Identification of ion channel-related genes as diagnostic markers and potential therapeutic targets for osteoarthritis through bioinformatics and machine learning-based approaches.

BACKGROUND: Osteoarthritis (OA) is a debilitating joint disorder characterized by the progressive degeneration of articular cartilage. Although the role of ion channels in OA pathogenesis is increasingly recognized, diagnostic markers and targeted therapies remain limited. METHODS: In this study, we analyzed the GSE48556 dataset to identify differentially expressed ion channel-related genes (DEGs) in OA and normal controls. We employed machine learning algorithms, least absolute shrinkage and selection operator(LASSO), and support vector machine recursive feature elimination(SVM-RFE) to select potential diagnostic markers. Then the gene set enrichment analysis (GSEA) and gene set variation analysis (GSVA) were performed to explore the potential diagnostic markers' involvement in biological pathways. Finally, weighted gene co-expression network analysis (WGCNA) was used to identify key genes associated with OA. RESULTS: We identified a total of 47 DEGs, with the majority involved in transient receptor potential (TRP) pathways. Seven genes (CHRNA4, GABRE, HTR3B, KCNG2, KCNJ2, LRRC8C, and TRPM5) were identified as the best characteristic genes for distinguishing OA from healthy samples. We performed clustering analysis and identified two distinct subtypes of OA, C1, and C2, with differential gene expression and immune cell infiltration profiles. Then we identified three key genes (PPP1R3D, ZNF101, and LOC651309) associated with OA. We constructed a prediction model using these genes and validated it using the GSE46750 dataset, demonstrating reasonable accuracy and specificity. CONCLUSIONS: Our findings provide novel insights into the role of ion channel-related genes in OA pathogenesis and offer potential diagnostic markers and therapeutic targets for the treatment of OA.

As society ages, the incidence of knee osteoarthritis continues to rise, bringing with it a series of social impacts and medical pressure. Despite the increasing recognition of the role of ion channels in the pathogenesis of OA, diagnostic markers and targeted therapies remain limited.This study investigated the role of TRP as possible diagnostic tools for OA.Seven TRP-related genes were identified as the best traits to distinguish OA from healthy samples, and then we constructed and validated risk scores for three key genes (PPP1R3D, ZNF101, and LOC651309) relevant to OA ion channel gene modules.Our findings provide novel insights into the role of ion channel-related genes in OA pathogenesis and offer a reference for further clinical diagnosis.

Kinematic and Dynamic Study on Traditional Chinese Cervical Manipulation / 医用生物力学

Objective To simultaneously collect and analyze the kinematic and dynamic parameters for two techniques of traditional Chinese cervical manipulation ( TCCM), and quantitatively describe its biomechanical characteristics. Methods A senior practitioner completed the TCCM (positioning and directional rotation pulling, lateral flexion, respectively) on 10 healthy subjects, and the fluorescent marker balls were pasted on the operator to capture manipulation movements. The dynamic parameters and the surface electromyography ( sEMG) signals were collected by pressure-sensitive gloves and wireless sEMG acquisition system. Results The upper arm muscle was the main force muscle during TCCM, and biceps brachii had the highest contribution rate. The range of motion (ROM), speed, pulling force, and time during cervical spine positioning and directional rotation pulling were all greater than those during cervical spine lateral flexion. The integrate electromyography ( iEMG) and root mean square (RMS) for each muscle of the operator during cervical spine positioning and directional rotation pulling were higher than those during cervical spine lateral flexion. Conclusions The overall ROM, three-dimensional (3D) motion angle, load intensity and time during CCTM have the characteristics of high speed, low amplitude and strong force, reflecting the biomechanical characteristics of ‘ cunjin ’ ( one-inch punch ) in traditional Chinese medicine. This study provides references for further standardizing manual teaching and training and improving clinical safety.

Clinical study of modified Shishi Niubangzi Decoction combined with strengthening tendons-waist exercise on lumbar disc herniation / 国际中医中药杂志

Objective:To observe the clinical effect of modified Shishi Niubangzi Decoction combined with strengthening muscle-waist exercise on lumbar disc herniation (LDH).Methods:Randomized controlled trial. A total of 60 patients with LDH admitted to the Pinggu Hospital, Beijing Traditional Chinese Medicine Hospital, were enrolled as the research objects between September 2020 and September 2021. According to the random number table, they were randomly divided into the treatment group and control group, 30 in each group. Both groups were given routine basic treatments (strengthening tendons-waist exercise and three-position six-step manipulation). On this basis, the treatment group was treated with modified Shishi Niubangzi Decoction, while the control group was treated with non-steroidal anti-inflammatory drugs (ibuprofen codeine sustained-release tablets). Both groups were treated for 4 weeks. The responsive rates, back pain intensity, leg pain and numbness by Visual Analogue Scale (VAS) and lumbar function by Oswestry Disability Index (ODI), and Japanese Orthopaedic Association (JOA) were compared between the two groups.Results:The response rate of treatment group was significantly higher than that of control group (93.3% vs. 73.3%; χ2=4.32, P=0.038). After treatment, scores of JOA (subjective symptoms, signs, activities of daily living) in the treatment group were significantly higher than those in the control group ( t=3.86, 2.71, 2.21, P<0.05). After treatment, scores of back pain (2.12±0.21 vs. 3.02±0.32, t=12.88), leg pain (2.04±0.64 vs. 2.64±0.66, t=3.58), lower limb numbness (1.75±0.24 vs. 2.41±0.70, t=4.89) in the treatment group were significantly lower than those in the control group ( P<0.01). At 1 week and 1 month after treatment, ODI scores in treatment group were significantly lower than those in control group ( t=10.22, 5.59; P<0.05). Conclusion:The modified Shishi Niubangzi Decoction combined with strengthening tendons-waist exercise can improve responsive rates, improve lumbar pain and function in LDH patients.

Clinical observation of acupuncture combined with sitting-position knee-adjustment manipulations for patellofemoral arthritis / 针灸推拿医学（英文版）

Objective: To observe the efficacy of acupuncture combined with sitting-position knee-adjustment manipulations in treating patellofemoral osteoarthritis. Methods: Ninety-two patients with patellofemoral osteoarthritis were randomized into an observation group and a medication group, with 46 cases in each group. The observation group received acupuncture and sitting-position knee-adjustment manipulations, and the medication group received oral celecoxib capsules. After 8-week treatment, changes in the short-form McGill pain questionnaire (SF-MPQ) and Lysholm knee scoring scale (LKSS) scores were observed, and the clinical efficacy was compared.Results: The total effective rate was 87.0％ in the observation group and 63.0％ in the control group; the between- group difference was statistically significant. Before treatment, there were no significant differences in the SF-MPQ score or LKSS score (P>0.05). After 8-week treatment, the SF-MPQ [including pain rating index (PRI), visual analog scale (VAS), and present pain intensity (PPI)] and LKSS scores showed notable changes in both groups (P<0.05); the SF-MPQ and LKSS scores in the observation group were significantly different from those in the control group (P<0.05).Conclusion: Combining acupuncture and sitting-position knee-adjustment manipulations can reduce pain and ameliorate joint function in patients with patellofemoral osteoarthritis, producing more significant efficacy than oral celecoxib capsules.

System Review of Acupotomy Treatment for Stenosing Tenovaginitis of Flexor Digitorum / 中国中医药信息杂志

Objective To systematically evaluate the efficacy of acupotomy treatment for the stenosing tenovaginitis of flexor digitorum.Methods Articles about RCTs of acupotomy treatment and local blocking for the treatment of stenosing tenovaginitis of flexor digitorum were retrieved from CBM, CNKI, VIP, Wanfang database, PubMed, Embase, and Cochrane library. And then according to the requirement of Cochrane Handbook for Systemetic Reviews 5.1.0, two reviewers independently chose tests, extracted data, had a cross check, accessed methodological qualities and finally used RevMan 5.3 software for Meta analysis. The total effective rate, cure rate and improvement rate of acupotomy treatment and local blocking in treating stenosing tenovaginitis of flexor digitorum were assessed by using odds ratio (OR).Results Totally 10 articles involving 1426 cases were included. The cure rate in acupotomy treatment group was higher than that of local blocking group [OR=13.11 (95% CI: 8.23–20.89),Z=10.83 (P<0.000 01)], funnel plot Begg’s testP=0.858, Egger’s testP=0.579; the improvement rate of local blocking rate was higher than that of acupotomy treatment group [OR=0.14 (95% CI: 0.10–0.19),Z=11.60 (P<0.000 01)], funnel plot Begg’s test P=1.000, Egger’s testP=0.926; the total effective rate of acupotomy treatment group was higher than that of local blocking group [OR=18.26 (95% CI: 9.95–33.50),Z=9.38 (P<0.000 01)], funnel plot Begg’s testP=0.592, Egger’s testP=0.936.Conclusion Acupotomy treatment for stenosing tenovaginitis of flexor digitorum is superior to local blocking injection. However, the quality of the RCTs is low, so more large-sample, multi-center, and high-quality clinical RCTs are required for further verification.

Establishment of a chondrocyte degeneration model by over-expression of human Wnt7b gene in 293ft cell line / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of human Wnt7b gene in rat chondrocyte degeneration.</p><p><b>METHODS</b>Wnt7b gene obtained by PCR was cloned to PCDH-GFP. 293ft cell line was transfected with PCDH-GFP and PCDH-Wnt7b, and the supernatant and transfected cells were collected. The expression level of Wnt7b in 293ft cells was detected by Western blotting. The first passage of chondrocytes were isolated from articular cartilages of newborn born (within 24 h) SD rats were cultured in the supernatants from the transfected cells (at 10- and 50-fold dilutions). The cell morphology of the rat chondrocytes was observed under inverted microscope, and the protein expressions of MMP13, MMP3 and type II collagen and mRNA expressions of A-can, ADAMTS5, Col X and Sox9 were examined by Western blotting or real-time PCR.</p><p><b>RESULTS</b>Human Wnt7b gene cloned to PCDH-GFP was expressed efficiently in 293ft cell line. Rat chndrocytes cultured for 24 h in the supernatants from PCDH-Wnt7b-transfected 293ft cells underwent changes from a polygonal to a spindle-shaped morphology. The protein expression levels of MMP13 and MMP3 increased while type II collagen decreased significantly, and the mRNA levels of A-can and Sox9 were down-regulated while Col X and ADMATS5 up-regulated in ratchondrocytes after incubation in supernatants from PCDH-Wnt7b-transfected 293ft cells.</p><p><b>CONCLUSION</b>Human Wnt7b gene can be expressed efficiently in 293ft cell line and can induce rat chondrocyte degeneration in vitro.</p>

Efficacy Observation of Tuina Therapy for Fibromyalgia Syndrome / 针灸推拿医学（英文版）

Objective: To observe the efficacy of tuina manipulations for fibromyalgia syndrome (FMS). ＜br> Methods:Sixty eligible FMS patients were randomized into two groups, 30 in each group. Patients in the observation group were intervened by tuina manipulations to the corresponding segment of the spine based on the detection of disorders of the spine-related bones and tendons;patients in the control group were directed to practice functional exercises. Before and after intervention, the fibromyalgia impact questionnaire (FIQ) and visual analogue scale (VAS) were adopted for evaluation. ＜br> Results:After intervention, the FIQ and VAS scores dropped significantly in both groups (all P＜0.05), and there were significant differences between the two groups (both P＜0.05). The total effective rate was 86.7%in the observation group versus 63.3% in the control group, and the difference was statistically significant (P＜0.05). ＜br> Conclusion:Spinal tuina manipulations are effective and safe in treating FMS, for significantly promoting the quality of life of the FMS patients.

Effect of Chinese Daoyingong method on knee osteoarthritis / 现代临床护理

ObjectiveTo explore the effect of Chinese Daoyingong method on knee osteoarthritis.Methods Eighty patients with knee osteoarthritis were randomly divided into the experiment group and control group in equal number.The control group was given the routine medicine combined with Chinese fumigation and the experiment group was treated with Chinese Daoyin method beside the treatment in the control group.The two groups were compared in terms of the score on visual analog scale of pain and the score on knee function before and after treatment.Results The visual analog scale of pain in the experiment group was significantly lower than that in the control group after intervention and the score on knee function was significantly higher than that of control group (P<0.001). Conclusion The intervention with Chinese Daoyingong method plus routine medication and Chinese herbal fumigation is effective in the treatment of knee osteoarthritis.

Advances of molecular targeted therapy based on Wnt signaling pathway in osteoporosis / 中华内分泌代谢杂志

The Wnt signaling pathway plays an important role in bone metabolism. Inducing the Wnt signaling pathway promotes bone formation while restraining it results in osteopenic states. Although the regulation of this signaling pathway may bring enormous therapeutic potential, it still requires cautious approach because of the risks of tumorigenesis. The role of the Wnt signaling pathway in bone metabolism and the molecular targets of therapeutic potential for osteoporosis are discussed in this review.

Development and validation of a CO-T1 three-dimensional finite element model of a healthy person under physiologic loads / 生物医学工程学杂志

A comprehensive, geometrically accurate, nonlinear C0-T1 three-dimensional finite element (FE) model was developed for the biomechanical study of human cervical spine and related disorders. The model was developed with anatomic detail from the computed tomography (CT) images of a 46-year-old female healthy volunteer, and applied the finite element model processing softwares such as MIMICS13.1, Hypermesh11.0, Abaqus 6.12-1, etc., for developing, preprocessing, calculating and analysing sequentially. The stress concentration region and the range of motion (ROM) of each vertebral level under axial rotation, flexion, extension, and lateral bending under physiologic static loadings were observed and recorded. The model was proven reliable, which was validated with the range of motion in previous published literatures. The model predicted the front and side parts of the foramen magnum and contralateral pedicle and facet was the stress concentration region under physiological loads of the upper spine and the lower spine, respectively. The development of this comprehensive, geometrically accurate, nonlinear cervical spine FE model could provide an ideal platform for theoretical biomechanical study of human cervical spine and related disorders.

Isolation, culture and osteogenic differentiation of adipose-derived stem cells from the abdominal cavity of rats / 中国组织工程研究

10.3969/j.issn.2095-4344.2013.23.008

Alendronate improves bone matrix structure of ovariectomized rats / 中国组织工程研究

BACKGROUND:Bisphosphonates that can increase bone density and inhibit bone resorption have been clinical y confirmed, but the structure of the bone matrix has been less studied. OBJECTIVE:To observe the effects of alendronate on bone structure and bone matrix metabolism, and then to investigate the control ing mechanism by which alendronate improves bone mass and increase bone intensity. METHODS:An ovariectomized rat model was prepared and intervened with alendronate as treatment group. Model and sham-surgery groups were set as controls. Alendronate effects on bone mineral density, bone metabolism, bone biomechanics, and bone structure were observed in bone loss rats using bone imaging, bone tissue pathology and biomechanical test and enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION:Alendronate intervention could fight against bone loss as compared with model group at weeks 4, 8, and 12 after treatment (P<0.05). Compared with the model group, the expression of urinary deoxypyridinoline and serum carboxyterminal propeptide of type Ⅰ procol agen was decreased significantly after alendronate intervention (P<0.05);the maximal load, maximal pressure and modelus on the lumbar vertebrae and femur were increased as wel as ratio of urinary pyridinoline/deoxypyridinoline of type Ⅰ procol agen (P<0.05). These findings suggest that alendronate intervention can inhibit bone loss in rats induced by estrogen deficiency, increase biomechanical properties, improve bone matrix structure, and meanwhile, recover the Ⅰ col agen crosslinking component due to ovariectomy.

Application of finite element analysis in Chinese cervical manipulation biomechanics / 生物医学工程学杂志

Clinical advantages of Chinese spinal manipulation therapy (CSMT) were recognized for spinal chronic lesions of soft tissues and bones, such as cervical spondylosis, etc. However, the security of CSMT and the hypotheses of practice mechanisms were questioned for lacking of the relevant basic researches. Researches have proved that these methods could be used to observe the dynamic effects of spine with application of finite element analysis (FEA) computer technology. Combining with other biomechanical experimental methods and applying advanced FEA technology for mechanical problems of CSMT, we may not only find the mechanisms of action and provide theoretical supports for the traditional Chinese therapy, but also standardize the key techniques and optimize the treatment options improving clinical outcomes, and even promote spreading of CSMT. Computer models are ideally suited for studying phenomena that cannot be satisfactorily investigated with other models. However, computer models of CSMT practice remain to be further refined. The results which had been acquired so far not only verified some of the traditional points of view, but also revised and specified some perspectives of the past. This paper intends to review FEA studies with Chinese cervical manipulation therapy (CCMT) for cervical spinal chronic lesions of soft tissues and bones, involving different effects for cervical spine joints (pulling/traction and thrusting) with practice techniques and cervical spine soft tissues (including vessels and its hemodynamics, muscles and fasciae, etc).

Inhibition effect of osthole on proliferation of rat chondrocytes / 中西医结合学报

To investigate the effects of osthole on chondrocyte proliferation in vitro.

Clinical Study on Combined Tuina with Qi Zheng Qing Peng Paste for Herniation of the Lumbar Intervertebral Disc / 针灸推拿医学（英文版）

Objective: To observe the clinical efficacy of combined tuina manipulations with Qi Zheng Qing Peng paste for herniation of the lumbar intervertebral disc. Method: All 568 cases were randomly allocated into an observation group (189 cases), control Ⅰ group (187 cases) and control Ⅱ group (192 cases). Combined tuina with Qi Zheng Qing Peng paste, combined tuina with diclofenac diethylamine emulsions and tuina alone were adopted in the above three groups respectively. The efficacies in the three groups were compared after 4 weeks of treatment. Results: The total effective rates in the observation group, control Ⅰ group and control Ⅱ group were 89.9%, 86.6% and 84.9% respectively, showing a significant difference (P ＜0.05) . Conclusion: Combined tuina manipulations with Qi Zheng Qing Peng paste works well for herniation of the lumbar intervertebral disc.

Morphological studies on osteoarthritis of the articular cartilage in C57 black mice and its relationship with etalloproteinase-3 and lnterleukln-1 in serum and synovial fluid / 中华风湿病学杂志

Objective To study the levels of Matrix Metalloproteinase-3 (MMP-3) and interleukin (IL-1) in the synovial fluid and plasma of C57 black mice with osteoarthritis (OA) and their relationships with the severity of pathological changes so as to investigate their effects and correlation with OA. Methods The C57 black mice with OA were enrolled for this study. Different levels of exercise were appicated based on their age. Knee joint pathological changes were examined for pathological severity of OA. ELISA sandwich method was used to measure the levels of MMP-3 and IL-1 in serum and synovial fluid. Correlation analysis was performed to demonstrate the relationship between the levels of MMP-3 and IL-1 in the serum and synovial fluid, and the pathological severity of OA. Results ①Morphological observations: C57 black mice were characterized by spontaneously developing OA and the incidence and the severity of osteoarthritis gradually increased with age and exercising burdens. ② The level of MMP-3 and IL-1 in the synovial fluid of exercising mice MMP-3 (84±6) ng/ml, IL-1 (48±3) ng/ml was higher than that in the aged ones [MMP-3 (84±6) ng/ml, IL-1 (71±5) ng/ml J, the difference was significant (P<0.01). The level of MMP-3 and IL-1 level in the serum had a linear correlation with that of the synovial fluid. At the same time, they also had linear correlation with the pathological severity of OA (All r>0.67, and all P<0.01). Conclusion The levels of MMP-3 and IL-1 in serum and synovial fluid can help to make early diagnosis of OA, especially elevated MMP-3 level.

Astragalus root injection regulates type I collagen expression of rat osteoblasts in vitro / 中国组织工程研究

BACKGROUND: Type Ⅰ collagen is a specific collagen secreted by in vitro cultured osteoblast, and the formed network is the basis of bone mineralization, which also reflects the ability of osteoblast bone formation. Studies have shown astragalus root increased osteoblast proliferation. However, the effect of astragalus root on improving type Ⅰ collagen expression of osteoblast remains poorly understood.OBJECTIVE: To evaluate the effect of astragalus root injection on the abilities of rat cranium-derived osteoblast proliferation and type Ⅰ collagen expression.METHODS: Rat osteoblast was cultured in vitro and divided into control group (MEM culture solution containing calf serum) and astragalus root groups (different concentrations). The effect on osteoblast proliferation was evaluated on days 1, 3, 5, 7, and 9 by MTT method. Moreover, the expression of type Ⅰ collagen protein was observed after 6 hours of treatment with astragalus root injection using in cell western-blot method. In addition, the gene expression of COLLal was investigated by real-time PCR method.RESULTS AND CONCLUSION: From days 3 to 9, the different concentrations of astragalus root injection improved osteoblast proliferation, respectively compared with control group (P ＜ 0.05), and this ascending trend peaked on day 7. Different concentretions of astragalus root injection improved COLLol mRNA expression, especially 15% astragalus root injection was the most effective. The type Ⅰ collagen protein expression of 15% and 10% astragalus root injection were significantly greater compared with the control group (P ＜ 0.05). Astragalus root injection improved in vitro cultured osteoblast proliferation and type Ⅰ collagen secretion in a certain dose-effect manner.

Comparison of the effect of glucosamine on the cartilage oligomeric matrix protein secretion in vitro chondrocytes and synoviocytes / 中华风湿病学杂志

Objective To compare the effect of glucosamine (Virtral-s) on the cartilage oligomeric matrix protein (COMP) secretion of chondrocytes and synoviocytes in vitro.Methods Chondrocytes and synoviocytes isolated from knee cartilage of osteoarthritic patients were cultured by phased enzymatic digestion.Sera containing Virtral-s of the experimental animals were obtained after orally administrated Virtral-s at the dosages that equal to human.Cells were cultured in the medium with Virtral-s containing sera.Super-natant COMP level was tested by enzyme-linked immunoabsorbent assays (ELISA).Results COMP conceu-tration of synoviocytes cultured in vitro was significantly higher than that of chondrocytes (P＜0.05).Virtral-s could significantly increase COMP secretion in cultured chondrocytes in vitro (P＜0.05),however,it had a weaker role on synoviocytes,ie,it could only mildly reduce COMP secretion of synoviocytes.Conclusion Glucosamine (Virtral-s)-containing serum can promote COMP secretion of chondrocytes in vitro,and it has no significant effect on synoviocytes in vitro.

Comparative study on effects of manipulation treatment and transcutaneous electrical nerve stimulation on patients with cervicogenic headache / 中西医结合学报

OBJECTIVE: To compare the effects of manipulation treatment and transcutaneous electrical nerve stimulation (TENS) on patients with cervicogenic headache. METHODS: Seventy patients with cervicogenic headache were randomly allocated to receive manipulation treatment and TENS treatment, which were given every other day respectively for total 40 days. The headache degree (numeric rating scale, NRS), frequency, lasting time and the range of motion (ROM) of the cervical spine 2 weeks before treatment and 4 weeks after treatment, were compared. RESULTS: Before treatment, there were no significant differences in the headache NRS scores , frequency, lasting time and the ROM scores between the two groups (P>0.05). After treatment, the headache NRS score, frequency, lasting time in the manipulation group decreased significantly (P0.05). The response rate of manipulation treatment is 94.5%, significantly higher than 64.5% of TENS treatment (P<0.05). CONCLUSION: The manipulation treatment is an effective method for patients with cervicogenic headache.

An transcranial Doppler ultrasonography and X-ray study of cervical vertigo patients treated by manipulation in supine position / 中西医结合学报

OBJECTIVE: To observe the transcranial Doppler ultrasonography and X-ray changes of cervical vertigo patients caused by cervical instability before and after the manipulation treatment in supine position. METHODS: Sixteen cervical vertigo patients who underwent supine manipulation treatment were reviewed retrospectively. After the treatment, the changes of transcranial Doppler ultrasonography of cervical vertebral artery and X-ray of cervical spine were observed. RESULTS: Before the treatment, the mean blood flow velocity of vertebral artery was (48.6+/-3.1) cm/s, much higher than that of the normal group (P<0.05), the means of the horizontal displacement of C3,4 and C4,5 were (3.18+/-0.42) mm and (3.46+/-0.26) mm respectively. After the treatment, the blood flow velocity and the C3,4 horizontal displacement decreased significantly (P<0.05). The effective rate of the manipulation treatment was 100%. CONCLUSION: The manipulation treatment is an effective method for cervical vertigo patients caused by cervical instability, and the effect is correlated with the improvement of the blood flow of cervical artery and the restoration of cervical stability.