Intranasal delivery of NS1-deleted influenza virus vectored COVID-19 vaccine restrains the SARS-CoV-2 inflammatory response

The emergence of SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus-2) variants and "anatomical escape" characteristics threaten the effectiveness of current coronavirus disease (COVID-19) vaccines. There is an urgent need to understand the immunological mechanism of broad-spectrum respiratory tract protection to guide broader vaccines development. In this study, we investigated immune responses induced by an NS1-deleted influenza virus vectored intranasal COVID-19 vaccine (dNS1-RBD) which provides broad-spectrum protection against SARS-CoV-2 variants. Intranasal delivery of dNS1-RBD induced innate immunity, trained immunity and tissue-resident memory T cells covering the upper and lower respiratory tract. It restrained the inflammatory response by suppressing early phase viral load post SARS-CoV-2 challenge and attenuating pro-inflammatory cytokine (IL-6, IL-1B, and IFN-{gamma}) levels, thereby reducing excess immune-induced tissue injury compared with the control group. By inducing local cellular immunity and trained immunity, intranasal delivery of NS1-deleted influenza virus vectored vaccine represents a broad-spectrum COVID-19 vaccine strategy to reduce disease burden.

The expression and prognostic significance of Aurora Kinase A in hepatocellular carcinoma / 中华肝胆外科杂志

Objective:To explore the expression of Aurora Kinase A (AURKA) in hepatocellular carcinoma (HCC) and its prognostic value.Methods:mRNA expression profiles and clinical data of HCC patients were downloaded from the Cancer Genome Atlas (TCGA) database. Expression of AURKA mRNA in HCC patients of TCGA database from normal liver tissue and all tumor tissues, normal tissues adjacent to cancer and matched tumor tissues were analyzed, and then expression of AURKA to was investigated in HCC tissues and normal liver tissues in the Human Protein Atlas (HPA) database. According to the TNM stage information of HCC patients in TCGA database, expression of AURKA in different stages was analyzed. Kaplan-Meier method was used to analyze whether the high and low expression of AURKA in HCC patients of TCGA database (with the median as the cut-off value) was significantly related to the length of survival. The RNA-seq expression profile data of HCC patients in the public resource platform of the Kaplan-Meier Plotter website was used for external verification. Cox univariate and multivariate analysis were performed on the age, sex, degree of differentiation, TNM stage, and AURKA mRNA expression of TCGA database patients.Results:374 cases of HCC tumor tissues and 50 cases of adjacent normal liver tissues in the TCGA database were included. All HCC tumor tissues in the TCGA database compared with the paired adjacent tissues mRNA level of AURKA was significantly increased, and the protein level was also increased, the difference was statistically significant ( P<0.05); With the tumor TNM stage increase of AURKA mRNA expression showed a gradual upward trend, and the difference was statistically significant ( P<0.05); in the TCGA database HCC cohort, high expression of AURKA mRNA was associated with poor HCC prognosis, and was obtained in Kaplan Meier Plotter database. The difference was statistically significant ( P<0.05); Cox multivariate regression analysis showed that TNM stage ( HR=1.69, 95% CI: 1.37-2.10) and AURKA mRNA expression level ( HR=1.03, 95% CI: 1.01-1.10) are the independent prognostic factors of HCC patients. Conclusions:AURKA is highly expressed in HCC, which is associated with the poor prognosis of HCC. AURKA is an independent prognostic factor of HCC.

Clinical efficacy of hyperthermic intraperitoneal chemotherapy plus cytoreductive surgery for peritoneal metastasis of gallbladder cancer / 中国肿瘤临床

Objective: To investigate the clinical efficacy of hyperthermic intraperitoneal chemotherapy (HIPEC) for gallbladder cancer with peritoneal metastasis. Methods: Data of 84 patients, who were admitted to Shanghai Eastern Hepatobiliary Surgery Hospital from January 2015 to January 2018, were retrospectively analyzed. Of the total patients, 31 received HIPEC combined with cytoreductive surgery (CRS) plus postoperative systemic chemotherapy one month after surgery as the study group (Group A), and the other 53 underwent CRS plus postoperative systemic chemotherapy one momth after surgery as the control group (Group B). The clinical effects and adverse reactions in the two groups were observed and compared. Results: The median survival time in the Group A was (21.72±2.96) months, significantly longer than that of (14.93±2.09) months in Group B (P0.05). Conclusions: HIPEC has significant clinical efficacy for gallbladder cancer with peritoneal metastasis. HIPEC can prolong the survival time and have less side effects.

The effects of targeted inhibition of hedgehog signaling pathway on proliferation, apoptosis and metastasis of hepatocellular carcinoma / 中华肝胆外科杂志

Objective To observe Hedgehog signaling pathways of liver cancer cell growth and the influence of the metastatic potential targeted inhibit Hedgehog.Methods Construction of Smo shRNA plasmid,The stable and low-expressed Smo-expressing HCC QGY-7701 cell line was screened after lipofection.The stable and low-expressed Smo-expressing HCC QGY-7701 cell line was screened,The cell cycle,apoptosis,invasion and metastasis of QGY-7701 cells were detected by Western blot,flow cytometry,CCK8 and transwell assay.Subcutaneous implantation of hepatocarcinoma cells in nude mice.Study on the growth and metastasis of hepatocarcinoma cells with low expression of Smo in.The ultrastructural changes of hepatoma cells with low expression of Smo were observed under electron microscope.Results RT-PCR and Western blot showed stable shR-Smo cell line was successfully constructed.Cell cycle test showed that compared with the control group,G0/G1 cells increased in shR-Smo,cells in S phase decreased;apoptosis,CCK8 and Transwell tests showed that Smo-gene silencing could significantly increase the apoptosis percentage of the hepatic cancer cells to (5.46％ ± 1.46％),proliferation activity decreasedand and the migration rates reduced to (7.82％ ±2.14)％;nude mice model showed that Smo-gene silencing could inhibit the growth of hepatocellular carcinoma cells in vivo,electron microscopy revealed that lysosomes increased significantly in Smo-gene silence cells.Conclusions Blocking Hh signaling pathways,liver cancer cells in vitro malignant degree of decline.Hedgehog in treating liver cancer have hidden meaning.

Application of sequential hepatic arterial and selective portal venous embolization prior to extended radical hepatectomy in patients with marginally resectable hepatocellular carcinoma / 中华肝胆外科杂志

Objective To evaluate the feasibility and efficacy of preoperative sequential transcatheter arterial chemoembolization (TACE) followed by selective portal venous embolization (PVE) in patients with marginally resectable hepatocellular carcinoma (HCC).The aim was to find out whether this combined procedure helped to increase the rate of extended radical liver resection.Methods From March 2009 to November 2016,29 patients with HCC which were marginally resectable underwent preoperative TACE combined with PVE were included into this study.All these patients were subsequently assessed to undergo radical hepatectomy.The complications,laboratory results,volume changes of each liver lobe and patient survival were analyzed.Results TACE combined with PVE was successful in all the 29 patients.There were no major complications.After the procedure,the volumes of the tumor and the part of the liver to be resected decreased to certain degree.The remnant liver volume (RLV) increased remarkably.The RLV were (395.4 ±58.7) cm3 and (599.2 ±75.2) cm3 before and after the procedure,respectively.The difference was significant (P ＜ 0.05).19 patients underwent radical hemihepatectomy or trisectionectomy,with a resection rate of 65.5％ (19/29).There were sufficient surgical margins in all the resected tumors.After operation,the 1-,3-,and 5-year survival rates were 58.8％,35.5％ and 17.6％,respectively.Conclusion For HCC patients who had marginally resectable HCC,preoperative TACE combined with PVE efficiently controlled the growth of the tumors,decreased the volume of the liver lobe with tumor,increased the RLV,and made it possible for a planned two-stage radical hepatectomy with sufficient surgical margin and better survival in a significant proportion of patients.

Influx of extracellular calcium participates in rituximab-enhanced ionizing radiation-induced apoptosis in Raji cells.

We have previously shown that rituximab has a significant radiosensitizing effect on Raji cells in vitro. To investigate whether calcium signals participate in rituximab- and radiation-induced cell death in Raji cells, confocal laser scanning microscopy was used to detect kinetic changes in intracellular free calcium concentration ([Ca2+]i). Cell survival, the rates of apoptosis in Raji cells and the kinetics of γ-H2AX foci induction and loss were also evaluated. X-irradiation of Raji cells induced an initial increase of [Ca2+]i in both the presence and absence of extracellular calcium, followed by a decrease in [Ca2+]i over time. Rituximab enhanced both the amplitude and the duration of intracellular calcium signals in the irradiated cells. EGTA significantly inhibited radiation- or radiation/rituximab combination treatment-induced apoptosis. However, the calcium chelators EGTA and BAPTA/AM conferred no survival advantage on the irradiated cells. Furthermore, although no significant difference was seen after 1h, the treatment of cells with a combination of irradiation and rituxiamb caused an increase of γ-H2AX foci when compared with irradiated cells after 8h. Both EGTA and BAPTA/AM suppressed the number of γ-H2AX foci induced by either radiation or radiation combined with rituximab. Our results suggest that rituximab increases the level of [Ca2+]i in irradiated Raji cells. The entry of calcium from the extracellular space plays an essential role in [Ca2+]i-dependent radiation-induced apoptosis in Raji cells. The calcium chelators inhibited the formation of γ-H2AX foci, which are thought to prevent the activation of Ca2+/Mg(2+)-dependent endonucleases and subsequent DNA fragmentation. The calcium chelators most likely modulate only particular features of apoptosis and fail to change the fates of cells that are already committed to die.

Rituximab sensitizes a Burkitt lymphoma cell line to cell killing by X-irradiation.

Clinical trials with rituximab in combination with chemotherapeutic regimens have shown promising results. Data on the effects of rituximab treatment in combination with irradiation are, however, limited and inconsistent. This study aims to investigate the effects of rituximab (R) on cell death induced by X-irradiation in Raji lymphoma cells and to evaluate its mechanisms. We found the cell growth inhibition by irradiation was enhanced by additional rituximab exposure both in cells precultured with rituximab followed by irradiation (R + irradiation) or in cells treated in the reverse sequence (irradiation + R). R + irradiation combination treatment induced more apoptotic cells than irradiation and irradiation + R treatment as early as 12 h after treatment. At 24 h, both combination treatments, R + irradiation and irradiation + R, showed apoptotic cells, which were significantly different from irradiation alone. G2/M cell cycle arrest was observed after irradiation alone and the combination treatment. The combination treatment revealed an elevation in reactive oxygen species (ROS) generation in a radiation dose-dependent manner. In addition, rituximab enhanced the cell growth inhibition and apoptotic cell death induced by the oxidative agent, H(2)O(2). We propose that rituximab mediates a significant in vitro radiosensitizing effect and induces cell cycle changes and apoptosis in Raji cells. ROS probably play an important role in these events.

Chromatid-type aberrations following irradiation in G0 lymphocytes with heavy ions.

We describe a low level of chromatid-type aberrations which included the relatively rare isochromatid/chromatid triradial in peripheral blood lymphocytes that were irradiated, ostensibly in G0, with accelerated heavy (12)C ions. These were produced only at the energies of 69 MeV/n (34.6 keV/microm), almost absent at the energy of either 58.6 MeV/n (46.07 keV/ microm) or 19.3 MeV/n (97 keV/microm), nor were they found after low-LET X-rays. Mechanisms potentially responsible for their formation are discussed.

Correlation between initial chromatid damage and survival of various cell lines exposed to heavy charged particles.

The biophysical characteristics of heavy ions make them a rational source of radiation for use in radiotherapy of malignant tumours. Prior to radiotherapy treatment, a therapeutic regimen must be precisely defined, and during this stage information on individual patient radiosensitivity would be of very great medical value. There are various methods to predict radiosensitivity, but some shortfalls are difficult to avoid. The present study investigated the induction of chromatid breaks in five different cell lines, including one normal liver cell line (L02), exposed to carbon ions accelerated by the heavy ion research facility in Lanzhou (HIRFL), using chemically induced premature chromosome condensation (PCC). Previous studies have reported the number of chromatid breaks to be linearly related to the radiation dose, but the relationship between cell survival and chromatid breaks is not clear. The major result of the present study is that cellular radiosensitivity, as measured by D0, is linearly correlated with the frequency of chromatid breaks per Gy in these five cell lines. We propose that PCC may be applied to predict radiosensitivity of tumour cells exposed to heavy ions.