Ruxolitinib Pharmacokinetics and Pharmacodynamics in Children with Acute and Chronic Graft-versus-Host Disease.

We evaluated the pharmacokinetics (PK) of oral ruxolitinib in children with steroid-refractory acute graft-versus-host disease (aGVHD) (age <12 years) and chronic GVHD (cGVHD) (age ≤18 years) using our published pediatric dosing. PK sampling was performed before and 2 hours after ruxolitinib administration in patients with established cGVHD. More extensive PK analyses were performed in patients with newly diagnosed aGVHD or cGVHD before and .5, 1, 2, 4, and 6 hours after ruxolitinib administration in patients weighing >10 kg and before, 3+, and 6+ hours in children weighing <10 kg. pSTAT1, pSTAT3, and pSTAT5 expression levels were measured on CD4+ and CD8+ T cells before and 2 hours after ruxolitinib administration as a pharmacodynamic marker of JAK/STAT inhibition. Thirteen patients were prospectively enrolled, including 8 with existing cGVHD (age 0 to ≤18 years), 4 with new-onset steroid-refractory aGVHD (age 0 to <12 years) and 1 with newly diagnosed steroid-refractory cGVHD. Great variability in PK was seen. Mean oral clearance (CL/F) was 7.76 ± 4.09 L/h (range, 3.1 to 15.3 L/h). The average elimination half-life was 2.32 ± 1.0 hours. Mean ruxolitinib clearance was higher in children age <2 years versus those age >2 years (12.1 ± 3.0 L/h versus 5.7 ± 2.8 L/h; P = .005) and was reduced with concurrent treatment with azoles and azithromycin. We saw a variable reduction in pSTAT1/3/5 expression on T cells at time of peak ruxolitinib absorption (2 hours after dosing). Children <10 kg had lower ruxolitinib exposure, possibly due to inherent increased drug clearance or variability in dosing methods, leading to decreased drug absorption.

Erratum: HIF-1α or HOTTIP/CTCF Promotes Head and Neck Squamous Cell Carcinoma Progression and Drug Resistance by Targeting HOXA9.

[This corrects the article DOI: 10.1016/j.omtn.2019.12.045.].

Test-dose pharmacokinetics guided melphalan dose adjustment in reduced intensity conditioning allogeneic transplant for non-malignant disorders.

AIMS: We studied melphalan pharmacokinetics (PK) and feasibility of melphalan full-dose adjustment based on test-dose PK in children and young adults with non-malignant disorders (NMD) undergoing allogeneic hematopoietic cell transplantation (HCT) using reduced intensity conditioning (RIC) containing alemtuzumab, fludarabine and melphalan. METHODS: Patients received test-dose melphalan (10% of planned full-dose) prior to conditioning. Blood samples for PK were obtained around test and full-dose melphalan (140 mg/m2 or 4.7 mg/kg in patients <10 kg). Melphalan concentration was measured by liquid chromatography electrospray ionization tandem mass-spectrometry assay and data were analysed using a population-PK model and Bayesian estimation. Test and full-dose melphalan clearance estimates were evaluated by pairwise Wilcoxon test and Bland-Altman plot. RESULTS: Twenty-four patients undergoing 25 transplants were included in the final analysis. Patients received standard full-dose melphalan in 17 transplants, with median area under the concentration-time curve (AUC) of 5.5 mg*h/L (range, 3.0-9.5 mg*h/L). Patients received test-dose melphalan in 23 transplants with a test-dose PK predicted full-dose AUC range of 2.9-16.8 mg*h/L. In seven transplants where patients had baseline organ impairment, test-dose PK predicted higher exposure for standard full-dose (median AUC 13.8 mg*h/L). Melphalan full-dose was adjusted in these patients, with achievement of desired target AUC (3.6-5.4 mg*h/L) and no excess toxicity. Mean ratio of test-dose clearance to full-dose clearance was 1.03. Twenty of 22 patients (91%) were within the 95% confidence intervals of the clearance ratio. CONCLUSION: Melphalan test-dose PK reliably predicts full-dose PK and allows for accurate adjustment of full-dose melphalan in RIC-HCT for NMD. This approach can avoid excess toxicity from increased systemic exposure, especially in patients with organ impairment.

Prognostic validity of the American joint committee on cancer eighth edition staging system for well-differentiated pancreatic neuroendocrine tumors.

BACKGROUND: The American Joint Committee on Cancer (AJCC) made improvements for staging pancreatic neuroendocrine tumors (pNETs) in its 8th Edition; however, multicenter studies were not included. METHODS: We collected multicenter datasets (n = 1,086, between 2004 and 2018) to validate the value of AJCC 8 and other coexisting staging systems through univariate and multivariate analysis for well-differentiated (G1/G2) pNETs. RESULTS: Compared to other coexisting staging systems, AJCC 7 only included 12 (1.1%) patients with stage III tumors. Patients with European Neuroendocrine Tumor Society (ENETS) stage IIB disease had a higher risk of death than patients with stage IIIA (hazard ratio [HR]: 4.376 vs. 4.322). For the modified ENETS staging system, patients with stage IIB disease had a higher risk of death than patients with stage III (HR: 6.078 vs. 5.341). According to AJCC 8, the proportions of patients with stage I, II, III, and IV were 25.7%, 40.3%, 23.6%, and 10.4%, respectively. As the stage advanced, the median survival time decreased (NA, 144.7, 100.8, 72.0 months, respectively), and the risk of death increased (HR: II = 3.145, III = 5.925, and IV = 8.762). CONCLUSION: These findings suggest that AJCC 8 had a more reasonable proportional distribution and the risk of death was better correlated with disease stage.

Alterations of the Oral Microbiota Profiles in Chinese Patient With Oral Cancer.

Oral cancer is the most common malignant tumor in the oral and maxillofacial region, of which more than 90% is squamous cell carcinoma. The incidence of oral cancer is on the rise worldwide. An imbalance between the microorganism composition and its host may lead to the occurrence of oral malignant tumors. Accumulating evidence suggests that the oral microbiota plays an important role in oral cancer; however, the association between oral microbiota and oral cancer has not yet been comprehensively studied. In this study, metagenomic sequencing was used to compare the microbial composition of three groups of samples from Chinese patients with oral cancer, patients with precancerous lesion, and normal individuals. In terms of microbiota richness, the oral microbiota of patients with precancerous lesions was richer than that of oral cancer patients and healthy controls, whereas in terms of microbiota diversity, there was little difference between the three groups. The three groups of samples exhibited statistically significant differences in microbiota composition and metabolic function at the family, genus, and species levels (P < 0.05). The differentially enriched phylum in oral cancer samples was Bacteroidetes (P < 0.05). At the genus level, the main differentially enriched taxa were Prevotella, Peptostreptococcus, Carnobacterium, and Diastella (P < 0.05). The species level was differentially enriched in Prevotella intermedia and Peptostreptococcus stomatis (p < 0.05). The prediction of microbiota function shows that oral cancer is mainly associated with coenzyme A biosynthesis, phosphopantothenic acid biosynthesis, inosine 5'-phosphate degradation, and riboflavin biosynthesis. Furthermore, the increase in C-reactive protein level in oral cancer patients was found to be closely related to P. intermedia. Overall, oral bacterial profiles showed significant differences between the oral cancer group and normal group. Hence, microbes can be employed as diagnostic markers and treatment targets for oral cancer.

New staging classification for pancreatic neuroendocrine neoplasms combining TNM stage and WHO grade classification [].

AJCC TNM stage and WHO grade (G) are two widely used staging systems to guide clinical management for pancreatic neuroendocrine neoplasms (panNENs), based on clinical staging and pathological grading information, respectively. We proposed to integrate TNM stage and G grade into one staging system (TNMG) and to evaluate its clinical application as a prognostic indicator for panNENs. Accordingly, 5254 patients diagnosed with panNENs were used to evaluate and to validate the applicability of TNMG to panNENs. The predictive accuracy of TNMG system was compared with that of each separate staging/grading system. We found that TNM stage and G grade were independent risk factors for survival in both the Surveillance, Epidemiology, and End Result (SEER) and multicenter series. The interaction effect between TNM stage and G grade was significant. Twelve subgroups combining the TNM stage and G grade were proposed in the TNMG stage, which were classified into five stages TNMG. According to the TNMG staging classification in the SEER series, the estimated median survival for stages I, II, III, IV, and V were 203, 174, 112, 61, and 8 months, respectively. The predictive accuracy of TNMG stage was higher than that of TNM stage and G grade used independently. The TNMG stage classification was more accurate in predicting panNEN patient's prognosis than either the TNM stage or G grade.

Ectopic fat in liver and skeletal muscle is associated with shorter overall survival in patients with colorectal liver metastases.

BACKGROUND: Myosteatosis has been associated with shorter overall survival in cancer patients. The increase in ectopic fat might not be limited to skeletal muscle only and might also extend to other sites such as the liver, resulting in non-alcoholic fatty liver disease (NAFLD). In this study, we assessed the relationship between myosteatosis and NAFLD and their association with overall survival in patients with colorectal liver metastases undergoing partial hepatectomy. METHODS: Patients were selected from a prospective cohort of 289 consecutive patients with colorectal liver metastases. All patients with a preoperative computed tomography (CT)-scan and liver biopsy obtained during surgery were included. If available a second pre-operative CT scan was used to calculate changes in body composition over time. Muscle radiation attenuation was defined as the average Hounsfield units on CT of all muscle tissue at the L3 level. Liver biopsies were graded by a liver pathologist using the steatosis, activity, and fibrosis scoring system for NAFLD. RESULTS: Two-hundred and eighteen patients had an available liver biopsy of which 131 patients had two available pre-operative CT scans with an average time interval of 3.2 months. One-hundred and thirty-five (62%) biopsies were classified as NAFLD. In multivariable Cox-regression analysis, NAFLD [hazard ratio (HR): 1.8, 95%-confidence interval (CI) 1.0-3.0, P = 0.037], increase in myosteatosis (HR 1.8, 95%-CI 1.1-2.9, P = 0.018), and skeletal muscle loss (HR 1.7, 95%-CI 1.0-2.9, P = 0.035) were independently associated with shorter overall survival while high visceral adipose tissue fat content was associated with longer overall survival (HR: 0.7, 95%-CI 0.5-0.9, P = 0.014). CONCLUSIONS: Ectopic fat content of liver as well as skeletal muscle tissue is independently associated with shorter overall survival in patients with colorectal liver metastases, while increased visceral adipose tissue fat content is associated with longer overall survival.

Long-term variations in solar radiation, diffuse radiation, and diffuse radiation fraction caused by aerosols in China during 1961-2016.

The effects of atmospheric aerosols on the terrestrial climate system are more regional than those of greenhouse gases, which are more global. Thus, it is necessary to examine the typical regional effects of how aerosols affect solar radiation in order to develop a more comprehensive understanding. In this study, we used global AErosol RObotic NETwork (AERONET) data and robust radiation observational evidence to investigate the impact of aerosols on total radiation, diffuse radiation, and the diffuse radiation fraction in China from 1961 to 2016. Our results showed that there were different temporal changes in the aerosol optical depth (AOD), total solar radiation, diffuse radiation and diffuse radiation fraction over the past 56 years. Specifically, the 550 nm AOD from 2005 to 2016 decreased significantly, with annual average AOD of 0.51. Meanwhile, the average total solar radiation reduced by 2.48%, while there was a slight increase in average diffuse radiation at a rate of 3.10 MJ·m-2·yr-1. Moreover, the spatial heterogeneities of AOD, total radiation, diffuse radiation, and the diffuse radiation fraction in China were significant. Aerosol particle emissions in the developed eastern and southern regions of China were more severe than those in the western regions, resulting in higher total radiation and diffuse radiation in the western plateau than in the eastern plain. In addition, aerosols were found to have negative effects on total radiation and sunshine hours, and positive effects on diffuse radiation and diffuse radiation fraction. Further, the diffuse radiation fraction was negatively correlated with sunshine hours. However, there was a positive correlation between AOD and sunshine hours. These results could be used to assess the impacts of climate change on terrestrial ecosystem productivity and carbon budgets.

Clinical characteristics and genetic analyses in a Chinese family affected by primary aldosteronism: a case report.

The clinical and aetiological characteristics of family-clustered primary aldosteronism (PA) are not fully understood and need further exploration. Our study reported a PA case with a family history accompanied by unusual concomitant disease and explored the genetic background of the affected family members, thus providing more evidence of the manifestation and pathogenesis of family-clustered PA. We studied a family with PA in which the proband and her maternal aunt were diagnosed with aldosterone-producing adenoma (APA) and primary adrenal hyperplasia (PAH), respectively. Apart from the diagnosis of APA, the proband also had a history of craniopharyngioma. Both patients achieved desirable blood pressure control and potassium levels after laparoscopic unilateral adrenalectomy. Multiple-gene panel analysis was applied in both resected adrenal lesions and peripheral blood of the proband to screen potential genetic variants. Then, the detected variant was verified by Sanger sequencing in her maternal aunt. No phenotype-related germline mutation was detected in the two affected patients. One somatic nonsense mutation (L168R) of KCNJ5 was detected in the DNA of resected APA from the proband, whereas her maternal aunt did not carry the same somatic mutation. Although no identical mutation was found in the two patients, it remains unknown whether certain unmeasured genetic or epigenetic factors are involved in the development of family-clustered PA. Further studies focused on PA cases with complex manifestations or with a family history will be needed to expand our knowledge of the pathogenesis of PA.

Paperspray Ionization Mass Spectrometry as a Tool for Predicting Real-Time Optimized Dosing of the Chemotherapeutic Drug Melphalan.

BACKGROUND: Melphalan, an important component of hematopoietic stem cell transplantation (HSCT) preparative regimens, is associated with significant toxicity and large between patient variability in pharmacokinetics making it difficult to calculate the optimal dose for pediatric patients. Paperspray (PS) ionization generates gas phase analyte ions directly from a dried blood spot without the need for prior sample preparation or chromatography. With these advantages, a validated PS-MS/MS assay was developed and applied to the 'real-time' determination of melphalan pharmacokinetics (PK). METHODS: Melphalan was quantified by stable-labeled isotope dilution analysis in whole blood by PS-MS/MS. Blood samples were obtained at timed intervals from patients during HSCT after administration of a very low (test) dose of melphalan to avoid toxicity. Pharmacokinetics parameters were calculated using WinNonlin v.6.4. From these data, the optimal therapeutic dose was estimated and full dose PK repeated. RESULTS: PS-MS/MS method was linear over a large dynamic range (25-50 000 ng/mL), intra- and interassay reproducibility of quality control samples was <15% CV. With essentially no prior sample preparation, PS-MS/MS measurement of blood melphalan concentrations showed excellent correlation (R2 = 0.959, n = 62) with a validated electrospray-LC-MS/MS method. Trapezoidal area under the curves calculated for 5 patients administered low dose melphalan showed a high linear correlation (R2 = 0.981) between the PS-MS/MS and LC-MS/MS methods. The faster PS approach permitted real-time PK evaluation of individual patients. CONCLUSIONS: A validated PS-MS/MS assay for melphalan in patients undergoing HSCT is described that facilitates pharmacokinetic-guided individualized precision dosing with immediate bedside dose adjustments to improve outcomes by balancing toxicity and efficacy of melphalan.

Analysis of Immune Microenvironment by Multiplex Immunohistochemistry Staining in Different Oral Diseases and Oral Squamous Cell Carcinoma.

PURPOSE: The aim is to investigate the impacts of using multiplex immunochemistry (mIHC) staining to analyses the co-expression of programmed death ligand-1 (PD-L1) and tumor infiltrating lymphocytes (TILs) [CD8+ T cells and Forkhead Box Protein 3 (FOXP3)+ regulatory T cells (Tregs)] in different oral diseases, and oral squamous cell carcinoma (OSCC). METHODS: Formalin fixed paraffin-embedded tissue sections from different oral diseases were stained with PD-L1 and TILs (CD8+ T cells and FOXP3+ Tregs) by mIHC staining simultaneously. The whole slide was scanned digitally to observe the cell phenotypes stained in the microenvironment. The contents of each slice were read using a computer-aided method to analyze and the cell densities were calculated using statistical software. RESULTS: We were able to characterize the tumor microenvironment (TME) of different oral diseases including oral leukoplakia (OLK), inflammatory gingiva (IG), oral lichen planus (OLP), and squamous cell carcinoma (SCC), with accurate visualization of various immune cells harboring complex immune phenotypes by mIHC staining. The results showed that PD-L1 was up-regulated in SCC tissues at different pathological stages, while CD8 and FOXP3 had no significant changes. The ratio of PD-L1/CD8 was also significantly up-regulated in SCC tissues compared with that of other oral diseases. In advanced stages of OSCC, the FOXP3/CD8 ratio increased, and immunosuppressive TME was more pronounced. In addition, we also found different immune phenotypes: the inflamed phenotype, immune-excluded phenotypes, and immune-desert phenotypes. By locating tumor epithelial cells, we found that PD-L1 expression is in both tumor cells and stromal cells. CONCLUSIONS: mIHC is useful for the visualization and evaluation of tumor microenvironment in immuno-oncology research. It allows single-cell imaging in situ and could effectively and quickly determine the immune phenotype of different oral diseases.

Study on clinical and biological characteristics of ameloblastic carcinoma.

BACKGROUND: Ameloblastic carcinoma (AC) is an odontogenic malignant tumor which is closely related to benign ameloblastoma. Because of its rarity, diagnosis and treatment are difficult. In this study, we summarized and analyzed the clinical and biological characteristics of AC. RESULTS: Fifteen patients with AC and a median age of 53 years were identified. Among of them, five patients who were tested carried a BRAF-V600E mutation. Two patients presented with cervical lymph nodes and lung metastases. Primary AC was more invasive, and the bone destruction ability of the primary type was more radical than that of the secondary type. CONCLUSIONS: This study revealed that the BRAF-V600E mutation was related to the aggressive behavior of AC, and early radical resection is crucial. Moreover, targeted therapy may be a new direction in the future.

Downregulation of MicroRNA-551b Correlates With Dissemination of Human Oral Squamous Cell Carcinoma.

PURPOSE: Altered expression of microRNAs contributes to invasion and metastasis of many human cancers; however, the importance of microRNAs in head and neck cancers remains to be elucidated. In this study, we examined whether altered microRNA (miR)-551b expression correlated with invasive phenotypes of human oral squamous cell carcinoma (OSCC) in vivo and in vitro. MATERIALS AND METHODS: Real-time polymerase chain reaction was used to detect the expression level of miR-551b in 71 OSCC tissues with lymph node metastasis and 50 nonmetastatic OSCC tissues. We also constructed miR-551b mimic-transfected cell lines HN4 and HN12. The effects of overexpressing miR-551b on the proliferation, migration, and invasion of OSCC cells were examined using Cell Counting Kit 8 (Dojindo, Kumamoto, Japan), plate clone formation, wound healing, and Transwell invasion experiments (Corning, Corning, NY). The association between clinical pathologic parameters and the expression level of miR-551b was analyzed using Kaplan-Meier survival analysis. RESULTS: The expression of miR-551b measured 0.33 ± 0.11 in the 71 OSCC tissues with lymph node metastasis versus 0.54 ± 0.06 in the 50 tissues with non-lymph node metastasis (P = .021). Regarding OSCC patients, the expression of miR-551b negatively correlated with patients' overall survival (P = .035). The ectopic expression of miR-551b inhibited the invasion and migration of OSCC cells. CONCLUSIONS: This is the first report showing that reduced miR-551b expression may be an event leading to OSCC invasion and metastasis.

HIF-1α or HOTTIP/CTCF Promotes Head and Neck Squamous Cell Carcinoma Progression and Drug Resistance by Targeting HOXA9.

Head and neck squamous cell carcinoma (HNSCC) is the sixth most frequently diagnosed cancer worldwide. However, the clinical outcomes remain unsatisfactory. The aim of this study is to unravel the functional role and regulatory mechanism of HOXA9 in HNSCC. A cohort of 25 HNSCC tumor tissues and normal tissue counterparts was collected. qRT-PCR and western blotting were performed to determine the levels of HOXA9 and epithelial-mesenchymal transition (EMT)-related markers. Cell Counting Kit-8 (CCK-8) and colony formation assays were conducted to monitor cell viability and cytotoxicity. Transwell and wound healing assays were used to determine cell migration and invasion. Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) staining was performed to detect cell apoptosis. Bioinformatic analysis, electrophoretic mobility shift assay and chromatin immunoprecipitation (ChIP) assays were performed to investigate the direct binding between HIF-1α or CCCTC binding factor (CTCF) and HOXA9. Glutathione S-transferase (GST) pull-down and RNA pull-down assays were used to validate the interaction between CTCF and HOTTIP. HOXA9 was upregulated in HNSCC tissues and cells. Knockdown of HOXA9 inhibited cell proliferation, migration, invasion, and chemoresistance but promoted apoptosis in CAL-27 and KB cells. Knockdown of HOXA9 also regulated EMT-related marker via targeting YAP1/ß-catenin. Silencing of HOTTIP or CTCF exerted similar tumor-suppressive effects in HNSCC. Mechanistically, HIF-1α or CTCF transcriptionally regulated HOXA9, and HOTTIP/CTCF cooperatively regulated HOXA9 in KB cells. HIF-1α or HOTTIP/CTCF transcriptionally modulates HOXA9 expression to regulate HNSCC progression and drug resistance.

Assessment of the radiation effect of aerosols on maize production in China.

With the recent increases in atmosphere aerosol concentration, its impact on agriculture in China is of considerable concern for scientific community. In this study, the effects that aerosols have on radiation and consequently the production of maize in China were investigated from 2002 to 2014 using the AErosol RObotic NETwork (AERONET) data, Second Simulation of a Satellite Signal in the Solar Spectrum radiative transfer (6S) model, and Agricultural Production Systems sIMulator (APSIM) model. Ten stations in the maize planting areas including Beijing, Xianghe, Taihu, Nanjing, Shanghai, Hefei, Baotou, Lanzhou, Qinghaihu, and Xuzhou stations were selected. The results showed that the APSIM-maize model, which was further calibrated, was able to simulate the interactions between maize and the climatic constraints in the maize planting areas of China. Our results indicated that aerosols obviously reduced the amount of solar radiation reaching the surface during the maize growing season in China. We also found that the aerosols have negative effects on both biomass and yield of maize in China at ten stations. The average annual maize biomass during the maize growing season from 2002 to 2014 decreased by 23.70%. The average yield of maize from 2002 to 2014 decreased by 15.10%. However, the influence of aerosol on different varieties of maize varied. We found the aerosols had greater negative impacts on summer maize than on spring maize. For spring maize, the average biomass and yield from 2002 to 2014 decreased by 10.36% and 5.16%, respectively. However, as for the summer maize, the average biomass and yield from 2002 to 2014 were reduced by 19.72% and 20.56%, respectively. Our findings can provide a useful method for estimating the effect of aerosols on crops at the national level, supporting local agricultural production in coping with the ongoing climate change.

Total laparoscopic pancreaticoduodenectomy versus open pancreaticoduodenectomy (TJDBPS01): study protocol for a multicentre, randomised controlled clinical trial.

INTRODUCTION: Pancreatoduodenectomy (PD) is one of the most complex abdominal operations to perform, and it is usually conducted for tumours of the periampullary region and chronic pancreatitis. Minimally invasive surgery has been progressively being developed for pancreatic surgery, first with the advent of hybrid-laparoscopy and recently with total laparoscopic surgery. Issues including the safety and efficacy of total laparoscopic pancreaticoduodenectomy (TLPD) and open pancreaticoduodenectomy (OPD) are currently being debated. Studies comparing these two surgical techniques are emerging, and large randomised controlled trials (RCTs) are lacking but are clearly required. METHODS AND ANALYSIS: TJDBPS01 is a multicentre, prospective, randomised controlled, parallel-group, superiority trial in 14 centres with pancreatic surgery experts who have performed ≥104 TLPDs and OPDs. A total of 656 patients who will undergo PD are randomly allocated to the TLPD group or OPD group in a 1:1 ratio. The trial hypothesis is that TLPD has superior or equivalent safety and advantages in postoperative recovery compared with OPD. The primary outcome is the postoperative length of stay. ETHICS AND DISSEMINATION: The Instituitional Review Board Approval of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology has approved this trial and will be routinely monitoring the trial at frequent intervals, as will an independent third-party organisation. Any results from this trial (publications, conference presentations) will be published in peer-reviewed journals and conference proceedings. TRIAL REGISTRATION NUMBER: NCT03138213.

Intrinsically Cancer-Mitochondria-Targeted Thermally Activated Delayed Fluorescence Nanoparticles for Two-Photon-Activated Fluorescence Imaging and Photodynamic Therapy.

A recent breakthrough in the discovery of thermally activated delayed fluorescence (TADF) emitters characterized by small single-triplet energy offsets (ΔEST) offers a wealth of new opportunities to exploit high-performance metal-free photosensitizers. In this report, two intrinsically cancer-mitochondria-targeted TADF emitters-based nanoparticles (TADF NPs) have been developed for two-photon-activated photodynamic therapy (PDT) and fluorescence imaging. The as-prepared TADF NPs integrate the merits of (1) high 1O2 quantum yield of 52%, (2) sufficient near-infrared light penetration depth due to two-photon activation, and (3) excellent structure-inherent mitochondria-targeting capabilities without extra chemical or physical modifications, inducing remarkable endogenous mitochondria-specific reactive oxygen species production and excellent cancer-cell-killing ability at an ultralow light irradiance. We believe that the development of such intrinsically multifunctional TADF NPs stemming from a single molecule will provide new insights into exploration of novel PDT agents with strong photosensitizing ability for various biomedical applications.

[Effect of long chain non-coding RNA H19 on the migration and invasion of oral cancer cells and its molecular mechanism].

OBJECTIVE: To investigate the effect of the long chain non-coding RNA H19 (lncRNA H19) on the invasion and migration of oral cancer cells and its related molecular mechanism. METHODS: The expression levels of lncRNA H19, miR-107, and cyclin-dependent kinase 6 (CDK6) in the immortalized oral epithelial cell line HIOEC and the oral cancer cell line CAL27 were detected by real-time quantitative polymerase chain reaction. CAL27 cells were transfected with siRNA H19, miR-107 mimics, pcDNA H19, or anti-miR-107, and the effects of H19 and miR-107 on the invasion and migration of cells were examined via Transwell assay. The TargetScan database predicted the targeting of H19, miR-107, and CDK6. Double luciferase reporter gene assay was performed to detect interactions among H19, miR-107, and CDK6. Western blot analysis was conducted to examine the effects of H19 and miR-107 on the protein level of the target gene CDK6. RESULTS: Compared with that in HIOEC cells, the expression of H19 was significantly increased in CAL27 cells (P<0.05). After transfection with siRNA H19, the expression of H19 decreased, and the invasion and migration ability of CAL27 cells were inhibited (P<0.05). H19 could bind specifically to the 3'-UTR of miR-107 to modulate the expression of miR-107. Compared with that in HIOEC cells, the expression of miR-107 significantly decreased in CAL27 cells (P<0.05). The expression of miR-107 increased after transfection with siRNA H19, and anti-mir-107 co-transfection could promote the invasion and migration ability of siRNA H19 in CAL27 cells (P<0.05). Compared with that in HIOEC cells, CDK6 expression significantly increased in CAL27 cells (P<0.05), and the expression level of the gene was coregulated by H19 and miR-107 (P<0.05). CONCLUSIONS: lncRNA H19 plays an important role in the development of oral cancer. It can regulate the invasion and migration of oral cancer cells by targeting the miR-107/CDK6 signaling axis.

Quercetin inhibits cell viability, migration and invasion by regulating miR-16/HOXA10 axis in oral cancer.

Oral cancer is a common tumor malignancy with high mortality and poor prognosis worldwide. Quercetin is one of the major flavonoids present in our daily diet, which is reported to have anti-proliferation and apoptotic effects in varying cancers, including oral cancer. The aim of the present study is to find the mechanism that underlies the role of quercetin in oral cancer. In this study, cell viability, migration and invasion were measured by MTT, trans-well or western blot assays in oral cancer cells. The levels of microRNA-16 (miR-16) and homeobox A10 (HOXA10) were measured in oral cancer tissues and cells by quantitative real-time polymerase chain reaction (qRT-PCR). The interaction between miR-16 and HOXA10 was probed by luciferase activity, RNA immunoprecipitation (RIP) and western blot. Results showed that quercetin suppressed cell viability, migration, invasion and abundances of metalloproteinase-9 (MMP-9) and MMP-2 in oral cancer cells. miR-16 was down-regulated and reversed by addition of quercetin. Moreover, overexpression of miR-16 also impaired cell viability, migration, invasion and abundances of MMP-9 and MMP-2 in oral cancer cells. Besides, HOXA10 was targeted by miR-16 and its restoration abated miR-16-mediated role in oral cancer. In addition, knockdown of miR-16 reversed the effect of quercetin on progression of oral cancer. Collectively, quercetin inhibited cell viability, migration and invasion by regulating miR-16 and HOXA10 in oral cancer cells. This finding indicated that quercetin might be promising for treatment of oral cancer.

Development and Characterization of MDR1 (Mdr1a/b) CRISPR/Cas9 Knockout Rat Model.

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein-9 nuclease (Cas9) technology is widely used as a tool for gene editing in rat genome site-specific engineering. Multidrug resistance 1 [MDR1 (also known as P-glycoprotein)] is a key efflux transporter that plays an important role not only in the transport of endogenous and exogenous substances, but also in tumor MDR. In this report, a novel MDR1 (Mdr1a/b) double-knockout (KO) rat model was generated by the CRISPR/Cas9 system without any off-target effect detected. Western blot results showed that MDR1 was completely absent in the liver, small intestine, brain, and kidney of KO rats. Further pharmacokinetic studies of digoxin, a typical substrate of MDR1, confirmed the deficiency of MDR1 in vivo. To determine the possible compensatory mechanism of Mdr1a/b (-/-) rats, the mRNA levels of the CYP3A subfamily and transporter-related genes were compared in the brain, liver, kidney, and small intestine of KO and wild-type rats. In general, a new Mdr1a/b (-/-) rat model has been successfully generated and characterized. This rat model is a useful tool for studying the function of MDR1 in drug absorption, tumor MDR, and drug target validation.