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OBJECTIVE@#To identify and characterize read-through RNAs and read-through circular RNAs (rt-circ-HS) derived from transcriptional read-through hypoxia inducible factor 1α (HIF1α) and small nuclear RNA activating complex polypeptide 1 (SNAPC1) the two adjacent genes located on chromosome 14q23, in renal carcinoma cells and renal carcinoma tissues, and to study the effects of rt-circ-HS on biological behavior of renal carcinoma cells and on regulation of HIF1α.@*METHODS@#Reverse transcription-polymerase chain reaction (RT-PCR) and Sanger sequencing were used to examine expression of read-through RNAs HIF1α-SNAPC1 and rt-circ-HS in different tumor cells. Tissue microarrays of 437 different types of renal cell carcinoma (RCC) were constructed, and chromogenic in situ hybridization (ISH) was used to investigate expression of rt-circ-HS in different RCC types. Small interference RNA (siRNA) and artificial overexpression plasmids were designed to examine the effects of rt-circ-HS on 786-O and A498 renal carcinoma cell proliferation, migration and invasiveness by cell counting kit 8 (CCK8), EdU incorporation and Transwell cell migration and invasion assays. RT-PCR and Western blot were used to exa-mine expression of HIF1α and SNAPC1 RNA and proteins after interference of rt-circ-HS with siRNA, respectively. The binding of rt-circ-HS with microRNA 539 (miR-539), and miR-539 with HIF1α 3' untranslated region (3' UTR), and the effects of these interactions were investigated by dual luciferase reporter gene assays.@*RESULTS@#We discovered a novel 1 144 nt rt-circ-HS, which was derived from read-through RNA HIF1α-SNAPC1 and consisted of HIF1α exon 2-6 and SNAPC1 exon 2-4. Expression of rt-circ-HS was significantly upregulated in 786-O renal carcinoma cells. ISH showed that the overall positive expression rate of rt-circ-HS in RCC tissue samples was 67.5% (295/437), and the expression was different in different types of RCCs. Mechanistically, rt-circ-HS promoted renal carcinoma cell proliferation, migration and invasiveness by functioning as a competitive endogenous inhibitor of miR-539, which we found to be a potent post-transcriptional suppressor of HIF1α, thus promoting expression of HIF1α.@*CONCLUSION@#The novel rt-circ-HS is highly expressed in different types of RCCs and acts as a competitive endogenous inhibitor of miR-539 to promote expression of its parental gene HIF1α and thus the proliferation, migration and invasion of renal cancer cells.
Subject(s)
Humans , Carcinoma, Renal Cell/pathology , Cell Proliferation , Hypoxia , Kidney Neoplasms , MicroRNAs/genetics , Neoplasm Invasiveness/genetics , RNA, Circular/metabolism , RNA, Small Interfering , Hypoxia-Inducible Factor 1, alpha Subunit/geneticsABSTRACT
Background@#Co-infections of the porcine reproductive and respiratory syndrome virus (PRRSV) and the Haemophilus parasuis (HPS) are severe in Chinese pigs, but the immune response genes against co-infected with 2 pathogens in the lungs have not been reported. @*Objectives@#To understand the effect of PRRSV and/or HPS infection on the genes expression associated with lung immune function. @*Methods@#The expression of the immune-related genes was analyzed using RNA-sequencing and bioinformatics. Differentially expressed genes (DEGs) were detected and identified by quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry (IHC) and western blotting assays. @*Results@#All experimental pigs showed clinical symptoms and lung lesions. RNA-seq analysis showed that 922 DEGs in co-challenged pigs were more than in the HPS group (709 DEGs) and the PRRSV group (676 DEGs). Eleven DEGs validated by qRT-PCR were consistent with the RNA sequencing results. Eleven common Kyoto Encyclopedia of Genes and Genomes pathways related to infection and immune were found in single-infected and co-challenged pigs, including autophagy, cytokine-cytokine receptor interaction, and antigen processing and presentation, involving different DEGs. A model of immune response to infection with PRRSV and HPS was predicted among the DEGs in the co-challenged pigs. Dual oxidase 1 (DUOX1) and interleukin-21 (IL21) were detected by IHC and western blot and showed significant differences between the co-challenged pigs and the controls. @*Conclusions@#These findings elucidated the transcriptome changes in the lungs after PRRSV and/or HPS infections, providing ideas for further study to inhibit ROS production and promote pulmonary fibrosis caused by co-challenging with PRRSV and HPS.
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Atypical polypoid adenomyoma (APA) is an uncommon type of polypoid characterized by fibroid stroma and endometrial glands. It occurs mostly in premenopausal women and rarely in postmenopausal women with irregular vaginal bleeding. In our current case, a 76-year-old woman presented with irregular vaginal bleeding. The final pathological diagnosis of the mass was APA. APA is not easy to diagnose before surgery. On the one hand, there was no obvious particularity in imaging features and clinical features, especially for uncomfortably identifying endometrial cancer. On the other hand, APA has a pedicle, attaching to any part of the uterine cavity, which can cause pseudocoel between the mass with the uterine cavity wall. So, when it comes to getting the pathological tissue in the absence of hysteroscopy, it is easy to access to the pseudocoel and obtain endometrial tissue rather than the pathological tissue of the mass. Therefore, preoperative imaging examination is of great significance diagnosis way of thinking to clinicians for APA. In the meantime, pathological tissue of APA can be obtained by hysteroscopy in visual conditions.
Subject(s)
Aged , Female , Humans , Pregnancy , Adenomyoma/pathology , Hysteroscopy , Magnetic Resonance Imaging , Uterine Hemorrhage , Uterine Neoplasms/diagnostic imagingABSTRACT
Objective:To investigate the influences of cardiovascular risk factors on left ventricular wall motion function in patients treated with epirubicin chemotherapy by layer-specific two-dimensional speckle tracking imaging (2D-STI).Methods:Sixty-four female patients with breast cancer treated with epirubicin and 36 controls from June 2018 to March 2019 in Renmin Hospital of Wuhan University were included. According to the numbers of cardiovascular risk factors, the patients were divided into group A (without cardiovascular risk factors, 31 cases), group B (single cardiovascular risk factor, 20 cases), and group C (multiple cardiovascular risk factors, 13 cases). All patients underwent echocardiography before and after epirubicin respectively. Global longitudinal strain, circumferential strain, radial strain (GLS, GCS, GRS), endocardial and epicardial layers of GLS and GCS (endoGLS, epiGLS, endoGCS, epiGCS) were obtained by EchoPAC software analysis. The differences of GLS, EndoGLS, EpiGLS, GCS, EndoGCS, EpiGCS and GRS before and after chemotheraphy were calculated as ΔGLS, ΔEndoGLS, ΔEpiGLS, ΔGCS, ΔEndoGCS, ΔEpiGCS and ΔGRS.Results:Compared with before chemotherapy, GLS, EndoGLS, EpiGLS, GCS, EndoGCS, EpiGCS and GRS were significantly reduced in group A, B and C after chemotherapy (all P<0.05). ΔEndoGLS and ΔEndoGCS showed an increasing trend in groups A, B and C, with statistical significance (all P<0.05). Correlation analysis showed that after chemotherapy with epirubicin, the numbers of cardiovascular risk factors were significantly correlated with strain parameters, and the correlation coefficient of EndoGLS was relatively higher ( r=-0.582, P<0.001). Conclusions:Layer-specific imaging can reliably detect the influences of cardiovascular risk factors on the left ventricular wall motion function in breast cancer patients treated with epirubicin. Parameters reflecting the endocardial longitudinal systolic function are sensitive indicators for the early detectin of myocardial damage and toxicity by epirubicin in patients with multiple cardiovascular risk factors.
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Objective@#To investigate the influences of cardiovascular risk factors on left ventricular wall motion function in patients treated with epirubicin chemotherapy by layer-specific two-dimensional speckle tracking imaging (2D-STI).@*Methods@#Sixty-four female patients with breast cancer treated with epirubicin and 36 controls from June 2018 to March 2019 in Renmin Hospital of Wuhan University were included. According to the numbers of cardiovascular risk factors, the patients were divided into group A (without cardiovascular risk factors, 31 cases), group B (single cardiovascular risk factor, 20 cases), and group C (multiple cardiovascular risk factors, 13 cases). All patients underwent echocardiography before and after epirubicin respectively. Global longitudinal strain, circumferential strain, radial strain (GLS, GCS, GRS), endocardial and epicardial layers of GLS and GCS (endoGLS, epiGLS, endoGCS, epiGCS) were obtained by EchoPAC software analysis. The differences of GLS, EndoGLS, EpiGLS, GCS, EndoGCS, EpiGCS and GRS before and after chemotheraphy were calculated as ΔGLS, ΔEndoGLS, ΔEpiGLS, ΔGCS, ΔEndoGCS, ΔEpiGCS and ΔGRS.@*Results@#Compared with before chemotherapy, GLS, EndoGLS, EpiGLS, GCS, EndoGCS, EpiGCS and GRS were significantly reduced in group A, B and C after chemotherapy (all P<0.05). ΔEndoGLS and ΔEndoGCS showed an increasing trend in groups A, B and C, with statistical significance (all P<0.05). Correlation analysis showed that after chemotherapy with epirubicin, the numbers of cardiovascular risk factors were significantly correlated with strain parameters, and the correlation coefficient of EndoGLS was relatively higher (r=-0.582, P<0.001).@*Conclusions@#Layer-specific imaging can reliably detect the influences of cardiovascular risk factors on the left ventricular wall motion function in breast cancer patients treated with epirubicin. Parameters reflecting the endocardial longitudinal systolic function are sensitive indicators for the early detectin of myocardial damage and toxicity by epirubicin in patients with multiple cardiovascular risk factors.
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Objective@#To construct the Z-score models for normal fetal brain fissures.@*Methods@#Four hundred and eighty-seven pregnant woman who underwent prenatal ultrasonography from January to August 2018 in the Second Xiangya Hospital of Central South University were collected randomly for analysis, and all the fetuses(18-38 weeks) were healthy. The calacarine fissure, pariet-ooccipital fissure, sylvian fissure of the fetuses were measured and they were chosed as the dependent variables, while the independent variable were the gestational age and biparietal diamete, as well as femur length. The regression analyses of the mean(M) and the standard deviation(SD) for each parameter were performed separately. Two observers randomly measured the brain fissures depth two hours apart to evaluate the interobserver repeatability.@*Results@#The brain fissure depth in normal fetuses during geatational age between 18 to 38 weeks can be evaluated by Z-score model.Positive correlations were found between the depth of calacarine fissure, pariet-ooccipital fissure, sylvian fissure and gestational age, biparietal diameter and femur length.@*Conclusions@#Z-score can intuitively reflect the development of brain fissure more accurate, and is valuable, and can be used as a powerful supplement to the traditional method.
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Objective To construct the Z-score models for normal fetal brain fissures.Methods Four hundred and eighty-seven pregnant woman who underwent prenatal ultrasonography from January to August 2018 in the Second Xiangya Hospital of Central South University were collected randomly for analysis,and all the fetuses(18-38 weeks) were healthy.The calacarine fissure,pariet-ooccipital fissure, sylvian fissure of the fetuses were measured and they were chosed as the dependent variables,while the independent variable were the gestational age and biparietal diamete,as well as femur length.The regression analyses of the mean(M) and the standard deviation(SD) for each parameter were performed separately. Two observers randomly measured the brain fissures depth two hours apart to evaluate the interobserver repeatability.Results The brain fissure depth in normal fetuses during geatational age between 18 to 38 weeks can be evaluated by Z-score model.Positive correlations were found between the depth of calacarine fissure,pariet-ooccipital fissure,sylvian fissure and gestational age,biparietal diameter and femur length. Conclusions Z-score can intuitively reflect the development of brain fissure more accurate,and is valuable, and can be used as a powerful supplement to the traditional method.
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Objective To study the changes of blood flow in posterior cerebral artery ( PCA ) in complete transposition of great arteries (CTGA) through the application of the pulsed Doppler . Methods Twenty CTGA fetuses ( CTGA group) and 20 healthy control fetuses ( control group) were involved ,the blood flow indexes peak systolic velocity ( Vs) ,end-diastolic velocity ( Vd) ,pulsatility index ( PI) ,resistance index ( RI) ,velocity-time integral ( VTI) of PCA-S1 ,PCA-S2 and MCA of the fetuses in the two groups were detected by pulsed Doppler . The differences in blood flow indexes between CTGA fetuses and healthy controls were analyzed by independent t -test . The rates of abnormal resistance in PCA-S1 and MCA in CTGA fetuses were compared through Chi-square test ( χ2 test) . Results Compared with control group ,the MCA-PI ,MCA-RI ,PCA-S1-PI and PCA-S1-RI of CTGA group decreased significantly( all P < 0 .05) ,MCA-VTI ,PCA-S1-VTI ,PCA-S2-VTI increased significantly ( all P < 0 .05) ,but no significant difference was found in PCA-S2-PI ,PCA-S2-RI ,Vs and Vd of the MCA and the PCA ( all P > 0 .05 ) . The rate of abnormal resistance in the MCA was significantly lower than that in the PCA-S1 in CTGA group ( P <0 .05) . Conclusions The pulsed Doppler can be used to study the changes of blood flow in PCA of CTGA fetuses and the differences of specific hemodynamic alterations may occured in different segments of the PCA in CTGA fetuses ,indicating a tendency to protect the PCA-supplying areas of the brain when ischemia and hypoxia .
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To study the blood flow parameters for 2 anatomical segments of posterior cerebral artery (PCA) in normal singleton fetal. Methods: The blood flow velocity parameters peak systolic velocity (PSV), end diastolic velocity (EDV), time-average maximum velocity (TAMAXV), velocity time integral (VTI) and resistance parameters systolic peak velocity and end diastolic velocity ratio (S/D), pulsatility index (PI), resistance index (RI) for 2 anatomical segments in PCA were recorded. The first segment of the PCA (PCAS1) was recorded between the origin of PCA and the proximal part of the posterior communicating artery. The second segment of the PCA (PCAS2) was recorded at the distal part of the posterior communicating artery. The blood parameters in both PCAS1 and PCAS2 were analyzed by using Pearson correlation and multiple curves fitting with gestational age (GA). Paired student's t test was performed to compare the difference in blood parameter between PCAS1 and PCAS2. Results: The blood flow velocity parameters in both PCAS1 and PCAS2 were increased with the GA (P0.05). Resistance parameters in PCAS1 were higher than those in PCAS2 (P<0.05). Conclusion: The blood flow velocity parameters in both PCAS1 and PCAS2 are increased with GA. The resistance parameters in both PCAS1 and PCAS2 do not change with GA. Study on the velocities and resistance in these 3 arterial branches provides a more comprehensive evaluation on the process of brain circulation.
Subject(s)
Female , Humans , Pregnancy , Blood Flow Velocity , Physiology , Diastole , Physiology , Gestational Age , Hemodynamics , Physiology , Posterior Cerebral Artery , Physiology , Systole , PhysiologyABSTRACT
To investigate hemodynamic parameters in 2 anatomical segments (S1 and S2) of anterior cerebral artery (ACA) in normal pregnancy during the second and third trimester of gestation. Methods: The peak systolic velocity (PSV), end diastolic velocity (EDV), time-average maximum velocity (TAMAXV), peak systolic velocity/end diastolic velocity (S/D), resistance index (RI), and pulsation index (PI) in S1 and S2 of fetal anterior cerebral artery (ACA) in 288 normal pregnant women were detected by power Doppler and pulsed Doppler. Multiple regression models were fitted to estimate the relation between Doppler variables and gestational age. The differences of hemodynamic parameters between ACAS1 and ACAS2 were compared. Results: The PSV, EDV, and TAMAXV of ACAS1 and ACAS2 were positively correlated with the weeks of pregnancy (P0.05). The PSV, TAMAXV, S/D, PI, and RI of ACAS1 were significantly higher than those of ACAS2, while EDV in ACAS1 was lower than that in ACAS2 (P<0.05). Conclusion: The velocity parameters (PSV, EDV, TAMAXV) of the 2 anatomical segments (ACAS1 and ACAS2) are increased with the increase of gestational age in normal pregnant fetus during the second and third trimester of gestation, and the resistance parameters (S/D, PI, RI) are not significantly correlated with gestational age. Distribution of blood flow is different in the blood supply territory between ACAS1 and ACAS2.
Subject(s)
Female , Humans , Pregnancy , Anterior Cerebral Artery , Physiology , Blood Flow Velocity , Fetus , Hemodynamics , Pregnancy Trimester, Third , Ultrasonography, PrenatalABSTRACT
AIM:To investigate the effect of CKLF1-C19 polypeptide (C19) on differentiation of human lung fibroblast (LFB) into myofibroblast (MFB) induced by TGF-β. METHODS:LFBs were cultured and identified. LFBs were treated with TGF-β(5 μg/L) to establish the cell model of LFB differentiate into MFB. The LFBs were divided into 6 experimental groups including control group,TGF-β group,and TGF-β plus different doses(1,0.1,0.01,0.001 mg/L) C19 groups. The cell morphology,cell proliferation rate, and the expression of α smooth muscle actin (α-SMA) and collagen Ⅰ were observed. RESULTS:Human primary LFB was successfully cultured and was confirmed by the method of immunofluorescence. TGF-β at 5 μg/L induced proliferation and differentiation of LFB. The mRNA levels of α-SMA and collagen Ⅰ in TGF-β group were higher than that in control group(P<0.05).The cell proliferation rates,mRNA levels of α-SMA and collagen Ⅰ, and the protein expression of α-SMA in 0.01 mg/L+TGF-β group and 0.001 mg/L+TGF-β group were markedly lower than those in TGF-β group(P<0.05). CONCLUSION:C19 at 0.01 mg/L and 0.001 mg/L effectively inhibits differentiation of LFB into MFB induced by TGF-β, thus inhibiting the process of airway remodeling and fibrosis to some extent.
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Objective To explore the acute toxicity effect of CKLF1-C19,a polypeptide of chemokine-like factor 1(CKLF1),on the KM mice. Methods A total of 40 KM mice,half male and half female,were randomly divided into 2 groups. The mice in the experimental group were injected with CKLF1-C19 at a dose of 25 mL/kg(100 μg/mL,1 mg/mL and 10 mg/mL)through the tail vein,and those in the control group received an equal volume of sterile saline solution. Changes in the body weight of the mice were recorded the day after treatment, and the general conditions of mice in the experimental group were observed closely and compared with the normal group. Then blood samples were taken from the abdominal aorta to measure liver and kidney function. Tissue samples of liver, kidney, spleen and lung were taken for histopathological examination by HE staining. Results In the maximum tolerance test,the mice of the two groups were in good condition, and their body weight was increased gradually, without significant difference between the experimental group and the control group(P > 0.05). There was no death within 14 days. The blood biochemical indexes of liver and kidney function showed no significant differences between the two groups(P > 0.05). The gross appearances of heart, liver,kidney,spleen and lung were normal in the two mouse groups, and the pathological examination with HE staining showed normal clear structure with no obvious changes in these organs of each group. Conclusions Our results demonstrate that CKLF1-C19 has no acute toxicity effect on mice.
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<p><b>OBJECTIVE</b>To study the clinical and laboratory features of the mild and severe hand-foot-mouth diseases (HFMD) in Shenzhen in 2008.</p><p><b>METHODS</b>145 cases were observed in East-Lake Hospital and Shenzhen Children's Hospital. Of the 145 cases, 124 mild cases and 21 severe cases were involved.All the clinical data and laboratory findings were collected and summarized. After collection of the acute and convalescent consecutive stools and peripheral bloods from the patients with HFMDI, EV71 genes were amplified from these samples by RT-PCR. Enterovirus 71 were cultured and isolated using Vero cell line and R&D cell line.</p><p><b>RESULTS</b>The WBC counts and blood glucose levels of the severe cases were significantly elevated, but the ages of the severe ones significantly decreased compared with those of the mild cases (P < 0.05). EV71 genes could be detected by RT-PCR with 35% positive rate in mild cases and 67% in severe cases. The EV71 gene detection rate of the severe cases was significantly increased in contrast to that of the mild ones. The EV71 were isolated and cultured from the stools of 9 patients, one specimens from the dead's stool. Two severe cases died of neurogenic pulmonary edema and brain-stem encephalitis.</p><p><b>CONCLUSIONS</b>EV71 mainly contributes to HFMD and is responsible for death of some severe cases. High fever, less rash, elevated white blood cell counts and blood glucose concentrations as well as age less than 4 years old should be used for prediction of severe cases.</p>
Subject(s)
Adult , Child , Female , Humans , Male , Blood Glucose , Physiology , Enterovirus , Enterovirus Infections , Blood , Pathology , Hand, Foot and Mouth Disease , Blood , Pathology , Virology , Laboratories , Leukocyte Count , Reverse Transcriptase Polymerase Chain Reaction , Severity of Illness IndexABSTRACT
Object To investigate the effects of Panax notoginseng saponins (PNS) on myocardial Gs? mRNA expression of seriously scalded rat. Methods A 30% skin full thickness scald model was produced by immersing rat in 95 ℃ water for 10 s. The effects of PNS on myocardial Gs? mRNA level were observed with dot blotting hybridization and in situ hybridization technique; effects on cAMP and adenyl cyclase (AC) activities were determined with radioimmunoassay. Results Myocardial Gs? mRNA, AC activity and cAMP content were reduced significantly 3 h after scalding. PNS (100, 200 mg/kg) could markedly increase the level of myocardial Gs? mRNA expression. The elevated quantity was correlated markedly with PNS dosage (r = 0.95, P