Blimp1 suppressed CD4+ T cells-induced activation of fibroblast-like synoviocytes by upregulating IL-10 via the rho pathway.

BACKGROUND: B lymphocyte-induced maturation protein 1 (Blimp1) is a risk allele for rheumatoid arthritis (RA), but its functional mechanism in RA remains to be further explored. METHODS: Flow cytometry was performed to detect CD4+ T cell differentiation. ELISA was used to measure inflammatory factor secretion. Lentivirus mediated Blimp1 overexpression vector (LV-Blimp1) or short hairpin RNA (sh-Blimp1) were used to infect CD4+ T cells stimulated by anti-CD28 and anti-CD3 mAbs. RA fibroblast-like synoviocytes (FLSs) were co-cultured with CD4+ T cells or T cell conditioned medium (CD4CM), and cell proliferation, invasion, and expression of adhesion molecules and cytokines in FLSs were evaluated. Mice were injected intradermally with type II collagen to establish a collagen-induced arthritis (CIA) mouse model, and the severity of CIA was evaluated with H&E and Safranin-O staining. RESULTS: Blimp1 knockdown increased pro-inflammatory factor secretion, but downregulated IL-10 concentration in activated CD4+ T cells. Blimp1 overexpression promoted regulatory T cells (Treg) CD4+ T cell differentiation and hindered T helper 1 (Th1) and T helper 17 (Th17) CD4+ T cell differentiation. Blimp1 overexpression suppressed the expression of pro-inflammatory factors and adhesion molecules in CD4+ T cells by upregulating IL-10. Moreover, Blimp1 overexpression impeded the enhanced effect of CD4+ T cells/CD4CM on cell adhesion, inflammation, proliferation, invasion and RhoA and Rac1 activities in FLSs by upregulating IL-10. Additionally, administration with LV-Blimp1 alleviated the severity of CIA. CONCLUSION: Blimp1 restrained CD4+ T cells-induced activation of FLSs by promoting the secretion of IL-10 in CD4+ T cells via the Rho signaling pathway.

Astragalus polysaccharides inhibits cell growth and pro-inflammatory response in IL-1ß-stimulated fibroblast-like synoviocytes by enhancement of autophagy via PI3K/AKT/mTOR inhibition.

The hyperplastic growth of rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLSs) and inflammatory response are pathological hallmarks of RA. It has been reported that Astragalus polysaccharides (APS) possess appreciable anti-inflammatory activity against adjuvant-induced arthritis. Nevertheless, little is known about the role and detailed mechanism underlying the therapeutic effects of APS in RA. This study demonstrated that administration of APS dose-dependently impaired cell viability, increased cell apoptosis by decreasing Bcl-2 expression, increasing Bax expression and Caspase3 activity in IL-1ß-stimulated RSC-364 cells and RA-FLS. Simultaneously, IL-1ß-induced production of pro-inflammatory cytokines IL-6 and TNF-α was significantly decreased after APS treatment. Furthermore, preconditioning with APS dramatically enhanced autophagy activity by increasing Beclin-1 and LC3II/LC3I expression coupled with decreasing p62 expression and augmenting the number of LC3 puncta in IL-1ß-stimulated RSC-364 cells. More importantly, autophagy inhibitor 3-methyladenine (3-MA) partly abolished APS-triggered inhibitory effects on cell growth and production of pro-inflammatory cytokines. APS also repressed the activation of PI3K/Akt/mTOR signaling pathway in IL-1ß-stimulated RSC-364 cells. Moreover, treatment with insulin-like growth factor-1 (IGF-1), an activator of PI3K/Akt signaling, partly reversed the therapeutic effects of APS in IL-1ß-stimulated RSC-364 cells. Collectively, we concluded that APS might attenuate the pathological progression of RA by exerting the pro-apoptotic and anti-inflammatory effects in IL-1ß-stimulated FLSs by regulating the PI3K/AKT/mTOR-autophagy pathway.

Recent perspectives of management of breast cancer metastasis - an update.

Breast cancer is the leading cause of cancer related deaths in women and one of the most common cancers globally. The major obstacle in the management of breast cancer, especially at advanced stages, is metastasis. Metastasis in the advanced stages of breast cancer could decrease survival to approximately 5 years. The reasons could include lack of targeted receptors or chemotherapeutic agents for the management of advanced-stage breast cancer metastasis. The new emerging avenues for the management of this deadly pathological state include local manipulations like radiofrequency ablation (RFA), microwave thermotherapy, cryosurgery (cryotherapy), chemoembolization, radioembolization, breast surgery, or metastasectomy. Few single-institution reports showed improved survival in selected patients like those with oligometastatic stage IV breast cancer. The present review article focused on these emerging new multimodality treatment approaches for a possible efficient management.

Report: Osteocyte enhancement function of bisphosphonates in prosthetic replacement.

Aseptic loosening after prosthetic replacement is the primary cause of shortened service life and lowered stability of prosthesis, and increased revision rate after joint replacement. Factors of causing the loosening of joint prosthesis include mechanical factors and biological factors. The mechanical effect of bisphosphonates (BP) is quite obvious, which can enhance osteocyte function, accelerate the generation of new bone and lower bone resorption activity of osteoclast and macrophage. In animal experiment and adjuvant therapy of patients after joint replacement, BP also shows up the functions of reducing osteolysis induced by wear debris, preventing stress shielding and interface fretting and enhancing bone density. This paper elaborated the mechanism of BP adjusting bone metabolism, and analyzed the action principle and the vital function of it in prosthetic replacement. It has proved that BP can effectively reduce the early peri-prosthesis bone absorption after total hip replacement and improve bone mass peri-prosthesis. It is currently the significant choice of preventing bone lose of peri-prosthesis after operation.

Zhuifeng Tougu capsules in the treatment of knee osteoarthritis (cold dampness obstruction syndrome): a randomized, double blind, multicenter clinical study.

BACKGROUND: In Traditional Chinese Medicine (TCM) theory, cold dampness obstruction is one of the common syndromes of osteoarthritis. Therefore, in clinical practice, the main treatment methods are to dispel wind, remove dampness, and dissipate cold, used to treat knee osteoarthritis (KOA). This report describes a mulitercenter clinical study to assess Zhuifeng Tougu Capsule's efficacy and safety in the treatment of patients who are cold dampness obstruction syndrome in KOA, and to provide evidence-based medical for the rational use of Zhuifeng Tougu Capsules in clinical practice. METHODS: This randomized, parallel group controlled, double-blind, double dummy trial will include a total of 215 KOA patients who meet the study criteria. 215 patients underwent 1:1 randomisation, with 107 cases assigned the experimental group (Zhuifeng Tougu Capsules + Glucosamine Sulfate Capsules Simulator) and 108 assigned the control group (Glucosamine Sulfate Capsules + Zhuifeng Tougu Capsules Simulator). After enrolment, patients received 12 weeks of treatment. The main efficacy measure is the Western Ontario and McMaster University Osteoarthritis Index (WOMAC) pain score. Visual analogue scale (VAS) pain score, Self-condition assessment VAS score, WOMAC KOA score, TCM syndrome score and TCM syndrome efficacy, ESR level, CRP level, suprapatellar bursa effusion depth, use of rescue drugs, and safety indicators are secondary efficacy indicators. RESULTS: Compared with before treatment, WOMAC pain score, VAS pain score, Self-condition assessment VAS score, WOMAC KOA score, and TCM syndrome score decreased significantly in both groups (P < 0.01). Also, the experimental group showed significant differences in the above indicators compared to control (P < 0.01). However, after treatment, no significant differences were showed in the ESR level, CRP level, and suprapatellar bursa effusion depth between the two groups (P > 0.05). No any serious adverse effects showed in the experimental group and control group. CONCLUSIONS: Zhuifeng Tougu Capsules can effectively improve knee joint function and significantly alleviate the pain of KOA. TRIAL REGISTRATION: Clinical trial registration was completed with the China Clinical Trial Registration Center for this research protocol (No. ChiCTR2000028750) on January 2, 2020.

China rheumatoid arthritis registry of patients with Chinese medicine (CERTAIN): Rationale, design, and baseline characteristics of the first 11,764 enrollees.

BACKGROUND: Chinese medicine (CM) has become a popular interventional treatment for rheumatoid arthritis (RA). However, limited knowledge about general characteristics and long-term clinical outcomes hampers the development of CM for RA. PURPOSE: The main objectives of the China Rheumatoid Arthritis Registry of Patients with Chinese Medicine (CERTAIN) were to describe the population of RA patients receiving CM treatment in multiple centers in China using different variables and compare these findings with internationally reported data. STUDY DESIGN: The CERTAIN is a prospective, multicenter, observational disease registry. METHODS: Adult RA patients who fulfilled the 2010 American College of Rheumatology/ European League Against Rheumatism classification criteria for RA and received CM treatment were recruited into the CERTAIN by rheumatologists from 145 hospitals across 30 provinces in China. Data on demographics, disease characteristics, comorbidities, treatments, and adverse events, with a 2-year follow-up, were collected and documented using a predefined protocol. RESULTS: In the 2 years since the study began in September 2019, 11,764 patients have been enrolled (enrolment is ongoing), and 13.10% of participants have completed the 6-month follow-up. We present the baseline characteristics of the first 11,764 enrollees. CONCLUSIONS: The CERTAIN is the first nationwide registry to document comprehensive data on CM treatment in patients with RA. The development of the CERTAIN resource is a significant step forward for Chinese RA patients, herbal medicine users, and research communities and will deepen our understanding of CM for RA. REGISTRATION: The study was registered at ClinicalTrials.gov (NCT05219214).

Paeoniflorin Suppresses Rheumatoid Arthritis Development via Modulating the Circ-FAM120A/miR-671-5p/MDM4 Axis.

Paeoniflorin is an active ingredient derived from Paeonia, which has an anti-inflammatory effect. However, the potential role and basis of paeoniflorin in rheumatoid arthritis (RA) are indistinct. Cell viability, cycle distribution, migration, and invasion were evaluated via Cell Counting Kit-8 (CCK-8), flow cytometry, and transwell assays. The contents of inflammatory cytokines were examined using enzyme-linked immunosorbent assay (ELISA). RNA expression levels were determined via qRT-PCR and western blot. The targeting relationship between miR-671-5p and circ-FAM120A (hsa_circ_0003972) or murine double minute 4 (MDM4) was validated via dual-luciferase reporter assay. Paeoniflorin restrained proliferation, migration, invasion, and inflammation and accelerated cell cycle arrest in RA fibroblast-like synoviocytes (RA-FLSs). Circ-FAM120A was boosted in RA synovial tissues and RA-FLSs. Circ-FAM120A upregulation, miR-671-5p knockdown, or MDM4 augmentation reversed the repressive effect of paeoniflorin on RA-FLS progression. Moreover, paeoniflorin attenuated RA-FLS progression by regulating the circ-FAM120A/miR-671-5p/MDM4 axis. Paeoniflorin inhibited RA-FLS proliferation, mobility, and inflammation and triggered cell cycle arrest via mediating the circ-FAM120A/miR-671-5p/MDM4 pathway.

Total glucosides of paeony protects THP-1 macrophages against monosodium urate-induced inflammation via MALAT1/miR-876-5p/NLRP3 signaling cascade in gouty arthritis.

BACKGROUND: Monosodium urate (MSU)-mediated inflammatory response is a crucial inducing factor in gouty arthritis. Here, we explored the underlying mechanism of total glucosides of paeony (TGP) in MSU-induced inflammation of THP-1 macrophages in gouty arthritis. METHODS: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to detect cell viability. Enzyme-linked immunosorbent assay (ELISA) was utilized to measure the production of interleukin 1ß (IL-1ß) and tumor necrosis factor α (TNF-α). Real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot assay were conducted to determine RNA and protein expression. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay and RNA pull down assay were used to confirm the interaction between miR-876-5p and MALAT1 or NLR family pyrin domain containing 3 (NLRP3). RESULTS: MSU-induced damage and inflammatory response in THP-1 macrophages were alleviated by the treatment of TGP in a dose-dependent manner. Overexpression of NLRP3 or MALAT1 reversed the protective effects of TGP in MSU-induced THP-1 macrophages. The binding relation between miR-876-5p and MALAT1 or NLRP3 was identified in THP-1 macrophages. MALAT1 up-regulated the expression of NLRP3 by sponging miR-876-5p in THP-1 macrophages. TGP suppressed MSU-induced inflammation in THP-1 macrophages through regulating MALAT1/miR-876-5p/NLRP3 axis. TGP suppressed MSU-induced activation of TLR4/MyD88/NF-κB pathway through regulating MALAT1/miR-876-5p/NLRP3 axis. CONCLUSION: In conclusion, TGP suppressed MSU-induced inflammation in THP-1 macrophages through regulating MALAT1/miR-876-5p/NLRP3 axis and TLR4/MyD88/NF-κB pathway, suggesting that TGP was a promising active ingredient for gouty arthritis treatment.

MEK inhibitor enhances sensitivity to chemotherapeutic drugs in multidrug resistant hepatocellular carcinoma cells.

The aim of the present study was to investigate the association between the mitogen-activated protein kinase (MAPK) signal transduction pathway and multidrug resistance in hepatocellular carcinoma cells. A Cell Counting Kit-8 assay was used to determine the drug sensitivity of HepG2 and HepG2/ADM hepatocellular carcinoma cell lines in combination with the MAPK/extracellular-signal-regulated kinase kinase (MEK) inhibitor U0126. Flow cytometry was used to analyze the rate of apoptosis. The reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to determine P-glycoprotein (P-gp) and multidrug resistance-associated protein 1 (MRP1) mRNA expression following treatment with various concentrations of U0126. P-gp and MRP1 expression levels were measured using Western blot analysis. The half-maximal inhibitory concentration was markedly decreased in combination with U0126. RT-qPCR results demonstrated that the expression of multidrug resistance 1 (MDR1) and MRP1 in HepG2/ADM cells was increased 5.37- and 6-14-fold compared with that in HepG2 cells. Furthermore, the expression levels in HepG2/ADM cells were decreased following U0126 treatment in a dose-dependent manner. The expression of P-gp and MRP1 in HepG2/ADM cells was increased 2.68- and 2.76-fold compared with that in HepG2 cells. Furthermore, the expression levels in HepG/ADM cells were decreased following U0126 treatment in a dose-dependent manner. The results of the present study indicate that the MEK inhibitor U0126 enhances sensitivity to chemotherapeutic drugs by downregulating P-gp and MRP1 expression in resistant hepatocellular carcinoma cells. The combination of MEK inhibitor and conventional chemotherapeutic drugs may provide novel therapeutic prospects for the treatment of drug-resistant hepatocellular carcinoma.

Histological transformation from gastric mucosa-associated lymphoid tissue lymphoma to gastric diffuse large B-cell lymphoma.

This study evaluated the clinical features, treatment and prognosis in Chinese patients with histological transformation (HT) from gastric mucosa-associated lymphoid tissue lymphoma to gastric diffuse large B-cell lymphoma. We reviewed the medical records of 71 patients diagnosed with HT between 2001 and 2013, retrospectively. Patients had a median age of 56 years. The ratio of sex (male:female) was 1.3:1. The clinical course was often insidious, lacking specific clinical presentation. Macroscopically, the antrum was the most commonly involved site. Thirty-one patients (45%) presented at stage I, and 25 (35%) presented with local (18/71, 25%) or distant (7/71, 10%) nodal involvement. There were also stage IIE (9/71, 12%) and stage IV (6/71, 8%) patients with advanced stages. For all 71 patients, the 5-year progression-free survival (PFS) and overall survival (OS) estimates were 50 and 56%, respectively. There was no statistical difference in 5-year PFS and OS estimates between patients receiving Helicobacter pylori (H. pylori) containing eradication (HPE) (p=0.189) and those receiving non-HPE (p=0.359). Upon the Cox regression model, advanced stages were the only independent prognostic factors associated with shorter PFS, and m-IPI was independently associated with shorter PFS and OS. There was no specific clinical manifestation for patients with HT. HPE is thus a promising therapeutic approach for such patients. Moreover, advanced stages and m-IPI significantly influenced patient outcome.

Laparoscopic jejunoileal side-to-side anastomosis for the treatment of type 2 diabetes mellitus in Chinese patients with a body mass index of 24-32 kg/m2.

OBJECTIVE: Laparoscopic jejunoileal side-to-side anastomosis (LJISSA) is an upcoming procedure that offers good metabolic improvement without causing significant malabsorption. The objective of this study was to evaluate the results of this novel procedure for the control of type 2 diabetes mellitus (T2DM) in patients with a body mass index (BMI) of 24-32 kg/m 2. MATERIALS AND METHODS: Fifty-seven patients with T2DM who underwent LJISSA between February 2010 and May 2013 were recruited in this study. Data collected included fasting blood glucose (FBG), 2 h postprandial blood glucose (2 h PBG), 1 h postprandial C peptide (1 h C-P), and glycosylated hemoglobin (HbA1c). RESULTS: Postoperatively, glycemic parameters (FBG and 2 h PBG, HbA1c and 1 h C-P) improved in all 57 patients. At 12 months, 34 patients had a remission of diabetes, and the remaining 23 patients showed a significantly decreased requirement for oral hypoglycemic agents. The patients with a BMI of 28-32 kg/m 2 had significant weight loss of between 7.8% and 20% (P < 0.05), whereas weight loss was not significant in those with a BMI of 24-28 kg/m 2. The group achieving remission had a higher BMI (28-32 kg/m 2), shorter duration of diabetes (<10 years), and higher stimulated C-P (>4 ng/mL). These three factors may be the predictors of diabetes resolution at 12 months. CONCLUSION: LJISSA seems to be a promising procedure for the control of T2DM. A multicenter study with a larger number of patients and a longer follow-up period is needed to substantiate our preliminary findings.

[Feasibility of terminal intestinal exteriorization in laparoscopic anterior resection for anterior cancer].

OBJECTIVE: To discuss the feasibility of terminal intestinal exteriorization (exteriorization without ileostomy) in laparoscopic anterior resection for rectal cancer. METHODS: Clinicopathological data of 77 patients undergoing laparoscopic anterior resection for low rectal cancer in our department from January 2011 to December 2013 were retrospectively analyzed. After laparoscopic rectal resection, 32 patients received terminal intestinal exteriorization (exteriorization group) and 45 patients received preventive ileostomy (ileostomy group). Anastomosis-related, stoma-related and intestinal stoma closure-related morbidity was compared between the two groups. RESULTS: There were no significant differences in operative time, blood loss and overall hospital stay between the two groups (all P>0.05). The total hospital cost was (5.39±1.74)×10(4) yuan in the exteriorization group, and (6.98±1.37)×10(4) yuan in the ileostomy group(P<0.01). The incidences of postoperative anastomotic fistula was not significantly different between the two groups(P>0.05). Three patients(9.4%) developed anastomotic leak in the exteriorization group and 2(4.4%) in the ileostomy group. The anastomotic leak was managed by opening the external intestinal wall and maturating an ileostomy under local anaesthesia. All these 5 patients were cured with nutritional support, antibiotics, continuous local drainage. In the exteriorization group, 5 patients had complications related to stoma and intestinal stoma closure operation(15.6%), which was lower than(42.2%) in the ileostomy group(P=0.013). CONCLUSION: Terminal intestinal exteriorization in laparoscopic anterior resection is a safe and feasible surgical procedure with little trauma and less hospital cost, which can be an alternative as a prophylactic treatment for patients with high risk of anastomotic leak.

The clinical value of Fufangkushen injection in the treatment of stomach cancer: a meta-analysis.

OBJECTIVE: The purpose of this meta-analysis was to evaluate the effects of Fufangkushen injection combined with chemotherapy in the treatment of stomach cancer. MATERIALS AND METHODS: The relevant clinical trials about Fufangkushen injection combined with chemotherapy in the treatment of stomach cancer were search in the data bases of Pubmed, EMBASE, Cochran and CNKI. The data related to objective response rate, Karnofsky (KPS) score and toxicity were extracted and pooled using the Stata 11.0 software. Dichotomous data was presented as risk ratio (RR) and its 95% confidence interval (95% CI). RESULTS: Thirteen relevant trials were included in this meta-analysis. Heterogeneity test indicated there was no statistical heterogeneity among the studies, thus the fixed effects mode was used to calculat the results. Pooled results indicated that the objective response rate (ORR) and KPS score improvement in Fufangkushen chemotherapy group was significant higher than that of control group (RR = 1.24, P < 0.05). Synthesis data also demonstrated the Fufangkushen injection can significantly decrease the risk of developing granulocytopenia in stomach cancer patients treated with chemotherapy (RR = 0.67,P < 0.05). CONCLUSION: Fufangkushen injection combined with chemotherapy can increase the objective response rate, improve the quality of life and decrease the risk of developing granulocytopenia in patients with stomach cancer.

Treatment of aganglionic megacolon mice via neural stem cell transplantation.

To explore a potential methodology for treating aganglionic megacolon, neural stem cells (NSCs) expressing engineered endothelin receptor type B (EDNRB) and glial cell-derived neurotrophic factor (GDNF) genes were transplanted into the aganglionic megacolon mice. After transplantation, the regeneration of neurons in the colon tissue was observed, and expression levels of differentiation-related genes were determined. Primary culture of NSCs was obtained from the cortex of postnatal mouse brain and infected with recombinant adenovirus expressing EDNRB and GDNF genes. The mouse model of aganglionic megacolon was developed by treating the colon tissue with 0.5 % benzalkonium chloride (BAC) to selectively remove the myenteric nerve plexus that resembles the pathological changes in the human congenital megacolon. The NSCs stably expressing the EDNRB and GDNF genes were transplanted into the benzalkonium chloride-induced mouse aganglionic colon. Survival and differentiation of the implanted stem cells were assessed after transplantation. Results showed that the EDNRB and GDNF genes were able to be expressed in primary culture of NSCs by adenovirus infection. One week after implantation, grafted NSCs survived and differentiated into neurons. Compared to the controls, elevated expression of EDNRB and GDNF was determined in BAC-induced aganglionic megacolon mice with partially improved intestinal function. Those founding indicated that the genes transfected into NSCs were expressed in vivo after transplantation. Also, this study provided favorable support for the therapeutic potential of multiple gene-modified NSC transplantation to treat Hirschsprung's disease, a congenital disorder of the colon in which ganglion cells are absent.