Effect of a self-developed fixation device on preventing endotracheal intubation-related pressure injury: a randomised controlled trial.

OBJECTIVE: To evaluate the effects of our self-developed endotracheal tube fixation device in mechanically ventilated patients. METHODS: In a dual-centre randomised controlled trial, patients who were expected to require mechanical ventilation for over 48 h were assigned to the observation group (using self-developed device) or the control group (using the traditional device). The primary endpoint was the incidence of endotracheal intubation-related pressure injury (EIRPI). RESULTS: Fifty-one patients in the observation group and 54 patients in the control group were analysed. The incidence of EIRPI was 7.8% in the observation group and 33.3% in the control group (p = 0.001). Lip pressure injury (PI) occurred in 0 versus 14 (25.9%) patients in the observation versus control groups (p < 0.001). Both oral-mucosal and facial PIs were similar between the two groups. CONCLUSIONS: The use of the novel device reduced the incidence of EIRPI, especially lip PI. Trial registration Chinese Clinical Trial Registry ChiCTR2300078132. Registered on 29 November 2023.

HPV prevalence and genotype distribution in 2,306 patients with cervical squamous cell carcinoma in central and eastern China.

Background: To explore the positivity rate and genotype distribution of human papillomavirus (HPV) in cervical squamous cell carcinoma (CSCC) tissues in central and eastern China and to provide theoretical basis for cervical cancer screening and prophylactic HPV vaccine development in China. Methods: DNA was extracted from paraffin-embedded tissues of CSCC samples and exfoliated cervical cells of cervical cancer screening populations. 23 HPV genotypes were detected by combining polymerase chain reaction (PCR) and reverse dot hybridized gene chip detection technology in 2,306 CSCC tissues and 10,245 cervical cancer screening populations. The genotype distribution of HPV infection was analyzed. Results: The overall infection rate of HPVs in 2,306 CSCC patients was 92.71%. The frequency of single-type HPV infection and multiple-type HPV infection were 86.48% and 13.51%, respectively. The most common HPV genotypes detected in Chinese CSCC tissues were HPV-16, HPV-18, HPV-31, HPV-33, HPV-45, HPV-52, HPV-58, and HPV-59. The overall positivity rate of these eight high-risk HPV (HR-HPV) genotypes in HPV-positive CSCC was as high as 96.91%. Of which the positivity rate of seven HR-HPV genotypes related to nine-valent HPV vaccines in HPV-positive CSCC was 95.09%. Meanwhile, the overall infection rates of HR-HPV and low-risk HPV (LR-HPV) in female aged 35-64 years who underwent cervical cancer screening were 13.16% and 1.32%, respectively. The high-frequency HR-HPV genotypes in cervical cancer screening women were HPV-52, HPV-58, HPV-16, HPV-53, HPV-68, HPV-39, HPV-51, and HPV-56, with positivity rates of 2.25%, 1.60%, 1.31%, 1.22%, 0.93%, 0.92%, 0.78%, and 0.74%, respectively. Conclusion: Among women screened for cervical cancer in China, detecting the 8 high-frequency HR-HPV genotypes can reduce technical difficulty and reagent costs, while also improving the efficiency and effectiveness of cervical cancer screening. HPV genotyping assists gynecologists in assessing the risk of HR-HPV-positive cervical intraepithelial neoplasia and guiding them in implementing appropriate interventions. Furthermore, HPV genotyping is helpful for doctors to follow up HR-HPV-positive women and to evaluate the protective effect of HPV vaccine.

Long Noncoding RNA NR2F1-AS1 Enhances the Migration and Invasion of Hepatocellular Carcinoma via Modulating miR-642a/DEK Pathway.

PURPOSE: Hepatocellular carcinoma (HCC), a malignant tumor that exists worldwide, has a high morbidity and mortality rate. Previous studies have reported that lncRNA NR2F1-AS1 plays a critical role in several cancers. Here, we aimed to investigate the biological function of NR2F1-AS1 and its molecular mechanism in the migration and invasion of HCC. METHODS: Quantitative real-time PCR (qRT-PCR) was performed to analyze NR2F1-AS1 expression in HCC. The biological function was investigated by transwell invasion and migration assays. The protein level was identified by Western blot. In addition, the downstream targets of NR2F1-AS1 and miR-642a were confirmed by luciferase reporter assays. RESULTS: NR2F1-AS1 was significantly upregulated in HCC and associated with the poor prognosis of HCC patients. Biological function experiments revealed that the silence of NR2F1-AS1 suppressed cell invasion and migration in HCC. More importantly, NR2F1-AS1 directly interacted with miR-642a and negatively regulated miR-642a. DEK was the target of miR-642a, and NR2F1-AS1 positively regulated DEK expression by suppressing miR-642a. CONCLUSION: Taken together, it is the first time we discovered the interaction of NR2F1-AS1 with miR-642a in modulating HCC cell invasion and migration.

Characterization of the complete chloroplast genome of Hedysarum polybotrys var. alaschanicum (Fabaceae) and its phylogeny.

Hedysarum polybotrys var. alaschanicum is an important medicinal plant and is widely used in traditional Chinese medicine. The complete chloroplast genome of H. polybotrys var. alaschanicum was assembled from Illumina pair-end sequence reads. The whole chloroplast genome is 122,933 bp in length and encodes a total of 110 genes, including 76 protein-coding genes, 30 tRNA genes and 4 rRNA genes. The overall GC content of the cp genome is 35.3%. A maximum likelihood (ML) phylogenetic analysis revealed that H. polybotrys var. alaschanicum was close to Hedysarum semenovii.

Longitudinal characteristics of lymphocyte responses and cytokine profiles in the peripheral blood of SARS-CoV-2 infected patients.

BACKGROUND: The dynamic changes of lymphocyte subsets and cytokines profiles of patients with novel coronavirus disease (COVID-19) and their correlation with the disease severity remain unclear. METHODS: Peripheral blood samples were longitudinally collected from 40 confirmed COVID-19 patients and examined for lymphocyte subsets by flow cytometry and cytokine profiles by specific immunoassays. FINDINGS: Of the 40 COVID-19 patients enrolled, 13 severe cases showed significant and sustained decreases in lymphocyte counts [0·6 (0·6-0·8)] but increases in neutrophil counts [4·7 (3·6-5·8)] than 27 mild cases [1.1 (0·8-1·4); 2·0 (1·5-2·9)]. Further analysis demonstrated significant decreases in the counts of T cells, especially CD8+ T cells, as well as increases in IL-6, IL-10, IL-2 and IFN-γ levels in the peripheral blood in the severe cases compared to those in the mild cases. T cell counts and cytokine levels in severe COVID-19 patients who survived the disease gradually recovered at later time points to levels that were comparable to those of the mild cases. Moreover, the neutrophil-to-lymphocyte ratio (NLR) (AUC=0·93) and neutrophil-to-CD8+ T cell ratio (N8R) (AUC =0·94) were identified as powerful prognostic factors affecting the prognosis for severe COVID-19. INTERPRETATION: The degree of lymphopenia and a proinflammatory cytokine storm is higher in severe COVID-19 patients than in mild cases, and is associated with the disease severity. N8R and NLR may serve as a useful prognostic factor for early identification of severe COVID-19 cases. FUNDING: The National Natural Science Foundation of China, the National Science and Technology Major Project, the Health Commission of Hubei Province, Huazhong University of Science and Technology, and the Medical Faculty of the University of Duisburg-Essen and Stiftung Universitaetsmedizin, Hospital Essen, Germany.

Linearly Patterned Programmed Cell Necrosis Induced by Chronic Hypoxia Plays a Role in Melanoma Angiogenesis.

BACKGROUND: Highly aggressive tumors are exposed to hypoxia and increased tumor interstitial fluid pressure (IFP) conditions which is resistant to blood supply. Physiological responses of the organism may reduce IFP through induction of orderly cell death. SPECIFIC AIMS: This study demonstrates that orderly cell death provided spatial structure for early angiogenesis in the hypoxic, high-IFP tumor microenvironment and the participation of linearly patterned programmed cell necrosis (LPPCN) in nascent melanoma angiogenesis. METHODS: Animal model, laser capture microdissection, wound healing and transwell assays, three-dimensional cultures, zymography assays, western-blotting analysis, immunohistochemistry and RT-PCR were performed. RESULTS: This study demonstrated a special form of cell death occurring in groups of malignant tumor cells which arrayed in lines. Both features of apoptosis and necrosis can be found in this cell death pattern and were termed as LPPCN. Its role as a stimulus of tumor angiogenesis was investigated using human melanoma samples and an animal model. Computer image analysis showed that LPPCN and tumor microvessels had identical spatial distributions. It can be induced by chronic hypoxia, high IFP and subsequent calcium influx. Higher number of tumor associated macrophages (TAM) and VEGF expression were found in the tumor with LPPCN. Based on the tumor-bearing animal model, it was found that block of caspase pathway inhibited LPPCN, microvessel density and vasculogenic mimicry (VM). CONCLUSIONS: LPPCN formation may play an important role in tumor angiogenesis due to stimulation of macrophage infiltration and HIF-1α regulation, and that inhibition of LPPCN may be a novel therapeutic strategy against tumor angiogenesis and metastasis.

Epithelial-mesenchymal transition regulated by EphA2 contributes to vasculogenic mimicry formation of head and neck squamous cell carcinoma.

PURPOSE: Vasculogenic mimicry (VM) was related to invasion and metastasis of head and neck squamous cell carcinoma (HNSCC) patients. This study was designed to investigate the role of EphA2 in VM formation of HNSCC. METHODS: The SiRNA technique was used to knock down the expression of EphA2 in vitro. The ability of cell migration and invasion were measured by transwell and wound healing assays; three-dimensional culture was used to detect the ability of channel-like structure formation; Western blot was used to detect the expression of epithelial-mesenchymal transition- (EMT-) related molecules in vitro. Further semiquantitative real-time RT-PCR assays and immunohistochemistry were used to demonstrate expression of EphA2 and EMT-related molecules according to VM presence or not in human tissue. RESULTS: Knocking down EphA2 in vitro leads to disabled channel-like structure formation, reduction of invasion and migration ability, and reverse of EMT-related markers. Both semiquantitative real-time RT-PCR and immunohistochemistry showed that expressions of EphA2, Twist, and Vimentin were higher in the VM-positive group than in the VM-negative group significantly, while expressions of E-cadherin, claudin4, and DSG-3 were reverse. CONCLUSIONS: EphA2 played a key role in VM formation of HNSCC through regulation of EMT.

Vasculogenic mimicry is a key prognostic factor for laryngeal squamous cell carcinoma: a new pattern of blood supply.

BACKGROUND: Recurrence and local lymph node metastasis affected the prognosis of patients with laryngeal cancer. The aim of this study was to analyze clinical and pathological significance of vasculogenic mimicry (VM) in laryngeal squamous cell carcinoma (LSCC) and evaluate its contribution to prognosis. METHODS: Data of 168 cases of LSCC were reviewed retrospectively to reveal clinical pathology and prognostic significance of VM. CD31 and periodic acid-Schiff double staining was used to identify VM. RESULTS: VM in LSCC contributed to lymph node metastasis (P = 0.003) and clinical progression. VM correlated to histopathology grade (P = 0.001) of LSCC. VM was an adverse prognostic factor for both disease-specific survival (P = 0.039) and metastasis-free survival (P = 0.042) by univariate survival analyses. And it was an independent prognostic factor for only disease-specific survival (P = 0.003) by multivariate survival analyses. CONCLUSIONS: VM existed in LSCC. LSCC with VM has more potential to invasion and metastasis.

[Role of vasculogenic mimicry and endothelium-dependent vessel in metastasis of laryngeal cancer].

OBJECTIVE: To investigate the contribution of vasculogenic mimicry (VM) and endothelium-dependent vessel (EDV) to invasion and metastasis of laryngeal squamous cell carcinoma (LSCC). METHODS: A total of 203 cases with LSCC was reviewed and followed up. VM and EDV in LSCC tissues were assessed by double staining with anti-CD31 immunohistochemistry and periodic acid-schiff. Kruskal-Wallis test and one-way ANOVA were used to analyze the relationship between VM, EDV and clinical pathology parameters of LSCC. Kaplan-Meier analysis was used to evaluate overall survival (OS) of patients with LSCC. RESULTS: VM related to pTNM stage, lymph node metastasis and pathology grade of LSCC, while EDV related to primary sites, pTNM stage, T stage and distant metastasis of LSCC. Univariate analysis showed VM (P = 0.014), pTNM stage (P = 0.009), T stage (P = 0.013), nodal status (P = 0.013), histopathology grade (P = 0.038), tumor size (P = 0.028), and radiotherapy (P < 0.0001) related to OS. VM (P = 0.011), primary sites (P = 0.049), tumor size (P = 0.001) and radiotherapy (P < 0.0001) related to disease free survival. Multivariate analysis indicated that VM was an adverse predictor for both OS and disease free survival. CONCLUSIONS: Both VM and EDV existed in LSCC. VM contributed to progression of LSCC through promoting lymph node metastasis. VM is an independent predictor for the prognosis of LSCC.

Overexpression of microtubule-associated protein-1 light chain 3 is associated with melanoma metastasis and vasculogenic mimicry.

Vasculogenic mimicry (VM) is an alternative type of blood supplement and is responsible for aggressive tumor biology and increased tumor-related mortality. Tumor cells obtain oxygen and nutriment through VM channels in the early, rapid-growth stage when blood vessels are insufficient. VM channels are characterized by tubular structures with tumor cells. Autophagy is a catabolic process, by which the cell digests damaged components or organelles of its own cytoplasm in response to nutrient deprivation, hypoxia, and the presence of non-functional protein aggregates. In fact, autophagy plays an important role in normal cell growth, development, and homeostasis. However, it is still controversial whether autophagy is also involved in cell death or cell survival in malignancy. In the present study, we therefore investigated the expression levels of two autophagy-related proteins, microtubule-associated protein-1 light chain 3 (LC3) and beclin-1, with respect to melanoma metastasis and vasculogenic mimicry. Melanoma is characterized by rapid growth, high-metastasis rate, and unpredictable behavior. A total of 70 human melanoma tissues were analyzed, showing that VM was present in 31 melanoma specimens (44.3%). Melanoma cells displayed high levels of autophagy when VM was present. Real-time quantitative PCR and immunohistochemical analyses showed that the expression levels of beclin-1 and LC3 mRNAs and proteins were both higher in the VM-positive melanoma than those in the VM-negative melanoma (p<0.05). Moreover, the expression of LC3, rather than beclin-1, was strongly associated with metastasis and poor clinical prognosis of human melanoma. Therefore, the enhanced autophagic activity may be related to VM and metastasis of melanoma.

The effect of SYT-SSX and extracellular signal-regulated kinase (ERK) on cell proliferation in synovial sarcoma.

The character of Synovial sarcoma is the chromosomal translocation t(X; 18)(p11.2;q11.2), which results in the fusion of the SYT gene with a SSX gene. There is little study that could fully elucidate the mechanism of pathogenesis of this fusion transcript. This study is designed to gain more insight into the function of this fusion gene. We evaluated the whole genome expression in SYO-1 cells inhibited as a result of specific small interfering RNA for SYT-SSX. Cell proliferation and apoptosis were analyzed by flow cytometer and MTT. The proteins correlated with proliferation were also detected using western blot. TUNEL and Immunohistochemical stain assessment were also carried out on TMA of SS tissues. The mRNA level reduced over 90% caused by SYT-SSX specific siRNA. Five pathways were employed, that ERK1/2 pathway was differential significantly (p = 0.043218). Meanwhile, down-regulation of SYT-SSX fusion gene expression would inhibit the proliferation of SS cell and the survival rate decreased (34.1%), while apoptotic rate increased (10.92%). After transfected with SYT-SSX-specific siRNA it caused a block in G1/G0 phase (31.99%) of SYO-1 cells compared with control cells. The protein level of ERK1/2, p-ERK, and cyclin D1 altered in same trend with expression of SYT-SSX. In TMA stain assessment, SYT-SSX positive group with high ki-67 LI expressed more cyclin D1and CDK4 than the SYT-SSX negative group. High ki-67 LI was detected in cases with p-ERK expression. Meanwhile, cyclin D1 and CDK4 were shown to be more expressed in tumor cells with p-ERK expression. Our results suggest that the fusion gene SYT-SSX should be considered to play important role on SS cell growth via ERK pathway. This study may be valuable for understanding the pathogenic role and molecular mechanism of the fusion gene SYT-SSX in synovial sarcoma through the proposed genome-wide approach. Furthermore, the research would open up the possibility of using SYT-SSX and ERK as a therapeutic target.

Vasculogenic mimicry contributes to lymph node metastasis of laryngeal squamous cell carcinoma.

BACKGROUND: Survival of laryngeal squamous cell carcinoma (LSCC) patients has remained unchanged over recent years due to its uncontrolled recurrence and local lymph node metastasis. Vasculogenic mimicry (VM) is an alternative type of blood supplement related to more aggressive tumor biology and increased tumor-related mortality. This study aimed to investigate the unique role of VM in the progression of LSCC. METHODS: We reviewed clinical pathological data of 203 cases of LSCC both prospectively and retrospectively. VM and endothelium-dependent vessel (EDV) were detected by immunohistochemistry and double staining to compare their different clinical pathological significance in LSCC. Survival analyses were performed to assess their prognostic significance as well. RESULTS: Both VM and EDV existed in LSCC type of blood supply. VM is related to pTNM stage, lymph node metastasis and pathology grade. In contrust, EDV related to location, pTNM stage, T stage and distant metastasis. Univariate analysis showed VM, pTNM stage, T classification, nodal status, histopathological grade, tumor size, and radiotherapy to be related to overall survival (OS). While, VM, location, tumor size and radiotherapy were found to relate to disease free survival (DFS). Multivariate analysis indicated that VM, but not EDV, was an adverse predictor for both OS and DFS. CONCLUSIONS: VM existed in LSCC. It contributed to the progression of LSCC by promoting lymph node metastasis. It is an independent predictors of a poor prognosis of LSCC.

A pilot study on morphology and the mechanism involved in linearly patterned programmed cell necrosis in melanoma.

Accumulating data provide evidence that autophagy contributes to programmed cell death (PCD) under certain circumstances. Immunohistochemistry and real-time PCR were performed to investigate the correlation the expression of autophagy-related proteins LC3 and Atg4B and linearly patterned programmed cell necrosis (LPPCN) in melanoma. LPPCN was recently reported to be a special PCD, which is similar to neither apoptosis nor conventional necrosis commonly observed in tumoral tissues. The mechanism involved in LPPCN remains unclear. Our data showed that the expression of LC3 and Atg4B in the LPPCN-positive group was significantly higher than that in the LPPCN-negative group regarding protein and mRNA levels (p<0.05). Based on morphological observation, immunohistochemistry and real-time PCR experiments in this study, it was concluded that autophagy may play a crucial role in the process of LPP+CN in melanoma. This study provides novel insights into the mechanism that regulates LPPCN in vivo during tumor development. We speculated that LPPCN may be an early stage event in tumoral neovascularization under hypoxic-microenvironmental conditions. Accordingly, LPPCN can be considered a novel target in the process of antiangiogenesis treatment, which can be expected to obtain a better clinical outcome in the future.

[Relationship between the expression of hypoxia-inducible factor-1alpha and epidermal growth factor receptor and micro vessel density and their clinical significance].

OBJECTIVE: Analyze the relationship between the expression of hypoxia-inducible factor-1alpha (HIF-1alpha) and epidermal growth factor receptor (EGFR) and CD105 micro vessel density (MVD) and their value in evaluating biologic behavior and prognosis in laryngeal cancer. METHODS: Ninety-one cases of laryngeal cancer were analyzed about their clinical and pathology data. In tumor tissue the expression of HIF-1alpha and EGFR was detected by immunohistochemistry and MVD was marked by CD105. RESULTS: The expression of HIF-1alpha was correlated with size, TNM stage, T stage, lymph node metastasis and histological grade (all P<0.05). The expression of EGFR was correlated with TNM stage, lymph node metastasis, histological grade and relapse (all P<0.05). MVD was correlated with type, TNM stage, T stage, lymph node metastasis, metastasis and histological grade (all P<0.05). The expression of HIF-1alpha and EGFR was correlated with MVD (F value was 7.644 and 5.197 respectively, P value was 0.001 and 0.025 respectively). The correlation between the expression of HIF-1alpha and EGFR was significant statistically (r= 0.238, P=0.007). The survival rate of patients of 3 years and 5 years were 56.1% and 44.2% respectively. Survival analysis by Log Rank showed that prognosis of laryngeal cancer patients was correlated with type, TNM stage and the expression of both HIF-1alpha and EGFR. While Cox multiple factors analysis demonstrated that TNM stage and expression of EGFR were independent prognostic factor of laryngeal cancer (P value was 0.049 and 0.041 respectively, RR was 1.300 and 2.417 respectively). CONCLUSIONS: HIF-1alpha and EGFR are key molecular event during development and progression of laryngeal cancer, which act in regulating tumor angiogenesis as well, and show intimate relationship with biological behavior and prognosis of laryngeal cancer.

[Inhibition of quercetin on liver fibrosis due to Schistosoma japonicum infection and on the expression of immediate early gene and metalloproteinase 1 inhibitor in liver tissue of mice].

Quercetin and praziquantel were used to treat mice with hepatic fibrosis due to Schistosoma japonicum infection. Quercetin treatment obviously relieved the degree of hepatic fibrosis, significantly reduced the expression of immediate early gene, tissue inhibitor of metalloproteinase 1 (TIMP 1), types I and III collagen compared to the control. The expression of c-jun mRNA, type I and type III collagen were reduced significantly compared to the group treated with praziquantel, whereas no difference in the expression of c-fos mRNA and TIMP1 between the two groups, indicating that quercetin may have better effect on schistosomal liver fibrosis than praziquantel in the long term.