G protein-coupled receptor kinase 2 (GRK2) participates in the phosphorylation and desensitization of G protein-coupled receptor (GPCR), impacting various biological processes such as inflammation and cell proliferation. Dysregulated expression and activity of GRK2 have been reported in multiple cells in rheumatoid arthritis (RA). However, whether and how GRK2 regulates synovial hyperplasia and fibroblast-like synoviocytes (FLSs) proliferation is poorly understood. In this study, we investigated the regulation of GRK2 and its biological function in RA. We found that GRK2 transmembrane activity was increased in FLSs of RA patients and collagen-induced arthritis (CIA) rats. Additionally, we noted a positive correlation between high GRK2 expression on the cell membrane and serological markers associated with RA and CIA. Immunoprecipitation-mass spectrometry and pull-down analyses revealed tumor necrosis factor receptor-associated factor 2 (TRAF2) as a novel substrate of GRK2. Furthermore, surface plasmon resonance (SPR) and molecular docking assays determined that the C-terminus of GRK2 binds to the C-terminus of TRAF2 at the Gln340 residue. GRK2 knockdown and the GRK2 inhibitor CP-25 attenuated synovial hyperplasia and FLS proliferation in CIA both in vitro and in vivo by decreasing GRK2 membrane expression and activity. Mechanistically, increased GRK2 transmembrane activity contributed to the recruitment of TRAF2 on the cell membrane, promoting GRK2-TRAF2 interactions that facilitate the recruitment of the E3 ubiquitin ligase TRIM47 to TRAF2. This enhanced TRAF2 Lys63 polyubiquitylation and induced nuclear factor (NF)-κB activation, leading to synovial hyperplasia and abnormal proliferation of FLSs. Our study provides a mechanistic and preclinical rationale for further evaluation of GRK2 as a therapeutic target for RA.

Objective:To investigate the association between plasma anti-Müllerian hormone(AMH) levels and ischemic stroke.Methods:In this case-control study, 93 ischemic stroke patients were randomly selected as the case group from a study on the prevention and treatment of metabolic syndrome, which was conducted in 2018-2019 in Changshu, Jiangsu Province, while 372 nonischemic stroke patients were selected as the control group according to the principle of 1∶4 matching.An enzyme-linked immunosorbent assay was used to measure plasma AMH levels.The conditional logistic regression model and restricted cubic spline were used to analyze the relationship between AMH levels and ischemic stroke.Results:A total of 465 subjects with an average age of (68.7±7.4)years were included in this study, of whom 215(46.2%)were men and 250(53.8%)were women.According to our conditional Logistic regression analysis, the risk of ischemic stroke was reduced by 44% for every unit increase in the log-AMH level( OR=0.56, 95% CI: 0.37-0.85)in the overall population after multivariate adjustment.Compared with the tertile with the lowest AMH level, the risk of ischemic stroke in the tertile with the highest AMH level decreased significantly( OR=0.37, 95% CI: 0.19-0.69). When subgrouped by sex, the tertiles with the highest AMH levels were associated with a 66% lower risk of ischemic stroke in men( OR=0.34, 95% CI: 0.13-0.88)and a 64% lower risk of ischemic stroke in women( OR=0.36, 95% CI: 0.15-0.87), compared with the tertiles with the lowest AMH levels.The results of restricted cubic spline analysis showed that there was a linear dose-response relationship between plasma AMH levels and ischemic stroke both in the general population and in male or female population( Pvalues for linear trends were 0.0002, 0.008 and 0.007, respectively). Conclusions:Higher plasma AMH levels decrease the risk of ischemic stroke with a dose-response pattern.

With the rapid development of nanotechnology, the research and development of nanomedicine has become one of the current development directions of drug innovation. The pharmacokinetic characteristics of nanomedicine are significantly different from general drugs because of the scale effect based on nanostructures, and pharmacokinetics studies of nanomedicine may be different from the general drugs. This article focuses on the research strategies and considerations on non-clinical pharmacokinetics of nanomedicine, including test agents, in vivo/in vitro assays, biological sample analysis, data evaluation and analysis etc., providing references for developers.

Tree shrew is a novel and high-quality experimental animal model. In this study, the real-time polymerase chain reaction methods were established to detect infection-related cytokines interleukin-6 (IL-6), IL-8, IL-10, IL-17A, interferon-γ (IFN-γ) and housekeeping gene glyceraldehyde-phosphate dehydrogenase ( ) of tree shrew. The results indicated that the establised methods had good specificity. The high point of the linear range of these reagents reached 1 × 10 copies, and the low points ranged from 10 copies (IL-6, IL-17A), 100 copies (IL-10, ) to 1 000 copies (IL-8, IFN-γ). In this interval, the linear correlation coefficient of each reagent was greater than 0.99. The lowest detectable values of IL-6, IL-8, IL-10, IL-17A, IFN-γ and were 8, 8, 4, 8, 128 and 4 copies, respectively. The results showed that the established detection methods had good specificity, sensitivity and wide linear range. The methods were suitable for detection of multiple concentration range samples, and could be used for the subsequent studies of tree shrew cytokines.
Animals ; Cytokines ; analysis ; Real-Time Polymerase Chain Reaction ; Shrews

Objective To investigate the association between ten single nucleotide polymorphisms (SNPs) in the peroxisome proliferator-activated receptors and pulse pressure (PP) as well as the relationships between gene-gene interaction between PPARα/δ/γ genes and PP.Methods A total of 820 subjects,with 550 females and 270 males,were recruited from a cohort study of “Prevention of Metabolic Syndrome and Multi-metabolic Disorders in Jiangsu Province of China Study (PMMJS)”.Ten SNPs of PPARα/δ/γ genes were selected.GMDR software (version 1.0.1) was used to evaluate the gene-gene interactions among PPARs SNPs associated with PP.Results The mean levels of PP in people with mutant genotype of rs1805192 in PPARγ genes (PA+AA) showed a significant increase by 1.341 mmHg (95％CI:0.431-2.252 mmHg) when compared to the persons with wild genotype (PP).In the subgroup of subjects with more than 30 mmHg levels of PP,a six-locus model comprised rs135539 of PPARα,rs2016520 of PPARδ,rs10865710,rs1805192,rs709158 and rs3856806 of PPARγshowed a highest level of prediction accuracy (0.577) and displayed a better cross-validation consistency (10/10).In the subgroup of subjects with less than 40 mmHg levels of PP,a two-locus model was statistically associated with PP with 0.628 of prediction accuracy and 10/10 of cross-validation consistency.Conclusion PPARγrs1805192 was associated with the occurrence of PP.Gene-gene interactions among rs135539 of PPARα,rs2016520 of PPARδ,rs10865710,rs1805192,rs709158 and rs3856806 of PPARγ were all significantly related to PP.

OBJECTIVETo investigate the therapeutical effects of visfatin and metformin on insulin resistance and reproductive endocrine disorder in rats with polycystic ovary syndrome (PCOS).

METHODSForty female Wistar rats were divided into 4 equal groups, and in groups A, B and C, the rats were injected subcutaneously with dehydroepiandrosterone (DHEA) for PCOS modeling, with group D as the blank control injected with soybean oil. Vaginal smears and serological testing were taken to assess the modeling. After the modeling, the rats in group A received 10 µg reorganized visfatin injection and those in group B were treated with metformin (14 mg/100 g) on a daily basis for 15 days. Serum levels of T, LH, FSH, FINS and blood glucose levels during OGTT were measured before and after the treatments, and HOMA-IR and LH to FSH ratio were calculated. The ovaries were then dissected for pathological examination.

RESULTSIn groups A and B, FINS, FPG, T, HOMA-IR and blood glucose levels during OGTT were significantly decreased after the treatments (P<0.05), which resulted in recovery of regular menses in 8 (80%) rats in group A and 7 (77.8%) rats in group B with the development of normal follicles. Visfatin and metformin produced equivalent therapeutic effects in improving the insulin resistance and hyperandrogenism in PCOS rats.

CONCLUSIONVisfatin and metformin have equivalent therapeutic effects in improving insulin resistance and hyperandrogenism and in promoting the recovery of regular menses and development of normal follicles in PCOS rats.

Animals ; Female ; Humans ; Insulin Resistance ; Metformin ; pharmacology ; Nicotinamide Phosphoribosyltransferase ; pharmacology ; Polycystic Ovary Syndrome ; complications ; drug therapy ; Rats ; Rats, Wistar

Objective To explore the correlation between the amount of residual undifferentiated embryonic stem cells (ESCs) in embryoid bodies and its tumorigenicity.Methods Mouse R1 ESCs were cultured in suspension to form embryoid bodies (EBs).Ten days later,EBs were digested into single cells and then re-plated in standard ESCs culture condition.The residual undifferentiated embryonic stem cells surface maker SSEA-1 was examined by flow cytometry in EBs.The morphology of residual undifferentiated cells in EBs were observed,meanwhile the surface marker SSEA-1 was examined by immunofluorescent staining.EBs were digested into single cells and grouped into 104,105,106,2×106,and then injected into limb muscle of nude mice.The correlation of the amount of cells and its tumorigenicity was observed.Results Residual undifferentiated ESCs were observed after EBs differentiated for 10 days,which displayed clonal morphology and expressed undifferentiated ceil markers of ESCs,such as SSEA-1.The expression rate of undifferentiated cells surface marker SSEA-1 was (13.5±0.75)％ in EBs differentiated for 10 days.Only two millions single cells harvested from EBs were able to form teratoma after being injected into muscle of nude mice for 6 weeks.Mature endoderm,mesoderm and ectoderm tissues could be found in teratoma.No teratoma formed in other groups.Conclusion A certain amount residual undifferentiated ESCs still exist after differentiation of ESCs into EBs.About 2.7× 105 undifferentiated cells are able to form teratoma by iniecting into muscle of nude mice.

BACKGROUND AND OBJECTIVEThe relationship between myeloperoxidase G-463A genetic polymorphisms and lung cancer susceptibility has been studied extensively. However, the outcomes are not consistent. The aim of this study is to evaluate the relationship between myeloperoxidase genetic polymorphisms and lung cancer susceptibility by meta analysis.

METHODSDocuments published were retrieved through databases associated with the study. Taking into account the possibilities of heterogeneity of the studies, a statistical test for heterngeneity was performed. The odds ratio and 95% CI were used to evaluate the risks. The meta analysis was applied with RevMan software 4.2, and the forest plot and funnel plot of meta analysis were worked out.

RESULTSA total of 5 381 cases and 5 827 controls from studies for Caucasian and a total of 1 558 cases and 1 755 controls from studies for East Asians were included. For Caucasian the pooled OR was 0.91 (95% CI: 0.81-1.02); For East Asians, the pooled OR is 0.83 (95% CI: 0.63-1.09). Publication bias exits in the study for Caucasian, but not for East Asians.

CONCLUSIONThe results of this study indicated that the polymorphism of myeloperoxidase G-463A was not significantly associated with the lung cancer risk for Caucasian or East Asians. However, further studies for the East Asians is needed for the few subjects.

Genetic Predisposition to Disease ; genetics ; Humans ; Lung Neoplasms ; epidemiology ; genetics ; Peroxidase ; genetics ; Polymorphism, Genetic ; genetics

OBJECTIVETo investigate the feasibility of using marrow stromal osteoblast-cancellous bone matrix compound artificial bone (MCCAB) as tissue-engineered bone, the osteogenesis of MCCAB in the cranial defect was observed in the experiment.

METHODSThe in vitro cultivated and induced marrow stromal cells of adult New Zealand rabbits were seeded into the alginate-cancellous bone matrix to form MCCAB. The MCCAB was then implanted into the cranial defect for 4 to 8 weeks. The cancellous bone matrix (CBM) alone or the marrow stromal osteoblasts (MSOs) alone was implanted as the control. The effectiveness of bone formation was assessed by histological and roentgenographic analysis.

RESULTSThe osteogenesis of MCCAB was better than CBM or MSOs and superior to the blank group.

CONCLUSIONMCCAB can effectively repair cranial defect. It could be used clinically to restore large bone defects.

Animals ; Bone Marrow Cells ; cytology ; physiology ; Bone Matrix ; cytology ; Cells, Cultured ; Feasibility Studies ; Male ; Osteoblasts ; cytology ; physiology ; Rabbits ; Skull ; abnormalities ; Stromal Cells ; cytology ; physiology

OBJECTIVETo investigate the feasibility of using marrow stromal osteoblast (MSO) as bone derived cell and using cancellous bone matrix (CBM) as scaffold for bone tissue engineering, the subcutaneous osteogenesis of MSO-CBM compound artificial bone (MCCAB) was observed in the experiment.

METHODSThe marrow stromal cells of adult New Zealand rabbits cultivated and induced in vitro were used to form MCCAB by mixing, seeding and solidifying methods assisted by alginate. The MCCABs were auto-transplanted subcutaneously into the rabbits for 4 to 8 weeks. The alginate-cancellous bone matrix composites or the cancellous bone matrix alone were implanted as control. The effectiveness of bone formation was assessed by means of roentgenography, histology and computerized histomorphometry.

RESULTSThe osteogenesis of MCCABs was better than that of the alginate-cancellous bone matrix composites and of the cancellous bone matrixes. In the MCCABs, both intramembranous and cartilaginous osteogeneses were seen but the former was obvious. In the control, only slight cartilaginous osteogeneses were seen.

CONCLUSIONSThe osteogeneses of the MCCABs constructed by using tissue engineering method were obvious when transplanted subcutaneously. The MSO and CBM can be used as good bone-derived cell and scaffold material respectively for tissue-engineered bone construction.

Animals ; Bone Marrow Transplantation ; Bone Matrix ; transplantation ; Bone Transplantation ; methods ; Male ; Osteoblasts ; transplantation ; Osteogenesis ; Rabbits ; Tissue Engineering