Objective To investigate the impacts of usnea acid(UA)on the proliferation of human ovarian cancer(OC)cells.Methods Ovarian adenocarcinoma cells SKOV3 were randomly divided into control group,L-UA group,M-UA group,H-UA group(with 10,20,and 50 μmol/L UA added respectively),pcDNA3.1-NC group and pcDNA3.1-PD-1 group.RT-qPCR method was applied to detect the expression of programmed cell death 1(PD-1)and programmed cell death ligand 1(PD-L1)in SKOV3 cells.CCK-8 assay and plate method were applied to detect the effect of UA on SKOV3 cell proliferation.The effect of UA on SKOV3 cells apoptosis was examined by flow cytometry.Mouse ovarian epithelial cancer cell line ID8 cells were collected to construct an OC mouse model,and the mass and volume of OC tumors were recorded.Immuno-histochemical microscopy was applied to detect the infiltration of PD-1,PD-L1,and CD8+T cells.Results The PD-1 mRNA,PD-L1 mRNA,colony count,A450 val-ue,PCNA protein,PD-1 protein,and PD-L1 protein in the L-UA,M-UA,and H-UA groups were lower than those in the control group,the apoptosis rate and Bax protein were higher in the control group(P<0.05).Com-pared with the H-UA group and the pcDNA3.1-NC group,the PD-1 mRNA,PD-L1 mRNA,colony count,A450 value,PCNA protein,PD-1 protein,and PD-L1 protein in the pcDNA-3.1-PD-1 group increased and the apoptosis rate and Bax protein decreased(P<0.05).The quality,volume,positivity rate of PD-1and PD-L1 of OC tumors in the UA group was lower than that in the control group,the counting number of CD8+T cell infiltration was higher than control group(P<0.05).Conclusions UA may inhibit progression of OC cell line SKOV3 by regula-ting cell proliferation,apoptosis,and immune escape.

Objective:To investigate the influence of trimethylamine N-oxide(TMAO)on the development of early neurological deterioration(END)in diabetic patients with acute ischemic stroke.Methods:In this cross-sectional study, 108 type 2 diabetes patients with acute ischemic stroke treated at the Department of Neurology in the Affiliated Wuxi People’s Hospital of Nanjing Medical University between October 2019 and November 2020 were consecutively recruited.END was defined as an increase in the National Institutes of Health Stroke Scale(NIHSS)≥ 2 points and exclusion of intracranial hemorrhage or bleeding transformation in cranial imaging evaluation within 5 days of initial deterioration of neurological dysfunction.The patients were divided into 2 groups, an END(n=36)group and a non-END group(n=72). Fasting plasma TMAO was measured using isotope dilution liquid chromatography coupled to tandem mass spectrometry.Results:Of the 108 patients, 36(33.3%)were diagnosed with END, and their plasma TMAO levels were significantly higher compared with patients without END( Z=-3.500, P<0.001). For prediction of END, the area under the ROC curve for plasma TMAO levels was 0.707(95% CI: 0.603-0.811, P<0.001). The frequencies of END in subjects grouped via tertiles of TMAO were 22.2%, 19.4% and 58.3%, respectively, with significant differences between the 3 groups( χ2=14.979, P=0.001). Univariate analysis showed that elevated TMAO( OR=1.160, 95% CI: 1.050-1.282, P=0.004)was associated with END.A multivariate logistic regression model further confirmed the association between TMAO and END( OR=1.145, 95% CI: 1.033-1.269, P=0.010). Conclusions:Increased plasma TMAO levels are associated with END in diabetic patients with acute ischemic stroke.

Clavicular crook plate is designed based on anatomic features of acromioclavicular joint,which matches clavicle appearance,and well attaches the shaft.Not passing through the joint,the internal fixation exhibits many advantages,such as convenient usage,reliable fixation and micromovement of fixed acromioclavicular joint.So the patients could move the joint early and avoid osteoarthritis.Allogenous tendon has abundant sources,simple sampling process,and short repairing duration and can reduce injury to patients.Cryopreserved allogenous tendon has low antigenicity and immunological refection,good histocompatibility.Reconstructed coracoclavicular ligament by allogenous tendon has good material intensity,and can meet the requirements of local stress,better substitute integrity and function of coracoclavicular ligament.Moreover,it has precise effect and simple operation.It acts as long-term fixator following plate removal,thus is important supplement of clavicular crook plate fixation.Clavicular crook plate fixation in combination with allogenous tendon transplantation reconstructs acromioclavicular joint stability,effectively provides early mechanical stability and subsequent biological stability,and maximizes restoration of acromioclavicular joint structure and function.