Serum metabolomics analysis of deficiency pattern and excess pattern in patients with rheumatoid arthritis.

BACKGROUND: Rheumatoid arthritis (RA) is a chronic and refractory autoimmune disease. Deficiency pattern (DP) and excess pattern (EP), as crucial types of Chinese medicine pattern diagnoses published by International Classification of Diseases 11th Revision (ICD-11), could provide new strategies for RA diagnosis. However, the biological basis of DP and EP of RA is not explicit. METHODS: 19 female RA DP patients, 41 female RA EP patients and 30 female healthy participants were included in the study. The serums of participants were collected and analyzed by metabolomics based on ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry to profile metabolic characteristics of RA DP and EP. Furthermore, bioinformatics analysis results were obtained by using Ingenuity Pathway Analysis (IPA) and statistical analysis was performed by SAS version 9.4 for further identification of potential biomarkers. RESULTS: Serum metabolic profiling revealed 25 and 24 differential metabolites in RA DP and EP respectively, and 19 metabolites were common to RA DP and EP. Compared with DP group, L-Homocysteic acid, LysoPE(P-16:0/0:0), N(omega)-Hydroxyarginine and LysoPC(16:0/0:0) decreased (P < 0.05), and Pyruvic acid, D-Ribose, Gamma-Glutamylserine, PE(22:0/24:1(15Z)), Inosinic acid increased (P < 0.05) in EP group. Menawhile, S-Nitrosoglutathione, 5-Thymidylic acid, SN38 glucuronide, PE(22:0/24:0), PC(24:0/24:1(15Z)) and Bisdiphosphoinositol tetrakisphosphate increased significantly in DP group compared to EP group (P < 0.05). For the unique metabolites, bioinformatics analysis results showed that 5-Methoxytryptamine involved in Melatonin Degradation II and Superpathway of Melatonin Degradation is the key metabolite to RA DP. Meanwhile, GABA is the key metabolite in EP group, which involved in Glutamate Dependent Acid Resistance, GABA Receptor Signaling, Glutamate Degradation III (via 4-aminobutyrate) and 4-aminobutyrate Degradation I. Bioinformatics analysis between unique metabolites of RA DP and EP groups with human target genes for RA showed that 5-methoxytryptamine and LysoPC(18:1(9Z)/0:0), the unique metabolites of RA DP, might participate in colorectal cancer metastasis signaling, tumor microenvironment pathway, apoptosis signaling, MYC mediated apoptosis signaling, erythropoietin signaling pathway and LXR/RXR activation. Simultaneously, GABA, LysoPA(18:1(9Z)/0:0) and L-Targinine, the unique metabolites of RA EP, might participate in neuroinflammation signaling pathway, osteoarthritis pathway, glucocorticoid receptor signaling, ILK signaling, IL-17 signaling and HIF1α signaling. CONCLUSIONS: The study indicates that serum metabolomics preliminarily revealed the biological basis of RA DP and EP. 5-methoxytryptamine, LysoPC(18:1(9Z)/0:0) and GABA, LysoPA(18:1(9Z)/0:0), L-Targinine might be the predictors to distinguish the DP and EP of RA respectively. These interesting results provide thoughts for further study of traditional medicine patterns of ICD-11. It also contributes to provide strategy for personalized precision treatment of RA and further validation is needed.

Effects and Safety of the Tripterygium Glycoside Adjuvant Methotrexate Therapy in Rheumatoid Arthritis: A Systematic Review and Meta-Analysis.

Objective: This study aimed to systematically review the efficacy and clinical safety of different courses and doses of tripterygium glycoside (TG) adjuvant methotrexate (MTX) therapy in the treatment of rheumatoid arthritis (RA). Methods: Randomized controlled trials (RCTs) of TG adjuvant MTX therapy in patients with RA were retrieved from SinoMed, China Network Knowledge Infrastructure, WanFang Data, PubMed, Cochrane Library, and Embase from inception to September 30, 2021. The effects and clinical safety evaluations were conducted using RevMan 5.3 software. Results: A total of 9 RCTs and 892 patients with RA were included in this study. In the meta-analysis, a total of 463 and 429 patients were enrolled into the TG adjuvant MTX therapy group and MTX monotherapy group, respectively. In comparison with MTX monotherapy, the results of the analyzed effects showed that the TG adjuvant MTX therapy can achieve 20%, 50%, and 70% improvements in American College of Rheumatology (ACR) criteria ACR20, ACR50, and ACR70 at P = 0.005, P = 0.0001, and P = 0.004, respectively. Simultaneously, the efficacy of the TG adjuvant MTX therapy was improved at either 30 or 60 mg/day over a six-month course compared to MTX monotherapy (P < 0.0001). There was no statistical difference in the effects between the doses of 30 and 60 mg/day after three months (P = 0.82). TG adjuvant MTX also reduced the expression rate of the swollen joint count, tender joint count, erythrocyte sedimentation rate, rheumatoid factor, and C-reactive protein in subgroup analyses with different courses and doses. In terms of hepatic adverse effects (P = 0.28), leukopenia (P = 0.78), gastrointestinal adverse effects (P = 0.17), cutaneous adverse effects (P = 0.94), and irregular menstruation adverse effects (P = 0.29), there was no statistically significant difference with TG adjuvant MTX therapy and MTX monotherapy with different courses and doses. Conclusions: TG adjuvant MTX therapy is more effective than MTX monotherapy and is a safe strategy for RA treatment in doses of 30 or 60 mg/day over a treatment course of six months. However, high-quality multicenter RCT studies with large sample sizes are still needed to confirm the effects and clinical safety of different courses and doses of TG adjuvant MTX therapy.

[Case-control study on manipulation following arthroscopic capsular release for the treatment of frozen shoulder].

OBJECTIVE: To compare therapeutic effects between the normal rehabilitation and combined with manipulative method after arthroscopic capsular release for the treatment of severe frozen shoulder, and to evaluate the application value of manipulationp. METHODS: From March 2007 to July 2010,arthroscopic capsular release was performed in 48 cases (48 shoulders, 23 left side, 25 right side). All the patients were divided into two groups: control group (11 males and 15 females) and manipulation group (9 males and 13 females). The patients in the control group were treated with conventional rehabilitation procedure, and the patients in the manipulation group were treated with additional manipulation procedure. From the 2nd day after operation, the manipulation was performed for 20 minutes every time, twice daily, and it continued for 10 days. All the cases were followed up and the scale of American Shoulder and Elbow Surgeons Standardized Assessment Form (ASES self-report section) and the range of motion (ROM) were recorded. RESULTS: The mean follow-up period was (12.54 +/- 5.78) months (ranging from 4 to 25 months). Both ASES scores and ROM in the manipulation group were better than those in the control group at the 1st month after operation, and the difference between the ASES scores and flexion of the shoulder were significant. However, there was no significan difference at the latest follow-up. CONCLUSION: Compared with the conventional rehabilitative procedure, manipulation following arthroscopic capsular release could promote the process of joint rehabilitation and help the patient back to normal life earlier, but there is no evidence of long term advantage.

[Effects of Chinese herbal medicine Qianggu Capsule on patients with rheumatoid arthritis-induced osteoporosis: a report of 82 cases].

BACKGROUND: Rheumatoid arthritis (RA) is a kind of chronic autoimmune disease and osteoporosis is one of its complications. OBJECTIVE: To explore the effects of Qianggu Capsule, a compound traditional Chinese herbal medicine, on bone mineral density (BMD) and osteoporosis in patients with RA. DESIGN, SETTING, PARTICIPANTS AND INTERVENTION: Eighty-two patients with rheumatoid arthritis and osteoporosis, who were treated in Shanghai Guanghua Hospital of Integrated Traditional Chinese and Western Medicine from January 2010 to December 2011, were divided into treatment group (42 cases) and control group (40 cases). The patients in the treatment group were administered with Qianggu Capsule and two disease-modifying antirheumatic drugs. The patients in the control group were administered with two common-used antirheumatic drugs. The course of treatment was 6 months. MAIN OUTCOME MEASURES: Blood levels of alkaline phosphatase (ALP), calcium, phosphorus, C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR) were determined before and after the treatment. BMD in the lumbar spine, femur and the left distal radius were also examined before and after the treatment. RESULTS: The ALP level, a bone metabolic parameter, was significantly increased in patients of the treatment group after treatment compared with before treatment. BMD values in the lumbar spine, femur and the radius were higher after treatment than before treatment (P<0.05). There were no changes in ALP level and BMD in the patients of the control group after the treatment when compared with before treatment. CONCLUSION: Treatment with Qianggu Capsule can increase BMD of RA patients, and then ameliorate their osteoporosis.

[Effects of early application of Tuina treatment on quadriceps surface myoelectricity in patients after total knee arthroplasty: a randomized controlled trial].

BACKGROUND: Total knee replacement surgery is commonly used in end-stage diseases of the knee. It is important for improving surgical efficacy and patient satisfaction by promoting early rehabilitation of patients and improving knee function. OBJECTIVE: To observe the effects of early application of Tuina treatment on quadriceps surface electromyography (EMG) in patients with rheumatoid arthritis having undergone total knee arthroplasty. DESIGN, SETTING, PARTICIPANTS AND INTERVENTION: The study was performed at the Orthopedic Department of Huashan Hospital, Fudan University, and the Shanghai Guanghua Hospital of Integrated Traditional Chinese and Western Medicine from June 2010 to September 2011. A total of 66 patients with rheumatoid arthritis who had undergone total knee replacement surgery were randomly divided into control group and observation group, 33 cases in each. The patients in the control group were administered with continuous passive training (CPM), and the patients in the observation group were treated with CPM combined with Tuina, from prior surgery to four weeks post-surgery. MAIN OUTCOME MEASURES: The knee function was evaluated using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) questionnaire at baseline and 4 weeks after the surgery. Quadriceps surface EMG was also detected at the same time points. RESULTS: After 4 weeks of Tuina and comprehensive rehabilitation intervention, the WOMAC questionnaire score of the observation group was decreased compared with the control group (P<0.01); median frequency and integrated electromyography of the rectus femoris and vastus medialis muscles, which were recorded by EMG, in the observation group were higher than those in the control group (P<0.01). CONCLUSION: Tuina can improve the recovery of patients who have undergone total knee replacement by increasing quadriceps EMG.

[Effect of Bushen Qianggu decoction on the proliferation of synovial fibroblasts and expression of PCNA and Bcl-2].

OBJECTIVE: To observe the effects of Bushen Qianggu decoction proliferation and PCNA and Bcl-2 expression. METHODS: Serum containing BQD was made and synovial fibroblasts were separated and cultured and passaged in vitro. Four groups were divided as 20% blank control group, serum containing 20% Tripterygium wilfordii multi-glycosides drug (TWMD), 20% of serum containing high and low of BQD, respectively. Serum containing drugs of different concentration were added into the synovial fibroblasts of the third generation, and then the synovial fibroblasts were cultured continued. The effects of different drugs on synovial fibroblasts and PCNA and Bcl-2 expression were observed. RESULTS: Compared with the control serum, BQD-containing serum promoted the apoptosis of synovial fibroblasts (P < 0.000 1); especially, high dose could inhibit proliferation. The expression of PCNA and Bcl-2 was significantly lower in BQD-containing serum (P < 0.000 1 vs control group). CONCLUSION: BQD can promote the apoptosis of synovial fibroblasts by improving of expression of PCNA and Bcl-2, which may be one of the mechanisms of BQD in preventing and treating osteoporosis of rheumatoid arthritis.

[Effects of Tuina and Chinese patent drug Shuxuetong injection on deep venous thrombosis and functional rehabilitation after total knee arthroplasty].

OBJECTIVE: To evaluate the efficacy of Tuina and Chinese patent drug Shuxuetong injection in preventing patients undergoing total knee arthroplasty from deep venous thrombosis and in functional rehabilitation. METHODS: A total of 120 patients with diagnosed rheumatoid arthritis in the Department of Orthopaedic Surgery, Shanghai Guanghua Hospital of Integrated Traditional Chinese and Western Medicine in China were enrolled for this study. The patients underwent total knee arthroplasty and were divided into treatment group (n=60) and control group (n=60) after surgery. Patients in the control group received conventional rehabilitation training, including using a continuous passive motion machine and training of muscle contractions of the lower limb. Patients in the treatment group were administered Shuxuetong injection and Tuina based on the conventional rehabilitation training. The course of treatment lasted for 2 weeks. Hospital for Special Surgery (HSS) knee score, rate of deep venous thrombosis and range of motion of the knee joint were evaluated before and after treatment. RESULTS: There was no significant difference in HSS knee score and range of motion as compared before and after treatment in two group (P>0.05). The rate of deep venous thrombosis of the treatment group was 13.33%, which was lower than 20% of the control group (P<0.05). CONCLUSION: Tuina combined with Shuxuetong injection treatment can prevent deep venous thrombosis in patients with rheumatoid arthritis after total knee arthroplasty.

[Study on estrogen receptor of adult human osteoblasts].

OBJECTIVE: To examine estrogen receptor (ER) in osteoblasts from adult human and to elucidate the mechanism of estrogen in modulating bone metabolism. METHODS: The cultured osteoblasts were harvested from bone chips by modified sequential digestive enzyme release and immunohistochemical assay of ER in osteoblasts were carried out in three groups of female adults: normal control (group 1), patients with moderate osteoporosis (group 2) and patients with serious osteoporosis (group 3). The percentages of ER-positive osteoblasts from the three groups were compared by t test. RESULTS: The brown marks that indicate ER were found in nuclei and plasma of the osteoblasts, and the percentages of ER-positive osteoblasts among three groups were significantly different. CONCLUSION: ERs exist in nuclei and plasma of the osteoblasts. Estrogen may modulate bone metabolism through binding ER in nuclei and plasma of the osteoblasts. The reduction of ER of osteoblasts may play an important role in the pathogenesis of postmenopausal osteoporosis.

[Culture of human osteoclasts].

OBJECTIVE: To establish a stable, useful culture system for human osteoclasts and to investigate the effect of osteoblasts on the differentiation, proliferation and activation of osteoclasts so as to provide a base for the studies on prevention and treatment of osteolysis and osteoporosis. METHODS: In the presence of 1,25-(OH)2D3, monocytes abstracted from human bone marrow were cultured in three groups: co-culture of monocytes and osteoblasts, monocytes alone, monocytes with conditional media (CM) of osteoblasts. Differentiation process of the cultured cells was observed under biological microscope. HE staining and tartrate-resistant acid phosphatase (Trap) staining were employed to assay the cultured cells. The resorption pits on bone slices, on which cells were cultured, were observed under scanning electronic microscope (SEM). RESULTS: In the group of co-culture of monocytes and osteoblasts, monocytes gradually fused to form multinucleated cells (MNCs), and the MNCs were also indicated in HE staining and Trap staining. The SEM showed a number of resorption pits on bone slices. In the other two groups, Trap-positive MNCs were not obtained, and resorption pits were not observed on bone slices. CONCLUSION: In this culture, monocytes obtained from human marrow fused to form multinucleated cells (MNCs) that express the main characteristics of the osteoclast phenotype, such as Trap-positive and the ability to form resorption lacunae when cultured on bone slices. Cell-to-cell contact with osteoblasts was necessary for the differentiation, proliferation and activation of osteoclasts.