Effect of circAGFG1 targeting miR-4429 on the proliferation, migration and invasion of cholangiocarcinoma QBC939 cells and its possible mechanism / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of circAGFG1 on the proliferation, migration and invasion of cholangiocarcinoma QBC939 cells and its possible mechanism. Methods: The tumor tissues and corresponding para-cancerous tissues of 33 patients with cholangiocarcinoma who underwent surgical resection in the 988th Hospital of the Joint Logistics Support Force from April 2017 to October 2019 were collected. qPCR was used to detect the expression level of circAGFG1 and miR-4429 in the tissues. Cholangiocarcinoma QBC939 cells were cultured in vitro and transfected with si-circAGFG1 or miR-4429 mimics, or co-transfected with si-circAGFG1 and anti-miR-4429. Then, cell proliferation was detected by CCK-8 method and clone formation test, cell migration and invasion were detected by scratch test and Transwell assay, and the protein expression of E-cadherin and N-cadherin in cells was determined by Western blotting. Dual-luciferase reporter gene experiment was adopted to verify the regulatory relationship between circAGFG1 and miR-4429. Results: The expression of circAGFG1 was higher (3.89±0.26 vs 1.00±0.08, P<0.05) while the expression of miR-4429 (0.28±0.03 vs 1.00±0.05, P<0.05) was lower in cholangiocarcinoma tissues than those in para-cancerous tissues. After the interference with circAGFG1 or over-expression of miR-4429, the cell proliferation level, number of clone formation, scratch healing rate, number of invaded cells, and the protein expression of N-cadherin in QBC939 cells were reduced (all P<0.05), but the protein expression of E-cadherin was elevated (P<0.05). circAGFG1 could targetedly bind with miR-4429, and interfering circAGFG1 promoted the expression of miR-4429 in QBC939 cells (all P<0.05). Down-regulation of miR-4429 reversed the effect of interfering circAGFG1 on the proliferation, migration and invasion of QBC939 cells (all P<0.05). Conclusion: The expression of circAGFG1 is up-regulated in cholangiocarcinoma tissues, which may promote the proliferation, migration and invasion of cholangiocarcinoma QBC939 cells by targetedly inhibiting the expression of miR-4429.

circ_0000615 regulates proliferation, migration and invasion of cholangiocarcinoma cells by targeting miR-432-5p / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探讨circ_0000615在胆管癌细胞中的表达及其对胆管癌细胞增殖、迁移和侵袭能力的影响和可能的调控机制。方法：通过qPCR检测circ_0000615在正常胆管上皮HIBEC细胞和胆管癌CCLP-1、QBC939、TFK-1和RBE细胞中的表达水平。双荧光素酶报告基因实验验证circ_0000615与miR-432-5p的靶向关系。将si-NC、si-circ_0000615（si-circ-1、si-circ-2）、inhibitor-NC和inhibitor-miR-432-5p转染至CCLP-1和QBC939细胞，转染细胞分为si-NC组、si-circ-1组、si-circ-2组、si-NC+inh-NC组、si-NC+inh-miR-432-5p组和si-circ-1+inh-miR-432-5p组，通过CCK-8、EdU和Transwell实验检测各转染组胆管癌细胞的增殖、迁移和侵袭能力。结果：在胆管癌细胞中circ_0000615呈高表达而miR-432-5p呈低表达（P<0.05或P<0.01）；双荧光素酶报告基因实验证实circ_0000615与miR-432-5p之间存在靶向关系（P<0.01）；敲减circ_0000615可明显抑制CCLP-1、QBC939细胞的增殖、侵袭和迁移能力（P<0.05或P<0.01）；下调miR-432-5p可部分逆转敲减circ_0000615对胆管癌CCLP-1和QBC939细胞增殖、迁移和侵袭的抑制作用（P<0.05或P<0.01）。结论：circ_0000615通过靶向miR-432-5p调控胆管癌细胞的增殖、侵袭和迁移。

Comparing the tested results of QBC Star and Sysmex XP-100 hematology analyzers and analyzing their operational performance / 国际检验医学杂志

Objective Both QBC Star and Sysmex XP-100 hematology analyzers are convenient to carry,which can be used nor-mally under the condition of the field(emergency).This study would compare their test results and operating performance,so to provide guidance for rational use of the instruments.Methods 100 fresh blood samples of 100 health soldiers anti-coagulated by EDTA-K2 were detected by QBC Star and Sysmex XP-100 haematology analyzers respectively,the results of two analyzers were comparatively analyzed and their test time and operating convenience were analyzed.Results There was no significant difference in the results of hemoglobin concentration (HGB),hematocrit (HCT)tested by the two methods (P >0.05).There were significant difference of the mean corpuscular hemoglobin concentration (MCHC),the sum of lymphocyte percent and middle type cells (LYM%+MID%),neutrophil percentage(NEUT%),white blood cell count(WBC),platelet count(PLT)tested by the two meth-ods(P <0.05).The values of MCHC and LYM%+MID% tested by the QBC Star were significantly lower than that detected by Sysmex XP-100(P <0.05),while the rest indicators tested by the former were higher than that of the latter.It took about 5 minutes to complete a blood sample analysis with QBC Star,while about 1 min was needed for Sysmex XP-100.Conclusion The test results of QBC Star and Sysmex XP-100 hematology analyzers couldn′t exchanged except for that of HCT and HGB.Under the condition of field(emergency),QBC Star hematology analyzer is suitable for individual medical examination,and Sysmex XP-100 hematology an-alyzer can be used for the batch medical examination.

Killing effect of CIK cells combined with Gemcitabine on QBC939 cells / 中国现代普通外科进展

Objective:To investigate the anti-tumor effects of cytokine-induced kill cells(CIK)methods combined with chemotherapeuticdrug Gemcitabine against Cholangiocarcinoma cancer cell lines QBC-939.Methods:Peripheral blood mononuclear cells(PBMC) of healthy people were stimulated by different cytokines,and were induced into CIK cells.CIK cells were cultured for 14 days as effector cells.The phenotype of CIK cells were analyzed by flow cytometer.QBC939 cells were cultured with the CIK cells at different effector-target ratio or various concentrations of Gemcitabine for 24 and 48 hours.The antitumor effects were measured by CCK8 methods.The expression of Bax was detected by using Western blot.Results:The CD3+CD4+,CD3+CD8+,CD3+HLA-DR+,CD3+CD56+,CD4+CD25+ double positive cel1 was up to 10.89％,60.27％,71.82％,9.03％ and 4.01％ after 14 days' cultivation.The killing effect of CIK and Gemcitabine increased with the increase of effector-target ratio and drug concentration or the extension of time.The killing effect of combination of CIK and Gemcitabine was obviously higher than that of each single factor.The protein levels detected hints of CIK cells and gemcitabine can up-regulated the expression of Bax,and the joint action of both is more significant.Conclusions:CIK cells have strong anti-tumor effect against QBC939 cells by inducing apoptosis of QBC939 cells,and it can enhance the anti-tumor effects of Gemcitabine against QBC939 cells when CIK and Gemcitabine are combined together.

Hemozoin Pigment: An Important Tool for Low Parasitemic Malarial Diagnosis

Low parasitemic condition in malaria remains a diagnostic challenge; as the available diagnostic methods failed to detect. Currently, hemozoin (Hz) pigment is gaining attention in the diagnosis of malaria. The major drawback is ease of detection of Hz in routine practice. A pilot study was conducted to evaluate the role of Hz pigment and to compare the performance of quantitative buffy coat assay (QBC) and PCR in such conditions. Clinically suspected cases of malaria were examined by both Giemsa stain and immunochromatographic test (ICT). Samples positive by ICT and negative by Giemsa stain were further examined by nested PCR targeting 18S rRNA and QBC for the presence of malaria parasites and pigments. Thirty blood samples fulfilled the inclusion criteria out of which 23 were Plasmodium vivax (Pv), 4 Plasmodium falciparum (Pf), and 3 mixed (Pv and Pf) by immunochromatographic test. Twenty-one out of 30 (70%) were positive by nested PCR in comparison to 25/30 (83%) by QBC. Samples containing both malaria parasites and Hz pigment by QBC completely showed concordance with the PCR result. However, 61% of total samples containing only Hz pigment were observed positive by PCR. Hz pigment remains an important tool for malaria diagnosis. Identification of leukocytes containing pigments by QBC not only indicates recent malarial infections but also puts light on severity of the disease. QBC assay is a rapid, highly sensitive, and cost-effective method to detect malaria parasites and Hz pigment especially in low parasitemic conditions.

Incidence, severity, prognostic significance of thrombocytopenia in malaria.

Background: Malaria is an infectious disease caused by plasmodium parasite. P. falciparum account for majority of morbidity and mortality. Thrombocytopenia and anaemia are the most frequently associated hematological complications in malaria. The low platelet count together with acute febrile syndrome emerged as the strongest predictor of malaria a finding that is frequent and present even before anemia and splenomegaly sets in. Severe thrombocytopenia is a good predictor of poor prognosis than mild and moderate thrombocytopenia. The aim is to study the incidence, severity, prognostic significance of thrombocytopenia in malaria. Methods: This was an observational and prospective study. The study enrolled 100 patients with thrombocytopenia and fever who were proven to have malaria either by peripheral smear or Quantitative Buffy Coat (QBC) test or malarial antigen assay were included in the study and patients with thrombocytopenia due to other causes were excluded from the study. Platelet count was estimated on a fully automated quantitative analyzer. All the 100 patients were followed during the hospital stay and upto discharge or till the outcome. Results: The incidence of thrombocytopenia was 73% indicating a common association in malaria. Complicated malaria was observed in 58.80% of P. falciparum infection whereas 66% of P. vivax infection was associated with uncomplicated malaria. Severe thrombocytopenia showed positive correlation with severity of malaria. Thrombocytopenic patients with effective anti-malarial treatment showed 95.90% recovery and 3 patients 4.10% had mortality. Patients with severe thrombocytopenia were 8.5 times more likely to have complicated malaria with P <0.001 according to student „t‟ test. Conclusion: Thrombocytopenia is the most common hematological finding in malaria. Severe thrombocytopenia showed positive correlation with complicated malaria and a good predictor of poor prognosis. Patients with classical malarial fever and thrombocytopenia who were negative for malaria parasite were not included in the study.

Assessment of Accuracy and Effectiveness of Rapid Diagnostic Test for Malaria Diagnosis at Primary Health Centres in Abeokuta Nigeria.

Malaria is a deadly disease that needs proper and prompt diagnosis in order to treat its symptoms as early as possible. Rapid diagnosis test is a pre-requisite for the effective treatment of malaria in other to reduce the mortality and morbidity of the disease especially at the Primary Health Centre (PHC) facilities. This study compares RDTs test results from PHCs with malaria Quantitative Buffy Coat (QBC) and microscopy test results. A total of 113 subjects with clinical signs of malaria were enrolled after obtaining consent of patients at the Primary Health Centres and questionnaires administered to assess awareness and use of RDTs kit. Storage and compliance to standards of usage of the Kits were observed. The results were analyzed using SPSS version 16.0. There was a significant difference (p<0.05) in sensitivity to malaria parasite between the three diagnostic methods as QBC was more sensitive compared with other diagnostic methods, while Microscopy was more sensitive compared with RDT kits. A total number of 86(76.1%), 28(24.8%) and 39(34.5%) malaria positive cases were detected by QBC, RDT and Microscopy respectively. Out of the 86(76.1%) blood samples confirmed positive by QBC, 27(31.4%) and 38(44.2%) positive cases were detectable by RDT and Microscopy respectively. Furthermore, Microcopy detected 15(53.6%) of the total positive cases detected by RDT, while RDT was able to detect 15(34.5%) of the total positive cases detected by microscopy. When compared with QBC, RDT shown a sensitivity and specificity of 32.56% (95% Cl= 22.84 – 43.52%) and 31.76% (95% Cl= 22.09% - 42.76%) respectively. On the other hand, 71.8% (95% Cl=55.12% - 84.98%) sensitivity and 87.1% (95% Cl= 78.02 – 93.35%) specificity was shown by RDT when microscopy was used as gold standard. Compliance to manufacturer’s instruction on RDT usage was poor as some of the health workers collected the blood sample directly from the pricked finger into the sample well rather than the designated capillary pipette method, while others did not comply with time before reading the results of the kits. The result from this study showed that the sensitivity and accuracy of RDTs kit is low and there is need for proper training of the health workers to avoid misuse of the kit.

Effects of Staurosporine on the Proliferation and Apoptosis of Human Cholangiocarcinoma QBC-939 Cells / 医药导报

Objective To study the effects of protein kinase C (PKC) inhibitor staurosporine (STS) on the proliferation and apoptosis of human cholangiocarcinoma QBC-939 cells and to explore its possible mechanism. Methods CCK-8 was used to detect the effects of PKC inhibitor STS on the proliferation of human cholangiocarcinoma QBC-939 cells. The effects of STS on the ultrastructural characteristics of QBC-939 cells were observed by routine transmission electron microscopy (TEM). The apoptosis rate and the cell cycle distribution of QBC-939 cells were detected by flow cytometry. The expression of cyclin B1,Cdk1 and p-Cdk1 in QBC-939 cells was detected by Western blotting. Results STS could significantly inhibit the proliferation of QBC-939 cells in a dose-dependent manner (P <0. 05) and the half inhibitory concentration ( IC50 ) of QBC-939 cells at 24th and 48th h was 334 nmol·L-1 and 118 nmol · L-1 , respectively. TEM observed that STS could induce typical apoptotic bodies and super-microstructural changes of QBC-939 cells. By Annexin V-FITC/ PI double labeling flow cytometry,we found that the apoptotic rate of QBC-939 cells after treatment with STS for 0,12,24 and 48 h was (10. 16±4. 52)% ,(22. 35±2. 19)% ,(34. 27±2. 30)% and (59. 70±5. 97)% ,respectively. By flow cytometry,compared with the control group,STS could significantly increase apoptosis rate of QBC-939 cells,decrease the percentage of cells in G0 / G1 phase and increase the percentage of cells in G2 / M phase (P<0. 05). Western blotting proved that the expression levels of cyclin B1 and Cdk1 proteins in the STS-treated QBC-939 cells were significantly decreased (P<0. 05),while the expression level of p-Cdk1 protein in the STS-treated QBC-939 cells was significantly increased ( P < 0. 05 ). Conclusion STS can significantly inhibit cell proliferation and induce apoptosis of human cholangiocarcinoma QBC-939 cells and the mechanism may be related to cell cycle arrest at G2 / M phase.

Proliferative activity of bile of patients undergoing transduodenal sphincteroplasty / 中国普通外科杂志

Objective To explore the effects of bile in patients undergoing transduodenal sphincteroplasty (TSB) on the growth of human cholangiocarcinoma cell line QBC939 and the potential relation between transduodenal sphincteroplasty and cholangiocarcinoma. Methods TSB and normal bile (NB) sample were used for this study. The proliferative effect of bile was measured by using methabenzthiazuron (MTT) assay, and cell cycle and apoptosis were analyzed by using flow cytometry. Results Compared with NB,TSB significantly promote the proliferation of human cholangiocarcinoma QBC939 cells (P

Tentativa de evidenciar o Trypanosoma cruzi no sangue periférico de pacientes com doença de Chagas, em fase crônica, por meio do Quantitative Buffy coat (QBC) / An attempt to detect Trypanosoma cruzi in the peripheral blood of patients with Chagas' disease, in chronic phase, using quantitative buffy coat (QBC)

Taking for granted the sensitivity of the Quantitative Buffy Coat (QBC) system, as documented in a murine experimental model, we assayed to detect Trypanosoma cruzi in the peripheral blood of 100 patients with Chagas disease in its chronic phase. By means of the method, no positivity occurred, evently as a consequence of small parasitemias, undetectable by this technique as assessed by the cases in consideration.

Valorizando a sensibilidade do sistema Quantitative Buffy Coat (QBC), documentada em modelo experimental murino, estando os animais com infecção aguda pelo Trypanosoma cruzi houve tentativa de evidenciar esse parasita no sangue periférico de 100 pacientes com doença de Chagas, em fase crônica. Com o emprego desse método, nenhuma positividade ocorreu, evidentemente em virtude das pequenas parasitemias, não reveláveis pela técnica, pelo menos conforme o verificado através da casuística considerada.

Effect of hydroxycamptothecine on PS_2 and COX-2 of the transplanted tumor in the nude mice by human bile duct carcinoma QBC939 cells / 西安交通大学学报(医学版)

Objective To investigate the effect of hydroxycam ptothecine on PS2 and COX-2 in nude mice by human bile duct carcinoma QBC939 cells in order to analyze the correlation between PS2 and COX-2 expressions and the development of the tumor.Methods After the model of the transplanted tumor of subcutaneous tissues located the nude mice was constructed by human bile duct carcinoma QBC939 cells,30 nude mice were randomly divided into 3 groups,namely,control group,hydroxycamptothecine low-dose group and high-dose group.The growth of the transplanted tumor was observed and immunohistochemical staining combined with image analysis was used to determine the expression of PS2 and COX-2 in the tumor tissues.Results Compared with that of the control group,the content of PS2 in the tumor tissues was increased in hydroxycamptothecine low-dose group and high-dose group.The content of COX-2 was decreased and the growth of the transplanted tumor was inhibited in both hydroxycamptothecine low-dose group and high-dose group(P