Abstract We have previously demonstrated the cardioprotective effects of exosomes derived from mesenchymal stem cells (MSCs). It is well known that the activation of Akt is involved in stem cell-induced cardioprotection. In the present study, we investigated whether exosomes released from Akt-overexpressing MSCs showed a beneficial effect on cardioprotection and angiogenesis. MSCs were collected from human umbilical cord (hucMSCs), and Akt was transfected into hucMSCs (Akt-hucMSCs) by using an adenovirus transfection system. Exosomes were isolated from control hucMSCs (Exo) and Akt-hucMSCs (Akt-Exo). An acute myocardial infarction model was created by ligation of the left anterior decedent coronary artery (LAD) in rats. Various source exosomes (400 µg of protein) were infused via the tail vein immediately after LAD ligation. The cardiac function was evaluated by using echocardiography after different treatments for 1 and 5 weeks, respectively. Endothelial cell proliferation, migration, and tube-like structure formation, as well as chick allantoic membrane assay, were used to evaluate the angiogenetic effects of Akt-Exo. The results indicated that cardiac function was significantly improved in the animals treated with Akt-Exo. In addition, Akt-Exo significantly accelerated endothelial cell proliferation and migration, tube-like structure formation in vitro, and blood vessel formation in vivo. The expression of platelet-derived growth factor D (PDGF-D) was significantly upregulated in Akt-Exo. However, the angiogenesis was abrogated in endothelial cells treated with the exosomes obtained from MSCs transfected with PDGF-D-siRNA. Our studies suggest that exosomes obtained from Akt-modified hucMSCs are more effective in myocardial infarction therapy through promoting angiogenesis. PDGF-D plays an important role in Akt-Exo-mediated angiogenesis.
Objective: To investigate whether the characteristics of human umbilical cord mesenchymal stem cells(hucMSCs) labeled with CM-Dil will be changed;and if CM-Dil labled hucMSCs can be traced in vivo.Methods: HucMSCs were isolated from human umbilical cord.The hucMSCs of passage 3 were selected for labeling in vitro.Using cell counting method to analyse growth curves of CM-Dil labeled hucMSCs;observed CM-Dil labeled hucMSCs in vivo with imaging instrument.Results: The efficiency of CM-Dil dye-labeled hucMSCs was high.The proliferation of labeled hucMSCs had no obvious change.CM-Dil dye-labeled hucMSCs can be observed in vivo.Conclusion: CM-Dil labeled hucMSCs can be traced in vivo.
Cellular immunotherapy exploits the capacity of the human immune system in self-protection and surveillance to achieve the anti-tumor effects. Natural killer (NK) cells are lymphocytes of innate immune system and they display a unique inherent ability to identify and eliminate tumor cells. In this review, we first introduce the basic characteristics of NK cells in the physiological and pathological milieus, followed by a discussion of their effector function and immunosuppression in the tumor microenvironment. Clinical strategies and reports regarding NK cellular therapy are analyzed in the context of tumor treatment, especially against solid tumors. Given the widely studied T-cell therapy in the recent years, particularly the chimeric antigen receptor (CAR) T-cell therapy, we compare the technical features of NK- and T-cell based tumor therapies at the clinical front. Finally, the technical challenges and potential solutions for both T and NK cell-based immunotherapies in treating tumor malignancies are delineated. By overviewing its clinical applications, we envision the NK-cell based immunotherapy as an up-and-comer in cancer therapeutics.
Objective: To construct green fluorescence protein(GFP)labeled recombinant adenovirus vector carrying TNFα-Tumstatin fusion gene and transfect it into hUCMSCs.Methods: The amplification products of TNFα-Tumstatin by PCR with a pair of primers with BglⅡand HindⅢ restriction endonuclease sites and signal peptide sequence of GM-CSF were subcloned into shuttle plasmid pAdtrack-CMV after digesting with BglⅡand HindⅢ.The resultant plasmid,after linearized by digesting with restriction endonuclease PmeⅠ,was transformed into E.coli.BJ5183 that had been transformed by adenoviral backbone plasmid pAdEasy-1.Recombinant plasmid were obtained by alternation of kanamycin and then confirmed by PCR and restriction endonuclease analysis.The adenovirus was packaged and propagated in human embryonal kidney cells(293A cells)after being linearized by digesting with restriction endonuclease PacⅠand then was transfected into hUCMSCs.Fusion gene expression in infected hUCMSCs was detected by RT-PCR. Results: The results of PCR and restriction endonuclease assay indicated that target gene was inserted into recombinant adenovirus vector successfully.The sequence of fusion gene was the same as that of designed fragments.hUCMSCs infected by the recombinant adenovirus expressed GFP and fusion gene which could be demonstrated by fluorescence microscope and RT-PCR respectively. Conclusion: Recombinant adenovirus vector containing TNFα-Tumstatin has been constructed successfully and could transfected hUCMSCs efficiently,which laid a foundation for further investigation of anti-tumor effect of hUCMSCs modified with TNFα-Tumstatin.
Bats are important reservoirs for many kinds of emerging zoonotic viruses. In order to explore potential pathogens carried by bats and trace the source of adenovirus outbreaks on the southeastern coast of China, we took pharyngeal and anal swabs from a total of 552 bats (Rhinolophus pusillus) collected from various areas of Chinese southeastern coast. Adenoviruses were identified in 36 out of the 552 samples (6.5%) . Complete genome sequences of two adenovirus isolations from Vero E6 cells were obtained, which were further validated as identical strains via next-generation sequencing and were named Bat-Advcxc6. The cell culture inoculated with the two samples exhibited remarkable cytopathic changes. The full genome has 37,315 bp and owns 29 open reading frames. Phylogenetic analyses confirmed that Bat-Advcxc6 represented a novel bat adenovirus species in the genus Mastadenovirus. Transmission electron microgram showed clear virus particles. Bat-Advcxc6 shared similar characteristics of G + C contents with Bat mastadenovirus WIV11 (Bat mastadenovirus C) found in China in 2016, but differed from this serotype due to a <75% similarity with DNA polymerase amino acid sequences in WIV11. As it is a newly found adenovirus strain according to the international classification criteria, further analyses of virus dynamics, epithelial invasion, and immunization assays are required to explore its potential threats of cross-species transmission.
Background. The prognostic role of CRP (C-reactive protein) in gynecological tumors has been previously reported in individual studies, but whether CRP can be used as a separate potential prognostic factor has not been systematically reviewed. The purpose of this research is to determine if there is a link between CRP levels and the prognosis of gynecological cancer patients. Methods. A systematic search was carried out to find the literature evaluating the predictive role of CRP in the prognosis of gynecological cancer patients. For the purpose of determining the relationship between CRP and clinicopathological characteristics, the pooled odds ratio (OR) was calculated. A hazard ratio (HR) with a 95% confidence interval (CI) was used to determine differences in overall survival (OS), disease-free survival (DFS), or progression-free survival (PFS) between patients with low and high CRP levels. Results. A total of 19 studies, including 4062 patients, were analyzed retrospectively. The FIGO stage was related to the CRP level (OR = 0.43, 95% CI: 0.19–1.00). Age, lymph node metastasis, and histological grade were not associated with CRP level (OR = 0.93, 95% CI: 0.69–1.25; OR = 0.91, 95% CI: 0.65–1.28; OR = 0.74, 95% CI: 0.52–1.05). Worse OS (HR = 1.40, 95% CI: 1.23–1.57), DFS (HR = 1.20, 95% CI: 1.12–1.28), and PFS (HR = 1.57, 95%CI: 1.23–1.91) were associated with elevated CRP levels, as shown by the pooled results. Subgroup analysis was performed according to cancer type (endometrial cancer: HR = 1.15, 95% CI: 1.02–1.28; ovarian cancer: HR = 1.67, 95% CI: 1.03–2.31; cervical cancer: HR = 1.42, 95% CI: 1.19–1.64), multivariate value (HR = 1.22, 95% CI: 1.10–1.33), and age (HR = 1.50, 95% CI: 1.28–1.72). Significant correlations were observed between CRP and OS. Conclusions. CRP may be utilized as a prognostic indicator for a variety of gynecologic malignancies, including cervical cancer, ovarian cancer, endometrial cancer, and vulvar cancer.
Abstract Background: To date, there have been no reported trials that directly compare pembrolizumab/carrelizumab monotherapy versus pembrolizumab/carrelizumab and chemotherapy in the first-line treatment setting of advanced/metastatic non-small cell lung cancer (NSCLC). We performed a Bayesian network meta-analysis of randomized controlled trials (RCTs) to compare the efficacy and safety of pembrolizumab/carrelizumab versus pembrolizumab/carrelizumab and chemotherapy in previously treated patients with NSCLC. Methods: The following search terms would be used in PUBMED, Scopus, EMBASE, and Cochrane Library databases on July 20, 2021, as the search algorithm: (pembrolizumab) OR (carrelizumab) OR (programmed death-1) AND (non-small cell lung cancer) OR (NSCLC). All RCTs that reported the outcomes of pembrolizumab/carrelizumab with or without chemotherapy compared with those of pembrolizumab/carrelizumab alone for patients with NSCLC were considered eligible for inclusion in this meta-analysis. The primary outcomes of interest were overall survival, progression-free survival, objective response rate based on the Response Evaluation Criteria in Solid Tumors for complete and partial responses, and treatment-related adverse events including immune-related adverse events. Secondary outcomes included overall survival, progression-free survival, objective response rate, and treatment-related adverse events for the FDA-approved doses. Conclusions: The results of our review will be reported strictly following the PRISMA criteria and the review will add to the existing literature by showing compelling evidence and improved guidance in clinic settings. Ethical approval: As this study is on the basis of published or registered previous studies, ethical approval and informed consent of patients are not required.
Regulatory T cells (T(regs)) have potential applications in clinical disease therapy, such as autoimmune diseases and transplant rejection. However, their numbers are limited. Forkhead box protein 3 (FoxP3) is a key transcription factor that controls T(reg) development and function. Here, we generated a cell-permeable fusion protein, protein transduction domain (PTD)-conjugated mouse FoxP3 protein (PTD-mFoxP3), and evaluated whether PTD-mFoxp3 can alleviate rheumatoid arthritis (RA) in the collagen-induced arthritis (CIA) mouse model. As expected, PTD-mFoxP3 was transduced into cells effectively, and inhibited T cell activation and attenuated the cell proliferation. It decreased interleukin (IL) 2 and interferon (IFN)-γ expression, and increased IL-10 expression in activated CD4(+)CD25(-) T cells. PTD-mFoxP3-transduced CD4(+)CD25(-) T cells attenuated proliferation of activated CD4(+)CD25(-) T cells. In addition, PTD-mFoxP3 blocked the Th17 differentiation programme in vitro and down-regulated IL-17 production from T cells by modulating induction and levels of retinoid-related orphan receptor gamma t (RORγt). Intra-articular delivery of PTD-mFoxP3 delayed disease incidence remarkably and alleviated autoimmune symptoms of CIA mice. Moreover, protective effects of PTD-mFoxP3 were associated with regulating the balance of T helper type 17 (Th17) and T(regs). These results suggest that PTD-mFoxP3 may be a candidate for RA therapy.
To investigate multitissue engraftment of human primitive hematopoietic cells and their differentiation in goats, human CD34 + Lin − cord blood cells transduced with a GFP vector were transplanted into fetal goats at 45–55 days of gestation. GFP + cells were detected in hematopoietic and nonhematopoietic organs including blood, bone marrow, spleen, liver, kidney, muscle, lung, and heart of the recipient goats (1.2–36% of all cells examined). We identified human β2 microglobulin-positive cells in multiple tissues. GFP + cells sorted from the perfused liver of a transplant goat showed human insulin-like growth factor 1 gene sequences, indicating that the engrafted GFP + cells were of human origin. A substantial fraction of cells engrafted in goat livers expressed the human hepatocyte-specific antigen, proliferating cell nuclear antigen, albumin, hepatocyte nuclear factor, and GFP. DNA content analysis showed no evidence for cellular fusion. Long-term engraftment of GFP + cells could be detected in the blood of goats for up to 2 yr. Microarray analysis indicated that human genes from a variety of functional categories were expressed in chimeric livers and blood. The human/goat xenotransplant model provides a unique system to study the kinetics of hematopoietic stem cell engraftment, gene expression, and possible stem cell plasticity under noninjured conditions.
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