Rosuvastatin(RVS)is an excellent drug with anti-inflammatory and lipid-lowering properties in the aca-demic and medical fields.However,this drug faces a series of challenges when used to treat atherosclerosis caused by hyperhomocysteinemia(HHcy),including high oral dosage,poor targeting,and long-term toxic side effects.In this study,we applied nanotechnology to construct a biomimetic nano-delivery system,macrophage membrane(M?m)-coated RVS-loaded Prussian blue(PB)nanoparticles(MPR NPs),for improving the bioavailability and targeting capacity of RVS,specifically to the plaque lesions associated with HHcy-induced atherosclerosis.In vitro assays demonstrated that MPR NPs effectively inhibited the Toll-like receptor 4(TLR4)/hypoxia-inducible factor-1α(HIF-1α)/nucleotide-binding and oligomerization domain(NOD)-like receptor thermal protein domain associated protein 3(NLRP3)signaling pathways,reducing pyroptosis and inflammatory response in macrophages.Additionally,MPR NPs reversed the abnormal distribution of adenosine triphosphate(ATP)-binding cassette transporter A1(ABCA1)/ATP binding cassette transporter G1(ABCA1)/ATP binding cassette transporter G1(ABCG1)caused by HIF-1α,promoting cholesterol efflux and reducing lipid deposition.In vivo studies using apolipoprotein E knockout(ApoE-/-)mice confirmed the strong efficacy of MPR NPs in treating atherosclerosis with favorable bio-security,and the mechanism behind this efficacy is believed to involve the regulation of serum metabolism and the remodeling of gut microbes.These findings suggest that the synthesis of MPR NPs provides a promising nanosystem for the targeted therapy of HHcy-induced atherosclerosis.

Child ; Chronic Disease ; Epstein-Barr Virus Infections/complications* ; Familial Primary Pulmonary Hypertension/complications* ; Herpesvirus 4, Human ; Humans ; Persistent Infection ; Pulmonary Arterial Hypertension

Objective To investigate the effect of long non-coding RNA (IncRNA) nuclear-enriched abundant transcript 1 (NEATl) on hypoxia-reoxygenation (H/R) glial astrocyte injury, and to explore whether the mechanism was related to the regulation of micro RNA (miR)-761. Methods Rat cortical astrocytes were cultured to construct a H/R injury model. Astrocytes were divided into control group, model group, model+ small interfering RNA negative control (si-NC) group, model+ si-NEATl group, model+ miR-NC group, model + miR-761 group, model + si-NEATl + anti-miR-NC group, model+si-NEATl+anti-miR-761 group. Expression of NEATl and miR-761 were detected by Real-time PCR. The experiment was repeated 3 times. The content of malonaldefryde (MDA), and the activity of superoxide dismutase (SOD) and catalase (CAT) were detected by kits. Dual luciferase reporter experiment and Real-time PCR were used to analyze the targeting relationship between NEATl and miR-761. The experiment was repeated 3 times. Results Compared with the control group, the cell apoptosis rate and MDA content of the model group increased significantly, SOD and CAT activities decreased significantly, NEATl expression increased significantly, and miR-761 expression decreased significantly (P< 0. 05). Compared with the model+si-NC group, the apoptosis rate and MDA content of the model+si-NEATl group reduced significantly, and SOD and CAT activities increased significantly (P < 0 . 0 5) . Compared with the model + miR-NC group, the apoptosis rate and MDA content of the model + miR-761 group reduced significantly, and SOD and CAT activities increased significantly (P < 0 . 0 5) . MiR-761 was the target gene of N E A T l, and NEATl negatively regulated miR-761 expression. Compared with the model+si-NEATl+anti-miR-NC group, the apoptosis rate and MDA content of the model+siNEAT1+anti-miR-761 group increased significantly, and SOD and CAT activities decreased significantly (P < 0 . 0 5) . Conclusion Interference with NEATl expression can protect astrocytes from H / R injury by up-regulating miR-761.

OBJECTIVE: To analyze the subcellular localization of GTPase of immunity-associated protein 2 (GIMAP2) for the further functional study. METHODS: In the study, we first obtained the protein sequences of GTPase of immunity-associated protein 2 (GIMAP2) from National Center for Biotechnology Information (NCBI) database, and then performed a prediction analysis of its transmembrane structure, nuclear localization signal (NLS), nuclear export signal (NES) and subcellular localization through bioinformatics online tools. GIMAP2 gene amplified by PCR was inserted into the expression vector pQCXIP-mCherry-N1 and positive clones were selected by ampicillin resistance. After using methods to extract and purify, the sequenced recombinant plasmid pQCXIP-GIMAP2-mCherry, together with the retroviral packaging plasmids VSVG and Gag/pol, was transferred into HEK293FT cells by liposomes for virus packaging. The virus supernatant was collected 48 h after transfection and directly infected the human breast cancer cell line MDA-MB-436. Immunofluorescence staining was constructed to detect the localization of endogenous and exogenous GIMAP2 in MDA-MB-436 cells. Meanwhile, green fluorescent chemical dyes were used to label mitochondria, endoplasmic reticulum, and lipid droplets in living MDA-MB-436 cells stably expressing the GIMAP2-mCherry fusion protein. Images for the three dye-labeled organelles and GIMAP2-mCherry fusion protein were captured by super-resolution microscope N-SIM. RESULTS: Bioinformatics analysis data showed that GIMAP2 protein composed of 337 amino acids might contain two transmembrane helix (TM) structures at the carboxyl terminus, of which TMs were estimated to contain 40-41 expected amino acids, followed by the residual protein structures toward the cytoplasmic side. NES was located at the 279-281 amino acids of the carboxyl terminus whereas NLS was not found. GIMAP2 might locate in the lumen of the endoplasmic reticulum. Sequencing results indicated that the expression vector pQCXIP-GIMAP2-mCherry was successfully constructed. Fluorescent staining confirmed that GIMAP2-mCherry fusion protein, co-localized well with endogenous GIMAP2, expressed successfully in the endoplasmic reticulum and on the surface of lipid droplets in MDA-MB-436 cells. CONCLUSION GIMAP2 localizes in the endoplasmic reticulum and on the surface of LDs, suggesting potential involvement of GIMAP2 in lipid metabolism.
Amino Acid Sequence ; Cytoplasm ; GTP Phosphohydrolases ; Humans ; Membrane Proteins ; Nuclear Export Signals ; Nuclear Localization Signals ; Recombinant Fusion Proteins ; Transfection

Child ; Child Health ; Child, Preschool ; China ; Diet ; Diet Surveys ; Feeding Behavior ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Rural Population ; statistics & numerical data ; Vulnerable Populations ; statistics & numerical data

Tianma(the tuber of Gastrodia eleta) is a widely used and pricy Chinese herb. Its counterfeits are often found in herbal markets, which are the plant materials with similar macroscopic characteristics of Tianma. Moreover, the prices of Winter Tianma(cultivated Tianma) and Spring Tianma(mostly wild Tianma) have significant difference. However, it is difficult to identify the true or false, good or bad quality of Tianma samples. Thus, a total of 48 Tianma samples with different characteristics(including Winter Tianma, Spring Tianma, slice, powder, etc.) and 9 plant species 10 samples of Tianma counterfeits were collected and analyzed by HPLC-DAD-MS techniques. After optimizing the procedure of sample preparation, chromatographic and mass-spectral conditions, the HPLC chromatograms of all those samples were collected and compared. The similarities and Fisher discriminant analysis were further conducted between the HPLC chromatograms of Tianma and counterfeit, Winter Tianma and Spring Tianma. The results showed the HPLC chromatograms of 48 Tianma samples were similar at the correlation coefficient more than 0.848(n=48). Their mean chromatogram was simulated and used as Tianma HPLC fingerprint. There were 11 common peaks on the HPLC chromatograms of Tianma, in which 6 main peaks were chosen as characteristic peaks and identified as gastrodin, p-hydroxybenzyl alcohol, parishin A, parishin B, parishin C, parishin E, respectively by comparison of the retention time, UV and MS data with those of standard chemical compounds. All the six chemical compounds are bioactive in Tianma. However, the HPLC chromatograms of the 10 counterfeit samples were significantly different from Tianma fingerprint. The correlation coefficients between HPLC fingerprints of Tianma with the HPLC chromatograms of counterfeits were less than 0.042 and the characteristic peaks were not observed on the HPLC chromatograms of these counterfeit samples. It indicated the true or false Tianma can be identified by either the similarity or characteristic peaks on HPLC fingerprint. Comparing the Winter Tianma with Spring Tianma showed that the HPLC chromatograms of 15 winter Tianma samples and 11 spring Tianma samples were similar at the mean correlation coefficient of 0.908. But the intensity of the characteristic peaks were different between the two groups of Tianma samples, i.e. the intensity of gastrodin, paishin A and C in winter Tianma was lower than those in spring Tianma. The Winter Tianma and Spring Tianma could be discriminated by either the Fisher unstandardized discrimination function or Linear discriminant function, based on the peak areas of 11 common peaks on HPLC chromatograms as variate.

Acupuncture Points ; Adult ; Drugs, Chinese Herbal ; administration & dosage ; Ear ; anatomy & histology ; Female ; Humans ; Melanosis ; drug therapy ; Middle Aged ; Young Adult

Hepatobiliary cystadenomas are rare cystic neoplasms that often occur in middle aged women. The exact etiology of these tumors is unknown. Diagnosis is often delayed in these cases. However, misdiagnosis and inappropriate treatment may result in unfavorable outcome. We report a case of hepatobiliary cystadenoma with pleural effusion. We also review the literature and discuss the current diagnostic and treatment modalities.
Bile Duct Neoplasms ; diagnosis ; pathology ; Bile Ducts, Intrahepatic ; pathology ; Cystadenoma ; diagnosis ; pathology ; Female ; Humans ; Middle Aged ; Pleural Effusion ; diagnosis ; pathology

Objective To determine effects of T-2 toxin and selenium on expression of aggrecanase in human chondrocyte.Methods Human chondrocytes were treated with T-2 toxin(0,1,10,20 μg/L),and/or sodium selenite(final concentration of selenium 0,0.1 mg/L) for 5 days.Aggrecan expression was determined by Western blotting,aggrecanase-1 and aggrecanase-2 mRNA levels were measured by RT-PCR.ResultsSelenium and T-2 toxin had effects on both aggrecan protein levels and its aggrecanases(include aggrecanase-1 and aggrecanase-2 ) mRNA levels(F =0.294,27.71 for aggrecan,F =107.45,362.25 for aggrecanase-l,F =34.68,22.26 for aggrecanase-2,respectively,all P ＜ 0.05),and there was interaction between selenium and T-2 toxin on aggrecan,aggrecanase-1 and aggrecanase-2 expression(F =79.99,230.76,388.33,all P ＜ 0.05).Furthermore,selenium presented significant antagonism to T-2 toxin on aggrecan,aggrecanase-1 and aggrecanase-2 expression.Aggrecan expression levels(0.278 ± 0.015,0.235 ± 0.029,0.195 ± 0.028,0.399 ± 0.028,0.299 ± 0.020,0.263 ±0.019) induced by both 1,10,20 μg/L T-2 toxin and 0,0.1 mg/L selenium were significantly decreased than the levels(0.472 ± 0.0358,0.197 ± 0.018,all P ＜ 0.05) in control group(0 mg/L toxin).Selenium partially blocked the effects induced by 1,10,and 20 μg/L T-2 toxin(all P＜ 0.05).One,10,20 μg/L T-2 toxin and 0,0.1 mg/L selenium increased both aggrecanase-1 mRNA levels(0.535 ± 0.033,1.071 ± 0.043,1.454 ± 0.058,1.057 ±0.048,0.555 ± 0.036,0.902 ± 0.045) and aggrecanase-2 mRNA levels(0.596 ± 0.038,0.656 ± 0.033,0.949 ±0.049,0.600 ± 0.040,0.453 ± 0.031,0.164 ± 0.011),when compared with control(0.481 ± 0.023,0.346 ±0.020 for aggrecanase-1 and 0.387 ± 0.020,1.021 ± 0.046 for aggrecanase-2,respectively,all P ＜ 0.05).Selenium partially blocked 10,20 μg/L T-2 toxins induced upregulation of aggrecanase-1 (all P ＜ 0.05) and aggrecanase-2 (all P ＜ 0.05 ).Conclusions These data suggest a possible molecular mechanism that T-2 toxin could induce cartilage matrix degradation through the upregulation of aggrecanases expression and enzyme activities.Trace element selenium has some protective effect on cartilage proteoglycan degradation induced by T-2 toxins.

Objective To observe the activities of serum peroxidase capacity,and lipid peroxidation of children from Kaschin-Beck disease (KBD) areas of Xinghai county in Qinhai province,and to explore the relationship between antioxidant capacity and KBD.Methods Sixty four KBD and forty six health subjects without KBD were chosen from KBD endemic areas,which included primary schools of Tangnaihai,Xialujuan and Qushian of Xinghai county in Qinghai province,and fifty nine age-matched healthy control subjects without KBD were from a non-KBD endemic area,Nanfan primary school of Chang'an county in Shaanxi province.Twenty patients with KBD and twenty control subjects from KBD areas and non-KBD area were extracted by simple random sampling method.2,3-DAN fluorescence technique was used to test the hair and blood selenium.The biochemical techniques were used to test the indicators of oxidative stress including malondialdehyde(MDA),antioxidant enzyme activities,total antioxidant capacity(T-AOC),serum superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GSHPx).ResultsAll patients with KBD had significantly lower serum GSH-Px activities[ (59.53 ± 25.23)kU/L] and selenium levels in hair[ (67.64 ± 17.28)μg/L] and blood[(36.27 ± 13.29)μg/L],respectively,than that of control subjects from KBD areas [ ( 91.88 ± 22.99 ) kU/L,( 153.32 ± 24.31 ) μg/L,( 63.06 ± 13.66) μg/L ] and nonKBD areas[ ( 122.68 ± 41.74)kU/L,(242.35 ± 38.56)μg/L,(98.93 ± 17.18)μg/L,all P ＜ 0.05].Serum MDA levels in KBD patients[ (4.64 ± 1.11 )μmol/L] were significantly higher than that in control subjects from KBD [(3.31 ± 1.22)μmol/L] and non-KBD areas[ (3.43 ± 1.29)μmol/L,all P ＜ 0.05].On the other hand,T-AOC,SOD and CAT activities were significantly higher in both KBD[(19.80 ± 6.64),(55.80 ± 8.14),(16.45 ± 5.61 ) kU/L] and control subjects[ (21.71 ± 8.82),(57.45 ± 6.96),(15.63 ± 9.18)kU/L] from KBD areas than that of control subjects from non-KBD area[ (13.56 ± 5.38),(42.79 ± 8.10),(6.05 ± 2.71 )kU/L,all P ＜ 0.05 ].Hair selenium levels,blood selenium levels and GSH-Px activity of control subjects from KBD areas were,respectively,significantly lower than that in control subjects from non-KBD area(all P ＜ 0.05).Conclusions These findings strongly confirm the evidence that KBD patients are susceptible to oxidative stress.The results also show the increase in antioxidant enzymes,which could probably be due to adaptive response to pro-oxidant in KBD state.Hence,there seems to be an imbalance between oxidant and antioxidant systems in KBD patients.