Drug-resistant gene polymorphisms in Plasmodium falciparum isolated from Bioko Island, Equatorial Guinea in 2018 and 2019 / 中国血吸虫病防治杂志

Objective To investigate the genetic polymorphisms of Plasmodium falciparum multidrug resistance protein 1 (PfMDR1), chloroquine resistance transporter (PfCRT) and Kelch 13 (PfK13) genes in Bioko Island, Equatorial Guinea, so as to provide insights into the development of the malaria control strategy in local areas. Methods A total of 85 peripheral blood samples were collected from patients with Plasmodium falciparum infections in Bioko Island, Equatorial Guinea in 2018 and 2019, and genomic DNA was extracted. The PfMDR1, PfCRT and PfK13 genes were amplified using a nested PCR assay. The amplification products were sequenced, and the gene sequences were aligned. Results There were no mutations associated with artemisinin resistance in PfK13 gene in Bioko Island, Equatorial Guinea, while drug-resistant mutations were detected in PfMDR1 and PfCRT genes, and the proportions of PfMDR1_N86Y, PfMDR1_Y184F and PfCRT_K76T mutations were 35.29% (30/85), 72.94% (62/85) and 24.71% (21/85), respectively. Conclusion There are mutations in PfMDR1, PfCRT and PfK13 genes in P. falciparum isolates from Bioko Island, Equatorial Guinea.

Efficacy and Safety of Low-dose Amphotericin B in Different Antifungal Strategies for Treatment of Invasive Fungal Disease in Patients with Hematological Malignancies / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of low-dose amphotericin B (AmB) in different antifungal strategies for treatment of invasive fungal disease(IFD) in patients with hematologic malignancies. Metheds: The clinical dada of the patients were collected and analyzed retrospectively and the levels of creatinine (Cr), urea nitrogen (BUN) and potassium (K) before and after using low-dose AmB were compared and statistically analyzed.</p><p><b>RESULTS</b>Among 97 cases, 2 cases were diagnosed as invasive fungal disease (IFD), 11 cases were diagnosed as clinical probable IFD, 15 cases were diagnosed as possible IFD, 69 cases were undefined IFD. The response rate of all patients treated with low-dose AmB was 69.4%, the response rate for targed therapy was 72.7%, the response rate for diagnosis-driven therapy was 63.6%, the response rate of empirical therapy was 75%, the efficacy of the combination with other antibiotics was 50%, 66.7% and 75%. According to all the patients received AmB, only 7 cases was detected with higher level of Cr (7.2) than normal and this level come back to normal with in 7 days after drug withdrew. Although the Cr level in serum after 1 day of drug withdrew was higher than that before administration of drug(64.86±3.00 vs 58.76±1.67 µmol/L) and was with statistical difference(P<0.05), but did not show significant difference in comparison with the level after drug withdrew 7 days (58.43±1.68 µmol/L,P>0.05).</p><p><b>CONCLUSION</b>AmB injection is an effective and safe method in empirical therapy and diagnosis-driven antifungal therapy for neutropenic, febrile patients with hematological malignancies.</p>

Analysis of miRNA differential expression in peripheral blood cells of primary immune thrombocytopenia patients / 中国实验血液学杂志

This study was purposed to clarify the difference of microRNA (miRNA) expression in the peripheral blood cells of patients with primary immune thrombocytopenia (ITP) and normal controls. Exqion miRCURY(TM) microarray was used to investigate differentially expressed miRNA of peripheral blood cells obtained from affected ITP patients and the healthy controls. Cluster analysis was used to identify miRNA expression profile between the ITP patients and the healthy controls. Real-time PCR was used for validation. The results showed that a total of 159 miRNA were found to be differentially expressed in ITP patients compared to the controls, with 79 up-regulated and 80 down-regulated. Based on these differentially expressed miRNA, a tree with clear distinction between the controls and ITP patients was generated by cluster analysis. Real-time PCR confirmed microarray analysis results. It is concluded that differentially expressed miRNA were found in the peripheral blood cells from ITP patients, which may be potential novel biomarkers for ITP as well as help to elucidate pathogenic mechanisms of ITP.

Apoptosis of KBM5R cell line with T315I point mutation induced by arsenic trioxide / 中国实验血液学杂志

This study was aimed to investigate the inducing-apoptosis effect of arsenic trioxide (ATO) on imatinib (IM)-resistant chronic myeloid leukemia (CML) cell line KBM5R with T315I point mutation. CML cell line KBM5R with T315I point mutation and wild-type cell line KBM5 were selected for study. Resistance of KBM5R cells to IM and proliferation of KBM5 and KBM5R cells treated with ATO were detected by MTT; apoptosis of KBM5 and KBM5R cells were quantified by flow cytometry; the expression of apoptosis-related protein caspase-3, -8, -9 was determined by Western blot. The results showed that (1) IC(50) of KBM5R and KBM5 cells treated with IM were 12.66 ± 0.565 µmol/L and 0.303 ± 0.031 µmol/L respectively, and significantly different from each other. (2) the proliferation of KBM5 and KBM5R cells treated with different concentrations of ATO was inhibited in dose- and time-dependent manners at 24, 48, 72, 96 hours, and inhibition of KBM5R cell proliferation was stronger than KBM5 in the same drug concentration and time. (3) the apoptosis rate of KBM5 and KBM5R cells treated with 2, 4, 8 µmol/L ATO for 48 hours increased in a concentration-dependent manner, and the apoptosis rate of KBM5R was higher than that of KBM5 cells in the same drug concentration. (4) the expression of cleaved caspase-3, -8, -9 protein in KBM5 and KBM5R cells treated with 4 µmol/L ATO for 24 hours significantly increased. It is concluded that KBM5R cells are significantly resistant to IM; ATO can inhibit the proliferation and induce the apoptosis of KBM5R and KBM5 cells. As compared with wild-type KBM5 cells, effect of ATO on inhibition of proliferation and induction of apoptosis in KBM5R cells are more stronger. ATO can induce the apoptosis of KBM5 and KBM5R cells through the activation of apoptosis-related caspase-3, -8, -9 protein.

Effect of Twist gene on the migration and invasion of gastric carcinoma cells / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of Twist gene on the migration and invasion of human gastric carcinoma cells.@*METHODS@#MKN28 cells, a human gastric carcinoma cell line, were transfected with PcDNA3.1-Twist plasmid by lipofectamine transfecting technique. The transfected cells were selected with geneticin. Expressions of Twist,ecadherin and vimentin protein were detected by Western blot in cells transfected Twist gene. Matrigel invision chambers were performed to analyse the cell migration and invasion.@*RESULTS@#MKN28 cells transfected with PcDNA3.1-Twist plasmid showed stronger intracellular expression of Twist protein than MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. The expression of ecadherin protein in MKN28 cells transfected with PcDNA3.1-Twist plasmid was significantly decreased compared with that in MKN28 cells transfected with PcDNA3.1 and MKN28 cells without the transfection. However, The expression of vimentin protein in MKN28 cells transfected with PcDNA3.1-Twist plasmid was significantly increased compared with that in MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. The migration and invasion ability of Twist+ - MKN28 cells were stronger than that of MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection.@*CONCLUSION@#Twist gene may promote the migration and invasion ability of gastric carcinoma cells through epithelial mesenchymal transition.

Effect of EphA2 protein on the expression of VEGF and MMP9 proteins in HCT116 cells / 中南大学学报(医学版)

OBJECTIVE@#To determine the effect of EphA2 protein on the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 9 (MMP9) proteins in HCT116 cells.@*METHODS@#High expression of EphA2 protein in HCT116 cells was confirmed by Western blot. HCT116 cells were transfected with EphA2 antisense oligonucleotide. The expression of the transfection efficiency was analyzed by Western blot. VEGF proteins in the cell supernatants were detected by enzyme linked immunosorbent assay(ELISA), and the expressions of MMP9 in cell supernatants were examined by gelatin zymography.@*RESULTS@#EphA2 antisense oligonucleotide suppressed the expression of VEGF and MMP9 proteins in HCT116 cells.@*CONCLUSION@#EphA2 could decrease the invasion and metastasis of HCT116 cells by suppressing the expression of VEGF and MMP9.

Growth suppression of colon cancer cells in vitro by DPC4 gene expression and its mechanism / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the effect of DPC4 gene expression on the growth of colon cancer cells and its mechanism.</p><p><b>METHODS</b>Expression plasmid pcDNA3.1-DPC4 was constructed and transfected into the colon cancer cell line SW620 by use of lipofectamine gene transfer technique. DPC4 protein expression was detected by Western blot and immunocytochemistry. The effect of DPC4 gene on the growth of SW620 cells was monitored by population doubling time (PDT) and cloning efficiency. The influence of DPC4 expression on p21WAF1 transcription was investigated by RT-PCR to detect p21WAF1 mRNA.</p><p><b>RESULTS</b>Successful expression of DPC4 protein was detected in the transfected SW620 cells. Compared with the control cells, PDT (74 h) of the DPC4 expressing cells was prolonged and the cloning efficiency (21%) decreased. In addition, the mRNA level of p21(WAF1) in DPC4 transfected cells was increased.</p><p><b>CONCLUSIONS</b>Overexpression of DPC4 gene inhibits the growth of colon cancer in vitro, and induction of p21(WAF1) expression may be an important functional aspect of DPC4.</p>