The Correlation Between Decreased Ornithine Level and Alleviation of Rheumatoid Arthritis Patients Assessed by a Randomized,Placebo-Controlled,Double-Blind Clinical Trial of Sinomenine

Sinomenine(SIN)is commonly used as part of rheumatoid arthritis(RA)therapy in China,but there is still no published evidence of the efficacy of SIN monotherapy.This work investigates the efficacy and safety of SIN in treating RA patients and analyzes the correlation between ornithine level and the alleviation of disease activity in RA patients.In this 24 week,randomized,placebo-controlled,double-blind clinical trial,people with mild to moderate RA were randomly assigned(1:1:1,stratified by hospital)to receive SIN(120 mg,twice daily),methotrexate(MTX)(10 mg per week),or SIN+MTX therapy.The primary outcome was the proportion of patients who achieved a 50%improvement in the American College of Rheumatology(ACR50)criteria at week 24 and who showed improvement according to the clinical disease activity index(CDAI).In this prospective subgroup analysis,we also assessed whether the 24-week alterations of disease activity in the treatment group were significantly correlated to the levels of blood ornithine.Of the 135 enrolled participants,38,39,and 36 patients were treated with SIN,MTX,and SIN+MTX,respectively.In the SIN-treated group,52.63%of patients achieved ACR50 after 24-weeks of treatment,which was comparable to the results in the MTX-treated and SIN+MTX-treated groups.Hepatic and gastrointestinal disorders were the main adverse events;however,the ratio of patients suffering from hepatic disorder in the SIN group(1/38)was much lower than that in the MTX(10/39)and SIN+MTX(8/36)groups.A total of 221 serum samples were collected at the four follow-up time points in the three treatments,and the levels of ornithine,citrulline,and arginine were obtained through ultrahigh performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF/MS).The serum ornithine level decreased after the 24-week treatment along with a decrease in disease activity,and may reflect therapeutic responses with a sensitivity value of 80%.In conclusion,SIN revealed a comparable efficacy to MTX for treating RA patients,but with fewer side effects.In addition,the serum ornithine level was found for the first time to have a close correlation with the alleviation of RA,which shows the value of this measure as an assessment indicator of drugs in treating RA.

Chordin-like 2 influences the differentiation fate of retinal pigment epithelium cells by dynamically regulating BMP pathway

AIM:To explore the functions of Chordin-like 2,which is encoded by CHRDL2,in the process of retinal pigmented epithelium(RPE)differentiation and damage repair.METHODS:The fetal RPE cells(f RPE)was obtained from aborted fetus which obeyed medical ethics.Real-time quantitative polymerase chain reaction was used to measure expression quantity of CHRDL2 and other functional genes expression.Knocking down and overexpression was used to analyze the functions about Chordin-like 2.Enzyme-linked immunosorbent assay(ELISA)was used to detect the secretion of bone morphogenetic proteins 4(BMP4).Flow cytometry was used to analyze cell cycle.Cell morphology was observed by phase contrast microscope(PCM).RESULTS:In normal RPE cells,CHRDL2 was firstly upregulated and followed a downregulation but eventually,it was expressed higher than the cells which undergone epithelial-mesenchymal transition(EMT).After knocking down CHRDL2,the secretion of BMP4 was decreased,RPErelated genes(OTX2,MITF,RPE65)were downregulated while EMT-related genes(SNAI1,VIM)were upregulated.However,the expression of these related genes after overexpression of CHRDL2 had contrary results.Chordin-like 2 also regulated the cell cycle by regulating BMP pathway.When CHRDL2 was knocked down,more f RPE cells stayed in S phase of cell cycle,while adding BMP4 reduced the proportion of the cells in S phase.However,overexpression of CHRDL2 increased more BMP4 secretion,this effect decreased the number of cells in S phase,but exogenous BMP inhibitor also could change this effect.At last,in the process of RPE cells differentiation,adding BMP4 at early stage could intervene normal RPE differentiation.Compared with BMP4,inhibiting BMP pathway had no significant negative effect at early stage,but suppressed differentiation at late stage.CONCLUSION:BMP pathway can be activated in a correct temporal order,otherwise,the cells have incorrect differentiation orientation.And Chordin-like 2 plays a role in dynamic regulation of BMP pathway and it also regulates the differentiation of RPE cells.Therefore,this research enlightens a new direction to inhibit EMT and promote cell redifferentiation after injury.

Genetic profiles of familial late-onset Alzheimer’s disease in China:The Shanghai FLOAD study

Compared with early-onset familial AD(FAD),the heritability of most familial lateonset Alzheimer’s disease(FLOAD)cases still remains unclear.However,there are few reported genetic profiles of FLOAD to date.In the present study,targeted sequencing of selected candidate genes was conducted for each of 90 probands with FLOAD and 101 unrelated matched normal controls among Chinese Han population.Results show a significantly lower rate of mutation in APP and PSENs,and APOE e4 genetic risk is higher for FLOAD.Among the Chinese FLOAD population,the most frequent variant was CR1 rs116806486[5.6%,95%CI(1.8%,12.5%)],followed by coding variants of TREM2(4.4%,95%CI(1.2%,10.9%))and novel mutations of ACE[3.3%,95%CI(0.7%,9.4%)].Next,we found that novel pathogenic mutations in ACE including frame-shift and nonsense mutations were in association with FLOAD regardless of APOE e4 status.Evidence from the Alzheimer’s disease Neuroimaging Initiative(ADNI)database also supported this finding in different ethnicities.Results of in vitro analysis suggest that frame-shift and nonsense mutations in ACE may be involved in LOAD through decreased ACE protein levels without affecting direct processing of APP.