Objective:To explore the role and mechanisms of action of human circular RNA 0001400 (hsa_circ_0001400) and its linear transcript RELL1 in the development of Kaposi's sarcoma (KS) .Methods:KS-associated herpesvirus (KSHV) was induced and extracted from BCBL-1 cells by phorbol ester. Human umbilical vein endothelial cells (HUVECs) were then infected with KSHV, inverted microscopy and real-time fluorescence-based quantitative PCR (qRT-PCR) were performed to observe cellular morphology and determine the expression of KSHV lytic gene ORF50 and latent gene LANA, respectively, so as to verify whether the infection was successful. HUVECs in the logarithmic growth phase were divided into 3 groups: HUVEC group (uninfected HUVECs), KSHV + HUVEC group (HUVECs infected with KSHV at a multiplicity of infection [MOI] of 0.5), and mitogen-activated protein kinase (MAPK) inhibitor group (HUVECs infected with KSHV at a MOI of 0.5 for 6 hours, followed by the treatment with 1 μmol/L MAPK inhibitor). Cell counting kit (CCK8) assay and flow cytometry were performed to assess the cellular proliferative ability and detect apoptosis in the above 3 groups, respectively. qRT-PCR and Western blot analysis were conducted to determine the transcription and protein expression levels of hsa_circ_0001400, its linear transcript RELL1, and rat sarcoma viral oncogene homolog (RAS) /MAPK signaling pathway-related genes in the HUVEC group and KSHV + HUVEC group. Five pairs of KS tissues and paraneoplastic tissues were collected, and mRNA expression of the above genes was verified by qRT-PCR in the KS tissue samples. Statistical analyses were performed using two-way analysis of variance, one-way analysis of variance, and t test. Results:Compared with the uninfected HUVECs, the infected HUVECs became rounder and grew more densely at 48 hours after the infection with KSHV. The mRNA expression of ORF50 and LANA genes could be detected in the KSHV-infected HUVECs, and their mRNA expression levels were significantly higher than those in the uninfected HUVECs (both P < 0.001), indicating successful infection of HUVEC by KSHV. The cellular proliferative rate in the KSHV + HUVEC group gradually increased over time during 24 to 72 hours after the infection, while that in the MAPK inhibitor group was markedly inhibited. The total apoptosis rate at 48 hours significantly differed among the 3 groups ( F = 673.98, P < 0.001), and was significantly higher in the MAPK inhibitor group than in the KSHV + HUVEC group and the HUVEC group (both P < 0.001), while there was no significant difference between the KSHV + HUVEC group and the HUVEC group ( P > 0.05). qRT-PCR showed that the mRNA expression of hsa_circ_0001400, RELL1, Kirsten rat sarcoma viral oncogene homologue (KRAS), MAPK11, and extracellular signal-regulated kinase (ERK) -2 was significantly higher in the KSHV + HUVEC group than in the HUVEC group (all P < 0.05), while ERK1 mRNA expression did not significantly differ between the 2 groups ( t = 0.92, P = 0.410). Western blot assay showed that the protein expression of RELL1, KRAS, MAPK11, ERK1, and ERK2 was significantly higher in the KSHV + HUVEC group than in the HUVEC group (all P < 0.01). The mRNA expression of RELL1, ERK1, and ERK2 was significantly higher in the KS tissues than in the paraneoplastic tissues (all P < 0.05) . Conclusion:KSHV infection may regulate the occurrence and development of KS by inducing the expression of hsa_circ_0001400 and its linear transcript RELL1, as well as activating the MAPK signaling pathway.

Objective:To observe the occurrence of epithelial-mesenchymal transition (EMT) in cervical cancer cell line Siha irradiated by X-rays with clinical conventional fraction radiotherapy model and investigate the role of exosomes in this process.Methods:Siha cells were irradiated by 6 MV-X rays with 50 Gy in 25 fractions. EMT was evaluated by cell morphology, EMT biomarkers and cell migration and invasion ability. Exosomes released from cells were detected by transmission electron microscopy and nanoparticle tracking analysis (NTA), and its function in EMT was explored by using an exosome inhibitor GW4869 (10 μmol/L).Results:After irradiation, EMT phenomenon was induced in the survived Siha cells, including the incidence of mesenchymal phenotype, upregulation of epithelial marker E-cadherin ( t=9.66, P<0.05), downregulation of mesenchymal marker N-cadherin ( t=41.61, P<0.05), and increase of cell migration and invasion abilities ( t=6.11, 13.22; P<0.05). Meanwhile, the secretion of exosomes was also increased after irradiation ( t=7.51, P<0.05). When the cells were pre-treated with GW4869, radiation-induced exosome secretion was reduced ( t=7.28, P<0.05), so that radiation-induced EMT was reversed. Conclusions:Ionizing radiation with clinical conventional fraction radiotherapy model promotes EMT of cervical cancer cells through increasing the secretion of exosomes.

This study aimed to explore the role of miR-130a-3p in cardiomyocyte hypertrophy and its underlying mechanisms. Pressure-overload induced myocardial hypertrophy mice model was constructed by thoracic aortic constriction (TAC). , norepinephrine (NE) was used to stimulate neonatal rat cardiomyocytes (NRCMs) and H9c2 rat cardiomyocytes to induce hypertrophic phenotypes. The expression of miR-130a-3p was detected in mice hypertrophic myocardium, hypertrophic NRCMs and H9c2 cells. The mimics and inhibitors of miR-130a-3p were transfected into H9c2 cells to observe the role of miR-130a-3p on the hypertrophic phenotype change of cardiomyocytes separately. Furthermore, whether miR-130a-3p regulated hypertrophic related signaling pathways was explored. The results showed that the expression of miR-130a-3p was significantly decreased in hypertrophic myocardium, hypertrophic NRCMs and H9c2 cells. After transfection of miR-130a-3p mimics, the expression of hypertrophic marker genes, atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and β-myosin heavy chain (β-MHC), and the cell surface area were notably down-regulated compared with the control group (mimics N.C. + NE group). But after transfection of miR-130a-3p inhibitor, the expression of ANP, BNP and β-MHC in H9c2 cells increased significantly, and the cell area increased further. By Western blot, it was found that the protein phosphorylation level of Akt and mTOR were down-regulated after over-expression of miR-130a-3p. These results suggest that miR-130a-3p mimics may alleviate the degree of cardiomyocyte hypertrophy, meanwhile its inhibitor can further aggravate cardiomyocyte hypertrophy. Over-expression of miR-130a-3p may attenuate cardiomyocytes hypertrophy by affecting the Akt pathway.
Animals ; Atrial Natriuretic Factor ; Cardiomegaly ; Mice ; MicroRNAs ; genetics ; Myocardium ; pathology ; Myocytes, Cardiac ; pathology ; Myosin Heavy Chains ; Natriuretic Peptide, Brain ; Nonmuscle Myosin Type IIB ; Proto-Oncogene Proteins c-akt ; Rats

Objective:To evaluate the rapid diagnostic value of serum novel coronavirus (2019-nCoV) IgM/IgG detection in COVID-19, aiming to further improve the diagnostic and screening system of COVID-19.Methods:Blood samples were collected from 32 patients with COVID-19 (tested positive for 2019-nCoV nucleic acid by RT-PCR and presented with clinical symptoms) and 34 non-COVID-19 patients (tested negative for 2019-nCoV nucleic acid by RT-PCR and clinically confirmed as non-COVID-19 patients). Colloidal gold-based immunochromatography was used for rapid detection of 2019-nCoV IgM/IgG in these samples. The sensitivity and specificity of the test, and the correlation of serum 2019-nCoV IgM/IgG with disease course were analyzed.Results:Among the 32 COVID-19 patients, nine tested positive for 2019-nCoV IgM with a positive rate of 28.1% (9/32) and 25 positive for 2019-nCoV IgG with a positive rate of 78.1% (25/32). The total positive rate was 84.4% (27/32). Two of the 34 non-COVID-19 patients tested positive for 2019-nCoV IgG with a positive rate of 5.9% (2/34), while none of them was positive for 2019-nCoV IgM. The positive rates of serum IgM were 42.9% (3/7), 30.8% (4/13) and 16.7% (2/12) at 10-20 d, 21-30 d and 31-40 d after the patients developed the symptoms of COVID-19, respectively, which showed a decreasing tread with prolonged disease course. The positive rates of serum IgG in COVID-19 patients were 57.1% (4/7), 84.6% (11/13) and 83.3% (10/12) at 10-20 d, 21-30 d and 30-40 d after symptom onset. The rate showed an increasing trend with prolonged disease course and reached the peak in about 21-30 d.Conclusions:Serum 2019-nCoV IgM/IgG detection (using colloidal gold method) had high sensitivity (84.4%) and strong specificity (94.1%) in the diagnosis of 2019-nCoV infection. It had a great value in the diagnosis and screening of COVID-19 and could be used as a valuable complementary method to the COVID-19 diagnostic system due to its advantages of flexibility, rapidity and simplicity.

Objective: To investigate the differences in frequency and function of natural killer cells (NK) between chronic hepatitis B (CHB) and acute hepatitis B (AHB). Methods: Patients with AHB and those with CHB in immune active (IA) phase were enrolled. The frequencies of NK, CD56^dimNK, CD56^brightNK and the expression of functional molecules IFNAR2 and NKp46 on the surface of NK cells were detected respectively among patients with CHB in IA phase, patients with AHB, and those recovered from AHB. At the same time, their correlations with ALT, HBV DNA and HBV markers were analyzed. Results: Between IA and AHB, the frequencies of NK cells and NKp46^dim NK cells in AHB cases were significantly lower than those in IA cases, but the frequency of NKp46^high NK cells in AHB was higher than that in IA. For patients who recovered from AHB, the frequency of NK cells and NKp46^dim NK cells increased; the varied ranges of frequencies of CD56^dimNK, IFNAR2⁺ NK and NKp46⁺ NK cells were on the rise, while the frequency of NKp46^high NK cells decreased after the recovery from AHB, and the varied ranges of CD56^brightNK and IFNAR2MFI, NKp46MFI decreased. In AHB, HBVDNA loads were positively correlated with ALT levels. Before and after the recovery of AHB: ΔHBV DNA and ΔALT, Δ NK/LY (%) were positively correlated; ΔALT and ΔNKp46^highNK/NK(%), ΔNKp46MFI, ΔIFNAR2MFI were positively correlated. Conclusions In CHB immune active phase, the activity of peripheral blood NK cells was too weak to remove the virus, but NK cells play an important role in eliminating the viruses and mediating liver tissue inflammation in AHB.

Objective To investigate the effect of surgical treatment on survival in colorectal carcinoma patients with synchronous hepatic metastasis.Methods The retrospective case-control study was done on 953 consecutive patients with synchronous colorectal hepatic metastasesl from January 2003 to December 2013.Results Median survival time (46.7 months)and 5-year survival rate (32％) for patients with resected hepatic metastases was significantly superior to that of with nonoperative treatment (17 months,4％).Expanded criteria for hepatic metastases resection raised resection rates (31％ vs.13.6％,P ＜0.05).For patients with resectable hepatic metastases,the inhospital cost for simultaneous resection group was lower than that in the staged resection group (36 698 vs.45 134 RMB,P ＜ 0.05).For patients of asymptomatic primary tumor with unresectable hepatic metastases,resection of the primary tumor was associated with an improved median survival (18.0 vs.15.0 months,P ＜ 0.05) Conclusions Expanding indications of hepatic metastases resection can improve survival in patients with synchronous colorectal hepatic metastases.Simultaneous resection of primary tumor and hepatic metastases were indicated in patients with resectable synchronous colorectal hepatic metastases.Resection of primary tumor was recommended for asymptomatic patients with unresectable hepatic metastases.

Objective To analyze CT imaging findings and pathological features of the thymic cyst.Methods The CT findings of 20 cases with thymic cyst confirmed by pathology were analyzed retrospectively.The analysis included the location,shape ,size ,density ,contour, unilocular or multilocular,with or without walls,enhancement and lesion periphery.The pathology results were compared to CT findings.Results All lesions were located in the anterior mediastinum,in which the location of left side in 12 cases,right side in 6 cases,median mediastinum in 2 cases.All lesions had clear border and unilocular,including round shape in 4 cases,oval shape in 8 cases,and irregular shape in 8 cases with triangular shape in 1 case,the size range was 2-6 cm.4 cases had obvious cyst wall.The CT value of the cyst had range of 4-53 HU,with 10 cases less than 10 HU and 10 cases greater than 10 HU.There was no enhancement within the cysts of 20 cases and the cyst wall had enhancement in 4 cases.All cases of pathological findings were confirmed as thymus cyst.Gross sample showed gray red or isabeling.The lesion was cystic in 18 cases,soft stereocyst in 2 cases,bright yellow liquid within the capsule in 10 cases,butter fat in 5 cases,chylomicrons in 3 cases,unilocular in 19 cases and multilocular in 1 case (butter fat).Microscope showed the fibrous capsule wall in all cases,lined with squamous epithelium,ciliated columnar epithelium,monolayer columnar epithelium,in which could be seen normal thymus tissue.The thickness of the wall was no more than 0.5 cm,and multilocular wall smooth,no obvious inflammatory response in the surrounding.Conclusion CT image findings can clearly show the thymic cyst site,shape,size,density and boundaries,unilocular or multilocular,with or without walls.The CT image can reflect pathological features of the thymic cyst,and can provide accurate and reliable information for the clinic.

Objective To identify the effect of both high glucose and high insulin on miR-145 level.Methods The human vascular smooth muscle cells ( VSMCs) were cultured and the proliferation of VSMCs was induced by high glucose and high insulin medium.The samples were divided into 4 groups: normal group, high glu-cose group (25 mmol/L), high insulin group (300 mU/L), high glucose and high insulin group.The ex-pression of miR-145 in VSMCs was assayed by real-time PCR.Proliferation of VSMCs was determined by MTT method After 72 h cultivation.The migration of VSMCs was analyzed by cell scratch test .VSMCs in each group was transfected by miR-145 virus ( lentiviral vector ) .Proliferation and migration were assayed after 48 h transfection.Results The expression of miR-145 in VSMCs of other three groups was decreased ( P<0.05 ) , especially the expression in the high glucose and insulin group was the lowest ( P<0.01 ) .Prolifera-tion and migration of VSMCs was promoted by high glucose and/or high insulin medium.Under fluorescent, transfection rate of VSMCs was about 80%after 48 h transfection.Proliferation and migration of VSMCs in each group after transfection were significantly lower than before ( P<0.05 ) .Conclusions High glucose and high in-sulin could decrease the expression of miR-145 in VSMCs, The overexpression of miR-145 may inhibit the prolif-eration and migration of VSMCs .

Objective The pathogenesis of HBV infection is the result of a complex interactions between the host immune system and the virus, the host immune system involves innate immune and adaptive immune. Now, it is thought that host immune responses to viral particles and proteins are important factors that determine whether HBV is cleared or persists and hepatocytes injured. Innate immune system is the first defending line of host against viral infection. However, many studies have shown that HBV can develop tactics to escape innate immune recognition and interfere with innate immune signaling pathways and induce immunosuppression. It is necessary to analysis the functions and status of host innate immunity in HBV infection which may contribute to find novel approaches to eliminate HBV. This review will present the current understanding of innate immune components including pattern-recognition receptors(PRRs)、dendritic cells(DCs)、natural killer(NK)/natural killer T(NKT)cells、T regulatory cells(Tregs) and interferons(IFNs).

Objective: To elucidate the functions of peripheral blood NK cells in chronic hepatitis B patients treated with interferon. Methods: Venous whole blood samples were obtained from patients in the immune clearance (IC) phase treated with peg-interferon-alpha-2a (Peg-IFNα-2a) at baseline (t=0), 12 weeks (t=12) and 24 weeks (t=24). The frequencyies of peripheral blood CD3^-CD56⁺ NK cells, CD56^brightNK cells and CD56^dimNK cells, the expression level of IFNAR2 and NKp46 on NK cells were detected by flow cytometry. The levels of serum HBV DNA, HBsAg, HBeAg and ALT were detected by Ditan Hospital clinical laboratory. Results: Forty-one patients in the IC phase treated with Peg-IFNα-2a, including 21 poor response (PR) patients and 20 good response (GR) patients, were recruited for this study. Theresult were as follows: The frequency of peripheral blood CD3^-CD56⁺ NK cells was increased at week 24 in GR compared with the baseline(11.74, 5.69%-18.15% vs 13.7, 9.36-20.18%, P>0.05), and it also was increased in PR(8.94, 6.26%-14.15% vs 12.5, 7.64-16.55%, P>0.05). The frequency of peripheral blood CD56^brightNK cells was increased at week 24 in GR compared with the baseline(9.49, 6.2%-12.48% vs 12.98, 7.75%-20.93%, P>0.05), and it also was increased in PR(11.45, 8.27%-19.13% vs 17.52, 12.3%-22.42%, P=0.0239). The expression level of NKp46 on NK cells was significantly increased at week 24 in GR compared with the baseline(90.55, 83.8-94.78 vs 93.8, 92.28-96.4, P=0.0263), but it was not increased in PR(95, 90.6-96.15 vs 94.3, 92.1-95.6, P>0.05). The expression level of NKp46^high on NK cells was significantly increased at week 24 in GR compared with the baseline(12.4, 8.58-19.08 vs 39.3, 23.15-49.3, P=0.0011), at that range of the increase was significantly higher than PR(14.2, 9.78-17.65 vs 27.58, 19.13-36.56, P=0.006). Conclusions The frequencies of peripheral blood CD3^-CD56⁺ NK cells and CD56^brightNK cells were increased from patients in the IC phase treated with Peg-IFNα-2a. The expression level of NKp46 on NK cells was increased in GR patients treated with Peg-IFNα-2a. The expression level of NKp46^high on NK cells was significantly increased, especially in GR patients treated with Peg-IFNα-2a.