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Cytokine‐induced killer (CIK) cells are a group of highly heterogeneous T cells expressing diverse T‐cell antigen receptors (TCRs), but possessing wide major histocompatibility complex (MHC)‐unrestricted antitumour activity, which have been used in the treatment of many tumours. However, the clonal diversity of different T‐cell subsets and whether there is a certain clonal preference during CIK culture remains to be clarified. We found that the proportion of oligoclones seen in some subfamilies of freshly isolated T cells decreased in varying degrees after culturing. This indicates that the diversity of T‐cell clones had been restored to a certain extent. The diversity differed among different T‐cell subsets, as some TCR subfamily oligoclones mainly appeared in CD3+CD56+ cells, which also indicates the heterogeneity of the CIK cell composition. [ABSTRACT FROM AUTHOR]

Highlights: [•] We isolated CD8+TCM from healthy donors. [•] We than transferred TCM with the gene encoding TCRs specific for tumor antigen. [•] TE derived from TCM exhibited improved survival and reacquired the TCM phenotype. [•] TCR gene transfer effectively promoted the specific killing of TCM. [•] TCM were more effective than CD8+T cells in effector function after gene transfer. [ABSTRACT FROM AUTHOR]

Context: The recent developments in non-viral gene therapy and DNA vaccine have fostered the development of efficient plasmid DNA (pDNA) purification processes. Objectives: This work aimed to establish a cost-effective purification process for the large-scale production of plasmid DNA for gene therapy and DNA vaccine. Materials and methods: E. coli DH5α harboring pCDNA3.1- GFP (7200 base pairs) was used as a model plasmid. Hydrophobic-interaction chromatography (HIC) was employed to purify supercoiled plasmid DNA (sc pDNA). Results: With this method, not only host contaminants, but also open circular plasmid DNA (oc pDNA) could be removed from sc pDNA. Anion-exchange HPLC analysis proved that the recovery of HIC could reach 75%. The plasmid DNA exhibited high purity with supercoiled percentage of 98 ± 1.2% and undetectable residual endotoxins, genomic DNA, RNA and protein. The purity of pDNA had nothing to do with the flow rate in the range at least up to 400 cm/h. Liposomes transfection experiment prove that the purified pDNA in this article had higher transfection efficiency than the control pDNA. Discussion and conclusion: In the present work, we confirmed the possibility of separation of sc pDNA from oc pDNA and other host contaminants using a single HIC chromatography. [ABSTRACT FROM AUTHOR]

Most of the malignant melanomas are already in the middle and advanced stages when they are diagnosed, which is often accompanied by the metastasis and spread of other organs. Besides, the prognosis of patients is bleak. The characteristics of the local immune microenvironment in metastatic melanoma have important implications for both tumor progression and tumor treatment. In this study, data on patients with metastatic melanoma from the TCGA and GEO datasets were selected for immune, stromal, and estimate scores, and overlapping differentially expressed genes were screened. A nine-IRGs prognostic model (ALOX5AP, ARHGAP15, CCL8, FCER1G, GBP4, HCK, MMP9, RARRES2 and TRIM22) was established by univariate COX regression, LASSO and multivariate COX regression. Receiver operating characteristic curves were used to test the predictive accuracy of the model. Immune infiltration was analyzed by using CIBERSORT and Xcell in high-risk and low-risk groups. The immune infiltration of the high-risk group was significantly lower than that of the low-risk group. Immune checkpoint analysis revealed that the expression of PDCD1, CTLA4, TIGIT, CD274, HAVR2 and LAG3 demonstrated the visible difference in groups with different levels of risk scores. WGCNA analysis found that the yellow-green module contained seven genes from the nine-IRG prognostic model, and the yellow-green module had the highest correlation with risk scores. The results of GO and KEGG suggested that the genes in the yellow-green module were mainly enriched in immune-related biological processes. Finally, the expression characteristics of ALOX5AP, ARHGAP15, CCL8, FCER1G, GBP4, HCK, MMP9, RARRES2 and TRIM22 were analyzed between metastatic melanoma and normal samples. Overall, a prognostic model for metastatic melanoma based on the tumor immune microenvironment characteristics was established, which left plenty of space for further studies. It could function well in helping people to understand characteristics of the immune microenvironment in metastatic melanoma.

Alternative splicing produces multiple mRNA variants of TP53 which have diverse biologic functions. In this study, we identified a novel splice variant of TP53 lacking a 200 nt portion of exon 4 (p53ΔE4p) from a human leukemia T cell line. No protein product of p53ΔE4p was identifiable by western blot; however, forced expression of the variant in HEK-293T cells expressing wild-type p53 could inhibit cell proliferation and promote cell death. Interestingly, this novel variant also significantly enhances the expression of reporter genes. Moreover, transcriptome analysis showed that genes related to DNA binding and regulation of transcription by RNA polymerase II function were significantly upregulated following p53ΔE4p transfection, suggesting a role for this variant in the regulation of gene expression., Competing Interests: None., (IJCEP Copyright © 2020.)

Objective: To evaluate the method, safety and short-term efficacy of ultrasound-guided percutaneous laser ablation (PLA) for benign solid thyroid nodule (BSTN)., Methods: The treatment group consisting of 12 patients with single BSTN were treated with ultrasound-guided PLA. After treatment, the size and blood flow signals in conventional ultrasonography, the ablation extent of lesions in contrast-enhanced ultrasonography (CEUS), and the related complications and thyroid function were assessed in comparison with the the baseline. A control group including 12 untreated patients with single BSTN was followed-up in the same manner as in the treatment group., Results: In the treatment group, all the 12 patients completed the treatment successfully. During the follow-up, the volume of the nodules was reduced gradually (P<0.05), and blood flow signals in the nodules disappeared. Eleven nodules were non-enhanced in CEUS immediately after the treatment, and the complete ablation rate was 91.7%. During the operation, 3 patients complained of intolerable pain, which was ameliorated or totally relieved after appropriate treatment. The thyroid function showed no significant changes after the treatment (P>0.05). The control group followed up for 6 months showed no significant changes in the nodule volume or thyroid function in comparison with the baseline (P>0.05)., Conclusion: Ultrasound-guided PLA is a safe and effective minimally invasive treatment of BSTN without obvious adverse effect on thyroid function.

Objective: To explore the appropriate acoustic parameters for augmenting the permeability of tumor cells using microbubble-enhanced ultrasound., Methods: Ninety intramuscular VX2 tumors grown in New Zealand white rabbits were randomly divided into 4 parts by 4 factors, namely sound intensity, pulse width, insonation time and microbubble dose, each at different levels. Therapeutic ultrasound was delivered directly to the tumor surface during intravenous infusion of microbubbles. The tumor was excised after the treatment and sliced to investigate the change in tumor cell permeability using lanthanum nitrate electron microscopy. The appropriate acoustic parameters of the 4 factors were investigated., Results: Sonoporation induced by microbubble-enhanced ultrasound augmented the permeability of the tumor cells, and the appropriate sound intensity, pulse width, insonation time and microbubble dose were 0.43 W/cm(2), 2.42 ms, 10 min, and 0.50 ml/kg, respectively., Conclusion: Sonoporation induced by microbubble-enhanced ultrasound can induce pores in tumor cell membrane, which may potential increase the efficacy of tumor chemotherapy.

Objective: To assess the clinical value of transoral carotid ultrasonography (TOCU) in the detection of the blood flow parameters of the distal extracranial internal carotid artery (ICA) in patients with moyamoya disease., Methods: Fourteen patients with the diagnosis of moyamoya disease confirmed by digital subtractive angiography (DSA) underwent examination with conventional carotid ultrasonography of the proximal internal carotid artery (ICA). TOCU was performed to observe the distal extracranial ICA and the internal diameter, peak systolic velocity (PSV) and end-diastolic velocity (EDV) were measured., Results: All the patients had unilateral moyamoya disease. Compared with the contralateral ICA, distal extracranial ICA showed a significantly reduced internal diameter, PSV and EDV (P<0.05). No significant differences were found in the internal diameter, PSV or EDV between the distal and proximal ICA on the diseased side. The internal diameter of the bilateral common carotid arteries (CCA) was similar, but the PSV and EDV were significantly lower in the contralateral CCA., Conclusion: TOCU can provide information of the blood flow in the distal extracranial ICA to facilitate an accurate diagnosis of moyamoya disease.

Objective: To study angiogenesis and the expression of nm23-H(1) tumor metastatic suppressor gene in primary breast carcinoma and their relationship with axillary lymph node metastasis., Methods: Intratumoral vascularization in 80 cases of breast cancer was observed preoperatively by power doppler imaging (PDI) and analysed with computer-assisted quantative assessment, and the expression of microvessel density (MVD) and nm23 protein was determined by immunohistochemical technique., Results: Blood flow signals and blood vessels postitive area within masses were more in axillary node positive (LN+) group than in axillary node negative (LN-) group (t = 7.07, P < 0.01), MVD expression were higher in the LN+ group than in the LN- group (t = 6.34, P < 0.01). Meanwhile the expression of nm23-H(1) protein was lower in the LN+ group than in the LN- group, and significant correlation was found between angiogenesis and the expression of nm23-H(1) protein., Conclusions: Angiogenesis and the expression of nm23-H(1) protein may play an important role in the lymphatic metastasis process of breast cancer. Furthermore, angiogenesis may correlates with the expression of nm23-H(1) protein in that progress.