Evaluation of recombinant baculovirus clearance during rAAV production in Sf9 cells using a newly developed fluorescent-TCID50 assay.

Introduction: Recombinant adeno-associated virus (rAAV) vectors provide a safe and efficient means for in vivo gene delivery, although its large-scale production remains challenging. Featuring high manufacturing speed, flexible product design, and inherent safety and scalability, the baculovirus/Sf9 cell system offers a practical solution to the production of rAAV vectors in large quantities and high purity. Nonetheless, removal and inactivation of recombinant baculoviruses during downstream purification of rAAV vectors remain critical prior to clinical application. Methods: The present study utilized a newly developed fluorescent-TCID50 (F-TCID50) assay to determine the infectious titer of recombinant baculovirus (rBV) stock after baculovirus removal and inactivation, and to evaluate the impact of various reagents and solutions on rBV infectivity. Results and discussion: The results showed that a combination of sodium lauryl sulfate (SLS) and Triton X-100 lysis, AAVx affinity chromatography, low pH hold (pH3.0), CsCl ultracentrifugation, and NFR filtration led to effective removal and/or inactivation of recombinant baculoviruses, and achieved a log reduction value (LRV) of more than 18.9 for the entire AAV purification process. In summary, this study establishes a standard protocol for downstream baculovirus removal and inactivation and a reliable F-TCID50 assay to detect rBV infectivity, which can be widely applied in AAV manufacturing using the baculovirus system.

Systematic comparison of rAAV vectors manufactured using large-scale suspension cultures of Sf9 and HEK293 cells.

Recombinant adeno-associated virus (rAAV) vectors could be manufactured by plasmid transfection into human embryonic kidney 293 (HEK293) cells or baculovirus infection of Spodoptera frugiperda (Sf9) insect cells. However, systematic comparisons between these systems using large-scale, high-quality AAV vectors are lacking. rAAV from Sf9 cells (Sf9-rAAV) at 2-50 L and HEK293 cells (HEK-rAAV) at 2-200 L scales were characterized. HEK-rAAV had ∼40-fold lower yields but ∼10-fold more host cell DNA measured by droplet digital PCR and next-generation sequencing, respectively. The electron microscope observed a lower full/empty capsid ratio in HEK-rAAV (70.8%) than Sf9-rAAV (93.2%), while dynamic light scattering and high-performance liquid chromatography analysis showed that HEK-rAAV had more aggregation. Liquid chromatography tandem mass spectrometry identified different post-translational modification profiles between Sf9-rAAV and HEK-rAAV. Furthermore, Sf9-rAAV had a higher tissue culture infectious dose/viral genome than HEK-rAAV, indicating better infectivity. Additionally, Sf9-rAAV achieved higher in vitro transgene expression, as measured by ELISA. Finally, after intravitreal dosing into a mouse laser choroidal neovascularization model, Sf9-rAAV and HEK-rAAV achieved similar efficacy. Overall, this study detected notable differences in the physiochemical characteristics of HEK-rAAV and Sf9-rAAV. However, the in vitro and in vivo biological functions of the rAAV from these systems were highly comparable. Sf9-rAAV may be preferred over HEK293-rAAV for advantages in yields, full/empty ratio, scalability, and cost.

The function role of HIGD1A in nonalcoholic steatohepatitis from chronic hepatitis B.

BACKGROUND: Accompanied by the growing prevalence of nonalcoholic fatty liver disease (NAFLD), the coexistence of chronic hepatitis B (CHB) and NAFLD has increased. In the context of CHB, there is limited understanding of the factors that influence the development of NASH. METHODS: We enrolled CHB combined NAFLD patients who had liver biopsy and divided them to NASH vs. non-NASH groups. A whole transcriptome chip was used to examine the expression profiles of long noncoding RNAs (lncRNAs) and mRNA in biopsied liver tissues. The function analysis of HIGD1A were performed. We knocked down or overexpressed HIGD1A in HepG2.2.15 cells by transient transfection of siRNA-HIGD1A or pcDNA-HIGD1A. In vivo investigations were conducted using hepatitis B virus (HBV) transgenic mice. RESULTS: In 65 patients with CHB and NAFLD, 28 were patients with NASH, and 37 were those without NASH. After screening 582 differentially expressed mRNAs, GO analysis revealed differentially expressed mRNAs acting on nicotinamide adenine dinucleotide phosphate (NADPH), which influenced redox enzyme activity. KEGG analysis also shown that they were involved in the NAFLD signaling pathway. The function analysis revealed that HIGD1A was associated with the mitochondrion. Then, both in vivo and in vitro CHB model, HIGD1A was significantly higher in the NASH group than in the non-NASH group. HIGD1A knockdown impaired mitochondrial transmembrane potential and induced cell apoptosis in HepG2.2.15 cells added oleic acid and palmitate. On the contrary, hepatic HIGD1A overexpression ameliorated free fatty acids-induced apoptosis and oxidative stress. Furthermore, HIGD1A reduced reactive oxygen species (ROS) level by increasing glutathione (GSH) expression, but Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK)/Acetyl-CoA carboxylase (ACC) pathway was not involved. CONCLUSION: Both in vivo and in vitro CHB model, an upward trend of HIGD1A was observed in the NASH-related inflammatory response. HIGDIA played a protective role in cells against oxidative stress. Our data suggested that HIGD1A may be a positive regulator of NASH within the CHB context.

Mechanism and Progress of Natural Products in the Treatment of NAFLD-Related Fibrosis.

Nonalcoholic fatty liver disease (NAFLD) has emerged as the most prevalent chronic liver disorder worldwide, with liver fibrosis (LF) serving as a pivotal juncture in NAFLD progression. Natural products have demonstrated substantial antifibrotic properties, ushering in novel avenues for NAFLD treatment. This study provides a comprehensive review of the potential of natural products as antifibrotic agents, including flavonoids, polyphenol compounds, and terpenoids, with specific emphasis on the role of Baicalin in NAFLD-associated fibrosis. Mechanistically, these natural products have exhibited the capacity to target a multitude of signaling pathways, including Hedgehog, Wnt/ß-catenin, TGF-ß1, and NF-κB. Moreover, they can augment the activities of antioxidant enzymes, inhibit pro-fibrotic factors, and diminish fibrosis markers. In conclusion, this review underscores the considerable potential of natural products in addressing NAFLD-related liver fibrosis through multifaceted mechanisms. Nonetheless, it underscores the imperative need for further clinical investigation to authenticate their effectiveness, offering invaluable insights for future therapeutic advancements in this domain.

Immune checkpoint inhibitors in cancer patients with COVID-19.

Immune checkpoint inhibitors (ICIs) are widely used to treat a variety of cancers and common infectious diseases with high efficacy. During the coronavirus disease 2019 (COVID-19) pandemic, studies suggested that COVID-19 patients may benefit from ICI immunotherapy. However, clinical studies on the safety and efficacy of ICI in COVID-19 patients are still being conducted. Currently, it is not clear whether cancer patients undergoing ICI immunotherapy should adjust their treatment strategy after infection with SARS-CoV-2 and whether ICI can reduce the viral load of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In this study, reports of patients with different types of tumors infected with SARS-CoV-2 under ICI immunotherapy were classified and sorted, including lung cancer, melanoma, squamous cell carcinoma of the head and neck, and hematologic malignances. The safety and efficacy of ICI in antitumor and anti-SARS-CoV-2 therapies were compared and further discussed to provide more reference materials for the application of ICI treatment. In a word, COVID-19 has changed the ICI treatment strategy for cancer patients indeed, and ICI treatment may be a "double-edged sword" for cancer patients complicated with COVID-19.

Clinical Features of Non-Alcoholic Fatty Liver Disease in the Non-Lean Population.

INTRODUCTION: The prevalence of non-alcoholic fatty liver disease (NAFLD) in non-lean patients is significantly increased, and obesity significantly increases the risk of cirrhosis and HCC in NAFLD patients. However, whether there is a difference in clinical manifestations of NAFLD between overweight and obesity remains unclear. The objective of this study was to assess the clinical and histological features of NAFLD among a non-lean population. METHODS: Current study enrolled consecutive non-lean (body mass index [BMI] >23 kg/m2) patients with NAFLD and available liver biopsy results. Patients were stratified by BMI into two groups for the comparison of their clinical and histological variables, which included the overweight (BMI 23∼<28 kg/m2) and the obese (BMI ≥28 kg/m2). Risk factors for moderate to severe fibrosis (stage >1) were also analyzed through the logistic regression model. RESULTS: Among 184 non-lean patients with metabolic-associated fatty liver disease enrolled, 65 and 119 were overweight and obese, respectively. Patients in the obesity group had a significantly lower level of gamma-glutamyl transpeptidase, higher levels of platelet, glucose, prothrombin time, and more common of moderate to severe inflammatory activity when compared to those in the overweight group. However, a significant low frequency of moderate to severe fibrosis was found in the obesity group versus the overweight group (19.33% vs. 40.00%, p = 0.002). Binary logistics regression analysis of fibrosis found that aspartate transaminase (AST), BMI, alanine transaminase (ALT), and cholesterol (CHOL) were independent predictors for moderate to severe fibrosis in non-lean patients with NAFLD. Compared with the traditional fibrosis-4 (AUC = 0.77) and aminotransferase to platelet ratio index (AUC = 0.79) indexes, the combined index based on AST, BMI, ALT, and CHOL was more accurate in predicting moderate to severe fibrosis in non-lean patients with NAFLD (AUC = 0.87). CONCLUSIONS: Clinical and histological features differed between obesity and overweight patients with NAFLD. When compared to the traditional serum markers, the combination index including AST, BMI, ALT, and CHOL provided a better model to predict moderate to severe fibrosis in non-lean patients with NAFLD.

The application of salvage surgery improves the quality of life and overall survival of extensively recurrent head and neck cancer after multiple operation plus radiotherapy.

Objectives: The prognosis, choice of reconstruction and the quality of life (QOL) after salvage surgery (SS) for extensively locoregional recurrent/metastatic head and neck cancer (R/M HNC) is an important issue, but there are few reports at present. Materials and methods: We analyzed extensively locoregional R/M HNC patients from March 1, 2015, to December 31, 2021 who underwent SS with latissimus dorsi or pectoralis major musculocutaneous flaps. QOL were accessed using QLQ-H&N35 and UW-QOL questionnaire. Wilcoxon signed-rank test was used to compare difference between pre- and post-QOL and Kaplan-Meier curves were used in estimate overall survival (OS) and disease-free survival (DFS). The literature review summarized recent 10 years clinical trials of nonoperative treatment in R/M head and neck cancer. Results: 1362 patients were identified and 25 patients were analyzed after screened. Median age at surgery was 59 years (range 43-77), 15/25(60%) were male and 22/25(88%) chose latissimus dorsi flap. Better mean pain score after applying massive soft tissue flaps revealed relief of severe pain(p<0.001) which strongly associated with improvement of QOL. The improved mean overall QOL score after surgery revealed a better QOL(p<0.001). As of June 1, 2022, 11/25 (44%) of the patients were alive. The 1-year, 2-year OS after SS was 58.4% and 37.2%, while the 1-year, 2-year DFS was 26.2% and 20.9%. The median OS of our study was better than nonoperative treatment of 11 included clinical trials. Conclusions: R/M HNC patients underwent SS can obtain survival benefit. The application of massive soft tissue flap in SS could significantly enhance the QOL for patients with extensively locoregional R/M HNC, especially by relieving severe pain.

The role of magnetic resonance imaging in assessing the extent of tongue squamous cell carcinoma: A prospective cohort study.

PURPOSE: To assess the false-positive and false-negative MRI results in evaluating the extent of tongue squamous cell carcinoma. METHODS: A prospective cohort series of 165 patients was enrolled to assess the false-positive and false-negative MRI results in evaluating the extent of tongue squamous cell carcinoma by comparing intraoperative tumor profile images and postoperative pathological sections. The differences between two-dimensional tumor margins were analyzed using Mimics 15.0 and Geomagic Control 16.0. A paired-samples t-test was used to analyze the agreement among MRI, intraoperative and pathological findings regarding the extent of tongue tumors. Multiple linear regression analysis was used to analyze associated factors. RESULTS: The mean and maximum false-positive values of pathological specimens was 1.95±1.39 mm (95% limit of agreement (LoA) 1.70-2.14) and 3.21 mm, respectively; the false-negative value was 0.44±0.49 mm. The false-positive value of intraoperative specimens was 1.52±0.87 mm (95% LoA 1.36-1.64); the false-negative value was 0.35±0.20 mm. Tumor morphology (ulcer type) (p<0.01) and depth of invasion (DOI) (≤5 mm) (p<0.01) were significantly correlated with the false-positive values of intraoperative and pathology specimens. CONCLUSION: The false-positive values are important when judging the invasion margin of tongue cancer and forming MRI-based operative plans; the false-negative value was almost negligible.

Diagnostic value of magnetic resonance imaging in cervical lymph node metastasis of oral squamous cell carcinoma.

OBJECTIVE: The objective of this study was to investigate the correlation between magnetic resonance imaging (MRI) characteristics of cervical lymph nodes and the pathologically confirmed status of cervical lymph node metastasis (LNM) in oral squamous cell carcinoma (OSCC) and to provide imaging evaluation parameters for the clinical diagnosis of cervical lymph node status in OSCC. STUDY DESIGN: In a retrospective analysis, 79 patients who were first pathologically diagnosed with OSCC were included. The MRI-derived imaging parameters of the cervical lymph nodes were evaluated and the pathological status of lymph nodes in neck dissection specimens was reviewed. The relationship between the imaging parameters and cervical LNM was analyzed. RESULTS: The MRI-derived imaging parameters of 4419 lymph nodes were evaluated, and the pathological status of 2463 lymph nodes was reviewed. The MRI-derived shortest axial diameter (SAD) and unclear boundary of the cervical lymph node were significantly related to LNM. The cutoff value of SAD that enabled identification of LNM was 3.6 mm, and it was 4.2 and 4.1 mm for the prediction of overall survival and disease-specific survival, respectively. CONCLUSIONS: The MRI-derived parameters SAD and unclear boundary of the cervical lymph node correlated with LNM in OSCC. MRI-derived SAD larger than 3 mm warrants simultaneous neck dissection at initial surgery.

Accuracy of Magnetic Resonance Imaging in Evaluating the Depth and Level of Invasion of Buccal Carcinoma: A Prospective Cohort Study.

PURPOSE: This study evaluated the accuracy of magnetic resonance imaging (MRI) in determining the depth and level of invasion of buccal carcinoma. METHODS: Patients with buccal squamous cell carcinoma diagnosed pathologically from July 2016 to December 2019 were included. The depth of invasion (DOI) and level of invasion (LOI) were evaluated by MRI, intraoperative specimens and pathological sections. Statistical analyses were performed using IBM SPSS software version 25.0 (IBM Corp., Armonk, NY). RESULTS: Forty-nine patients were ultimately included. The overall difference in DOI between MRI and pathological sections (DMP) was 5.55 ± 2.40 mm, and T category correlated with the differences in DOI measurement and LOI assessment. The threshold value of DOI by MRI to identify lymph node metastasis was 8.5 mm, and that for overall survival (OS) and disease-specific survival (DSS) was 14.1 mm for both. Buccinator invasion on MRI correlated with OS and DSS. CONCLUSION: Tumors with MRI-derived DOI larger than 8.5 mm deserve simultaneous neck dissection at initial surgery. Buccinator invasion was found to be an independent prognostic factor for buccal carcinoma patients.

The identify role and molecular mechanism of the MALAT1/hsa-mir-20b-5p/TXNIP axis in liver inflammation caused by CHB in patients with chronic HBV infection complicated with NAFLD.

BACKGROUND/AIMS: To identify the inflammatory damage caused by chronic hepatitis B (CHB) in patients of chronic hepatitis B virus (HBV) infection complicated with non-alcoholic fatty liver disease (NAFLD), then guiding clinicians to carry out antiviral treatment. METHODS: According to the pathological features of liver biopsy, treatment-naïve obese patients of chronic HBV infection complicated with NAFLD who had elevated alanine transaminase (ALT) were divided into CHB group and NASH group. Transcriptome chips were used to analyze the expression profiles of long non-coding RNA (lncRNA) and mRNA in liver puncture tissues from the two groups. The chip data of CHB and NASH groups were analyzed for differential expression analysis, gene function analysis, signal pathway analysis, target gene prediction and competing endogenous RNAs (ceRNA) network analysis. RESULTS: By comparing CHB group with NASH group, a total of 44 differentially expressed lncRNAs and 567 differentially expressed mRNAs were screened. GO analysis predicted that the differentially expressed mRNAs may affect monooxygenase activity and oxidoreductase activity. KEGG analysis predicted that the differentially expressed mRNAs may be related to signaling pathways involved in oxidative phosphorylation, phagosomes, and NAFLD. Differential analysis of lncRNA shown that the expression of metastasis associated in lung adenocarcinoma transcript 1 (MALAT1) in CHB group was significantly upregulated. Subsequently, through target gene prediction and ceRNA network analysis, we found thioredoxin interacting protein (TXNIP), which was significantly upregulated in the CHB group and had a ceRNA relationship with MALAT1. It is predicted that there may be a ceRNA regulation relationship of MALAT1/hsa-miR- 20b-5p/TXNIP. CONCLUSION: The MALAT1/hsa-miR-20b-5p/TXNIP axis may mediate CHB-induced inflammatory damage in chronic HBV infection complicated with NAFLD, and the mechanism may be related to the activation of NLRP3 inflammatory bodies and downstream inflammatory responses.

Clinical features of familial clustering in patients infected with 2019 novel coronavirus in Wuhan, China.

An epidemic caused by SARS-Coronavirus-2 (SARS-CoV-2) infection has appeared in Wuhan City in December 2019. The disease has shown a "clustering epidemic" pattern, and family-clustered onset has been the main characteristic. We collected data about 130 cases from 35 cluster-onset families (COFs) and 41 cases from 16 solitary-onset families (SOFs). The incidence of 2019 coronavirus disease (COVID-19) in COFs was significantly higher than that of SOFs. Our study also showed that patients with exposure to high-risk factors (respiratory droplets and close contact), advanced age, and comorbidities were more likely to develop COVID-19 in the COFs. In addition, advanced age and elevated neutrophil/lymphocyte ratio (NLR) were risk factors for death in patients with SARS-CoV-2 infection in the COFs.

Is it necessary to receive radiation for pT3-4N0 oral cancer without other adverse risk features?

OBJECTIVE: The purpose of this study was to explore the necessity of adjuvant radiotherapy for well-differentiated pT3-4aN0M0 OSCC without other negative features histologically. PATIENTS AND METHODS: This is a double-center, ambispective cohort study enrolling 250 patients with well-differentiated pT3-4aN0M0 OSCC. RESULTS: A total of 250 patients were enrolled in the double-center study, 155(62.0%) men and 95 (38.0%) women, and the mean age was 60.1 ± 11.1 years. T staging was classified as follows: T3 (n = 99, 39.6%) and T4a (n = 151, 60.4%). Kaplan-Meier analysis showed that there was no significant difference in the DSS between patients who received adjuvant radiotherapy (72.2%) and those who did not (77.4%) (p = .615). Specifically, no significant difference was found in the DSS of pT3N0M0 or pT4aN0M0 patients who received adjuvant radiotherapy compared with those who did not (pT3N0M0: 71.9% vs. 75.8%, p = .993; pT4aN0M0: 72.4% vs. 78.5%, p = .491). The Cox proportional hazards regression models showed that no factor was independent prognostic factor for pT3-4aN0M0 patients, or pT3N0M0 subgroup or pT4aN0M0 subgroup in DSS. And no independent prognostic factor was found for the surgery-alone subgroup and adjuvant radiotherapy subgroup. CONCLUSIONS: The results showed that adjuvant radiotherapy did not obviously improve the prognosis of pT3-4aN0M0 well-differentiated OSCC without other negative features.

Disruption of TP63-miR-27a* Feedback Loop by Mutant TP53 in Head and Neck Cancer.

BACKGROUND: Alterations in the epidermal growth factor receptor and PI3K pathways in head and neck squamous cell carcinomas (HNSCCs) are frequent events that promote tumor progression. Ectopic expression of the epidermal growth factor receptor-targeting microRNA (miR), miR-27a* (miR-27a-5p), inhibits tumor growth. We sought to identify mechanisms mediating repression of miR-27a* in HNSCC, which have not been previously identified. METHODS: We quantified miR-27a* in 47 oral cavity squamous cell carcinoma patient samples along with analysis of miR-27a* in 73 oropharyngeal and 66 human papillomavirus-positive (HPV+) samples from The Cancer Genome Atlas. In vivo and in vitro TP53 models engineered to express mutant TP53, along with promoter analysis using chromatin immunoprecipitation and luciferase assays, were used to identify the role of TP53 and TP63 in miR-27a* transcription. An HNSCC cell line engineered to conditionally express miR-27a* was used in vitro to determine effects of miR-27a* on target genes and tumor cells. RESULTS: miR-27a* expression was repressed in 47 oral cavity tumor samples vs matched normal tissue (mean log2 difference = -0.023, 95% confidence interval = -0.044 to -0.002; two-sided paired t test, P = .03), and low miR-27a* levels were associated with poor survival in HPV+ and oropharyngeal HNSCC samples. Binding of ΔNp63α to the promoter led to an upregulation of miR-27a*. In vitro and in vivo findings showed that mutant TP53 represses the miR-27a* promoter, downregulating miR-27a* levels. ΔNp63α and nucleoporin 62, a protein involved in ΔNP63α transport, were validated as novel targets of miR-27a*. CONCLUSION: Our results characterize a negative feedback loop between TP63 and miR-27a*. Genetic alterations in TP53, a frequent event in HNSCC, disrupt this regulatory loop by repressing miR-27a* expression, promoting tumor survival.

Prognosis of Pediatric Patients With Mucoepidermoid Carcinoma of the Parotid Gland.

OBJECTIVE: To summarize the prognosis of pediatric patients with mucoepidermoid carcinoma (MEC) of the parotid gland. METHODS: Pediatric patients with MEC of parotid gland who were surgically treated at the Capital Medical University School of Stomatology from 2000 to 2014 were retrospectively analyzed. Clinical characteristics, pathology reports, and operation records were reviewed and analyzed. RESULTS: In total, 33 patients with an average age of 13.2 years were enrolled. The 5-year overall survival and disease-free survival were 95.8% and 84.4%, respectively. The disease-free survival and overall survival rates were lower in the under-10 age group (75.0 versus 87.7% and 83.3% versus 100%), though no statistically significant difference was found (P = 0.279 and P = 0.075). The patients who underwent complete resection all had a good prognosis without any recurrence or death regardless of whether the cut margin was 1.0 cm, 0.5 cm, or only extracapsular. One patient experienced 3 recurrences within 18 months and eventually died of disease. CONCLUSION: Good outcomes were achieved in pediatric patients with MEC of the parotid gland. Radical resection ensured a good prognosis regardless of the extent of resection. Frequent recurrence in a short period was associated with a poor prognosis. TRIAL REGISTRATION: None.

Prognostic implications of aberrantly expressed methylation­driven genes in hepatocellular carcinoma: A study based on The Cancer Genome Atlas.

RNA­Sequencing and methylation data for hepatocellular carcinoma (HCC) were downloaded from The Cancer Genome Atlas (TCGA). The aberrantly expressed methylation­driven genes in HCC and normal tissues were identified using the Limma package and the MethylMix algorithm. The Database for Annotation, Visualization and Integrated Discovery and ConsensusPathDB were used for Gene Ontology (GO) enrichment and pathway analysis. Univariate and multivariate Cox regression analyses were used to construct a prognostic risk model of HCC. Survival curve and receiver operating characteristic (ROC) curves were applied to evaluate the clinical utility of the risk model. A total of 238 methylation­driven genes were successfully identified from cancer and normal tissues. GO enrichment analysis indicated that these genes functioned in the extracellular space, interfering with lipid metabolism in hepatocytes and regulating adaptive immune responses. In total, 14 relevant pathways were identified. The following prognostic risk model was generated: Risk score=CALML3 (degree of methylation) x (­4.860) + CCNI2 x (2.071) + TNFRSF12A x (­3.369) + IFITM1 x (1.203) + ENPP7P13 x (­1.366) + DDT x (2.139) + RASAL2­AS1 x (­1.384) + ANKRD22 x (­3.215). The median risk score (0.970) derived from this model was set as cutoff value for assigning patients to high­ or low­risk group. The 5­year survival rate was 35.8% [95% confidence interval (CI)=27.1­47.4%] in the high­risk group and 61.7% (95% CI=51.4­74.2%) in the low­risk group (P<0.0001). The ROC curve showed an area under the curve of 0.742, indicating that this model is appropriate for predicting the survival rate of patients. Furthermore, the methylation and expression levels of two key genes, tumor necrosis factor superfamily member 12A and D­dopachrome decarboxylase, were significantly associated with prognosis and were correlated with cg00510447, cg26808293, cg11060661 and cg16132339 methylation. In conclusion, a prognostic risk model for HCC is proposed based on the bioinformatic analysis of methylation­driven genes. The findings of the present study may improve understanding of the pathogenesis and prognosis of HCC.

Logistic regression analysis of clinical and computed tomography features of pulmonary abscesses and risk factors for pulmonary abscess-related empyema.

OBJECTIVES: This study was conducted to investigate the risk factors for pulmonary abscess-related empyema by investigating the clinical characteristics and chest computed tomography imaging features of patients with pulmonary abscesses. METHODS: We retrospectively analyzed the chest computed tomography findings and clinical features of 101 cases of pulmonary abscess, including 25 cases with empyema (the experimental group) and 76 cases with no empyema (the control group). The potential risk factors for pulmonary abscess-related empyema were compared between the groups by using univariate and multivariate logistic regression analyses. RESULTS: The incidence of pulmonary abscess-related empyema was 24.8% (25/101). Univariate analysis showed that male gender, diabetes, pleuritic symptoms, white blood cells >10×109/L, albumin level <25 g/L, and positive sputum cultures were potential clinical-related risk factors and that an abscess >5 cm in diameter and transpulmonary fissure abscesses were potential computed tomography imaging-related risk factors for pulmonary abscess-related empyema. Multivariate logistic regression analysis showed that transpulmonary fissure abscesses (odds ratio=9.102, p=0.003), diabetes (odds ratio=9.066, p=0.003), an abscess >5 cm in diameter (odds ratio=8.998, p=0.002), and pleuritic symptoms (odds ratio=5.395, p=0.015) were independent risk factors for pulmonary abscess-related empyema. CONCLUSIONS: Transpulmonary fissure abscesses, diabetes, giant pulmonary abscesses, and pleuritic symptoms increased the risk of empyema among patients with pulmonary abscesses.

Accuracy of magnetic resonance imaging in evaluating the depth of invasion of tongue cancer. A prospective cohort study.

OBJECTIVES: This study compared the consistency of depth of invasion (DOI) measurements by magnetic resonance imaging (MRI) and intraoperative and postoperative pathological sections due to a lack of large sample studies. MATERIALS AND METHODS: From April 2015 to December 2017, patients with squamous cell carcinoma of the tongue were included in the study. Different invasion depths were measured by MRI and on intraoperative and postoperative pathological sections. The differences between two-dimensional tumor margins were analyzed using Mimics 15.0 and Geomagic Control 16.0. Statistical analyses were performed using IBM SPSS software version 25.0 (IBM Corp., Armonk, NY). RESULTS: This study included 150 patients, the overall difference between MRI and postoperative pathological sections (DMP) and the overall difference between intraoperative and postoperative pathological sections (DIP) based on pathological specimens were 2.32 ±â€¯1.68 mm and 0.68 ±â€¯0.99 mm. The overall difference between MRI and intraoperative pathological sections (DMI) based on intraoperative specimens was 1.64 ±â€¯1.32 mm. The tumor growth pattern and T stage were significantly correlated with measurement differences. The cutoff value of MRI depth that could identify nodal metastasis was 8 mm, and were both 11 mm for OS and DSS. CONCLUSION: Clinicians performing T staging on patients with tongue cancer based on MRI measurements must consider the false-positive mean depth of 2.3 mm as well as the growth pattern and specific infiltration depth. The prognostic MRI depths that enabled the identification of nodal metastasis, OS and DSS were 8 mm, 11 mm and 11 mm, respectively. CLINICAL TRIAL REGISTRATION: Name: A Prospective, Observational, Real-world Study Based on the Register System of Oral and Maxillofacial Malignant Tumors. (ClinicalTrials.gov ID: NCT02395367).

A pilot study of modified resection for anterior floor of the mouth squamous cell carcinoma without infiltration of the mandible.

OBJECTIVE: To explore the application of modified resection compared with traditional segmental resection of the mandible for patients with anterior floor of the mouth and tongue squamous cell carcinoma (SCC) without infiltration of the mandible. SUBJECTS AND METHODS: This is a retrospective study including 36 eligible patients with anterior floor of the mouth SCC(9 patients received modified mandibulectomy, and 27 patients received segmental mandibulectomy). RESULTS: No patients in the modified mandibulectomy group developed recurrence in the floor of the mouth, and all of the patients survived. Only one patient developed osteoradionecrosis. When the modified mandibulectomy group was compared with the segmental mandibulectomy group, the former exhibited a lower recurrence rate in the floor of the mouth (0.0% vs. 14.8%), less blood loss (516.7 ± 70.7 ml vs. 533.3 ± 93.0 ml), shorter durations of gastric tube placement (11.4 ± 4.5 days vs. 20.7 ± 11.9 days) and tracheostomy (6.9 ± 0.6 days vs. 8.5 ± 1.6 days), a lower postoperative infection rate (11.1% vs. 18.5%), and a shorter postoperative hospital stay (13.7 ± 3.8 days vs. 15.9 ± 5.1 days). CONCLUSION: This modified mandibulectomy method is safe and feasible and is recommended for further prospective study in a clinical setting.

The impact of age on oral squamous cell carcinoma: A longitudinal cohort study of 2,782 patients.

BACKGROUND: The purposes of this study were to assess the influence of age on oral squamous cell carcinoma patients and sought to analyze the reasons that may contribute to this difference. METHODS: This study enrolled 2,782 patients included 2,443 patients in a retrospective cohort to find the influence of age and 339 patients in a prospective cohort to testify these findings. The patients were divided into young age-group (≤40 years old), moderate age-group (41-75 years old), and advanced age-group (>75 years old). All patients were diagnosed as oral squamous cell carcinoma and were surgically treated in our hospital. Chi-square test, Kaplan-Meier analysis, and Cox proportional-hazards regression model were performed for statistical analysis. RESULTS: Younger patients started smoking (p < 0.001) and drinking (p < 0.001) earlier than the older patients and consumed more tobacco (p = 0.005) and alcohol (p = 0.156). Patients with advanced age had worse outcomes in both recurrence (p = 0.002) and survival (p < 0.001). They also had more severe comorbidity (p < 0.001) and were more likely to receive conservative treatment (p = 0.011). CONCLUSIONS: Compared with young patients, older patients had worse prognosis, and it was related with their more severe comorbidity and received more conservative treatment. Young adults smoking and drinking earlier and heavier than old patients, it may relate with their occurrence of oral squamous cell carcinoma.