Vitamin C-induced competitive binding of HIF-1α and p53 to ubiquitin E3 ligase CBL contributes to anti-breast cancer progression through p53 deacetylation.

Vitamin C (VC), in regard to its effectiveness against tumors, has had a controversial history in cancer treatment. However, the anticancer mechanisms of VC are not fully understood. Here, we reported that VC exerted an anticancer effect on cancer cell and xenograft models via inhibiting HIF-1α-dependent cell proliferation and promoting p53-dependent cell apoptosis. To be specific, VC modulated the competitive binding of HIF-1α and p53 to their common E3 ubiquitin ligase CBL, thereby inhibiting tumorigenesis. Moreover, VC treatment activated SIRT1, resulting in p53 deacetylation and CBL-p53 complex dissociation, which in turn facilitated CBL recruitment of HIF-1α for ubiquitination in a proteasome-dependent manner. Altogether, our results provided a mechanistic rationale for exploring the therapeutic use of VC in cancer therapy.

Therapy of genomic unstable solid tumours (WHO grade 3/4）in clinical stage III/IV using individualised neoantigen tumour peptides-INP trial (individualised neoantigen tumour peptides immunotherapy): study protocol for an open-label, non-randomised, prospective, single-arm trial.

INTRODUCTION: Neoantigens derived from tumour somatic mutations are recognised as ideal vaccine targets. Tumour neoantigens have been studied in a wide range of tumours. Most of research on neoantigens has focused just on a unique tumour and a single mutated gene. Currently, a few studies have reported using a mixture of neoantigen peptides derived from multiple genetic mutation sites in the treatment of genomic unstable advanced solid malignancies. The trial aims to evaluate the safety and efficacy of individualised tumour neoantigen peptide mixtures in the treatment of genomic unstable advanced solid malignant tumours. METHODS AND ANALYSIS: This is a prospective, non-randomised, open, single-centre, single-arm, phase I trial. Patients with genomic unstable advanced solid malignancies are eligible for study participations. 20 patients will be included in the trial. Through the whole exome and transcriptome sequencing analysis of the fresh blood and tumour tissues of the enrolled patients, the 20 25-33aa antigen peptides with the highest mutation scores of the patients will be screened out, and the corresponding new antigen peptides will be synthesised and prepared. Patients will be treated with their own individualised neoantigen polypeptide combined with a polypeptide adjuvant (human granulocyte-macrophage colony-stimulating factor). The primary endpoint is safety indicators, including general and specific adverse events which will be monitored continuously. Secondary endpoints are progression-free survival, objective response rate, objective duration of remission, 1-year survival rate and overall survival. ETHICS AND DISSEMINATION: This study has received approval from the Ethics Committee of Chongqing University Cancer Hospital on 21 November 2019 (207/2019). The findings of this trial will be disseminated through national and international presentations and peer-reviewed publications. TRIAL REGISTRATION NUMBER: ChiCTR1900025364.

Compare the Efficacy and Safety of Modified Combined Short and Long Axis Method versus Oblique Axis Method for Right Internal Jugular Vein Catheterization in Adult Patients (The MCSLOA Trial): Study Protocol of a Randomized Controlled Trial.

Background: Ultrasound-guided internal jugular vein (IJV) catheterization has become a standard procedure as it yields a higher success rate and fewer mechanical complications compared with an anatomical landmark technique. There are several common methods for ultrasound guidance IJV catheterization, such as short-axis out-of-plane, long-axis in-plane and oblique axis in-plane, but these technologies are still developing. It is important to further study the application of different ultrasound-guided IJV puncture techniques and find an effective and safe ultrasound-guided puncture technique. Methods: A China randomized, open-label, parallel, single center, positive-controlled, non-inferiority clinical trial will evaluate 190 adult patients undergoing elective surgery and need right jugular vein catheterization. Study participants randomized in a 1:1 ratio into control and experimental groups. The control group will take the oblique axis in-plane method for IJV catheterization. The experimental group will take the Modified combined short and long axis method. The primary endpoint of the trial is the rate of one-time successful guidewire insertion without posterior wall puncture (PWP). Secondary endpoints are the number of needle insertion attempts, the total success rate, the procedure time, and mechanical complications. Conclusion: This randomized controlled trial will evaluate the effectiveness and safety of Modified combined short and long axis method and oblique axis in-plane method for right IJV catheterization in adult patients.

Construction and Validation of an Immune-Related lncRNA Prognosis Model for Thyroid Cancer.

BACKGROUND: Immune-related long noncoding RNAs (lncRNAs) play an important role in the development of cancer. This study aimed to identify immune-related lncRNAs in thyroid cancer (THCA) and develop a prognostic model for THCA. METHODS: We downloaded immune-related gene sets from the Gene Set Enrichment Analysis (GSEA) website and obtained THCA gene expression and clinical data from The Cancer Genome Atlas (TCGA) database. Immune-related lncRNAs were then obtained by performing correlation analysis on the expression of lncRNAs and immune-related genes. A prognostic model for THCA immune-related lncRNAs was developed through univariate Cox regression and multiple Cox regression analyses. We confirmed the results in clinical samples using quantitative real-time PCR. RESULTS: A total of 26 immune-related lncRNAs in THCA were obtained. Then we constructed a prognosis model composed of seven lncRNAs (LINC01614, AC017074.1, LINC01184, LINC00667, ACVR2B-AS1, AC090673.1, and LINC00900). Our model can be used as an independent prognostic factor. Principal component analysis displayed that the lncRNAs in the model can distinguish between high and low-risk groups. Clinical correlation analysis showed that the expression levels of AC090673.1 (P<0.05), LINC01184 (P<0.001), and LINC01614 (P<0.001) were related to disease stage, and LINC00900 (P<0.001) and LINC01614 (P<0.001) were related to T stage. We validated this model in cancer and paracancerous tissues from 24 THCA patients. CONCLUSION: We identified and experimentally validated seven immune-related lncRNAs that can serve as potential biomarkers for THCA prognosis.

Nucleocapsid mutations R203K/G204R increase the infectivity, fitness, and virulence of SARS-CoV-2.

Previous work found that the co-occurring mutations R203K/G204R on the SARS-CoV-2 nucleocapsid (N) protein are increasing in frequency among emerging variants of concern or interest. Through a combination of in silico analyses, this study demonstrates that R203K/G204R are adaptive, while large-scale phylogenetic analyses indicate that R203K/G204R associate with the emergence of the high-transmissibility SARS-CoV-2 lineage B.1.1.7. Competition experiments suggest that the 203K/204R variants possess a replication advantage over the preceding R203/G204 variants, possibly related to ribonucleocapsid (RNP) assembly. Moreover, the 203K/204R virus shows increased infectivity in human lung cells and hamsters. Accordingly, we observe a positive association between increased COVID-19 severity and sample frequency of 203K/204R. Our work suggests that the 203K/204R mutations contribute to the increased transmission and virulence of select SARS-CoV-2 variants. In addition to mutations in the spike protein, mutations in the nucleocapsid protein are important for viral spreading during the pandemic.

Palindromic-assisted self-annealing transcription amplification for reliable genotyping of epidermal growth factor receptor exon mutations.

Reliable discrimination of specific epidermal growth factor receptor (EGFR) gene mutations plays a critical role in guiding lung cancer therapeutics. Until now, convenient and accurate recognition of the specific deletion of EGFR exons has remained particularly challenging. Herein, we propose a palindromic-assisted self-annealing transcription amplification (PASTA) strategy for the reliable detection of circulating EGFR exon mutations. We designed a palindromic DNA hairpin nanorobot consisting of a palindromic tail, a T7 promoter, a target recognition region, and a transcription template. The nanorobot enabled prompt self-assembly into a target-hairpin/hairpin-target dimer in the presence of single-stranded DNA target and further triggered in vitro transcription. In a proof-of-concept experiment for detecting circulating 15n-del EGFR mutation, a detection limit of 0.8 fM and a linear detection range of 1 fM to 100 pM was achieved, and an accuracy of 100% was reached in clinical validation by analyzing 20 samples from clinical lung cancer patients. Empowered by the intrinsic sensitivity and selectivity, the proposed PASTA approach will lead to the development of a universal platform for reliable molecular subtyping.

Asymptomatic carbon dioxide embolism during transoral vestibular thyroidectomy: A case report.

BACKGROUND: Endoscopic thyroidectomy has obvious advantages over conventional surgical techniques in terms of postoperative cosmetic outcome. Although the incidence of carbon dioxide embolism (CDE) during endoscopic thyroidectomy is very low, it is potentially fatal. The clinical manifestations of CDE vary, and more attention should be paid to this disorder. CASE SUMMARY: A 27-year-old man was scheduled for thyroidectomy by the transoral vestibular approach. The patient had no other diseases or surgical history. During the operation, he developed a CDE following inadvertent injury of the anterior jugular vein. The clinical manifestation in this patient was a transient sharp rise in end-tidal carbon dioxide, and his remaining vital signs were stable. In addition, loud coarse systolic and diastolic murmurs were heard over the precordium. The patient was discharged on day 4 after surgery without complications. CONCLUSION: A transient sharp rise in end-tidal carbon dioxide is considered a helpful early sign of CDE during endoscopic thyroidectomy.

Aspiration pneumonia during general anesthesia induction after esophagectomy: A case report.

BACKGROUND: Esophageal cancer is a common malignant tumor of the digestive system. At present, surgery is the most important treatment strategy. After esophagectomy and gastric esophagoplasty, the patients are prone to regurgitation. However, these patients currently do not receive much attention, especially from anesthesiologists. CASE SUMMARY: A 55-year-old woman was scheduled for right lower lung lobectomy. The patient had undergone radical surgery for esophageal cancer under general anesthesia 6 mo prior. Although the patient had fasted for > 17 h, unexpected aspiration still occurred during induction of general anesthesia. Throughout the operation, oxygen saturation was 98%-100%, but the airway pressure was high (35 cmH2O at double lung ventilation). The patient was sent to the intensive care unit after surgery. Bedside chest radiography was performed, which showed exudative lesions in both lungs compared with the preoperative image. After surgery, antibiotics were given to prevent lung infection. On day 2 in the intensive care unit, the patient was extubated and discharged on postoperative day 7 without complications related to aspiration pneumonia. CONCLUSION: After esophagectomy, patients are prone to regurgitation. We recommend nasogastric tube placement followed by rapid sequence induction or conscious intubation.

New combined microRNA and protein plasmatic biomarker panel for pancreatic cancer.

INTRODUCTION: Lack of diagnostic makers results in loss of operation opportunity in that most patients are diagnosed at the late stage. Pancreatic cancer (PC) has been regarded as a fatal disease with a 5-year survival rate below 10%. Therefore, the development of diagnostic biomarkers for PC is in urgent need to control the mortality of the disease. MATERIALS AND METHODS: This is a case-control study including 640 plasma samples from healthy controls (HC), patients with benign pancreatic diseases (BPD), patients with PC; and patients with other gastrointestinal (GI) cancers. Eight biomarker candidates, including miR-20a, miR-21, miR-25, miR-155, miR-196a, miR-210, Macrophage Inhibitory Cytokine-1(MIC-1) and CA19-9, were evaluated to establish two diagnostic indexes in this study. RESULTS: The plasma level of the six miRNAs and MIC-1, CA19-9 were elevated in PC patients compared with those of healthy controls (P<0.001). Among them, miR-20a, miR-21, miR-25, MIC-1 and CA19-9 could distinguish PC patients from those with other GI cancers or BPD. With multivariable logistic regression, we established two specific indexes for diagnosis of PC(Index1 contains miR-21, MIC-1 and CA19-9; Index2 contains miR-25, MIC-1 and CA19-9). In a randomized setting of 260 HC, 168 PC, 132 other GI cancers and 80 BPD patients, both indexes performed not only better sensitivity for PC but also better specificity to distinguish PC from other GI cancers than CA19-9 and individual biomarkers. CONCLUSIONS: These results indicated that combination of biomarkers as a panel could improve diagnostic values compared with using a single marker. Such panels as illustrated in this study could provide novel plasmatic biomarker for PC diagnosis.

[Expression of microRNA-100 and its relation with prognosis of colorectal cancer].

OBJECTIVE: The aim of this study was to investigate the expression of microRNA-100 (miR-100) and its relation with prognosis in colorectal cancer (CRC). METHODS: The expression of miR-100 was analyzed by quantitative real-time PCR (qRT-PCR) in 172 CRC tissue samples. The relation of miR-100 expression patterns with clinical pathological significance in CRC was analyzed. The effects of alterations of miR-100 expression and its consequences on CRC cell proliferation, apoptosis and migration were demonstrated in cells cultured in vitro. RESULTS: The relative expression of miR-100 in CRC tissues and peritumoral tissues were -6.185 ± 1.921 and -3.698 ± 1.786, respectively, with a significant difference between the two groups (P<0.01). There was a significant difference between the relative expression of miR-100 in CRC with lymph node metastasis (-5.706 ± 1.809) and without lymph node metastasis (-6.775 ± 1.902, P<0.01). The relative expression of miR-100 in tumors of different TNM stages were -7.267 ± 1.888 in stage I, -6.443 ± 1.859 in stage II, -5.923 ± 1.796 in stage III, and -4.639 ± 1.516 in stage IV, with a significant difference among them (P<0.01). Different differentiation grades showed different expression of miR-100, i.e. -7.389 ± 1.828 in well differentiated tumors, -6.095 ± 1.843 in moderately differentiated tumors, and -5.476 ± 2.088 in poorly differentiated tumors (P<0.01). There was no significant correlation between miR-100 expression and overall survival rates of the CRC patients (P=0.179). Overexpression of miR-100 in the CRC cell line HCT-8 inhibited cell proliferation, but promoted cell apoptosis and migration. CONCLUSIONS: The expression of miR-100 is correlated with lymph node metastasis, TNM stage and differentiation grade, and may be a potential biomarker indicating the development of CRC.

Up-regulation of microRNA-145 associates with lymph node metastasis in colorectal cancer.

Metastasis is the main cause of mortality in patients with solid tumours. Identifying the exact molecules associated with CRC metastasis may be crucial to understand the process, which might also be translated to the diagnosis and treatment of CRC. In this study, we investigate the association of microRNA expression patterns with the lymph node metastasis of colorectal cancer. Among these candidate miRNAs, the expression of miRNA-145 was significantly related to lymph node metastasis of CRC. Both in vitro and in vivo study demonstrated that up-regulation of miR-145 could improve the ability of migration and invasion of colorectal cancer cell, while no effect on proliferation was observed. The mechanism of this promotion is associated with the stabilization of Hsp-27, a protein which plays an important role in the promotion of metastasis. These results may be crucial to understanding CRC metastasis and may be translated to the diagnosis and treatment of CRC.

[Expression and clinical significance of plasma small RNA in patients with pancreatic cancer].

OBJECTIVE: The aim of this study was to identify six miRNAs expressed in plasma of patients with pancreatic cancer (PCa) and analyze their value as a diagnostic index of pancreatic cancer. METHODS: Plasma total RNAs were extracted from 30 PCa patients and 26 normal controls, and the abundance of six microRNAs was measured using real-time PCR. The possibility to combine them with CA19-9 as diagnostic biomarkers was analyzed. RESULTS: The expression level of miR-21, miR-210, miR-155, miR-20a, miR-25 and miR-196a in plasma of patients with pancreatic cancer were 1.65×10(6), 5.98×10(4), 2.83×10(3), 3.47×10(6), 2.76×10(6), and 1.03×10(3) (copies/µl), while the normal controls were 4.08×10(5), 2.54×10(4), 8.55×10(2), 1.79×10(6), 9.32×10(5), and 4.67×10(2) (copies/µl), respectively, with a significant difference between the two groups (P < 0.05). The areas under the ROC curve of miR-21, miR-210, miR-155, miR-20a, miR-25 and miR-196a were 0.893, 0.810, 0.820, 0.766, 0.816 and 0.729, respectively. MiR-21 had the highest diagnostic value when it was used as diagnostic marker alone. The combination of miR-155 and miR-25 was more effective to distinguish PCa from normal than to be used alone, and the area under the ROC curve was 0.913 (95%CI 0.838-0.988) .When CA199 associated with miR -210 and miR-25, respectively, the areas under the ROC curves were 0.96 (95%CI was 0-1.0) and 0.942 (95% CI was 0.876-1.0), which were higher than CA199 alone (0.862, 95%CI was 0.748-0.975). There was a high improvement in diagnostic sensitivity and accuracy when miR-210 and miR-25 were combined with CA19-9, respectively. CONCLUSIONS: Plasma miR-21, miR-155, miR-25, miR-210 have diagnostic value for pancreatic cancer, and deserve further study.

[Therapeutic study of leukemia by pegylated liposomal daunorubicin].

OBJECTIVE: To explore the antitumor effect and toxicity of pegylated liposomal daunorubicin (PL-DNR) on leukemia. METHODS: PL-DNR was prepared by dry lipid hydration and remote loading, and its physicochemical indexes were analyzed. The inhibiting effect of PL-DNR on leukemia cells was observed in terms of in vitro cytotoxicity experiment. The therapeutic effect in vivo was assessed by tumor inhibition in leukemia L1210-bearing mice. Apoptosis in cardiomyocytes was detected using the terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method (TUNEL staining). RESULTS: The average diameter of PL-DNR was (110 ± 10)nm and the encapsulation efficiency was 94.21%. The in vitro cytotoxicity experiment showed that the inhibiting ability of PL-DNR in the treatment groups was continuously enhanced as the experiment proceeded. The in vivo pharmacodynamic experiment also indicated obvious tumor-inhibiting effect of PL-DNR. At the end of the experiment, the tumor volume of the PL-DNR group was (433.71 ± 234.77)mm(3), significantly smaller than that of (1 293.77 ± 381.26) mm(3) in the DNR group (P < 0.05). Moreover, the tumor weight of the PL-DNR group was (0.66 ± 0.29)g and that of the DNR group was (1.25 ± 0.43)g (P < 0.05). The myocardial toxicity experiment showed that the median apoptosis index of cardiomyocytes in the PL-DNR group was 13.83%, significantly lower than that of 42.67% in the DNR group (P < 0.05), indicating a lower toxicity of PL-DNR to the myocardium. CONCLUSION: Compared with the free DNR, PL-DNR can improve the therapeutic effect on leukemia and reduce the.