Characteristics of local extension based on tumor distribution in nasopharyngeal carcinoma and proposed clinical target volume delineation.

OBJECTIVES: To summarize the characteristics of local extension of eccentric and central nasopharyngeal carcinoma (NPC) by magnetic resonance imaging (MRI) and to improve clinical target volume (CTV) delineation. METHODS: MRI of 870 newly diagnosed NPC patients were reviewed. According to tumor distribution features, the NPCs were divided into eccentric and central lesions. RESULTS: All local invasions presented as continuous invasion from gross lesions and structures adjacent to the nasopharynx were more likely to be invaded. There were 240 (27.6%) and 630 (72.4%) cases with central and eccentric lesions, respectively. The spread of eccentric lesions was centered on the ipsilateral Rosenmüller's fossa; and most anatomic sites had significantly higher invasion rates in the ipsilateral side than the contralateral side (P < 0.05). However, they were at low risk of concurrent bilateral tumor invasion (<10%), except the prevertebral muscle (15.4%) and nasal cavity (13.8%). The extension of central NPCs was centered on the nasopharyngeal superior-posterior wall and was more common in the superior-posterior direction. Furthermore, bilateral tumor invasion into the anatomical sites was common. CONCLUSION: Local invasion of NPC was characterized by continuous invasion from proximal to distal sites. The eccentric and central lesions showed different invasion features. Individual CTV delineation should be based on the distribution characteristics of tumors. The eccentric lesions had a very low probability of invasion into the contralateral tissue; thus routine prophylactic radiation of contralateral parapharyngeal space and skull base foramina may not be necessary.

Individualized clinical target volume delineation and efficacy analysis in unilateral nasopharyngeal carcinoma treated with intensity-modulated radiotherapy (IMRT): 10-year summary.

PURPOSE: To evaluate the long-term local control, failure patterns, and toxicities after individualized clinical target volume (CTV) delineation in unilateral nasopharyngeal carcinoma (NPC) treated with intensity-modulated radiotherapy (IMRT). METHODS: Unilateral NPC was defined as a nasopharyngeal mass confined to one side of the nasopharynx and did not exceed the midline. From November 2003 to December 2017, 95 patients were retrospectively included. All patients received IMRT. The CTVs were determined based on the distance from the gross tumor. The contralateral para-pharyngeal space and skull base orifices were spared from irradiation. RESULTS: There were three local recurrences and eight regional recurrences in 10 patients during an 84-month follow-up. All local recurrences were within PGTVnx, and all in-field recurrences. No recurrences were found in traditional high-risk areas including contralateral the para-pharyngeal space and skull base orifices. The 10-year local-recurrence-free survival, regional-recurrence-free survival and overall survival were 96.2%, 90.5% and 84.7%, respectively. The dosimetry parameters of the tumor-contralateral organs were all lower than the values of the tumor-ipsilateral side (P < 0.05). The late toxicities occurred mainly in the tumor-ipsilateral organs, including radiation-induced temporal lobe injury, impaired visuality, hearing loss and subcutaneous fibrosis. CONCLUSION: Individualized CTV delineation in unilateral NPC could yield excellent long-term local control with limited out-of-field recurrences, reduced dose to tumor- contralateral organs and mild late toxicities, which is worthy of further exploration.

Primary Exploration of the Clinical Application of 3D-Printed Complete Dentures.

PURPOSE: To explore the applications of 3D scanning and 3D printing techniques in the restorative treatment of edentulous patients. MATERIALS AND METHODS: A total of 30 edentulous patients (Atwood classes 1 to 4) who visited The 960th Hospital of the People's Liberation Army, Jinan, China, from March 1, 2018 to May 1, 2020 were selected, and the patients were randomly divided into two groups: a traditional complete denture group (group A) and a 3D-printed complete denture group (group B). Each group comprised 15 patients. In group A, the traditional method was used to fabricate complete dentures. In group B, 3D scanning, computer-aided design (CAD), 3D printing, and the duplicate denture technique were used to fabricate the dentures. A single-blinded method was used. Patient satisfaction was measured with a 0-10 visual analog scale (VAS) at four time points: immediately and 1 month, 3 months, and 6 months after denture delivery. SPSS version 22.0 software was used to analyze the data. RESULTS: The ability to speak, ability to chew, and comfort in the two groups gradually improved at the first three time points. VAS scores increased to a satisfactory level after 3 months. The esthetics and stability of the two groups were scored high after the initial delivery. The VAS scores of the two groups regarding esthetics, ability to speak, ability to chew, stability, and comfort were not significantly different (P > .05) at any time point. The number of visits in the 3D-printed complete denture group were significantly decreased in comparison to the traditional group. CONCLUSION: The use of 3D printing for manufacturing complete dentures can rapidly restore edentulous patients and meet patient demands regarding esthetics and function.

Exosomal microRNA-15a from ACHN cells aggravates clear cell renal cell carcinoma via the BTG2/PI3K/AKT axis.

Accumulating studies have indicated that exosomal microRNAs (miRNAs/miRs) can mediate clear cell renal cell carcinoma (ccRCC) at the early stage, but the mechanisms remain to be specified. Here, we investigated the mechanism of exosomal miR-15a in ccRCC. After successful isolation of exosomes from RCC cells, we found that miR-15a was upregulated in ccRCC cells. Moreover, upregulation of miR-15a by pre-miR-15a promoted the proliferation, migration, invasion, and epithelial-mesenchymal transition of ccRCC cells. A luciferase assay revealed that B-cell translocation gene 2 (BTG2) was a target gene of miR-15a and negatively correlated with miR-15a expression. BTG2 was poorly expressed in ccRCC, which reduced the proliferation of ccRCC cells. In addition, overexpression of BTG2 could reverse the promotive effects of miR-15a on ccRCC. Furthermore, BTG2 reduced PI3K/AKT pathway activity. Our results collectively indicated that exosomal miR-15a from RCC cells accelerated cell viability by downregulating BTG2 and promoting the activity of the PI3K/AKT signaling pathway. We demonstrated a novel mechanism by which exosomal miR-15a exerted pro-proliferatory effects on ccRCC, highlighting the potential of exosomal miR-15a as a target for ccRCC prognosis.

A novel low systemic diacylglycerol acyltransferase 1 inhibitor, Yhhu2407, improves lipid metabolism.

Diacylglycerol acyltransferase 1 (DGAT1) plays a pivotal role in lipid metabolism by catalyzing the committed step in triglyceride (TG) synthesis and has been considered as a potential therapeutic target of multiple metabolic diseases, including dyslipidemia, obesity and type 2 diabetes. Here we report a novel DGAT1 inhibitor, Yhhu2407, which showed a stronger DGAT1 inhibitory activity (IC50 = 18.24 ± 4.72 nM) than LCQ908 (IC50 = 78.24 ± 8.16 nM) in an enzymatic assay and led to a significant reduction in plasma TG after an acute lipid challenge in mice. Pharmacokinetic studies illustrated that Yhhu2407 displayed a low systemic, liver- and intestine-targeted distribution pattern, which is consistent with the preferential tissue expression pattern of DGAT1 and therefore might help to maximize the beneficial pharmacological effects and prevent the occurrence of side effects. Cell-based investigations demonstrated that Yhhu2407 inhibited free fatty acid (FFA)-induced TG accumulation and apolipoprotein B (ApoB)-100 secretion in HepG2 cells. In vivo study also disclosed that Yhhu2407 exerted a beneficial effect on regulating plasma TG and lipoprotein levels in rats, and effectively ameliorated high-fat diet (HFD)-induced dyslipidemia in hamsters. In conclusion, we identified Yhhu2407 as a novel DGAT1 inhibitor with potent efficacy on improving lipid metabolism in rats and HFD-fed hamsters without causing obvious adverse effects.

Establishing a Predictive Nomogram for Cervical Lymph Node Metastasis in Patients With Papillary Thyroid Carcinoma.

OBJECTIVES: The purpose of this study was to establish a nomogram for predicting cervical lymph node metastasis (CLNM) in patients with papillary thyroid carcinoma (PTC). MATERIALS AND METHODS: A total of 418 patients with papillary thyroid carcinoma undergoing total thyroidectomy with cervical lymph node dissection were enrolled in the retrospective study from January 2016 to September 2019. Univariate and multivariate Logistic regression analysis were performed to screen the clinicopathologic, laboratory and ultrasound (US) parameters influencing cervical lymph nodes metastasis and develop the predicting model. RESULTS: CLNM was proved in 34.4% (144/418) of patients. In the multivariate regression analysis, Male, Age < 45 years, Tumor size > 20mm, multifocality, ambiguous boundary, extracapsular invasion and US-suggested lymph nodes metastasis were independent risk factors of CLNM (p < 0.05). Prediction nomogram showed an excellent discriminative ability, with a C-index of 0.940 (95% confidence interval [CI], 0.888-0.991), and a good calibration. CONCLUSION: The established nomogram showed a good prediction of CLNM in patients with PTC. It is conveniently used and should be considered in the determination of surgical procedures.

Transcriptomics and Prognosis Analysis to Identify Critical Biomarkers in Invasive Breast Carcinoma.

OBJECTIVE: Invasive breast cancer (BRCA) is one of the prevalent types of invasive tumors with high mortality worldwide. Due to the lack of effective treatment to control the recurrence of distant metastases, the prognosis of BRCA is still very unsatisfactory. We aimed to find some biomarkers by bioinformatics analysis for survival prediction. METHODS: Differentially expressed genes (DEGs) were screened out based on tumor group and normal group. Then, the weighted gene correlation network analysis (WGCNA) was employed to identify the clinically associated gene sets. Meanwhile, the enrichment analyses were performed for the functional annotation of the critical genes. The Kaplan Meier analysis calculated the essential genes' prognostic value. RESULTS: After threshold screening, 1655 DEGs were obtained for subsequent analysis. 51 out of 1655 DEGs were significantly associated with BRCA patients' estrogen receptor status via WGCNA. Three genes (FABP7, CXCL3, and LOC284578) out of the 51 genes were associated with overall survival, and 3 genes were relapse-free survival associated. Finally, we obtained 5 essential prognostic associated genes (FABP7, CXCL3, LOC284578, CAPN6, and NRG2), which could be used as prognostic factors for BRCA. CONCLUSION: Our findings obtained a gene module associated with BRCA clinical trait and several key genes that acted as essential components in the prognostic of cancer, which may improve its treatment.

Patterns of supraclavicular area failure after mastectomy in breast cancer patients: implications for target volume delineation.

PURPOSE: To characterize the pattern of post-mastectomy supraclavicular lymph node (LN) metastases in patients with breast cancer (BC) and to provide insights for individualized clinical target volume delineation for radiotherapy. METHODS: We retrospectively analyzed 88 patients with BC who developed post-mastectomy regional LN metastases. The affected regional LNs were categorized as the ipsilateral medial supraclavicular LN area (IMSC-LN), ipsilateral lateral supraclavicular LN area (ILSC-LN), ipsilateral infraclavicular LN area (IIC-LN), and ≥2 groups in the ipsilateral clavicular LN area (MMIC-LN). Clinical characteristics were included in a multivariate analysis to identify risk factors for clavicular LN metastases. RESULTS: The ILSC-LNs (68.2%) were the most common metastatic site. IMSC-LN metastases showed a significant association with estrogen-receptor (ER) negative status, left-sided BC, and positive axillary LNs. Tumor size ≥2.4 cm and Her2 type were predictors of ILSC-LN metastases. Additionally, tumor size ≥2.4 cm, and level I ipsilateral axillary metastases were associated with MMIC-LN metastasis. CONCLUSION: ILSC-LN was the most frequently affected group of supraclavicular lymph nodes. ER-negative status, left-sided BC, tumor size, and positive ipsilateral axillary LNs are potentially associated with the pattern of supraclavicular LN metastatic involvement.

High Biologically Effective Dose Radiotherapy for Brain Metastases May Improve Survival and Decrease Risk for Local Relapse Among Patients With Small-Cell Lung Cancer: A Propensity-Matching Analysis.

To evaluate whether high biologically effective dose (BED) radiotherapy improves local control and survival outcomes for patients with brain metastases (BMs) from small-cell lung cancer (SCLC) and to determine possible prognostic factors. From January 1998 to June 2018, 250 patients with BM from SCLC were retrospectively analyzed. The Cutoff Finder program was used to classify patients by BED. Overall survival (OS) and BM progression-free survival (BM-PFS) were analyzed using the Kaplan-Meier method and log-rank test. A Cox regression model was used to calculate the hazard ratio and 95% CI for prognostic factors for OS among the study population and propensity score (PS)-matched patients. A BED of 47.4 was taken as the optimal cutoff value. Both OS and BM-PFS were significantly improved in the high-BED (>47.4 Gy) than in the low-BED (≤47.4 Gy) group (median OS: 17.5 months vs 9.5 months, P < .001, median BM-PFS: 14.4 months vs 8.3 months, P < .001). Biologically effective dose (P < .001), Eastern Cooperative Oncology Group performance status (P = .047), smoking (P = .005), and pleural effusion (P = .004) were independent prognostic factors for OS. Propensity score matching with a ratio of 1:2 resulted in 57 patients in the high-BED group and 106 patients in the low-BED group. In the PS-matched cohort, OS and BM-PFS were significantly prolonged in the high-BED group compared with the low-BED group (P < .001). Biologically effective dose >47.4 Gy improves survival among patients with BM from SCLC. Eastern Cooperative Oncology Group score, smoking, and pleural effusion independently affect OS of SCLC patients with BM.

A rapid and convenient derivatization method for quantitation of short-chain fatty acids in human feces by ultra-performance liquid chromatography/tandem mass spectrometry.

RATIONALE: Short-chain fatty acids (SCFAs) are associated with intestinal microbiota and diseases in humans. SCFAs have a low response in mass spectrometry, and in order to increase sensitivity, reduce sample consumption, shorten analysis time, and simplify sample preparation steps, a derivatization method was developed. METHODS: We converted seven SCFAs into amide derivatives with 4-aminomethylquinoline. The reaction occurred for 20 min at room temperature. The analytes were separated on a reversed-phase C18 column and quantitated in the positive ion electrospray ionization mode using multiple reaction monitoring. Acetic acid-d4 was used as the stable-isotope-labeled surrogate analyte for acetic acid in the working solutions, while the other stable-isotope-labeled standards were used as internal standards (ISs). RESULTS: Method validation showed that the intra-day and inter-day precision of quantitation for the seven SCFAs over the whole concentration range was ≤3.8% (n = 6). The quantitation accuracy ranged from 85.5% to 104.3% (n = 6). Most important, the collected feces were vortexed immediately with ethanol. CONCLUSIONS: This study provides a new derivatization method for a precise, accurate, and rapid quantitation of SCFAs in human feces using ultra-performance liquid chromatography/tandem mass spectrometry. This method successfully determined the concentration of SCFAs in human feces and could assist in the exploration of intestinal microbiota and diseases.

Role of postoperative radiotherapy in pT3N0 rectal cancer: A risk-stratification system based on population analyses.

The impact of adjuvant radiotherapy in pT3N0 rectal cancer is controversial. We aimed to determine the risk factors for cancer-specific survival (CSS) among these patients and to develop a risk-stratification system to identify which of these patients would benefit from adjuvant radiotherapy. In this review of the Surveillance, Epidemiology, and End Results database (2010-2014), we analyzed the data of pT3N0 rectal cancer patients who had not undergone neoadjuvant radiotherapy. Prognostic factors were identified using the Cox proportional hazards model, and risk scores were derived according to the ß regression coefficient. A total of 1021 patients were identified from the database search. The overall 5-year CSS was 86.31%. Multivariate analysis showed that age (P < 0.001), tumor differentiation (P = 0.044), number of nodes resected (P = 0.032), marital status (P = 0.005), and radiotherapy (P = 0.006) were independent prognostic factors for CSS. A risk-stratification system composed of age, tumor differentiation, and number of nodes resected was generated. Low-risk patients had better CSS than high-risk patients (92.13% vs 72.55%, P < 0.001). The addition of radiotherapy to surgery doubled the CSS among the high-risk patients (42.06% vs 91.26%, P = 0.001) but produced no survival benefit among the low-risk patients (93.36% vs 96.38%, P = 0.182). Our risk-stratification model based on age, tumor differentiation, and number of nodes resected predicted the outcomes of pT3N0 rectal cancer patients. This model could help identify patients who may benefit from adjuvant radiotherapy.

[Simultaneous determination of gallic acid and hesperidin contained in Xiaogu capsule by HPLC].

OBJECTIVE: To develop an HPLC method for simultaneous determination of gallic acid and hesperidin in Xiaogu capsule, in order to provide a simple, rapid and accurate method for quality control of the compound preparation of traditional Chinese medicine. METHOD: Xiaogu capsule was extracted with methanol heating reflux method. Synergi 4 mu Hydro-RP 80A (4.6 mm x 250 mm, 5 microm) was adopted as the chromatographic column, with acetonitrile--0.04 mol x L(-1) phosphate monobasic sodium solution (20: 80) as the mobile phase. The flow rate was 1.0 mL x min(-1), the detection wavelength was 283 nm, and the column temperature was 25 degrees C. RESULT: Under the conditions, gallic acid and hesperidin reached the baseline resolved peak, with a good linearity within the range of 21.6-216.0 mg x L(-1) (r = 0.999 93) for gallic acid, and 4.5-45.0 mg x L(-1) (r = 0.999 95) for hesperidin, respectively. Their average recoveries (n = 9) were 101.5% (RSD 3.7%) and 94.7% (RSD 2.7%), respectively. The average contents of gallic acid and hesperidin contained in Xiaogu capsule were detected to 5.10% and 0.091 1%, respectively. CONCLUSION: The method established in this study can determine the content of gallic acid and hesperidin contained in Xiaogu capsule in a rapid and accurate manner, which provided reference for quality evaluation of the medicine.

[Outgrowth of neuronal axons on adipose-derived stem cell transplanting for treatment of cerebral infarction in rats].

AIM: To investigate the effects of adipose-derived stem cell (ADSC) transplanting on the outgrowth of neuronal axons and the expressions of GFAP, Neuritin, NF-200 in the brain post focal cerebral ischemia in rats. METHODS: 54 male adult Sprague-Dawley rats were randomly divided into 3 groups: sham-operated group, middle cerebral artery occlusion (MCAO) group and MCAO+ADSC-treated group (n=18 in each group). A permenant focal cerebal ischemia model was established using modified Longa's method ADSC was labeled by DAPI before the transplantation. One day after MCAO, 30 µL of cell suspension containing 1×10(6); cells were injected into the lateral ventricle of MCAO+ADSC-treated group. At 7 d, 14 d and 28 d after MCAO, the expressions of GFAP, Neuritin and NF-200 were detected in ischemic region by Western blot and Immunofluorescence analysis. RESULTS: DAPI staining positive cells were observed around the cerebral infarcted area in the ADSC group. The expressions of Neuritin, NF200 were higher, but GFAP was lower than that of the MCAO group at 7 d, 14 d and 28 d (P<0.05). CONCLUSION: The transplantation of ADSC can induce regeneration and repairment of impaired neuronal axons in rat brain after cerebral ischemia, partly by inhibiting the expression of GFAP and enhancing the expressions of Neuritin, NF-200 in the brain.

[Effects of the transplantation of adipose-derived stem cell on the expression of Notch1-Dll4 signaling pathway in brain of rats with focal cerebral ischemia].

OBJECTIVE: To investigate the effects of the transplantation of adipose-derived stem cells (ADSC) on the expression of Notch1-Dll4 signaling pathway in brains of rats with focal cerebral ischemia. METHODS: Sixty-five male adult Sprague-Dawley rats were randomly divided into 4 groups: sham-operated group, MCAO (occlusion of middle cerebral artery) group, ADSC-treated group and ADSC & DAPT-treated group. A permanent model of focal cerebral ischemia was established by modified Zea-Longa's method. At 24 hours post-MCAO, 1×10(6) DAPT-labeled ADSC were injected into the lateral ventricle of rats in the ADSC-treated group and the same dose of ADSC with DAPT (γ secretase inhibitor, N-[N-(3,5-difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester) to the rats in the ADSC & DAPT-treated group. Rats are sacrificed at 4, 7, 14 and 28 d post-MCAO. The amount of microvessels was quantified. And the levels of Notch1, Dll4 and Hes1 were detected by Western blot and immunohistochemistry. RESULTS: The density of microvessels significantly increased in the ADSC group (13.93 ± 0.50, 17.90 ± 0.62, 20.78 ± 0.80, 17.28 ± 1.65) versus the MCAO group (7.03 ± 0.22, 10.83 ± 0.63, 16.35 ± 0.54, 13.80 ± 2.38) (P < 0.05) and the ADSC + DAPT group (5.73 ± 0.30, 7.58 ± 0.52, 7.65 ± 0.45, 6.48 ± 1.47) (P < 0.05). And compared with the MCAO group (1.29 ± 0.07, 2.13 ± 0.21, 1.92 ± 0.03) and the ADSC + DAPT group (1.162 ± 0.099, 1.684 ± 0.180, 1.041 ± 0.040), the expressions of Notch1, Dll4 and Hes1 proteins were significantly up-regulated at 14d in the ADSC group (2.52 ± 0.22, 4.52 ± 0.36, 2.62 ± 0.05) (P < 0.05). CONCLUSION: The transplantation of ADSC can improve angiogenesis by up-regulating the post-MCAO expression of Notch1-Dll4 signaling pathway in rats.

Fast determination of the release degree of compound sulfamethoxazole tablets by using circulation release system combined with ultraviolet spectral net analyte signal method.

Based on ultraviolet spectral net analyte signals, this research has studied the determination of the two effective components, sulfamethoxazole (SMZ) and trimethoprim (TMP), contained in compound sulfamethoxazole tablets in acid environment. The linear ranges of SMZ and TMP were 0.48-7.84 microg/ml (the regression correlation coefficient r = 0.9981) and 0.12-1.5 microg/ml (r = 0.9986), the corresponding average recoveries were 99.5 and 101.0%, respectively, and the relative standard deviations were 1.87 and 3.60%, respectively. The method was simple, fast and accurate. A circulation release system for the determination of the tablet release degree has been built, and the on-line filtering and small-volume sampling could thus be carried out. Thanks to the combination of the circulation release system and the determination method of SMZ and TMP in acid environment, the determination of release degree for compound sulfamethoxazole tablets was fast and accurate, especially for the early time of rapid release.