Dry eye disease (DED) is a prevalent and intractable ocular disease induced by a variety of causes. Elevated sphingomyelin (SM) levels and pro-inflammatory cytokines were detected on the ocular surface of DED patients, particularly in the meibomian glands. Sphingomyelin synthase 2 (SMS2), one of the proteins involved in SM synthesis, would light a novel way of developing a DED therapy strategy. Herein, we report the design and optimization of a series of novel thiophene carboxamide derivatives to afford 14l with an improved highly potent inhibitory activity on SM synthesis (IC_{50, SMS2} = 28 nmol/L). Moreover, 14l exhibited a notable protective effect of anti-inflammation and anti-apoptosis on human corneal epithelial cells (HCEC) under TNF-α-hyperosmotic stress conditions in vitro, with an acceptable ocular specific distribution (corneas and meibomian glands) and pharmacokinetics (PK) profiles (t _{1/2, cornea} = 1.11 h; t _{1/2, meibomian glands} = 4.32 h) in rats. Furthermore, 14l alleviated the dry eye symptoms including corneal fluorescein staining scores and tear secretion in a dose-dependent manner in mice. Mechanically, 14l reduced the mRNA expression of Tnf-α, Il-1β and Mmp-9 in corneas, as well as the proportion of very long chain SM in meibomian glands. Our findings provide a new strategy for DED therapy based on selective SMS2 inhibitors.

Objective:To investigate the influence of frailty on post-treatment outcomes in elderly heart failure patients with reduced ejection fraction treated with Sacubitril/Valsartan.Methods:The 231 heart failure patients aged 60 years or over with reduced ejection fraction were enrolled from October 2017 to October 2018 in Department of Geriatric Medicine, Henan Provincial People's Hospital.Patients were divided into the frailty group(n=116)and the control group(n=115). Frailty diagnosis was made by five indexes suggested by LP Fried.Both groups were treated with sacubitril/valsartan(49/51 mg)for 1 year.The left ventricular ejection(LVEF), estimated glomerular filtration rate(eGFR), N-terminal pro B-type natriuretic peptide(NT-proBNP)and other clinical and laboratory indexes were detected before and after treatment and compared between the frailty group and the control group.Results:16 subjects in the frailty group and 11 subjects in the control group dropped out of the study.The frailty group versus the control group showed a higher mortality rate of cardiovascular causes(13.0% or 13/100 vs.6.7% or 7/104, χ2=6.437, P=0.027), a higher first re-hospitalization rate(18.0% or 18/100 vs.11.5% or 12/104, χ2=4.458, P=0.043)and a higher all-cause mortality(16.0% or 16/100 vs.8.6% or 9/104, χ2=3.875, P=0.039). In the frailty group, levels of serum NT-proBNP and creatinine were higher and eGFR was lower after treatment than before treatment[(2 253±144) ng/L vs.(2 094±136) ng/L, (137±24) μmol/L vs.(125±23) μmol/L, (49.2±5.9) ml·min -1·1.73 m -2vs.(56.7±6.3) ml·min -1·1.73 m -2, t=3.674, 2.893 and 2.068, P=0.017, 0.026 and 0.029]. In the control group, serum NT-proBNP levels were lower after treatment than before treatment[(1 828±123) ng/L vs.(1 945±128) ng/L, t=1.896, P=0.043], while serum creatinine levels[(120±22) μmol/L vs.(117±19) μmol/L, t=2.099, P=0.650]and eGFR[(59.8±6.5) ml·min -1·1.73 m -2vs.(61.6±6.8) ml·min -1·1.73 m -2, t=2.444, P=0.173]had no significant difference between post-treatment and pre-treatment. Conclusions:Frailty has adverse affects on the mortality, re-hospitalization rate and renal function in elderly heart failure patients with reduced ejection fraction treated with Sacubitril/Valsartan.

Objective To investigate the thawing modes on stability of coagulation control products after frozen,looking for a new theoretical basis for cost control and the quality and safety of laboratory.Methods Using ACL TOP 700 automated co-agulation analyzer and supporting the same batch of reagents and quality control materials conduct of the study:after daily QC,recycled the remaining control materials immediately and dispensed into two EP tubes and frozen at-40℃,respectively thawed by room-temperature and 37℃water bath after 24 hours,and examined together with the date of quality control ma-terial,got 20 pairs of data for analysis the financial impact of two alternate ways on coagulation QC parameters.Results For the room-temperature thawing group,FIB high value increasedby an average 0.23 g/L (t=4.026 9,P<0.05);TT normal value average reduction of 0.46 s (t=-3.813 8,P<0.05),TT high value reduced by an average 0.41s (t=-3.972 8,P<0.05);D-Dimer low-value increased by an average of 14.75 ng/ml FEU (t=2.281 6,P<0.05),while APTT,PT normal and high value,FIB normal value,D-Dimer high value were no significant difference after thawing (P>0.05).For the 37℃water bath group,both normal and high value of APTT,PT,FIB,TT and D-Dimer were no significant difference after tha-wing (P>0.05).Conclusion The commercialization of coagulation control materials can be for the second QC,just follow the principle of rapid after melting and timely detection,other laboratories can be used as a reference.

Objective To investigate the diagnostic value of D-dimer age-adjusted threshold in elderly patients with Pulmonary thromboembolism(PTE).Methods Collected patients older than 50 years of suspected PTE,who visited Yan′an People′s Hospital and the Hospital Affiliated to Yan′an University from June 2015 to September 2016,using the revised Geneva criteria for clinical probability assessment firstly.The low-risk group was excluded from the study;the patients with moderate-to-high risk were performed D-dimer detection and CTPA.All patients determined both by D-dimer age-adjusted thresholds and traditional thresholds,comparing the diagnostic efficacy of the two methods subsequently.Results 163 patients were diagnosed with PTE by CTPA among the 549 subjects,the positive rate was 29.69％.The diagnostic sensitivity of plasma D-dimer was 83.44％,78.53％,the specificity was 17.88％,31.61％ respectively,the missed diagnosis rate was 16.56％,21.47％,respectively.The misdiagnosis rates were 82.12％ and 68.39％,respectively,and the Younden index was 0.013 2 and 0.101 4 respectively.All subjects were divided into four groups according to age:50-60 years,60-70 years,70-80 years,and equal or greater than 80 years age group.Compared with the traditional threshold,the misdiagnosis rates of the four groups of age-adjusted thresholds decreased by 4.00％,15.70％,21.36％ and 17.39％,respectively and the specificity was increased by 1.23,1.65,2.56 and 3.00 times,respectively.Conclusion The diagnostic accuracy of the age-corrected threshold is higher than the traditional threshold;combined with clinical practice,the optimal threshold is determined by the ROC curve,the clinician can serve as a reference.

Objective To study the expression of microRNA-301 in pancreatic carcinoma andvalidate the significance of miR-301 in invasion and metastasis of pancreatic carcinoma.Methods miR-301 expression were detected by FQ-PCR in 5 pancreatic cancer eell lines(PANC-1,PaCa-2,AsPC-1,Hs766T.BxPC-3).Further immunohistochemistry in pancreatic cancer tissue microarrays was detected miR-301 expression,which contained 60 pancreatic cancer specimens along with 10 normal adjacent tissues and 10 normal pancreas tissues.After high expression of miR-301 in pancreatic carcinoma being confirmed.the clinical significance of high expression of miR-301 in invasion and metastasis of pancreatic carcinoma were studed.Pancreatic cancer cell lines(PANC-1.PaCa-2)were transfected by 100 nmoml/L miR-301 inhibitor(anti-miR-301)or negative eontrol(Anti-miR~(TM) Negative Control#1).COX-2 and MMP-2 protein expression in pancreatic cancer cell lines were detected by WB.and cell migration assays were performed using transwell technology.Results FQ-PCR resuhs indicated that miR-301 expression was higher in pancreatic cancer cell lines than normal pancreatic cells.The relative level of miR-301 in 5 pancreatic cancer cell lines(PANC-1,PaCa-2,AsPC-1,Hs-766T,BxPC-3)and normal pancreatic cell were 33.09± 4.21,30.76±3.18,47.57±3.56,20.20 ±1.21,76.75±13.51 and 1.00±0.08 respectively.The miR-301 level in all 5 pancreatic cancer cells were significantly higher than those of normal pancreatic cell(t=8.86,9.53,6.39,6.77,11.18,P<0.01).Immunohistochemistry results also showed miR-301 expression was higher in pancreatic carcinoma tissues than those in the cancer adjacent tissues and normal pancreatic tissues.The relative levels of miR-301 in pancreatic carcinoma tissues.normal adjacent tissues and normal pancreas tissues were 0.88±0.09,0.22±0.04 and 0.14±0.05 respectively.The miR-301 levels in pancreatic carcinoma tissues were significantly higher than those of normal adjacent tissues and normal pancreatic tissues(t=15.1,10.6,P<0.01).There was no significant difference between normal adjacent tissues and normal pancreas tissues(t=1.32,P=0.22).After miR-301 inhibitor was introduced into pancreatic cancer cells PANC-1 and PaCa-2.miR-301 levels were reduced while the protein levels of COX-2 and MMP-2.which were invasion and metastasis related factors,were down-regulated.The cell migration assay indicated the numbers of PANC-1 and PaCa-2 cells,which migrated to lower chamber.were 587±27 and 363±13 respectively after miR-301 inhibitor was applied.The numbers of migrated cells were 1091 4-15.737±44 when the netative control was applied.The cell invasion ability was decreased significantly in the inhibitor group compared with the negative group(t=7.89,7.56,P<0.01).Conclusions miR-301 is highly expressed in pancreatic cancer cell lines and pancreatic cancer tissues.Inhibition of miR-301 expression can effectively supress the invasion of pancreatic cancer cells.miR-301 may serve as a new biomarker for early detection of pancreatic cancer and molecular target for early treatment of pancreatic cancer.

Osteoarthritis is mainly caused by the degenerative changes of cartilage and cartilage extracellular matrix,while Aggrecanases degradate Proteoglycans which are the major components of cartilage.This review includes three aspects:(1) We have concluded the major enzymes(ADAMTS-4 and ADAMTS-5) which regulate the metabolism of cartilage extracellular matrix.Meanwhile,we have summarized the structure of aggrecanases(ADAMTS-4 and ADAMTS-5) and introduced the function of each regional structure;(2) We have concluded the way cytokines and glycosaminoglycans regulate the metabolism of aggrecanases,and discussed the regulation and control principle of cytokines and glycosaminoglycan;(3) We have summarized the majority of inhibitors to the aggrecanases,introduced the endogenic inhibitors,and put our emphasis on the extrinsic inhibitors(chelating agents,polypeptides and so on).Through deeper research on the enzymes,it will help us further understand the pathogenesis of osteoarthritis,and open up new avenues to clinical treatment.Abstract:SUMM ARY Osteoarthritis is mainly caused by the degenerative changes of cartilage and cartilage extra-cellular matrix,while Aggrecanases degradate Proteoglycanswhich are the major components of cartilage.This review includes three aspects:(1) W e have concluded the major enzymes(ADAMTS-4 and ADAMTS-5) which regulate the metabolism of cartilage extracellular matrix.Meanwhile,we have sum-marized the structure of aggrecanases(ADAMTS-4 and ADAMTS-5) and introduced the function of each regional structure;(2) W e have concluded the way cytokines and glycosam inoglycans regulate the metab-olism of aggrecanases,and d iscussed the regulation and control principle of cytokines and glycosam inogly-can;(3) W e have summarized the majority of inhibitors to the aggrecanases,introduced the endogenic inhibitors,and put our emphasis on the extrinsic inhibitors(chelating agents,polypeptides and so on).Through deeper research on the enzymes,it will help us further understand the pathogenesis of osteoar-thritis,and open up new avenues to clinical treatment.