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Objective:To analyze the expression of circular RNA (circRNA) in peripheral blood mononuclear cells (PBMCs) of patients with gout and to explore the possible mechanism of circRNA in the pathogenesis of gout.Methods:Peripheral blood samples of 24 patients with acute gout (AG), 24 patients with intermittent gout (IG) and 24 healthy control subjects (HC) were collected. Three cases of AG, IG, and HC were randomly selected, and the differentially expressed circRNA in PBMCs was screened by human circNA microarrays. The 6 circRNAs with large differences between the two comparison groups were selected, and the relative expression levels of 6 circRNAs in all the collected 72 PBMCs of the study subjects were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The significantly differentially ex-pressed circRNA (fold change>1.5, P<0.05) was analyzed by GO analysis, Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis, and its interaction with microRNA (miRNA) was predicted. The median (interquartile range) was used to describe the data, and the Mann-Whitney U test was used for comparison between groups. Results:(1) The microarray analysis results showed that compared with the HC group, the AG group and the IG group had 116 and 41 significantly differently expressed circRNAs, respectively; com-pared with the IG group, the AG group had 105 significantly differently expressed circRNAs. (2) Among the 6 circRNAs verified by PT-qPCR, the expression trends of 5 were consistent with the microarray results. The expression of hsa_circRNA_105034 in the AG group [5.17(4.60)] was statistically significantly different com-pared to the IG [1.68(2.39)] and HC [0.90(0.73)] groups (AG vs IG: Z=-4.413, P<0.01; AG vs HC Z=-5.052, P<0.01). (3) Bioinformatics analysis: ① GO analysis found that differential circRNA swere mainly involved in DNA transcriptional regulation, positive cell regulation and protein modification, etc. ② KEGG pathway analysis revealed that differential circRNA might be involved in the immune response mediated by the mitogen-activated protein kinase signaling pathway. ③ CircRNA might affect its inflammatory response by targeting molecules such as miRNA-146a, miRNA-302b and miRNA-23a. Conclusion:There are differentially expressed circRNAs in PBMCs of patients with gout, which may be closely related to the occurrence and development of gout.
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@#Acute lung injury is one of the common and serious complications of acute aortic dissection, and it greatly affects the recovery of patients. Old age, overweight, hypoxemia, smoking history, hypotension, extensive involvement of dissection and pleural effusion are possible risk factors for the acute lung injury before operation. In addition, deep hypothermia circulatory arrest and blood product infusion can further aggravate the acute lung injury during operation. In this paper, researches on risk factors, prediction model, prevention and treatment of acute aortic dissection with acute lung injury were reviewed, in order to provide assistance for clinical diagnosis and treatment.
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Objective To observe the clinical therapeutic effect of Bushen Huoxue formula for treatment of diabetic nephropathy at G3a stage. Methods Sixty patients with stage G3a diabetic nephropathy were admitted to the Department of Nephrology of Affiliated Hospital of Tianjin Institute of Traditional Chinese Medicine from June 2017 to June 2018, and according to difference in treatment, they were divided into an integrated traditional Chinese and western medicine treatment group and a western medicine treatment group with 30 cases in each group. The two groups were treated with Huangkui capsule 2.5 g/time, 3 times a day; the integrated traditional Chinese and western medicine treatment group was additionally given Bushen Huoxue formula, one dose daily, twice a day taken orally; 2 months for 1 course. The changes of 24-hour urinary protein, serum creatinine (SCr) and urea nitrogen (BUN) were observed after 3 courses of treatment. Results After 3 courses of treatment, the levels of 24-hour urinary protein, SCr and BUN of the integrated traditional Chinese and western medicine treatment group were lower than those of the western medicine treatment group [24-hour urinary protein quantification (g): 1.45±0.26 vs. 2.11±0.35, SCr (μmol/L): 105.15±12.31 vs. 158.32±17.26, BUN (mmol/L): 7.26±2.41 vs. 12.87±3.24], the differences being statistically significant (all P < 0.05). The total effective rate of integrated traditional Chinese and western medicine treatment group was significantly higher than that of western medicine treatment group [86.67% (26/30) vs. 53.33% (16/30), P < 0.05]. Conclusion Bushen Huoxue formula in the treatment of patients with diabetic nephropathy at stage G3a can decrease their urinary protein and SCr, BUN significantly.
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Objective:To investigate the expressions of miR23b and Sp1 in ovarian endometriosis and their clinic significance.Methods:qPCR was used to detect the expression of miR23b and Sp1 mRNA in paired ectopic/eutopic and normal endometrium.Immunohistochemistry and Western bolt were used to determine the expression and distribution of Sp1 in paired ectopic/eutopic and normal endometrium.The association ofmiR23b and Sp1 with the endometriosis was analyzed.Results:MiR23b mRNA expression in paired ectopic/eutopic and normal endometrium was gradually increased (P＜0.05).Sp1 protein mainly distributed in the nucleus of endometrial glandular epithelial and stromal cells,with a little or without expression in cytoplasm.Spl mRNA and protein expression in paired ectopic/eutopic and normal endometrium was gradually reduced (P＜0.05).Pearson correlation analysis showed that miR23b was negatively correlated with Sp 1 (r=-0.526,P＜0.05).Conclusion:MiR23b and Sp1 are involved in the pathogenesis of ovarian endometriosis,which may facilitate the formation of ectopic lesions.
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Objective To compare the rates of major adverse cardiovascular events(MACE)and bleeding events of three different antiplatelet strategies during temporary withdrawal of antiplatelet therapy for non-cardiac surgery within 1 year after drug-eluting stent (DES)implantation.Methods Retrospectively analyzed 42 patients who had accepted non-cardiac surgery and required temporary withdrawal of antiplatelet therapy within 1 year after drug-eluting stent implantation. The patients were divided into three groups according to the bridging antiplatelet strategies they received.All patients discontinued clopidogrel 5 to 7 days before the non-cardiac surgery. The tirofiban group was treated with intravenous tirofiban 0.4ug/kg·min in the first 30 min followed 0.1μg/(kg·min). The dosage was reduced by half for patients whose Creatinine clearance were less than 30 ml/min.The low molecular weight heparin group was treated with subcutaneous enoxaparin (Clexane 4000 AxaIU, once per day) .The asprin group was given only oral asprin(100 mg, once per day) . Tirofiban and low molecular weight heparin were continued until clopidogrel was resured. Perioperative cardiovascular events and serious bleeding were recorded. Results The rates of major adverse cardiac events in the tirofiban and the low molecular weight heparin group were lower than the aspirin group. Acute myocardial infarction caused by confirmed in-stent thrombosis was diagnosed in one patient in the aspirin group. One case of asymptomatic ST-T changes was found in the low molecular weight the aspirin group. 3 cases in the aspirin group presented ST-T changes on ECG and among them 1 case was STEMI due to LAD thrombosis requiring primary and 2 other cases were agina pectoris.There were no significant differences in bleeding events among the three groups.Conclusions Potential for the perioperative management with tirofiban or low molecular weight heparin is safe and feasible for patients who had recently undergone DES implantation and required noncardiac surgery with the interruption of antiplatelet therapies.
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Objective To investigate the effects of baicalein (Bai) on acute lung injury (ALI) induced by intestinal ischemia/reperfusion (I/R) and its mechanism in mice.Methods Twenty-four male C57BL/6J mice were divided into three groups by random number table:namely sham group,I/R group and Bai+I/R group,with 8 mice in each group.Intestinal I/R induced lung injury model was reproduced by clamping superior mesenteric artery for 90 minutes,followed by reperfusion.Bai (100 mg/kg) was intraperitoneally injected 1 hour before ischemic challenge in the Bai+I/Rgroup.The mice in sham group underwent the similar procedure with I/R group but without vascular occlusion.All mice were sacrificed at 4 hours of reperfusion,and blood was collected from inferior vena cava and lung tissues were harvested.Lung tissues were stained with hematoxylin-eosin (HE),and histological changes were examined under light microscope for pathological score.Lung wet/dry (W/D) ratio was calculated.Lung cell apoptosis was determined by TdT-mediated dUTP nick end labeling (TUNEL) technique.Serum levels of tumor necrosis factor-α (TNF-α) and interleukin-6(IL-6) were determined by enzyme-linked immunosorbent assay (ELISA).The mRNA expressions of TNF-α and IL-6 in lung tissues were determined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR).The protein expression levels of cytoplasmic inhibitory factor-α of nuclear factor-κB (IκB-α) and nucleus NF-κB were determined by Western Blot.Results Under light microscope,a normal lung tissue structure was shown in the sham group and no evidence of obvious lung injury was found.In the I/R group,the alveolar structure was seriously damaged.The alveolar wall was widened and there was significant interstitial edema and leukocytes infiltration.In the Bai+I/R group,pathological damage was significantly decreased as indicated by reduced lung tissue edema and leukocytes infiltration.Compared with the sham group,the lung pathological scores,W/D ratio and cellular apoptosis in the I/R group were significantly increased.Bothserum TNF-α and IL-6 contents and lung TNF-α and IL-6 mRNA expressions were significantly increased.Furthermore,I/R significantly resulted in a decrease of IκB-α in the cytoplasm and an increase of NF-κB in the nucleus.Notably,Bai treatment significantly attenuated ALI induced by intestinal I/R injury.Compared with the I/R group,the lung pathological scores and W/D ratio in the Bai+I/R group were significantly decreased (lung pathological score:4.59±1.17 vs.6.27±1.34,W/D ratio:3.79±0.28 vs.4.32±0.57),cellular apoptosis was significantly decreased [(4.85 ± 2.47)％ vs.(8.15 ± 2.33)％],both serum TNF-α and IL-6 contents and lung TNF-α and IL-6 mRNA expressions were significantly decreased [serum TNF-α (pg/L):124.18±30.49 vs.167.72 ± 38.65,IL-6 (ng/L):1.65 ± 0.69 vs.2.43 ± 0.57;lung TNF-α mRNA (2-△△Ct:4.75 ± 2.38 vs.7.69 ± 2.32,IL-6 mRNA (2-△△ Ct):16.45 ±4.39 vs.27.69 ± 6.82],additionally,Bai pretreatment significantly increased cytoplasmic IκB-α protein expression (gray value:0.47 ± 0.11 vs.0.27 ± 0.09),while decreased nuclear NF-κB protein expression (gray value:0.57 ± 0.13 vs.1.07 ± 0.14,all P ＜ 0.05).Conclusion Bai could attenuate intestinal I/R injury induced ALI via the inhibition of inflammation and apoptosis.
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Objective To investigate the risk factors and prognosis of perioperative myocardial infarction in the patients undergoing noncardiac surgery. Methods Clinical data of 562 patients who had accepted non-cardiac surgery was collected and retrospectively analyzed. The risk factors, treatments and outcomes of all these patients were recorded and analyzed. Results A total of 19 out of the 562 patients had perioperative myocardial infarction ( PMI) . The incidence was 3. 4% . The mean occurrence time was (43. 5 ± 12. 7)h after operation. Eleven PMI patients (11 ∕ 19) were non-ST-segment elevation myocardial infarction and eight patients (8 ∕ 19) were ST-segment elevation myocardial infarction. Thirteen PMI patients were left coronary artery occlusion and six patients were right coronary artery occlusion. Advanced age, history of myocardial infarction, unstable angina, change of ST-T segment on electrocardiography (ECG), multivessel diseases, diabetes,hypertension,and high risk non-cardiac surgery were the risk factors of PMI and positively correlated to PMI. Sixteen PMI (16 ∕ 19) patients accepted PCI treatment and three patients (3 ∕ 19) accepted drug conservative treatment. Two patients had unstable angina attack after treatment and one patient had arrhythmia. The heart function in two patients decreased by one or more than one class within the follow up of 1 year. No patient had recurrent acute myocardial infarction or deceased during follow-up. Conclusions Many factors could lead to PMI. Making preoperative assessment, recognizing patients of high risks and dealing with patients who had PMI in time was necessary.
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Objective To evaluate the remission situation of early re-induction with priming low dose regimen containing G-CSF in treating acute myeloid leukemia (AML).Methods Ninety-seven AML patients in our hospital from March 2015 to January 2017 were retrospectively analyzed.All cases adopted the standard DA regimen for conducting the induction chemotherapy,among them,38 cases had significant residual disease on 14 d of induction chemotherapy,21 cases adopted the low dose priming regimen for conducting the early re-induction chemotherapy,17 cases adopted the tandard DA gregimen for conducting the re-induction chemotherapy.The complete remission(CR) rate and and adverse reactions were compared between two groups.Results The total CR rate in all 97 cases was 60.8％;among 38 cases needing re-induction chemotherapy,the CR rate in the priming regimen re-induction group was 76.2 ％,which was significantly higher than 41.2 ％ in the DA regimen re-induction group,the difference was statistically significant (P=0.028);the occurrence rates of side effects such as infection and cytopenia during re-induction chemotherapy process had no difference between two groups(P＞0.05).Conclusion For AML patients with obvious residual disease on 14 d of induction chemotherapy,adopting low dose priming regimen in re-duction chemotherapy has higher CR,which is superior to the standard DA regimen.
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This study aims to knock out the goat β-lactoglobulin (BLG) gene using CRISPR-Cas9 system and knock in human lactoferrin (hLF) at the BLG locus, and further study the effect of RAD51 stimulatory compound (RS-1) on homologous recombination efficiency. First, we designed an sgRNA targeting the first exon of goat BLG gene and constructed a co-expression vector pCas9-sgBLG. This sgRNA vector was then transfected into goat ear fibroblasts (GEFs), and the target region was examined by T7EN1 assay and sequencing. Second, we constructed a targeting vector pBHA-hLF-NIE including NEO and EGFP genes based on BLG gene locus. This targeting vector together with pCas9-sgBLG expression vector was co-transfected into GEFs. Transfected cells were then treated with 0, 5, 10 and 20 μmol/L RS-1 for 72 h to analyse the EGFP expression efficiency. Next, we used 800 μg/mL G418 to screen G418-resistent cell clones, and studied hLF site-specific knock-in cell clones by PCR and sequencing. The editing efficiency of sgBLG was between 25% and 31%. The EGFP expression efficiency indicated that the gene knock-in efficiency was improved by RS-1 in a dose-dependent manner, which could reach 3.5-fold compared to the control group. The percentage of positive cells with hLF knock-in was increased to 32.61% when 10 μmol/L RS-1 was used. However, when the concentration of RS-1 increased to 20 μmol/L, the percentage of positive cells decreased to 22.22% and resulted in an increase of senescent cell clone number. These results suggested that hLF knock-in and BLG knock-out in GEFs were achieved by using CRISPR/Cas9 system, and optimum concentration of RS-1 could improve knock-in efficiency, which provides a reference for efficiently obtaining gene knock-in cells using CRISPR/Cas9 in the future.
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<p><b>OBJECTIVE</b>To investigate the clinical value of serum anti-Ku86 in early detection of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Expression levels of Ku86 protein in HCC and adjacent normal liver tissues were detected by Western blotting. Serum anti-Ku86 level in 83 patients with early HCC and 124 patients with liver cirrhosis were detected by enzyme-linked immunosorbent assay (ELISA). Chemiluminescence was used to measure the serum level of α-fetoprotein (AFP).</p><p><b>RESULTS</b>Expression of Ku86 protein in HCC was increased when compared with the adjacent normal liver tissues (0.21 ± 0.05 vs. 0.08 ± 0.02, P < 0.01). Serum anti-Ku86 level was significantly elevated in HCC patients compared with that in liver cirrhosis patients (0.47 ± 0.22 vs. 0.22 ± 0.06 Abs at 450 nm, P < 0.01), but there was no significant difference between HBV infection and HCV infection in HCC patients (0.51 ± 0.19 vs. 0.47 ± 0.24, P = 0.267). Of note, serum anti-Ku86 level was significantly decreased after surgical resection of the tumors in the 30 HCC cases tested (P < 0.01). The results of ROC analysis indicated a better performance of anti-Ku86 (0.857) than AFP (0.739) for early detection of HCC. In 83 HCC patients, the positive rate of anti-Ku86 was 61.4% (51/83), significantly higher than that of the AFP positive rate (27.7%, 23/83). The anti-Ku86 level was positive in 37 of 60 HCC cases with negative AFP. Combination assay of AFP and anti-Ku86 could detect 60 of 83 HCC cases (72.3%, 60/83). There was no significant correlation of anti-Ku86 and AFP (r = 0.156, P = 0.161).</p><p><b>CONCLUSIONS</b>Serum anti-Ku86 level is significantly elevated and is not related to HBV and HCV infection in HCC patients. Serum anti-Ku86 antibody may be a potential biomarker for early detection of HCC, and can be used in combination with AFP in clinics.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Antigens, Nuclear , Allergy and Immunology , Autoantibodies , Blood , Biomarkers, Tumor , Blood , Carcinoma, Hepatocellular , Blood , Diagnosis , Virology , DNA-Binding Proteins , Allergy and Immunology , Early Detection of Cancer , Hepatitis B , Blood , Hepatitis C , Blood , Ku Autoantigen , Liver Cirrhosis , Blood , Liver Neoplasms , Blood , Diagnosis , Virology , ROC Curve , alpha-Fetoproteins , MetabolismABSTRACT
Objective To explore issues of the human tissue biobank, ranging from its establishment, collection and preservation of samples, quality control, management and application. Methods Development of standardized operational procedures, collection of such samples as fresh frozen tissues from the surgery, paraffin-embedded tissues, whole blood, serum, plasma, and cerebrospinal fluid. Specimens were classified and made aliquots according to different requirements, and then stored at room temperature, -80℃ refrigerator or in liquid nitrogen. Microsoft Access database system was used in the management of these specimens. Results From September 2004 to September 2008, a total of 11 872 samples from patients with benign and malignant diseases were collected and preserved. Among them, 4360 tubes of fresh frozen tissue samples from 2500 cases were provided within and beyond the province. These samples were also applied to DNA, RNA, protein extraction and tissue microarray, and immunohistochemistry-related research. Conclusion Human tissue biobank is highly useful in sharing human tissue resources effectively, as it can provide high quality specimens from benign and malignant diseases and normal control. In addition, it plays a very important role in exploring pathogenesis, developing new technologies for early detection of disease and new therapeutic strategies.