pH-sensitive fluorescent proteins (FPs) are highly advantageous for the non-invasive monitoring of exocytosis events. Superecliptic pHluorin (SEP), a green pH-sensitive FP, has been widely used for ...imaging single-vesicle exocytosis. However, the docking step cannot be visualized using this FP, since the fluorescence signal inside vesicles is too low to be observed during docking process. Among the available red pH-sensitive FPs, none is comparable to SEP for practical applications due to unoptimized pH-sensitivity and fluorescence brightness or severe photochromic behavior. In this study, we engineer a bright and photostable red pH-sensitive FP, named pHmScarlet, which compared to other red FPs has higher pH sensitivity and enables the simultaneous detection of vesicle docking and fusion. pHmScarlet can also be combined with SEP for dual-color imaging of two individual secretory events. Furthermore, although the emission wavelength of pHmScarlet is red-shifted compared to that of SEP, its spatial resolution is high enough to show the ring structure of vesicle fusion pores using Hessian structured illumination microscopy (Hessian-SIM).
The tumor microenvironment (TME) is the internal environment of malignant tumor progression, and the host antitumor immune response and normal tissue destruction occur in the TME. Tumor-infiltrating ...lymphocytes (TIL) is a crucial component of the TME and reflect the host antitumor immune response. The purpose of this study was to discuss the methodology for TIL evaluation and assess the prognostic value of TIL in gastric cancer. In total, we reviewed 1,033 gastrectomy cases between 2002 and 2008 at the Third Affiliated Hospital of Soochow University. To understand the prognostic value of TIL in gastric cancer (GC), TIL were assessed by optical microscopy, and verified by immunohistochemistry. There is no current consensus on TIL scoring in GC. In this study, we discussed a TIL evaluation system that includes an analysis of the amount and percentage of TIL in a tumor. Ultimately, 439 (52.7%) cases showed high levels of TIL and 394 (47.3%) cases had low levels. There was a statistically significant relationship among TIL, tumor size, histological grade, LN metastasis, nerve invasion, tumor thrombus, pTN stage, and WHO subtypes (
< 0.001, respectively). TIL
was a positive significant predictor of overall survival (OS) in Kaplan-Meier survival analysis (
< 0.001) and multivariate Cox regression analysis (
= 0.431, 95% CI: 0.347-0.534,
< 0.001). After surgery, patients with malignant tumors underwent chemoradiotherapy according to standard therapeutic guidelines based on TNM stage. The TNM scoring system cannot reflect the full information of TME; therefore, TIL can be used as a diagnostic supplement. We constructed a nomogram model that showed more predictive accuracy for OS than pTN stage. In summary, this study proves that high levels of TIL are associated with a positive prognosis and that TIL reflect the protective host antitumor immune response.
Engineered tissue constructs require the fabrication of highly perfusable and mature vascular networks for effective repair and regeneration. In tissue engineering, stem cells are widely employed to ...create mature vascularized tissues in vitro. Pericytes are key to the maturity of these vascular networks, and therefore the ability of stem cells to differentiate into pericyte-like lineages should be understood. To date, there is limited information regarding the ability of stem cells from the different tissue sources to differentiate into pericytes and form microvascular capillaries in vitro. Therefore, here we tested the ability of the stem cells derived from bone marrow (BMSC), dental pulp (DPSC) and dental apical papilla (SCAP) to engineer pericyte-supported vascular capillaries when encapsulated along with human umbilical vein endothelial cells (HUVECs) in gelatin methacrylate (GelMA) hydrogel. Our results show that the pericyte differentiation capacity of BMSC was greater with high expression of α-SMA and NG2 positive cells. DPSC had α-SMA positive cells but showed very few NG2 positive cells. Further, SCAP cells were positive for α-SMA while they completely lacked NG2 positive cells. We found the pericyte differentiation ability of these stem cells to be different, and this significantly affected the vasculogenic ability and quality of the vessel networks. In summary, we conclude that, among stem cells from different craniofacial regions, BMSCs appear more suitable for engineering of mature vascularized networks than DPSCs or SCAPs.
Background/Aims: The role of Tumor-infiltrating lymphocytes (TILs) in the prognosis of patients with lung cancer is still controversial. We performed a meta-analysis to evaluate the prognostic role ...of TILs in lung cancer. Methods: Studies were recruited by searching PubMed, Embase and the Cochrane Library and assessed by further quality evaluation. The pooled hazard ratios (HRs) with 95% confidence intervals (CIs) were calculated to investigate the association between TIL subsets and lung cancer patients' outcome. Results: A total of 29 articles including 8,600 patients were enrolled into the meta-analysis. Our results indicated that high level of CD8+ cells infiltration in tumor stroma (TS) or tumor nest (TN) was associated with better OS in lung cancer patients (HR = 0.76, 95% CI 0.62-0.93, P = 0.006; HR = 0.80, 95% CI 0.67-0.96; P = 0.018, respectively). Similar results could be also observed in CD3+ T cells infiltration. High CD4+ T lymphocytes infiltration in TS was explicitly accompanied by better OS (HR = 0.65, 95% CI 0.46-0.91; P = 0.013), rather than in TN. In contrast, high density of FOXP3+ T cells infiltration in TS showed a poor PFS (HR = 2.67, 95% CI, 1.74-4.08, P < 0.001). Conclusion: This meta-analysis clarified that high level of CD8+ and CD3+ T cells infiltration in TS or TN, and high CD4+ T lymphocytes infiltration in TS showed better OS in lung cancer patients, whereas high density of FOXP3+ T cells infiltration in TS could be recognized as a negative prognostic factor.
Macrophages constitute a major component of tumor-infiltrating immune cells. M2 macrophages have been reported to promote tumor progression through promoting tumor angiogenesis and metastasis and ...regulating T-cell function. Here, we identified a protumorigenic subset of macrophages that constitutively expressed programmed cell death 1 (PD1) and accumulated in advanced-stage gastric cancer (GC). These PD1
tumor-associated macrophages (TAMs) exhibited an M2-like surface profile, with a significant increase in the expression of CD206, IL-10, and CCL1, and a clear decrease in the expression of MHC class II, CD64, and IL-12 and the ability to phagocytose ovalbumin. Moreover, PD1
TAMs can suppress CD8
T-cell function and this immunosuppressive activity can effectively be enhanced upon triggering PD1 signal. GC-derived exosomes effectively educated monocytes to differentiate into PD1
TAMs with M2 phenotypic and functional characteristics. Together, our results are the first to show that GC-derived exosomes can effectively induce PD1
TAM generation, and these cells can produce a large number of IL-10, impair CD8
T-cell function, and thereby create conditions that promote GC progression. Thus, methods in which immunotherapy is combined with targeting PD1
TAMs and tumor-derived exosomes should be used to restore immune function in GC patients.
Super-resolution correlative light and electron microscopy (SR-CLEM) is a powerful approach for imaging specific molecules at the nanoscale in the context of the cellular ultrastructure. Epon epoxy ...resin embedding offers advantages for SR-CLEM, including ultrastructural preservation and high quality sectioning. However, Epon embedding eliminates fluorescence from most fluorescent proteins. We describe a photocontrollable fluorescent protein, mEosEM, that can survive Epon embedding after osmium tetroxide (OsO
) treatment for improved SR-CLEM.
Background
An increasing number of studies have found that long non-coding RNAs (lncRNAs) play an important role in carcinogenesis and tumor progression, whereas their molecular mechanisms of ...function remain largely unknown.
Aims
This study was aimed to explore the biological function and underlying mechanism of a new lncRNA LINC00200 in gastric cancer (GC).
Methods
qRT-PCR analysis was conducted to examine the LINC00200 expression level in both GC tissues and cell lines. Functional assays were carried out to detect the effect of LINC00200 on GC cell proliferation, invasion and migration. The interaction between LINC00200 and miR-143-3p was confirmed by luciferase reporter assays. Rescue assays were performed to confirm the influence of LINC00200-miR-143-3p-SERPINE1 axis on GC development.
Results
LINC00200 was found to be upregulated in GC tissues and cell lines. Moreover, knockdown of LINC00200 suppressed GC cell proliferation, invasion and migration in vitro and inhibited tumorigenesis in mouse xenografts. Finally, mechanism research indicated that LINC00200 functioned as a ceRNA to sponge for miR-143-3p, thus leading to the disinhibition of its target gene SERPINE1.
Conclusions
LINC00200 is significantly overexpressed in GC and accelerates GC progression through regulating miR-143-3p/SERPINE1 axis. Our results may provide a potential diagnostic biomarker and therapeutic target for the management of GC patients.
Background
PD-L1 expression in tumor cells can predict the efficacy of PD-1/PD-L1 inhibitors and prognosis in patients. However, the correlation between the PD-L1 expression and the novel lung ...adenocarcinoma classification are obscure.
Methods
94 lung adenocarcinoma cases were reviewed in the Third Affiliated Hospital of Soochow University from January to December 2019. PD-L1 (DAKO 22C3) was used to test the PD-L1 expression in lung cancer tissue.
Result
TPS was used to interpret the PD-L1 expression. The negative, low positive and high positive of PD-L1 were 52 cases (55.30%), 29 cases (30.90%) and 13 cases (13.80%). The subtype ratio of acinar, lepidic and solid in adenocarcinoma were correlation with the PD-L1 TPS (
r
= − 0.37,
P
< 0.001;
r
= − 0.22,
P
= 0.013;
r
= 0.68,
P
< 0.001). The results of
χ
2
test showed the PD-L1 expression had the significant difference with gender (
P
= 0.027), age (
P
= 0.018), smoking history (
P
= 0.021), lymph node metastasis (
P
= 0.001), TNM stage (
P
= 0.002), acinar structure (
P
= 0.017) and solid structure (
P
< 0.001). Multi-factor linear regression results suggested that solid structure, TNM stage and smoking history were associated with PD-L1 expression (
P
< 0.05). The solid structure showed more capability to PD-L1 expression (
β
= 0.398).
Conclusion
PD-L1 expression was heterogeneity in lung adenocarcinoma. The solid structure, TNM stage and smoking history were correlation to up-regulation of PD-L1 expression, and solid structure was the most importance factor.
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•Quasi-columnar (Gd0.9Yb0.1)2Zr2O7 (GYbZ)/yttria-stabilized zirconia (YSZ) coating experiences delamination failure under burner rig test.•Delamination initiates at particle-embedded ...regions beneath taller GYbZ column tips, followed by lower regions affected by shorter tapered columns.•The discontinuities of the strain tolerance lead to multiple out-of-plane stress concentration spots in the quasi-columnar coating.•Various strain energy release rate trends at different stress concentration spots reveal delamination mechanisms of quasi-columnar GYbZ/YSZ coatings.
A double-ceramic-layer (DCL) thermal barrier coating consisting of quasi-columnar structured (Gd0.9Yb0.1)2Zr2O7 (GYbZ) on top and yttria-stabilized zirconia (YSZ) beneath was deposited by plasma spray-physical vapor deposition (PS-PVD). The as-deposited GYbZ/YSZ coating is composed of a dense continuous layer at the bottom and a columnar layer growing on it. The columnar layer comprises taller coarse columns, shorter tapered columns, and particles filling in the gaps between the columns. The thermal shock behavior of the coating is evaluated in a burner rig at 1300 °C, during which the coating mainly fails in the form of delamination. The coating initially delaminates in the regions where the particles are embedded beneath the taller GYbZ columns tips. Subsequently, delamination occurs in the lower region of GYbZ coating and eventually fails at the TGO/YSZ interface. A finite element model tailored to the microstructural characteristics of the DCL coating was established. Stress distributions of the coating indicate that the discontinuities of the strain tolerance caused by abrupt microstructural or componential changes lead to multiple out-of-plane stress (σ22) concentration spots. The position and procedure of the delamination of GYbZ coating obtained based on the energy release rate of each σ22 concentration spot are consistent with experimental results.
•GO was deposited on the surface of sintered NdFeB by electrophoretic deposition.•EPD-GO coating is reduced partially.•EPD-GO coating provides a corrosion barrier to sintered NdFeB from NaCl aqueous ...solution.•EPD-GO is more stable than GO in NaCl solution owing to the reduction.
Graphene oxide (GO) was deposited uniformly on the surface of permanent magnet material NdFeB by electrophoretic deposition (EPD). Electrophoretic deposited graphene oxide (EPD-GO) coating was reduced partially after EPD process, owing to the removal of oxygen functional groups. And EPD-GO coating showed excellent adhesion to the NdFeB matrix. According to the results of electrochemical tests, the decrease in corrosion current density and the positive shift in corrosion potential have both demonstrated that EPD-GO coating served as a corrosion inhibitor, protecting NdFeB from NaCl aqueous solution.
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