Characterization of CD103+ CD8+ tissue-resident T cells in esophageal squamous cell carcinoma: may be tumor reactive and resurrected by anti-PD-1 blockade.

Though therapy that promotes anti-tumor response about CD8+ tumor-infiltrating lymphocytes (TILs) has shown great potential, clinical responses to CD8+ TILs immunotherapy vary considerably, largely because of different subpopulation of CD8+ TILs exhibiting different biological characters. To define the relationship between subpopulation of CD8+ TILs and the outcome of antitumor reaction, the phenotype and function of CD103+ CD8+ TILs in esophageal squamous cell carcinoma (ESCC) were investigated. CD103+ CD8+ TILs were presented in ESCC, which displayed phenotype of tissue-resident memory T cells and exhibited high expression of immune checkpoints (PD-1, TIM-3). CD103+ CD8+ TILs were positively associated with the overall survivals of ESCC patients. This population of cells elicited potent proliferation and cytotoxic cytokine secretion potential. In addition, CD103+ CD8+ TILs were elicited potent anti-tumor immunity after anti-PD-1 blockade and were not affected by chemotherapy. This study emphasized the feature of CD103+ CD8+ TILs in immune response and identified potentially new targets in ESCC patients.

miR-1306-3p targets FBXL5 to promote metastasis of hepatocellular carcinoma through suppressing snail degradation.

This study aimed to elucidate the effect of miR-1306-3p on metastasis of hepatocellular carcinoma (HCC) and potential mechanism involved. miR-1306-3p promoted migration and invasion of HCC in vivo and in vitro. Moreover, miR-1306-3p inhibited snail to enhance its expression via directly targeting FBXL5, thus inducing the epithelial-mesenchymal transition (EMT) in HCC. Intriguingly, miR-1306-3p expression was transcriptionally enhanced by FoxM1. Consistently, miR-1306-3p was upregulated in HCC compared with paracarcinoma and correlated with poor prognosis of HCC patients. Our researches suggest that miR-1306-3p is a tumor enhancer in regulating of HCC metastasis, and miR-1306-3p may be clinically utilized as a factor for the clinical diagnosis and prognosis of HCC.

Blocking of the PD-1/PD-L1 Interaction by a D-Peptide Antagonist for Cancer Immunotherapy.

Blockade of the protein-protein interaction between the transmembrane protein programmed cell death protein 1 (PD-1) and its ligand PD-L1 has emerged as a promising immunotherapy for treating cancers. Using the technology of mirror-image phage display, we developed the first hydrolysis-resistant D-peptide antagonists to target the PD-1/PD-L1 pathway. The optimized compound (D) PPA-1 could bind PD-L1 at an affinity of 0.51 µM in vitro. A blockade assay at the cellular level and tumor-bearing mice experiments indicated that (D) PPA-1 could also effectively disrupt the PD-1/PD-L1 interaction in vivo. Thus D-peptide antagonists may provide novel low-molecular-weight drug candidates for cancer immunotherapy.

The immunogenicity of a novel cytotoxic T lymphocyte epitope from tumor antigen PL2L60 could be enhanced by 4-chlorophenylalanine substitution at position 1.

PIWIL2, a member of PIWI/AGO family, is expressed in germline stem cells and precancerous stem cells, but not in adult somatic cells. PIWIL2 plays an important role in tumor development. It is considered as a cancer­testis antigen (CT80). It has been reported that the spliced fragment of PIWIL2, PL2L60, was widely expressed in cancer cell lines. In this study, HLA-A2-restricted epitopes from PL2L60 were predicted by online tools. To improve the activity of the native epitope, a candidate peptide P281 with potent binding affinity was chosen to investigate the modification strategy. A series of aromatic amino acids were introduced to substitute the first residue of P281. Then, we tested the binding affinity and stability of the peptide analogs and their ability to elicit specific immune responses both in vitro and in vivo. Our results indicated that the cytotoxic T lymphocytes (CTLs) induced by [4-Cl-Phe1]P281 could elicit more potent activities than that of P281 and other analogs. The CTLs induced by this analog could lyze target cells in HLA-A2-restricted and antigen-specific manners. [4-Cl-Phe1]P281 also showed the best resistance against degradation in human serum. In conclusion, the introduction of the unnatural amino acid, 4-Cl-Phe, into the first position could enhance the activity of the native epitope to induce cytotoxic T lymphocytes. It might be a good strategy to modify other promising native epitopes. The novel epitopes identified in this study could be used as novel candidates to the immunotherapy of HLA-A2 positive patients with tumors expressing PL2L60.

Lewis acid-catalyzed, copper(II)-mediated synthesis of heteroaryl thioethers under base-free conditions.

A Lewis acid (Ag(I), Ni(II), or Fe(II)) catalyzed, Cu(II)-mediated thiolation reaction between heteroarenes and thiols was achieved with good yield under base-free conditions. DMSO could serve as an effective methylthiolation reagent for the synthesis of heterocyclic methyl thioethers.

In vitro and in vivo identification of a novel cytotoxic T lymphocyte epitope from Rv3425 of Mycobacterium tuberculosis.

The identification of novel cytotoxic T lymphocyte (CTL) epitopes is important to analysis of the involvement of CD8(+) T cells in Mycobacterium tuberculosis infection as well as to the development of peptide vaccines. In this study, a novel CTL epitope from region of difference 11 encoded antigen Rv3425 was identified. Epitopes were predicted by the reversal immunology approach. Rv3425-p118 (LIASNVAGV) was identified as having relatively strong binding affinity and stability towards the HLA-A*0201 molecule. Peripheral blood mononuclear cells pulsed by this peptide were able to release interferon-γ in healthy donors (HLA-A*02(+) purified protein derivative(+)). In cytotoxicity assays in vitro and in vivo, Rv3425-p118 induced CTLs to specifically lyse the target cells. Therefore, this epitope could provide a subunit component for designing vaccines against Mycobacterium tuberculosis.

Combined sevoflurane-dexmedetomidine and nerve blockade on post-surgical serum oxidative stress biomarker levels in thyroid cancer patients.

BACKGROUND: The incidence of thyroid cancer is increasing annually. Clinical routine thyroid surgery can be performed under a cervical plexus block, but cannot mediate the stress response during the surgery. If thyroid surgery is performed under nerve block, an inappropriate level of blockade may occur. Similarly, the stress response caused by surgery is more serious than that caused by conventional anesthesia. Therefore, it is important to combine blockade with more effective anesthesia methods. AIM: To investigate the effects of combining sevoflurane-dexmedetomidine inhalation general anesthesia with the cervical plexus nerve block on the post-surgical levels of the serum oxidative stress biomarkers levels in thyroid cancer patients. METHODS: We enrolled 96 thyroid cancer patients admitted to the hospital between January 2019 and December 2020. Participants were divided into a control group (n = 47) and an experimental group (n = 49). The experimental group received a combination of inhaled sevoflurane-dexmedetomidine and cervical plexus block, while the control group received conventional general anesthesia. The groups were compared for serum levels of monocyte chemotactic protein-1 (MCP-1) and glutathione peroxidase (GSH-Px) before and after surgery, and the adrenocorticotropic hormone (ACTH) and norepinephrine (NE) levels at 1 and 12 h post-surgery. The Bispectral index (BIS) and the incidence of anesthesia side effects were also compared. RESULTS: Following surgery, MCP-1 was significantly lower in the experimental group compared to the control group, whereas GSH-Px was significantly higher than that in the control group (P < 0.001). The serum ACTH and NE levels were significantly lower in the experimental group than those the control group at 1 and 12 h post-surgery (P < 0.001). BIS was significantly lower in the experimental group than that in the control group at 20 minutes into the operation, but the direction of the difference was reversed at eye opening (P < 0.001). The incidence of side effects was 10.20% (5/49) and 12.76% (6/47) in the experimental and control groups, respectively, the difference being non-significant. CONCLUSION: Sevoflurane-dexmedetomidine inhalation general anesthesia combined with cervical plexus nerve block can reduce the postoperative stress and inflammatory responses in thyroid cancer patients, while maintaining high anesthesia effectiveness and safety.

Continuous positive airway pressure versus mandibular advancement device in the treatment of obstructive sleep apnea: a systematic review and meta-analysis.

BACKGROUND: The goal of this study was to review relevant randomized controlled trials in order to determine the efficacy of continuous positive airway pressure (CPAP) versus mandibular advancement device (MAD) in the treatment of obstructive sleep apnea (OSA). METHODS: Using appropriate keywords, we identified relevant studies using PubMed, the Cochrane Library, and Embase. Key pertinent sources in the literature were also reviewed, and all articles published through October 2019 were considered for inclusion. For each study, we used odds ratios (ORs), mean difference (MD), and 95% confidence interval (95% CI) to assess and synthesize outcomes. RESULTS: We included 14 RCTs, for a total of 249 patients in the CPAP group and 247 in the MAD group. Compared with MAD, CPAP significantly decreased apnea hypopnea index (AHI) (WMD: -7.08, 95%CI: -9.06∼-5.10) and the percentage of stage 1 and 2 after therapy (WMD: -3.728, 95%CI: -6.912∼-0.543). However, compared with MAD, CPAP significantly decreased the SF-36-social function score (WMD: -3.381, 95%CI: -6.607∼-0.154).There was no significant difference in Epworth sleepiness scale score after therapy between the two groups. CONCLUSION: CPAP has better therapeutic efficacy in OSA patients than MAD.

Follow-up study of the pulmonary function and related physiological characteristics of COVID-19 survivors three months after recovery.

BACKGROUND: The long-term pulmonary function and related physiological characteristics of COVID-19 survivors have not been studied in depth, thus many aspects are not understood. METHODS: COVID-19 survivors were recruited for high resolution computed tomography (HRCT) of the thorax, lung function and serum levels of SARS-CoV-2 IgG antibody tests 3 months after discharge. The relationship between the clinical characteristics and the pulmonary function or CT scores were investigated. FINDINGS: Fifty-five recovered patients participated in this study. SARS-CoV-2 infection related symptoms were detected in 35 of them and different degrees of radiological abnormalities were detected in 39 patients. Urea nitrogen concentration at admission was associated with the presence of CT abnormalities (P = 0.046, OR 7.149, 95% CI 1.038 to 49.216). Lung function abnormalities were detected in 14 patients and the measurement of D-dimer levels at admission may be useful for prediction of impaired diffusion defect (P = 0.031, OR 1.066, 95% CI 1.006 to 1.129). Of all the subjects, 47 of 55 patients tested positive for SARS-CoV-2 IgG in serum, among which the generation of Immunoglobulin G (IgG) antibody in female patients was stronger than male patients in infection rehabilitation phase. INTERPRETATION: Radiological and physiological abnormalities were still found in a considerable proportion of COVID-19 survivors without critical cases 3 months after discharge. Higher level of D-dimer on admission could effectively predict impaired DLCO after 3 months discharge. It is necessary to follow up the COVID-19 patients to appropriately manage any persistent or emerging long-term sequelae. FUNDING: Key Scientific Research Projects of Henan Higher Education Institutions.

The association of preoperative atrial fibrillation with post-cardiopulmonary bypass hyperfibrinolysis in rheumatic valvular heart disease patients.

OBJECTIVE: The purpose of this study was to assess the fibrinolytic status after cardiopulmonary bypass in rheumatic valvular heart disease patients, and detect the associated factors of post-cardiopulmonary bypass hyperfibrinolysis. METHODS: According to the fibrinolytic status after cardiopulmonary bypass, 203 rheumatic valvular heart disease patients were divided into two groups: hyperfibrinolysis group (H group, n = 78) and non-hyperfibrinolysis group (NH group, n = 125). The demographic characteristics, operative variables, and postoperative follow-ups were compared between these two groups. RESULTS: The incidence of hyperfibrinolysis was 38.4% after cardiopulmonary bypass. Patients in the H group had a significant higher incidence of preoperative atrial fibrillation than patients in the NH group (92.3% vs. 55.2%, P < 0.01). Furthermore, postoperative daily drainage (655.3 ±â€¯131.5 ml vs. 535.4 ±â€¯161.4 ml, P < 0.01), transfusion volume of fresh frozen plasma (621.8 ±â€¯220.2 ml vs. 455.2 ±â€¯208.5 ml, P < 0.01), and red blood cells (5.9 ±â€¯2.2 u vs. 4.7 ±â€¯2.8 u, P < 0.01) was greater in the H group than in the NH group. Moreover, the logistic regression analysis revealed that preoperative atrial fibrillation was associated with post-cardiopulmonary bypass hyperfibrinolysis (OR = 19.691, 95% CI = 6.849-56.612; P < 0.05). CONCLUSION: Preoperative artial fibrillation is associated with post-cardiopulmonary bypass hyperfibrinolysis in rheumatic valvular heart disease patients.

Structure-activity relationships of the dimeric analogues of endomorphin-2 with different lengths of spacers.

In the present study, seven novel dimeric analogues of endomorphin-2 with longer spacers were designed and synthesized. Through dimerization, their affinity for delta-opioid receptor was mostly increased, especially the delta-opioid receptor preferred dimeric analogue, DEM(12). The results were confirmed by the in vitro bioassay. The structure-activity relationships were also discussed.

Positive Correlation Between Regional Cerebral Oxygen Saturation and Mixed Venous Oxygen Saturation During Off-Pump Coronary Artery Bypass Surgery.

BACKGROUND: The balance of oxygen delivery and consumption is essential in patients who are critically ill. Mixed venous oxygen saturation (Sv̄O2 ) is a standard method to evaluate oxygen delivery and consumption during anesthesia. However, Sv̄O2 is monitored through a pulmonary artery catheter, which is invasive. Regional cerebral oxygenation (rScO2) reflects oxygen saturation in a small region of the frontal lobes and is monitored noninvasively through near-infrared spectroscopy. In the present study, the correlation between rScO2 and Sv̄O2 was calculated during off-pump coronary artery bypass grafting surgery to determine whether a positive correlation exists between rScO2 and Sv̄O2 . METHODS: A total of 56 subjects were consecutively enrolled in the study. Then rScO2 and Sv̄O2 were simultaneously monitored. The parameters were recorded at 5 time points: T1, 10 min after intubation (1.0 FIO2 for 10 min); T2, 20 min after intubation (0.60 FIO2 for 10 min); T3, at the end of the revascularization of the left anterior descending artery (0.60 FIO2 ); T4, after protamine infusion (0.60 FIO2 ); and T5, 10 min after protamine infusion (1.0 FIO2 for 10 min). The correlation between rScO2 and Sv̄O2 and the variation trend between rScO2 and Sv̄O2 when FIO2 increased from 0.60 to 1.0 were analyzed. RESULTS: There was a significant positive correlation between rScO2 and Sv̄O2 at these 5 time points (r 2 = 0.77, 0.81, 0.70, 0.83, and 0.92, respectively). There also was a significant positive correlation between Δ rScO2 and Δ Sv̄O2 (n = 112, r 2 = 0.72, P < .001). Linear regression analysis revealed that Sv̄O2 had a positive correlation with rScO2 and cardiac output (r 2 = 0.68, P = .013). CONCLUSIONS: There was a positive correlation between rScO2 and Sv̄O2 during off-pump coronary artery bypass grafting surgery, and there also was a positive correlation in the variation trend between rScO2 and Sv̄O2 .

Use of epoprostenol to treat severe pulmonary vasoconstriction induced by protamine in cardiac surgery.

Since there were a few articles to report the treatment of severe pulmonary vasoconstriction induced by protamine in cardiac surgery, we described the use of epoprostenol to reverse this condition.A total of 5 cases of severe pulmonary vasoconstriction induced by protamine in cardiac surgery were reviewed. The demographic, clinical data and treatment process were obtained. All the patients were followed up.Severe pulmonary vasoconstriction was occurred 4 to 10 minutes after protamine infusion. The primary sign was sudden hypotension, the pulmonary artery pressure was increased gradually, the arterial oxygen partial pressure was decreased in all the patients. Epoprostenol was infused via pulmonary artery catheter at dosage of 20 to 40 ng/kg·min in all the patients, 2 patients were underwent re-cardiac pulmonary bypass assistance. The hemodynamic instability status lasted 40 to 65 minutes respectively. All the patients were recovered uneventfully.All physicians should alert to the incidence of severe pulmonary vasoconstriction induced by protamine in cardiac surgery. Use epoprostenol through pulmonary artery catheter could treat pulmonary artery vasoconstriction effectively and safely.

Intrinsic Expression of Immune Checkpoint Molecule TIGIT Could Help Tumor Growth in vivo by Suppressing the Function of NK and CD8+ T Cells.

TIGIT, an immune checkpoint molecule widely expressed on NK cells, activated T cells and Tregs, has been involved in delivering inhibitory signals through the interaction with PVR. The blockade of TIGIT/PVR interaction is a promising approach in cancer immunotherapy. Here, we unexpectedly discovered the expression of TIGIT in murine tumor cells. To elucidate the mechanism of such intrinsic expression, TIGIT knockout murine colorectal CT26 and MC38 cell lines were generated by using CRISPR/Cas9 system. Although TIGIT knockout showed no effects on proliferation and colony formation of tumor cells in vitro, the tumor growth in mice was considerably inhibited. TIGIT knockout led to the increase of IFN-γ secretion by NK and CD8+ T cells. Further, in BABL/c nude mice, CD8+ T cells depleting mice and NK cells depleting nude mice, the promotion of tumor growth was significantly diminished, suggesting that both NK cells and CD8+ T cells were involved in the tumor promoting process mediated by intrinsic TIGIT. In addition, blocking TIGIT/PVR interaction by the antibody or recombinant PVR protein could elicit anti-tumor effects by facilitating the tumor infiltration and restoring the function of CD8+ T cells, and the antibody-mediate TIGIT blockade could inhibit MC38 tumor growth through blocking TIGIT expressed on tumor cells. We therefore propose a novel TIGIT/PVR interaction mode that tumor intrinsic TIGIT delivers inhibitory signals to CD8+ T cells and NK cells by engaging with PVR.

Roles of nitric oxide synthase inhibitor on antinociceptive effects of mu-opioid agonist in mice.

In the present study, it was found that intraperitoneal (i.p.) pre-injection of N(G)-nitro-L-arginine methyl ester (L-NAME) significantly influenced the endomorphin-1 (EM-1) and endomorphin-2 (EM-2) induced antinociception. These effects could be inhibited or reversed by L-Arg or naloxone. Our results suggest that the modulatory effect of NO system on the mu-receptor evoked analgesia is different between the two mu receptor subtypes.

Polycondensation and carbonization of phenolic resin on structured nano/chiral silicas: reactions, morphologies and properties.

In this research, we extended a bioinspired and templated synthesis way for SiO2 to carbonaceous materials, with the success in morphology control and inducing chirality at the nano-scale. The biopolymer-analogue polyamine, i.e., polyethyleneimine (PEI) was employed as a catalytic template for SiO2 formation, and the as-formed PEI@SiO2 hybrids, which combine the rigidity of SiO2 and the chemical activity of PEI, were further used as hard-templates and basic catalysts for the deposition of phenolic resin on PEI@SiO2 under mild conditions. Through further carbonization and etching SiO2, SiO2/carbon composites and carbonaceous materials were produced, respectively. After characterization of these products by SEM, TEM, XPS, Raman spectroscopy, FT-IR, and TG-DTA, it was demonstrated that the morphologies were well transmitted in these successive steps. By taking advantage of the diverse modulation ways on the morphologies and structures of initial PEI templates, it is easy to achieve SiO2/carbon and carbonaceous products with different morphologies, including nanofibrils, nanobelts, and nanotubes. Moreover, this process could also fulfill a steady chirality transmission. When PEI complexed with chiral tartaric acid, the resulting chiral complex could function both as a template and chirality source, and finally chiral nanostructured carbonaceous products were obtained.

The Association of Peroxisome Proliferator-Activated Receptor δ and Additional Gene-Gene Interaction with C-Reactive Protein in Chinese Population.

Aims. To examine the association between 4 single nucleotide polymorphisms (SNPs) of peroxisome proliferator-activated receptors δ (PPARδ) polymorphisms and C-reactive protein (CRP) level and additional gene-gene interaction. Methods. Line regression analysis was performed to verify polymorphism association between SNP and CRP levels. Generalized multifactor dimensionality reduction (GMDR) was employed to analyze the interaction. Results. A total of 1028 subjects (538 men, 490 women) were selected. The carriers of the C allele (TC or CC) of rs2016520 were associated with a significant decreased level of CRP, regression coefficients was -0.338, and standard error was 0.104 (p = 0.001). The carriers of the G allele (CG or GG) of rs9794 were also significantly associated with decreased level of CRP, regression coefficients was -0.219, and standard error was 0.114 (p = 0.012). We also found a potential gene-gene interaction between rs2016520 and rs9794. Subjects with rs2016520-TC or CC, rs9794-CG or GG genotypes have lowest CRP level, difference (95% CI) = -0.50 (-0.69 to -0.21) (p < 0.001), compared to subjects with rs2016520-TT and rs9794-CC genotypes. Conclusions. rs2016520 and rs9794 minor allele of PPARδ and combined effect between the two SNP were associated with decreased CRP level.

LncSox4 promotes the self-renewal of liver tumour-initiating cells through Stat3-mediated Sox4 expression.

Liver cancer has a tendency to develop asymptomatically in patients, so most patients are diagnosed at a later stage. Accumulating evidence implicates that liver tumour-initiating cells (TICs) as being responsible for liver cancer initiation and recurrence. However, the molecular mechanism of liver TIC self-renewal is poorly understood. Here we discover that a long noncoding RNA (lncRNA) termed LncSox4 is highly expressed in hepatocellular carcinoma (HCC) tissues and in liver TICs. We find that LncSox4 is required for liver TIC self-renewal and tumour initiation. LncSox4 interacts with and recruits Stat3 to the Sox4 promoter to initiate the expression of Sox4, which is highly expressed in liver TICs and required for liver TIC self-renewal. The expression level of Sox4 correlates with HCC development, clinical severity and prognosis of patients. Altogether, we find that LncSox4 is highly expressed in liver TICs and is required for their self-renewal.

Endomorphins, endogenous opioid peptides, provide antioxidant defense in the brain against free radical-induced damage.

Oxidative stress has been considered to be a major cause of cellular injuries in a variety of chronic health problems, such as carcinogenesis and neurodegenerative disorders. The brain appears to be more susceptible to oxidative damage than other organs. Therefore, the existence of antioxidants may be essential in brain protective systems. The antioxidative and free radical scavenging effects of endomorphin 1 (EM1) and endomorphin 2 (EM2), endogenous opioid peptides in the brain, have been investigated in vitro. The oxidative damage was initiated by a water-soluble initiator 2,2'-azobis(2-amidinopropane hydrocholoride) (AAPH) and hydrogen peroxide (H2O2). The linoleic acid peroxidation, DNA and protein damage were monitored by formation of hydroperoxides, by plasmid pBR 322 DNA nicking assay and single-cell alkaline electrophoresis, and by SDS-polyacrylamide gel electrophoresis. Endomorphins can inhibit lipid peroxidation, DNA strand breakage, and protein fragmentation induced by free radical. Endomorphins also reacted with galvinoxyl radicals in homogeneous solution, and the pseudo-first-order rate constants were determined spectrophotometrically by following the disappearance of galvinoxyl radicals. In all assay systems, EM1 was more potent than EM2 and GSH, a major intracellular water-soluble antioxidant. We propose that endomorphins are one of the protective systems against free radical-induced damage in the brain.

Novel epitopes identified from efflux pumps of Mycobacterium tuberculosis could induce cytotoxic T lymphocyte response.

Overcoming drug-resistance is one of the major challenges to control tuberculosis (TB). The up-regulation of efflux pumps is one common mechanism that leads to drug-resistance. Therefore, immunotherapy targeting these efflux pump antigens could be promising strategy to be combined with current chemotherapy. Considering that CD8+ cytotoxic T lymphocytes (CTLs) induced by antigenic peptides (epitopes) could elicit HLA-restricted anti-TB immune response, efflux pumps from classical ABC family (Mycobacterium tuberculosis, Mtb) were chosen as target antigens to identify CTL epitopes. HLA-A2 restricted candidate peptides from Rv2937, Rv2686c and Rv2687c of Mycobacterium tuberculosis were predicted, synthesized and tested. Five peptides could induce IFN-γ release and cytotoxic activity in PBMCs from HLA-A2(+) PPD(+) donors. Results from HLA-A2/K(b) transgenic mice immunization assay suggested that four peptides Rv2937-p168, Rv2937-p266, Rv2686c-p151, and Rv2686c-p181 could induce significant CTL response in vivo. These results suggested that these novel epitopes could be used as immunotherapy candidates to TB drug-resistance.