harvest-primary-culture-leptomeningeal-lymphatic-endothelial

Harvest and Primary Culture of Leptomeningeal Lymphatic Endothelial Cells

Leptomeningeal lymphatic endothelial cells (LLECs) are a recently discovered intracranial cellular population with a unique distribution clearly distinct ...

The First Affiliated Hospital of Kunming Medical University

HPV accounts for most of incidence of cervical cancer. Genetic variations of E6 and E7 may be associated with the development of cervical cancer in specific geographic regions. HPV-58 has been found to be a relatively prevalent high-risk HPV among southwest Chinese women. To explore gene intratypic variations and polymorphisms of HPV-58 E6 and E7 genes originating in Southwest China, a total of 2000 scraped cell samples were collected for DNA extraction and HPV typing. Then, the E6 and E7 genes of HPV-58 (n=22) were sequenced and compared to others submitted to GenBank, followed by an analysis of the diversity of secondary structure by DNASTAR software. Phylogenetic trees were then constructed by Neighbor-Joining and the Kimura 2-parameters methods, followed by an analysis of selection pressures acting on the E6/E7 genes by PAML software. 22 were HPV-58 positive among 215 high-risk types' samples. The nucleotide variation rate of E6 was 86.36% (19/22) among the 22 HPV-58 E6 sequences studied. 4 single nucleotide changes were identified among the E6 sequences with 3/4 synonymous mutations (C187T, A260C, C307T) and 1/4 non-synonymous mutations (A388C, from Lys to Asn, in alpha helix). The most common mutations of E6 genes are the C307T and A388C. 8 single nucleotide changes were identified among the HPV-58 E7 sequences with 2/8 synonymous mutations (T726C, T744G) and 6/8 non-synonymous mutations (G599A, C632T, G694A, G760A, G761A, T803C). The nucleotide variation rate of E7 was 72.73% (17/22). The most common mutations of E7 genes are C632T, G694A, T744G, G760A (from Gly to Ser, in turn), G761A and T803C. The phylogenetic analyses demonstrate that all HPV-58 E6/E7 variants identified belonged to the Southeast Asia lineage. There was no evidence of positive selection in the sequence alignment of HPV-58 E6 and E7 genes.

Genetic variability of HPV-58 E6 and E7 genes in Southwest China.

The advent of disease evaluation by means of multi-slice spiral computed tomography (MSCT) and magnetic resonance imaging (MRI) represents a continually emerging role in the evaluation of various diseases; however, its role is yet to be adequately defined. Thus, the aim of the study was to compare the diagnostic value of MSCT and MRI in the diagnosis of peritoneal metastasis in primary ovarian carcinoma. Between January 2013 and December 2015, MSCT or MRI data were collected from 42 patients who had been previously diagnosed with peritoneal metastasis of ovarian carcinoma at the First Affiliated Hospital of Kunming Medical University. The tumor location, size, edge, and shape were all evaluated independently by three qualified imaging physicians using a double-blind method to confirm whether the patients were indeed suffering from peritoneal metastasis, as well as to rank the metastatic lesions recorded on a five-point scale. It was hypothesized that MRI and MSCT were comparable in the evaluation of ovarian carcinoma. Therefore, a receiver operating characteristics (ROC) curve was used to analyze the results and also to directly compare the respective diagnostic values of MSCT and MRI. In total, 165 metastatic lesions were confirmed by means of surgical operation. MSCT revealed 131 metastatic lesions, while MRI confirmed 154 metastatic lesions. The metastatic sites were primarily located on the subphrenic, epiploon, and gastrocolic ligaments and were further confirmed by either MRI or CT. In regard to MSCT, the most common site of underdiagnoses was in the vicinity of the uterus-rectum-fossa. MRI displayed a high detection rate in every site. The omission diagnostic rate of MSCT and MRI were 20.61% and 6.67%, respectively, while the accuracy rates were 79.39% and 93.33%, respectively. The obtained results revealed that the MSCT value of area under the ROC curve was smaller than that for MRI. Our findings provided evidence asserting that MRI, in comparison to MSCT, was more accurate in diagnosing peritoneal metastasis in patients with ovarian carcinoma.

Comparison between multi-slice spiral CT and magnetic resonance imaging in the diagnosis of peritoneal metastasis in primary ovarian carcinoma.

To investigate the regulatory mechanism of miR-152 on proliferation, metastasis and tumorigenesis of human acute monocytic leukemia SHI-1 cells.

[Regulatorg Mchanism of MiR-152 on Proliferation, Metastasis and Tumorigenicity of SHI-1 Cells].

To investigate the biological effects and mechanism of WNK1 on K562 cells through regulating MAPK7.

[Effects of WNK1 on Human Chronic Myeloid Leukemia K562 Cells via MAPK7 Phosphorylation and Its Relative Mechanism].

Although chimeric antigen receptor T-cell (CAR-T) therapy development for B-cell malignancies has made significant progress in the last decade, broadening the success to treating T-cell acute lymphoblastic leukemia (T-ALL) has been limited. We conducted two clinical trials to verify the safety and efficacy of GC027, an "off-the-shelf" allogeneic CAR-T product targeting T-cell antigen, CD7. Here, we report 2 patients as case reports with relapsed/refractory T-ALL who were treated with GC027.

Eradication of T-ALL Cells by CD7-targeted Universal CAR-T Cells and Initial Test of Ruxolitinib-based CRS Management.

The Warburg effect is involved in drug resistance and recurrence of cancer, and poses a challenge for the treatment of chronic myelogenous leukemia (CML). Hypoxia-inducible factor 1α (HIF-1α) plays a key role in the Warburg effect. microRNAs (miRs) targeting HIF-1α have potential of regulating such aberrant metabolic process. The present study demonstrated that miR-18a-5p was expressed at a low level in K562/ADM cells via reverse transcription-quantitative PCR (RT-qPCR). The results of the luciferase reporter assay indicated that miR-18a-5p could specifically bind the 3'-untranslated region of HIF-1α. Through RT-qPCR and western blotting, it was revealed that miR-18a-5p downregulated the expression of HIF-1α. By inhibiting HIF-1α, miR-18a-5p suppressed aerobic glycolysis in K562/ADM cells, according to the results produced by glucose uptake, lactate production, pyruvate level and ATP synthesis measurement, along with the results obtained from extracellular acidification rate and oxygen consumption rate assays. These results provided new evidence that miR-18a-5p may suppress the Warburg effect by targeting HIF-1α. Furthermore, via CCK-8 and flow cytometry assays, cells transfected with miR-18a-5p mimics were more sensitive to Adriamycin (AMD) compared with AMD group. Reversing the Warburg effect by miR-30a-5p might provide a potential therapeutic strategy for CML.

MicroRNA-18a-5p regulates the Warburg effect by targeting hypoxia-inducible factor 1α in the K562/ADM cell line.

To investigate the regulatory effect and mechanism of DNA methyltransferase 3A (DNMT3a) in hydroquinone-induced hematopoietic stem cell toxicity.

[Role of DNMT3a in Hydroquinone-Induced Hematopoietic Stem Cell Toxicity].

Evodia rutaecarpa has multiple pharmacological effects and is widely used in the prevention and treatment of migraine, diabetes, cardiovascular disease, cancer, and other chronic diseases; however, the pharmacological effects of its active compound evodiamine (Evo) have not been thoroughly investigated. The purpose of this study was to investigate the effects of Evo on antiplatelet activation and thrombosis. We discovered that Evo effectively inhibited collagen-induced platelet activation but had no effect on platelet aggregation caused by activators such as thrombin, ADP, and U46619. Second, we found that Evo effectively inhibited the release of platelet granules induced by collagen. Finally, evodiamine inhibits the transduction of the SFKs/Syk/Akt/PLC2 activation pathway in platelets. According to in vivo studies, Evo significantly prolonged the mesenteric thromboembolism induced by ferric chloride and had no discernible effect on the coagulation function of mice. In conclusion, the antiplatelet and thrombotic effects of Evo discovered in this study provide an experimental basis for the investigation of the pharmacological mechanisms of Evo and the development of antiplatelet drugs.

Effect of Evodiamine on Collagen-Induced Platelet Activation and Thrombosis.

The occurrence and development of malignancies are closely related to abnormal cell cycle regulation. Myeloid leukemia factor 1 (MLF1) is a small nucleocytoplasmic shuttling protein associated with cell cycle exit, apoptosis, and certain immune functions. Therefore, it is pertinent to explore the role of MLF1 in health and diseases. Studies to date have suggested that MLF1 could act as a double-edged sword, regulating biochemical activities directly or indirectly. In hematopoietic cells, it serves as a protective factor for the development of lineages, and in malignancies, it serves as an oncogenesis factor. The diversity of its functions depends on the binding partners, including tumor inhibitors, scaffolding molecules, mitochondrial membrane proteins, and transcription factors. Emerging evidence indicates that MLF1 influences immune responses as well. This paper reviews the structure, biological function, and research progress on MLF1 in health and diseases to provide new insights for future research.

Kun Wu

Department of Clinical Laboratory

The First Affiliated Hospital of Kunming Medical University

Harvest and Primary Culture of Leptomeningeal Lymphatic Endothelial Cells

Kun Wu

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Clinical Laboratory

The First Affiliated Hospital of Kunming Medical University

Harvest and Primary Culture of Leptomeningeal Lymphatic Endothelial Cells

Department of Clinical Laboratory