BACKGROUND:Hypoxic exercise can promote the degradation of body fat,and changes in the external environment can affect the circadian rhythm of animals,but the mechanisms by which changes in circadian rhythm regulate adipose tissue browning and fat degradation are unclear. OBJECTIVE:To elucidate the mechanism of clock gene regulation on epididymal adipose tissue Browning in obese rats undergoing hypoxia exercise. METHODS:Forty obese rats were randomly selected and divided into four groups(n=10 per group):normoxic sedentary group,hypoxic sedentary group,normoxic exercise group,and hypoxic exercise group for 4 weeks of intervention.The rats in the sedentary groups were not intervened,while those in the hypoxic groups lived in a hypoxic chamber with an oxygen concentration of 13.6%for the whole day.In the exercise groups,adaptive training was performed in the 1st week,and the speed and length of training remained unchanged for the last 3 weeks.The body mass,body length and perirenal fat mass of obese rats were measured.Serum levels of triacylglycerol,total cholesterol,low-density lipoprotein cholesterol,and high-density lipoprotein cholesterol in obese rats were detected by a biochemical assay kit.Liver fat content was observed by oil red O staining.Hematoxylin-eosin staining was used to evaluate the browning of epididymal adipose tissue of rats in different groups.RNA sequencing combined with bioinformatics analysis was used to analyze transcriptome changes in adipose tissue.The mRNA expressions of PGC-1α,Beclin 1,KLF 2 and Perilipin 1 in epididymal adipose tissue were detected by RT-PCR. RESULTS AND CONCLUSION:Hypoxic exercise intervention significantly decreased body mass,body fat percentage,Lee's index,serum triacylglycerol,total cholesterol,and low-density lipoprotein cholesterol levels(P＜0.01),and significantly increased high-density lipoprotein cholesterol level(P＜0.01).Oil red O staining and hematoxylin-eosin staining results showed that hypoxic exercise was more effective in promoting fat mobilization in liver tissue and promoting the browning of parepididymal adipose tissue compared with normoxic sedentary group,hypoxic sedentary group,and normoxic exercise group.RNA-seq results showed that hypoxic exercise significantly upregulated the expression of clock genes Dbp,Nr1d1,Sik1 and adipose tissue browning gene Ppargc1a(PGC-1α)and downregulated the expression of Arntl(Bmal1),accompanied by the enhanced expression of genes related to substance metabolism.qRT-PCR indicated that hypoxic exercise significantly increased the mRNA expression levels of PGC-1α and Perilipin1(P＜0.01).Therefore,these findings indicate that clock genes play an important role in promoting adipose tissue browning during hypoxic exercise.

Objective:To investigate the effect of Qideng Mingmu capsule on the formation and remodeling of retinal neovascularization in mice with oxygen-induced retinopathy (OIR).Methods:Thirty-six postnatal day 7 (P7)SPF grade C57BL/6J pups were divided into normal group, OIR group, Qideng Mingmu capsule group and apatinib group by random number table method, with 9 mice in each group.The mice in the normal group were raised in normal environment.The mice in the other three groups were fed in hyperoxic environment of (75±2)% oxygen concentration for 5 days from P7 to P12 and then were fed in normal environment for 5 days from P12 to P17 to establish the OIR model.From P12, mice in Qideng Mingmu capsule group and apatinib group were given intragastric administration of Qideng Mingmu capsule (900 mg/kg) and vascular endothelial growth factor receptor 2 inhibitor apatinib (70 mg/kg) respectively, once a day for 5 consecutive days.On P17, paraffin sections of mouse eyeballs were made and stained with hematoxylin-eosin to count the number of vascular endothelial cells that broke through the internal limiting membrane.The retinal slices were prepared and stained with FITC-dextran to quantify the retinal non-perfusion area, neovascularization density and total vascular density.The distribution and fluorescence intensity of retinal vascular endothelial cell marker CD31 and pericyte marker α-smooth muscle actin (α-SMA) were observed by double immunofluorescence staining.Immunohistochemical staining was used to detect the expression and distribution of retinal hypoxia inducible factor-1α (HIF-1α) and vascular endothelial cadherin (VE-cadherin).The use and care of animals were in accordance with the Regulations on the Management of Laboratory Animals issued by the Ministry of Science and Technology.This study was approved by the Animal Ethics Committee of Chengdu University of Traditional Chinese Medicine (No.2019-30).Results:The number of vascular endothelial cells breaking through the internal limiting membrane in normal group, OIR group, Qideng Mingmu capsule group and apatinib group were (2.83±4.40), (37.33±5.43), (23.83±6.79) and (14.00±9.34), respectively, with a statistically significant overall difference ( F=28.313, P<0.001).There were more vascular endothelial cells breaking through internal limiting membrane in OIR group than in normal group, Qideng Mingmu capsule group and apatinib group, showing statistically significant differences (all at P<0.05).In the observation of mouse retinal slices, there were large non-perfusion areas, neovascularization buds and disordered distribution of blood vessels in OIR group.The distribution of blood vessels was more uniform and the areas of non-perfusion and neovascularization were smaller in Qideng Mingmu capsule group and apatinib group than in OIR group.The relative area of central retinal non-perfusion area and neovascularization density were significantly lower in normal group, Qideng Mingmu capsule group and apatinib group than in OIR group (all at P<0.05).The immunofluorescence intensity of CD31 and the absorbance value of HIF-1α were significantly lower, and the immunofluorescence intensity of α-SMA and the absorbance value of VE-cadherin were significantly higher in normal group, Qideng Mingmu capsule group and apatinib group than in OIR group (all at P<0.05). Conclusions:Qideng Mingmu capsule can inhibit retinal neovascularization formation, increase vascular pericyte coverage, relieve retinal hypoxia and increase vascular integrity in OIR mice.It can protect the retinal vessels of OIR mice.

Accurate segmentation of ground glass nodule (GGN) is important in clinical. But it is a tough work to segment the GGN, as the GGN in the computed tomography images show blur boundary, irregular shape, and uneven intensity. This paper aims to segment GGN by proposing a fully convolutional residual network, i.e., residual network based on atrous spatial pyramid pooling structure and attention mechanism (ResAANet). The network uses atrous spatial pyramid pooling (ASPP) structure to expand the feature map receptive field and extract more sufficient features, and utilizes attention mechanism, residual connection, long skip connection to fully retain sensitive features, which is extracted by the convolutional layer. First, we employ 565 GGN provided by Shanghai Chest Hospital to train and validate ResAANet, so as to obtain a stable model. Then, two groups of data selected from clinical examinations (84 GGN) and lung image database consortium (LIDC) dataset (145 GGN) were employed to validate and evaluate the performance of the proposed method. Finally, we apply the best threshold method to remove false positive regions and obtain optimized results. The average dice similarity coefficient (DSC) of the proposed algorithm on the clinical dataset and LIDC dataset reached 83.46%, 83.26% respectively, the average Jaccard index (IoU) reached 72.39%, 71.56% respectively, and the speed of segmentation reached 0.1 seconds per image. Comparing with other reported methods, our new method could segment GGN accurately, quickly and robustly. It could provide doctors with important information such as nodule size or density, which assist doctors in subsequent diagnosis and treatment.
Algorithms ; China ; Disease Progression ; Humans ; Multiple Pulmonary Nodules ; Neural Networks, Computer ; Tomography, X-Ray Computed/methods*

Objective: To establish a method for isolation and culture of primary endothelial cells from non-human primate coronary arteries, and to provide a cell model for the study of human coronary endothelial cells. Methods: The coronary arteries of macaca mulattas were separated aseptically. The primary endothelial cells were separatedviatissue adhesion after collagenase digestion. CD31 positive cells were detected and sorted by flow cytometry to determine the purity of endothelial cells. After stimulation with prostaglandin E2(PGE2), the cellular viability and proliferation ability of primary coronary endothelial cells from macaca mulattas were evaluated by high-content cell imaging and CCK-8 assay, and the migration ability and tube function of primary coronary endothelial cells from macaca mulattas were measured by Transwell method and Matrigel glue method, respectively. Results: The confluence percentage of primary coronary artery cells of macaca mulattas was about 80% after 10-14 daysin vitroculture, and the cellular morphology was irregular polygons and paver shape. The purity of endothelial cells was about 31.7% by flow cytometry. After sorting, the purity of endothelial cells was confirmed by flow cytometry, which was more than 95%. PGE2could significantly up-regulate the proliferation, migration and tube formation abilities of primary coronary endothelial cells of macaca mulattas. Conclusion This study successfully established the isolation and culture method of primary coronary endothelial cells from macaca mulattas, and proved that it could be used as anin vitrocell model to simulate human coronary endothelial cells through functional studies.

Objective: To compare the efficiency of different methods for extracting rhesus monkey lung fibroblasts and their effects on functions, so as to provide a method for obtaining primary lung fibroblasts that are closer to human fibroblasts. Methods: Two extraction methods for rhesus monkey lung fibroblasts were used, direct tissue block adhesion method and collagenase combined digestion with tissue block adhesion method. The cell morphology was observed with the inverted microscope, the purity of isolated rhesus monkey lung fibroblasts was identified by immunofluorescence, cell viability was detected by CCK-8, the expression of α-SMA was detected by flow cytometry and the effect of long-term in vitro culture on cell apoptosis was detected by apoptosis kit. Western blot was used to detect the expression of α-SMA protein. Results: The combined digestion with collagenase and tissue block adhesion method could see small and bright cells crawling out in 24 hours, and cells could be seen crawling out in a large area after 48 hours. The cells were in a long spindle shape, after 4 days to 5 days, a single layer of cells could be formed. Identified by immunofluorescence, all cells expressed α-SMA. Tissue adhesion method showed small and bright cells crawling out after 72 hours. After 4 days to 5 days, the cells crawled out in a small area and showed a long spindle shape. After a week, the cells crawled out in a large area and formed a single layer of cells and the cells are all expressed α-SMA by immunofluorescence. The experimental results showed that the cell viability of the cells crawled out by the collagenase digestion method was significantly higher than that of the tissue adhesion method. After TGF-β1 stimulates the cells, the cells extracted by collagenase digestion method proliferated faster and expressed α-SMA more obviously. Conclusion Both methods can isolate rhesus monkey lung fibroblasts in vitro, but the collagenase digestion method extracts cells in a shorter time and in better condition. The expression of related proteins is more stable after stimulation by stimulants, which is an effective method to obtain rhesus monkey lung fibroblasts, and it is also an effective method to obtain primary lung fibroblasts that are closer to human.

Objective To investigate the effects and related risk factors of different vascular access types on new atrial fibrillation in maintenance hemodialysis (MHD) patients.Methods This was a single-center prospective cohort study.Patients who established long-term dialysis access and were voluntarily followed up in the Second Hospital of Tianjin Medical University from January 1,2013 to June 30,2013 were enrolled to follow-up for 5 years.Patients were divided into fistula group (patients with autogenous arteriovenous fistula) and catheter group (patients with tunneled cuffed internal jugular vein catheter).The incidences of new atrial fibrillation in the two groups were compared by Kaplan-Meier survival analysis.Cox regression analysis and receiver operator characteristic curve (ROC) were used to assess the risk factors of new atrial fibrillation.Results A total of 315 eligible patients were enrolled,including 150 males (47.62％).There were 189 patients (60.00％) in the fistula group,and 126 patients (40.00％) in the catheter group.Multivariate Cox regression analysis showed that older age (HR=1.021,95％CI 1.003-1.040),arteriovenous fistula (HR=1.899,95％CI 1.019-3.539),increased dialysis blood flow (HR=1.030,95％CI 1.010-1.051) and left atrial diameter (HR=1.097,95％CI 1.022-1.177) were independent risk factors for new atrial fibrillation in MHD patients (all P ＜ 0.05).Kaplan-Meier survival analysis showed that the incidence of new atrial fibrillation in fistula group was higher than that in catheter group (Log-rank A2=9.53,P=0.002).ROC curve analysis showed that age [the area under the curve (AUC)=0.608,P=0.008],arteriovenous fistula (AUC=0.594,P=0.021),dialysis blood flow (AUC=0.659,P＜0.001) and left atrial diameter (AUC=0.604,P=0.011) could predict the occurrence of new atrial fibrillation.Condusions Older age,arteriovenous fistula,increased blood flow during dialysis and left atrial diameter are independent risk factors for new atrial fibrillation in MHD patients,which can predict the occurrence of atrial fibrillation.The incidence of new atrial fibrillation in patients with arteriovenous fistula is higher than that in patients with catheter.

Objective To investigate the clinical value and safety of cedilanid, esmolol in the treatment of atrial fibrillation. Methods From August 2014 to August 2016 our Hospital from 117 patients with atrial fibrillation clinical data, according to the random number distribution principle, the patients were divided into observation group 59 cases and control group of 58 cases, all patients were given the treatment of primary disease, the clinical symptoms, the patients in the observation group were given oxygen, has given furosemide, cedilanid, after micro injection pump intravenous nitroglycerin, 5-20 g/min. Start after the injection of nitroglycerin, establish another vein channel, every 30 min to 0.2 mg patients, the treatment group were treated with intravenous injection of small dose esmolol. The clinical efficacy, ventricular rate, systolic blood pressure, diastolic blood pressure and adverse reactions were observed in two groups. Results The early and late effective rates of the two groups were not significantly different. The observation showed that after treatment, the ventricular rate, systolic pressure and diastolic pressure in the observation group were significantly higher than those in the control group (P<0.05), and the incidence of adverse reactions in the two groups was significantly different (P<0.05). Conclusion High dose cedilanid combined with small dose esmolol in treatment of atrial fibrillation, obvious curative effect, high safety, can choose the appropriate application.

OBJECTIVE:To excavate and evaluate the security alert signals of linezolid after marketing,and to provide refer-ence for rational drug use in clinic. METHODS:The reporting odds ratio(ROR)method was used to excavate the security alert signals from the adverse drug events(ADE)data in the OpenFDA platform of FDA during second quarter of 2004-2016. The low-er limit of 95%CI >1 was regarded as suggestive of ADE alert signal. RESULTS:A total of 6828534 reports were extracted, among which there were 7224 reports mainly induced by linezolid. Top 10 ADE reports were thrombocytopenia,drug interac-tion,thrombocytopenia,nausea,anaemia,serotonin syndrome,diarrhoea,pyrexia,drug ineffective,vomiting. After the detec-tion of top 200 ADE reports by ROR method,120 signals related to 18 system organ class(SOC)were identified. Top 5 ADE sig-nals in turn by SOC were medical examination,nervous system disorders,blood and lymphatic system disorders,metabolism and nutrition disorders,cardiac disorders. The high risk signals with clinical reference value included 42 cases of optic neuropathy (ROR=56.33),350 cases of serotonin syndrome(ROR=52.86),162 cases of lactic acidosis(ROR=18.30),31 cases of endo-carditis (ROR=15.38),566 cases of thrombocytopenia (ROR=14.29),122 cases of bone marrow failure (ROR=14.20),261 cases of panhematopenia (ROR=11.92),86 cases of disseminated intravascular coagulation (ROR=10.91),58 cases of toxic epidermal necrolysis(ROR=9.06),etc. As for ADE reports,male was slightly higher than female,and the patients in age group of ≥45 reported evidently more compared to younger age group. CONCLUSIONS:By detecting and evaluating alert signals of li-nezolid through OpenFDA platform,we could effectively lay the foundation for further research of pharmacovigilance.

Objective To investigate effects of exogenous sonic hedgehog (Shh) on proliferation of neural stem cells (NSCs) in ependymal area and recovery of motor function after spinal cord injury (SCI) in adult rats.Methods Fifty-five female SD rats were involved in the study:five were selected as normal control group and fifty as Shh group (n =25) and SCI group (n =25) after being subjected to SCI at T10 segment using the modified Allen' s method according to the random number table.At 1,3,7,14,and 28 days after operation,restoration of hindlimb motor function of SD rats was assessed with modified Tarlov scale and changes of double positive cells of Brdu and Nestin with double-stained immunofluorescence.Results Tarlov scale revealed statistical difference between Shh and SCI groups since days 7 postoperatively (P ＜ 0.05).In the double-staining test,number of double positive cells of Brdu and Nestin was greater in Shh Group than in SCI Group since day 3 postoperatively [(97.20 ± 18.23) vs (72.60± 15.60),(153.60 ±25.76) vs (112.20 ±23.63),(133.80 ±21.02) vs (94.20± 18.70),(89.80 ± 15.42) vs (43.40 ± 10.62),P ＜ 0.05].Conclusion Exogenous Shh is conducive to the proliferation of ependymal NSCs and the recovery of motor function in SCI rats.

Objective: To evaluate the hybrid of drug eluting stent (DES) with bare metal stent (BMS) and exclusive DES implantation for treating the patients with multi-lesion coronary disease.
Methods: A total of 6495 patients with multi-lesion coronary disease received elective PCI in our hospital from 2004-04 to 2006-10 were retrospectively studied. The patients were divided into 2 groups, Hybrid group, n=848 and Exclusive DES group, n=5647. With 1:1 propensity score matching, there were 823 pair of patients were ifnally studied. The clinical outcomes included 1, 2 years post-operative all cause death, myocardial infarction (MI), target lesion revascularization (TLR), target vessel revascularization (TVR), major adverse cardiac events (MACE) and in-stent thrombosis. The relative risks of all outcomes were assessed by Cox’s proportional-hazard model after propensity match.
Results: With propensity match, Cox’s proportional-hazard model analysis indicated that compared with Exclusive DES group, Hybrid group had the higher risks of TLR (HR 2.38, 95%CI 1.50-3.70), TVR (HR 1.61, 95%CI 1.15-2.27), MACE (HR 1.37, 95%CI 1.02-1.85), all P<0.01. The all cause death, MI and the ratio of all cause death/MI were similar between 2 groups in 1, 2 years follow-up period, all P>0.05.
Conclusion:Compared with exclusive DES, the hybrid of DES with BMS implantation had the higher risk of TLR, TVR and MACE for treating the patients with multi-lesion coronary disease.