNA and protein among the three groups (P < 0.05). Therefore, Yaobishu may alleviate the degeneration of the rabbit intervertebral disc by increasing the expression of AQP1 and AQP3.

Electroacupuncture down-regulates expression of ADAMTS-4 of intervertebral disc in rats with lumbar intervertebral disc degeneration / 针刺研究

OBJECTIVE: To observe the effect of electroacupuncture (EA) stimulation on the levels of a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4) protein in the lumbar intervertebral disc tissue and serum in prolapsed lumbar intervertebral disc degeneration (PLIDD) rats, so as to explore its mechanism underlying improvement of intervertebral disc degeneration (IDD). METHODS: A total of 48 Sprague-Dawley rats were divided into sham operation (n＝12), model (n＝18) and EA (n＝18) groups. The PLIDD model was established by puncturing the lumbar discs (L4-L5, L5-L6) with a gauge-22 syringe needle. After modeling, EA stimulation was applied to "Pangguangshu"(BL28), "Zusanli"(ST36) and "Zhijian"for 20 min, 6 times per week for 4 weeks. The lumbar intervertebral disc tissue and blood samples were collected at the 4th, 6th and 8th week after modeling, respectively. The expression of ADAMTS-4 in the lumbar intervertebral disc tissue was detected by immunohistochemistry and Western blot (WB), separately. The content of ADAMTS-4 in the serum was detected by ELISA. RESULTS: Both immunohistochemical stain and WB showed that the expression levels of lumbar ADAMTS-4 at the 4th, 6th and 8th week were significantly up-regulated in the model group relevant to the sham operation group (P<0.01). Following EA treatment, the expression levels of ADAMTS-4 on day 14 and 28 were notably lower in the EA group than in the model group (P<0.05). The serum ADAMTS-4 contents were significantly increased in the model group than in the sham operation group at the 3 time-points (P<0.05, P<0.01), and considerably decreased in the EA group than in the model group on day 28 after EA intervention(P<0.05)．. CONCLUSION: EA can down-regulate the ADAMTS-4 expression of lumbar intervertebral disc tissue in PLIDD rats, which may contribute to its effect in relieving lumbar intervertebral disc degeneration.

Serum Caspase-3 and Caspase-9 Activity in Patients with Lubar Intervertebral Disc Protrusion / 现代检验医学杂志

Objective To explore the relationship between apoptosis promoting effector molecules caspase-3,caspase-9 and lubar intcrvertebral disc protrusion.Methods 99 of operation patients with lubar intervertebral disc protrusion in their mater nity ward were recruited.Among them,single segment of lubar intervertehral disc protrusion were 70 (Group A),more than one segments of lubar intervertebral disc protrusion were 29(Group B).In addition,40 unrelated healthy people from physi cal examination center were enrolled as controls.Enzyme linked immunosorbent assay (ELISA) was used to examine serum caspase-3 and caspase-9 levels in lubar intervertebral disc protrusion patients.Results The level of caspase-3 in control group,group A and group B,respectively were 11.24±0.41,14.31±0.67 and 17.43±1.86 pmol/L.The caspase-3 activity in each group was statistically significant difference (F=8.47,P＜0.01).The level of caspase-9 in control group,group A and group B respectively were 18.54±2.19,30.57±3.63 and 43.68±5.15 pmol/L.The caspase-9 activity in each group was statistically significant difference (F=7.85,P=0.001).Compared with control group,the caspasc 3 and caspase-9 ac tivity in group A (q=3.08.3.29,all P＜0.05),group B (q=5.78,4.50,all P=0.014) was statistically significant differ ence.The caspase-3 and caspase-9 activity in group A,group B was statistically significant difference (q=3.21,3.22,all P＜0.05).Conclusion The augment of caspase-3 and caspase-9 promoted apoptosis of lubar intervertebral disc protrusion.It was connected with quantity of protrusive segments.The more segments of protrusion,the higher caspase-3 and caspase-9 levels of examination would be.

Relationship between Apoptosis Promoting Effector Molecules TFAR19, Apaf-1 and Lubar Intervertebral Disc Protrusion / 现代检验医学杂志

Objective To explore the relationship between apoptosis promoting effector molecules TFAR19,Apaf-1 and lubar intervertebral disc protrusion.Methods 99 of operation patients with lubar intervertebral disc protrusion in the First Affiliated Hospital of Guangxi Medical University Department of Spine and Osteoputhy were recruited.Among them,single segment of lubar intervertebral disc protrusion were 70 (Group A),more than one segments of lubar intervertebral disc protrusion were 29 (Group B).In addition,40 unrelated healthy people from physical examination center were enrolled as controls (Group C).Enzyme-linked immunosorbent assay (ELISA) was used to examine serum TFAR19,Apaf-1 levels in lubar intervertebral disc protrusion patients.Results The level of TFAR19 in Group A,Group B and Group C respectively were 1.85±0.14,2.33±0.25 and 1.30±0.09 ng/ml.The TFAR19 activity in each group was statistically significant difference (F=7.979,P＜0.01).Compared with Group C,the TFAR19 activity in Group B (q=5.59,P＜0.01),Group A (q=3.60,P=0.012) was statistically significant difference.The TFAR19 activity in lubar intervertebral disc protrusion patients subgroups (Group A,Group B) was statistically significant difference (q =2.93,P =0.012).The level of Apaf-1 in Group A,Group B and Group C respectively were 159.22±11.87,203.20±20.21 and 107.52±11.58 pg/ml.The Apaf-1 activity in each group was statistically significant difference (F=8.828,P＜0.01).Compared with Group C,the Apaf-1 activity in Group B (q=5.86,P＜0.01),Group A (q=3.89,P=0.007) was statistically significant difference.The Apaf-1 activity in lubar intervertebral disc protrusion patients subgroups (Group A,Group B) was statistically significant difference (q =2.97,P=0.037).Male/female ratio between each groups was not statistically significant difference (x2=0.229,P =0.892).Age between each groups was not statistically significant difference (F=0.091,P =0.91).Conclusion The augment of TFAR19 and Apaf-1 promotes apoptosis of lubar intervertebral disc protrusion.It is connected with quantity of protrusive segments.The more segments of protrusion,the higher TFAR19 and Apaf-1 level of examination will be.

Value of diffusion tensor imaging and T2 mapping to assess lumbar intervertebral disc degeneration / 实用放射学杂志

Objective To investigate the value of ADC and FA of diffusion tensor imaging(DTI)and T2 value of T2 mapping for assessing lumbar intervertebral disc degeneration.Methods 12 cases of healthy volunteers(8 males and 4 females),28 cases of patients with chronic low back pain(15 males and 13 females,19-70 years old)were performed lumbar spine MRI,DTI and T2 mapping to obtain ADC,FA and T2 value.Intervertebral discs were classified according to the Pfirrmann grading.The correlations of different degeneration grade with ADC,FA and T2 value were analyzed.The diagnostic value of ADC,FA and T2 values of lumbar intervertebral disc degeneration were compared. Results Both ADC value and T2 value were significantly negative correlated with lumbar intervertebral disc degeneratic Pfirrminn grading(r=-0.779,r=-0.708,P<0.001).FA value were positively correlated with Pfirrminn grading(r=0.474,P<0.001), the correlation was not closely.Conclusion DTI and T2 mapping can be effectively used to quantitatively evaluate the degeneration degree of lumbar intervertebral disc,the diagnostic value of ADC was the highest,followed by T2 ,and FA was the worst.

Study of iterative Dixon water-fat separation with echo asymmetric and least-squares estimation for evaluation of intervertebral disc degeneration / 国际生物医学工程杂志

Objective To analyze quantitatively the multiple parameters of iterative Dixon water-fat separation with echo asymmetric and least-squares estimation (IDEAL) MR imaging sequence, in order to realize safe, noninvasive and accurate observation of intervertebral disc degeneration, and to predict early degeneration in time, improve image quality, and to realize precision medical treatment by providing personalized examinations and optimized imaging protocols.Methods A total of 40 volunteers, 23 males and 17 females, with 20-84 years of age (mean (53.4±16.5) years), were participated in our study to accept lumbar spine MRI examination, including sequences of OSAG T2 FSE, OSAG T1 FSE, OAX T2 FSE, OSAG T2 FSE FS and OSAG IDEAL T2.Lumbar intervertebral disc degeneration was evaluated by Pfirrmann classification.The water signal fraction (WSF) of each disc was measured and calculated accurately with IDEAL in phase and IDEAL out phase images.The differences between Pfirrmann grade and WSF were analyzed statistically.The signal-to-noise ratio (SNR) measurements were carried out on equal size region of interest (ROI) from the same levels of vertebral body (L3) and background zone of T2 FSE FS and T2 IDEAL water image mid-sagittal planes, respectively, and the average value of vertebral body signal strength and the standard deviation of background signal were measured.SNRs of the two sequences were calculated, and the image qualities were compared statistically.Results Of all 200 lumbar discs that measured using Pfirrmann classification, 39 discs were classified as Pfirrmann grade Ⅰ, 42 as grade Ⅱ, 27 as grade Ⅲ, 53 as grade Ⅳ, and 39 as grade Ⅴ.The SNR of T2 FSE FS images was (11.42±6.17) dB, while that of the IDEAL water images was (20.63±9.44) dB, and there was a statistically significant difference between them (P＜0.05).The WSF decreased along with the increase of Pfirrmann grade.The significant differences of WSF were found in all grades (P＜0.05) except for the grade Ⅳ and Ⅴ (P＞0.05).Conclusions The SNR of the IDEAL water images is significantly higher than that of T2 FSE FS images, which bring about better image quality.The WSF calculated based on the IDEAL in phase and IDEAL out phase images will make possible the quantitative, noninvasive and accurate analysis of lumbar disc degeneration, especially on the early stage of disc degeneration.

Effect of Metalloproteinases Inhibitor on Expression of MMP-3 in Degenerated Lumbar Intervertebral Disc / 中国康复理论与实践

@# ObjectiveTo explore the effect of metalloproteinases (MMPs) inhibitor on expression of MMP-3 in degenerated lumbar intervertebral disc.MethodsThe animal model of degenerated lumbar intervertebral disc was established with rats. The 32 rats were randomly divided into the experimental group and control group with 16 animals in each group. From the fourth week after operation, the animals of the experimental group were injected with tetracycline 25 mg per day, those of the control group with saline. Two weeks late, the tissue of degenerated lumbar intervertebral disc was taken and the expression of MMP-3 was tested by immumohistochemistry and Western-bloting.ResultsThe expression of MMP-3 in the experimental group decreased and significantly different from the control group ( P<0.05).ConclusionMMPs inhibitor can decrease expression of MMP-3 in degenerated lumbar intervertebral disc.