Efficacy of knee-balancing manipulation plus heat-sensitive moxibustion for knee osteoarthritis and its influence on CTX-Ⅰ, TRACP-5b, ADAMTS-4, and MMP-3 / 针灸推拿医学（英文版）

Objective: To observe the efficacy of knee-balancing manipulation plus heat-sensitive moxibustion in treating knee osteoarthritis (KOA) and its impact on the expression of C-telopeptide of type Ⅰ collagen (CTX-Ⅰ), tartrate-resistant acid phosphatase 5b (TRACP-5b), A disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4), and matrix metalloproteinase 3 (MMP-3). Methods: A total of 134 unilateral KOA patients were randomized into a knee-balancing group, a heat-sensitive moxibustion group, and a joint intervention group. The knee-balancing group received knee-balancing Tuina (Chinese therapeutic massage) manipulation for treatment. The heat-sensitive moxibustion group received heat-sensitive moxibustion treatment. The joint intervention group received the heat-sensitive moxibustion in addition to the knee- balancing manipulation. The intervention period lasted for four weeks. After the treatment, and at the 2-week and 6-week follow-ups, the three groups were assessed using the visual analog scale (VAS) for knee joint pain and Western Ontario and McMaster Universities arthritis index (WOMAC), and clinical efficacy was also evaluated. The enzyme- linked immunosorbent assay was adopted to detect the expression levels of serum CTX-Ⅰ, TRACP-5b, ADAMTS-4, and MMP-3. Results: The knee-balancing group had 44 participants, but one dropped out; there was no dropout case among the 44 participants in the heat-sensitive moxibustion group; among the 46 participants in the joint intervention group, two cases dropped out. After the treatment, and at the 2-week and 6-week follow-ups, the total effective rate was found higher in the joint intervention group than in the knee-balancing and heat-sensitive moxibustion groups (P<0.05). Compared with the baseline, the VAS and WOMAC scores and the serum levels of CTX-Ⅰ, TRACP-5b, ADAMTS-4, and MMP-3 decreased significantly in all three groups after treatment and at the 2-week and 6-week follow-ups (P<0.05). At the same three time points, the VAS and WOMAC scores and serum levels of CTX-Ⅰ, TRACP-5b, ADAMTS-4, and MMP-3 were lower in the joint intervention group than in the knee-balancing and heat-sensitive moxibustion groups (P<0.001). Conclusion: Either used alone or combined, the knee-balancing manipulation and heat-sensitive moxibustion therapy can improve the symptoms and down-regulate the serum levels of CTX-Ⅰ, TRACP-5b, ADAMTS-4, and MMP-3 in KOA patients, producing durable efficacy; nevertheless, a more significant efficacy can be achieved by combining the two methods.

ADAMTS proteoglycanases downregulation with impaired oocyte quality in PCOS

ABSTRACT Objective: A disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4) and ADAMTS-5 normal expression levels are essential for ovulation and subsequent fertilization. The objective of the present study was to assess expression pattern of these genes in cumulus cells (CCs) taken from patients with polycystic ovary syndrome (PCOS) and to investigate any possible relationship with the oocyte quality. Subjects and methods: ADAMTS-4 and -5 expression levels within CCs containing oocytes at the metaphase II (MII) and germinal vesicle (GV) stages, taken from 35 patients with PCOS and 35 women with normal ovarian function, were investigated using RT-qPCR. Moreover, possible correlations between ADAMTS-4, ADAMTS-5, and progesterone receptors (PRs) expression as well as oocyte quality were evaluated. Results: ADAMTS-4 and -5 expression levels were dramatically diminished in the CCs of the PCOS patients when compared to the controls. ADAMTS-4 and -5 expression levels were correlated with each other and with the oocyte quality. Furthermore, lower expression levels of ADAMTS-4 and -5 in the PCOS patients were strongly correlated with the diminished PRs expression levels. Conclusions: Downregulation of ADAMTS-4 and -5 in the human CCs of the PCOS patients correlated with the decline in the PRs expression, and impaired oocyte quality may cause lower oocyte recovery, maturation, and fertilization rate.

ADAMTS4 is upregulated in colorectal cancer and could be a useful prognostic indicator of colorectal cancer

SUMMARY OBJECTIVE ADAMTS4 is a member of the ADAMTS4 family, which secretes proteinases. The mechanism of tumor metastasis may be correlated to its promotion of angiogenesis. It was determined whether ADAMTS4 participates in colorectal cancer progression. Methods The expression in clinical samples and CRC cell lines was investigated. Using immunohistochemistry (IHC), fluorescence in situ hybridization (FISH), and RT-PCR, the expression of ADAMTS4 was determined in colorectal tumors of different cancer stages and anatomic sites, and in three cell lines of different aggressiveness. Results The overexpression of ADAMTS4 was observed in tissue samples by IHC, and this was mainly located in the cytoplasm, as detected by FISH. The qRT-PCR and western blot analyses further supported the clinical sample findings. Conclusion The present data support the notion that the overexpression of ADAMTS4 in CRC might be useful as a non-invasive biomarker for detecting CRC in patients.

RESUMO OBJETIVO ADAMTS4 é um membro da família ADAMTS4, que secreta proteinases. O mecanismo da metástase do tumor pode ser correlacionado a sua promoção da angiogênese. Determinou-se se ADAMTS4 participa na progressão do câncer colorretal. Métodos A expressão em amostras clínicas e linhas de células CRC foi investigada. Usando a imuno-histoquímica (IHC), a hibridação fluorescente in situ (HFIS) e o RT-PCR, a expressão de ADAMTS4 foi determinada em tumores colorretais de diferentes estágios do câncer e locais anatômicos, e em três linhas de células de níveis de agressividade distintos. Resultados A superexpressão de ADAMTS4 foi observada em amostras de tecido por IHC, e esta foi localizada principalmente no citoplasma, como detectado pelo HFIS. O qRT-PCR e a análise de wester blot corroboraram os resultados clínicos da amostra. Conclusão Os dados atuais corroboram a noção de que a superexpressão de ADAMTS4 no CRC pode ser útil como um biomarcador não invasivo para a detecção de CRC em pacientes.

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.

Mechanism of thermosensitive moxibustion on knee osteoarthritis in rabbit models / 中国针灸

<p><b>OBJECTIVE</b>To observe the impacts of thermosensitive moxibustion (TSM) on the expressions of nitric oxide (NO), typeⅠdisintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), typeⅡcollagen and proteoglycan (PG) in the rabbit models of knee osteoarthritis (KOA) and explore the mechanism of TSM on KOA.</p><p><b>METHODS</b>A total of 42 Japanese long-eared male rabbits were divided into a blank group (6 rabbits), a model group (6 rabbits), a moxibustion group (24 rabbits) and a sham-operation group (6 rabbits) according to the random number table. In the blank group, the rabbits were fed normally. In the model and moxibustion groups, the papain injection was given to establish KOA models. The rabbits in the sham-operation group were treated with the intracavity injection of 0.9% NaCl solution. The rabbits were forced to move for 30 min every day, continuously for 15 days during modeling. At the end of modeling, in the moxibustion group, moxibusiton was applied at "Dubi" (ST 35), once a day, 40 min each time, for 14 days totally. According to the temperature changes during moxibustion, the rabbits were divided into a TSM group and a non-TSM group. 6 rabbits were collected randomly from the two groups. The usual feeding was given in the blank group, the model group and the sham-operation group, without any intervention. The body mass and behavioristics changes were observed in each group. At the end of treatment, the nitrate reduction method was adopted to determine NO expression in the serum. The real-time PCR was adopted to determine the expressions of ADAMTS-4, typeⅡcollagen and PG in the cartilage.</p><p><b>RESULTS</b>① After modeling, compared with the blank group, the body mass was all reduced and the Lequesne MG score was increased in the model group, TSM group, non-TSM group and sham-operation group (<0.05, <0.01). After intervention, compared with the blank group, the body mass was decreased and the Lequesne MG score was increased in the model and sham-operation groups (<0.05, <0.01). Compared with the model group, the body mass was increased and the lequesne MG score was decreased in the TSM, non-TSM, and sham-operation groups (<0.05, <0.01). Compared with the non-TSM group, the body mass in the TSM group was increased remarkably (<0.05), but the difference in Lequesne MG score was not statistically significant (>0.05). ② After intervention, compared with the blank group, the expressions of NO and ADAMTS-4 were all increased and the expressions of typeⅡcollagen and PG were decreased in the model group, TSM group, non-TSM group and sham-operation group (<0.05, <0.01). Compared with the model group, the expressions of NO and ADAMTS-4 were all remarkably lower and the expressions of typeⅡcollagen and PG were increased in the TSM group, non-TSM group and sham-operation group (<0.05, <0.01). Compared with the non-TSM group, the expressions of NO and ADAMTS-4 were all remarkably lower and the expressions of typeⅡcollagen and PG were increased in the TSM group after intervention (all <0.05).</p><p><b>CONCLUSION</b>The thermosensitive moxibustion alleviates the inflammatory reactions and protects the joint cartilage through inhibiting the expressions of NO and ADAMTS-4 to achieve the effects in the treatment of KOA.</p>

The effects of recombinant human interleukin-1 receptor antagonist on the cartilage repair in rat temporomandibular joint with osteoarthritis / 实用口腔医学杂志

Objective:To investigate the effects of recombinant human interluekin-1 receptor antogonist (rhIL-1Ra) on the cartilage repair in rat temporomandibular joint(TMJ) with osteoarthritis(OA).Methods:Collagenase-Ⅱ was injected into bilateral TMJs of 24 adult rats for the induction of bilateral TMJOA,1 week after injection,5μg rhIL-1Ra(diluted in 0.05 ml normal saline) was injected into each right TMJ and the left joint received the same amount of normal saline injection as the control.12 animals were sacrificed at 2and 4 weeks after the first injection respectively.HE staining,immunnohistochemical method and RT-PCR examination were conducted.Mankins scere was used to evaluate the TMJOA degree.1 adult SD rat was used as healthy control,and sacrificed at 2 weeks of the experiment.Results:The TMJs of both sides showed typical OA-related cartilage degradation 2 week after IL-1Ra treatment,the Mankin~ score of the IL-1Ra treated and control joints was 1.33±0.52 and 2.00±6.63 (P＞0.05),4 week after treatment that was 3.00± 0.63 and 6.50 ± 0.84 (P＜0.05),respectively.Lower expression of ADAMTS-4 and ADAMTS-5 was observed in the treated joints than in the controls (P＜0.05).Conclusion:Intra-articular injection of IL-1Ra into TMJ can alleviate the cartilage lesion,the mechanism may lie in the inhibition of the expression of ADAMTS-4 and ADAMTS-5.

The value of plasma ADAMTS4 in evaluating the stability of carotid plaque / 中华普通外科杂志

Objective To explore the relationship between the level of plasma ADAMTS4 and the vulnerability of carotid atherosclerotic plaque in patients with carotid artery stenosis and the clinical value of evaluating the vulnerability of the carotid atherosclerotic plaque.Methods 60 patients with carotid artery stenosis were divided into stable plaque group and vulnerable plaque group according to the histopathological typing after carotid endarterectomy (CEA).30 patients who had atherosclerosis but no carotid plaque formation served as control.Using ELISA to measure the level of plasma ADAMTS4.Results The level of plasma ADAMTS4 was significantly higher in the vulnerable-plaque group (112.74 ± 17.47) ng/ml compared with the stable plaque group (56.67 ± 22.14)ng/ml and the control group (56.67 ± 22.14)ng/ ml,P ＜ 0.001.Logistic analysis showed that high level of plasma ADAMTS4 was risk factor for vulnerability of carotid atherosclerotic plaque (P ＜ 0.05,OR =8.240,95 ％ CI:1.512-22.915).ROC curve showed that ADAMTS4 =100.935 ng/ml could accurately predict significant vulnerability with accuracy of 94.3％.Conclusion ADAMTS4 is an effective,sensitive and non-invasive biomarker to evaluate the vulnerability of carotid atherosclerotic plaque.

Expression of ADAMTS4 in the heart of myocardial infarction rat / 国际生物医学工程杂志

Objective To detect the expression of a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4),Versican,and Aggrecan in hearts of myocardial infarction rats.Methods Male Wistar rats were chosen to make myocardial infarction model.Fifteen rats were randomly assigned into the myocardial infarction group and five rats into the sham-operation group.Rats were killed 3 d after surgery and the left ventricles were taken to produce frozen section.ADAMTS4,Versican,and Aggrecan protein expressions were detected by imnunohistochemistry method.Results ADAMTS4 expressed in myocardial cells at marginal zone of the infarcted area.ADAMTS4 expressed weakly while Versican expressed strongly in capillary endothelial cells of marginal zone of the infarcted area.Aggrecan showed no expression in the cardiac tissue and vascular endothelium of both myocardial infarction group and sham-operation group.Conclusions ADAMTS4 is involved in the inflammation after myocardial infarction.Degradation of ADAMTS4 to Versican exists in the vessels after myocardial infarction.Aggrecan is not involved in the pathophysiological process of myocardial infarction.

Role of nuclear factor of activated T-cells in regulating ADAMTS-4 promoter activity in nucleus pulposus cells / 第二军医大学学报

Objective: To observe the regulatory effect of nuclear factor of activated T-cells (NFAT)-1, 4, 5 on ADAMTS-4. promoter activity in nucleus pulposus cells, so as to discuss the underlying molecular mechanism of intervertebral disc degeneration. Methods: The ADAMTS-4-pβ-gal-Basic was inserted into PGL3-Basic vector after double digestion by the restriction enzyme Xho I and Hind III. The purified PGL3-ADAMTS-4 plasmid was obtained through screening. Rat nucleus pulposus cells were cultured in vitro and, via Lipofectamine2000 transfection reagent, transfected with NFAT-1, NFAT-4, NFAT-5 or DN-TonEBP expressing plasmids with or without appropriate backbone vector and 175 ng ADAMTS-4 promoter. The normal cells were taken as controls and transfected cells were taken as treatment group. Dual-Luciferase™ reporter assay system was used to detect the effect of NFAT-1, 4, 5(TonEBP) on ADAMTS-4 promoter activity. Results: We successfully constructed PGL3-ADAMTS-4 plasmid and identified two NFAT-Bind elements in the promoter. Compared with the control group, ADAMTS-4 promoter activity was significantly inhibited in NFAT-1 group (P 0. 05). Conclusion: NFAT-1 can regulate ADAMTS-4 expression, which may play a role in modulating aggrecan content in the intervertebral disc physiology and/or intervertebral disc degeneration, providing a new strategy for biological treatment of intervertebral disc degeneration.

Effects of Different Magnitudes of Mechanical Stretch on Human Intervertebral Disc Cells / 天津医药

Objective To investigate the effects of different magnitudes of mechanical stress on human interverte-bral disc degeneration. Methods The human intervertebral disc cells were subjected to different magnitudes of mechanical stress (0, 6%, 12%, or 18%elongation) for 24 h using a Flexercell Strain Unit. The mRNA expressions of anabolic genes (col-lagen-1A1, collagen-2A1, aggrecan and versican) and catabolic genes (MMP-3, MMP-13, ADAMTS-4 and ADAMTS-5) were examined by real-time PCR and Western blot methods. Results The expression levels of collagen-1A1 and collagen-2A1 were increased at 12%of mechanical stress, and collagen-2A1 was decreased at 18%of mechanical stress compared with those of control. The mRNA expressions of catabolic genes, MMP-13 and ADAMTS-5, were increased at 12%and 18%of mechanical stress than those of control. The mechanical stretch induced a magnitude-dependent increase in ADAMTS-4 synthesis, which was finely tuned by stretching-triggered activation of distinct mitogen-activated protein kinase cascades. Specifically, an ERK1/2 specific inhibitor, U0126, significantly inhibited the stretching-induced ADAMTS-4 expression, whereas the inhibitors of p38 and JNK, SP6000125 and SB203580, showed only slightly effect on the stretching-induced ADAMTS-4 expression. Conclusion The different magnitudes of mechanical stretch exhibited different effects on the bio-logical behavior of intervertebral disc cells, which profoundly affects the intervertebral disc degeneration.

Correlation study on the ADAMTS-4 in early brain injury after experimental subarachnoid hemorrhage and blood-brain barrier permeability in rats / 重庆医学

Objective To establish the rat model of experimental subarachnoid hemorrhage (SAH) and to detect the expression of ADAMTS-4 in early brain injury(EBI) after SAH in rats .Methods Totally 72 adult male SD rats were randomly divided into 2 groups ,the control group and SAH group .The expression of ADAMTS-4 and the content of brain Evans Blue(EB) were detected . The ADAMTS-4 inhibitor α2-macroglobulin was used to study the relationship between the expression of ADAMTS-4 and blood-brain barrier permeability .Results The expression of the mRNA and protein of ADAMTS-4 increased dramatically at 24 h and the contents of EB increased progressively at 12 h after SAH .The protein expression of ADAMTS-4 and the content of brain EB was positive correlated(r=0 .917 ,P<0 .05) .The expression of ADAMTS-4 was inhibited by α2-macroglobulin(P<0 .05) and the con-tents of EB decreased dramatically (P<0 .05) .Conclusion ADAMTS-4 may be involved in the pathological progression of early brain injury after SAH .