Silencing of AJAP1 expression by promoter methylation activates the Wnt/ß-catenin signaling pathway to promote tumor proliferation and metastasis in salivary adenoid cystic carcinoma.

Background: Salivary adenoid cystic carcinoma (SACC) is a unique malignant tumor of the salivary gland with poor prognosis, which is not effective with chemotherapy and targeted drugs. Therefore, it is important to explore the molecular mechanism underlying SACC invasion and metastasis to develop novel therapeutic strategies and targets in clinical research. Methods: Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot (WB) were performed to detect the expression of Adherens Junctions Associated Protein 1 (AJAP1). Methylation-specific PCR was used to evaluate the methylation of the AJAP1 promoter. AJAP1 was overexpressed or knocked down by lentivirus-mediated transfection. Kaplan-Meier analysis was conducted to create a survival curve and the log-rank test was used to analyze the overall survival (OS). The prognostic correlation was assessed using univariate and multivariate Cox regression analyses. Co-immunoprecipitation (Co-IP) was utilized to pull down the possible binding protein of AJAP1 and laser scanning confocal microscopy was applied to detect the subcellular localization of AJAP1, E-cadherin, and ß-catenin. Cell viability, colony formation, wound healing, and Transwell invasion assays were performed to evaluate the function of AJAP1 in vitro. A subcutaneous xenograft assay in nude mice was performed to verify the function of AJAP1 in vivo. Results: AJAP1 was downregulated in SACC tumors and was closely related to SACC lymph node/distant metastasis, which was an independent risk factor for SACC prognosis. Methylation-specific PCR confirmed that high methylation of the AJAP1 promoter was the main cause of its silencing. Overexpression or knockdown of AJAP1 in SACC cells could significantly inhibit or promote the proliferation, invasion, and metastasis of SACC cells, respectively, in both the in vitro and in vivo experiments. Mechanically, we found that AJAP1 binds to E-cadherin and ß-catenin to form a complex in cytomembrane, reducing the nuclear translocation of ß-catenin and blocking the Wingless/Integrated/ß-catenin (Wnt/ß-catenin) signaling pathway to play a suppressive role in cancer. Conclusions: In conclusion, these results suggest that the downregulation of AJAP1 protein expression may play a certain role in progression and metastasis of SACC. Our study indicates that AJAP1 may be a potential prognostic molecular marker and therapeutic target for SACC.

Construction of nano-drug delivery and antitumor system of stimuli-responsive polypeptides.

The size of the nanoparticles is moderate and the dispersion is well, which will not be recognized nonspecifically and clearance by the endothelial reticular system. In this study, stimuli-responsive polypeptides nano-delivery system has been constructed, which can realize the response to various stimuli in the tumor microenvironment. Tertiary amine groups are grafted to the side chain of polypeptides as the point of charge reversal and particle expansion. In addition, a new kind of liquid crystal monomer was prepared by substituting cholesterol-cysteamine, which can promote polymers to realize the transformation of spatial conformation by adjusting the ordered arrangement of macromolecules. The introduction of hydrophobic elements greatly enhanced the self-assembly performance of polypeptides, which could effectively improve the drug loading and encapsulation rate of nanoparticles. Nanoparticles could achieve targeted aggregation in tumor tissues, and there were no toxicity and side effects on normal bodies during treatment, with good safety in vivo.

A Novel and Effective Model to Predict Skip Metastasis in Papillary Thyroid Carcinoma Based on a Support Vector Machine.

Introduction: Skip metastasis, referred to as lymph node metastases to the lateral neck compartment without involvement of the central compartment, is generally unpredictable in papillary thyroid carcinoma (PTC). This study aims to establish an effective predictive model for skip metastasis in PTC. Meterials and Methods: Retrospective analysis was performed of clinical samples from 18192 patients diagnosed with thyroid cancer between 2016 to 2020. The First Affiliated Hospital of Wenzhou Medical University. The lateral lymph node metastasis was occureed in the training set (630 PTC patients) and validation set (189 PTC patients). The univariate and multivariate analyses were performed to detect the predictors of skip metastasis and the support vector machine (SVM) was used to establish a model to predict skip metastasis. Results: The rate of skip metastasis was 13.3% (84/631). Tumor size (≤10 mm), upper location, Hashimoto's thyroiditis, extrathyroidal extension, absence of BRAFV600E mutation, and less number of central lymph node dissection were considered as independent predictors of skip metastasis in PTC. For the training set, these predictors performed with 91.7% accuracy, 86.4% sensitivity, 92.2% specificity, 45.2% positive predictive value (PPV), and 98.9% negative predictive value (NPV) in the model. Meanwhile, these predictors showed 91.5% accuracy,71.4% sensitivity, 93.1% specificity, 45.5% PPV, and 97.6% NPV in validation set. Conclusion: This study screened the predictors of the skip lateral lymph node metastasis and to establish an effective and economic predictive model for skip metastasis in PTC. The model can accurately distinguish the skip metastasis in PTC using a simple and affordable method, which may have potential for daily clinical application in the future.

A Systematic Pan-Cancer Analysis of CASP3 as a Potential Target for Immunotherapy.

CASP3 is the gene encoding caspase-3, a specific protease that cleaves substrates such as poly-ADP ribose polymerase and acetyl-DEVD-7-amino-4-methylcoumarin. This enzymatic activity leads to DNA fragmentation, which is a hallmark of apoptosis. Although recent studies have demonstrated that CASP3 plays a vital role in tumour suppression by promoting apoptosis, these reports did not consider systematic pan-cancer analyses. Therefore, we performed a specific pan-cancer analysis using The Cancer Genome Atlas and Genotype-Tissue Expression databases to analyse CASP3 expression in terms of cancer prognosis, DNA methylation status, tumour mutative burden (TMB), and microsatellite instability (MSI), as well as immune cell infiltration in different tumours and the molecular mechanisms underlying these. We found that CASP3 expression was significantly associated with the prognosis of most tumours. Additionally, promoter methylation status was associated with CASP3 expression in bladder urothelial carcinoma, oesophageal carcinoma, kidney renal clear cell carcinoma, kidney renal papillary cell carcinoma, lung squamous cell carcinoma, prostate adenocarcinoma, sarcoma, testicular germ cell tumours, and uterine corpus endometrial carcinoma. TMB and MSI were associated with CASP3 expression in 15 tumours. Moreover, CASP3 expression was correlated with the tumour microenvironment in nearly all tumour types. Further, we observed that in addition to apoptosis, CASP3 action plausibly involves B cell activation, antigen presentation, immune responses, chemokine receptors, and inflammatory function. Our study thus provides a relatively comprehensive understanding of the carcinogenicity of CASP3 in different tumours and suggests that CASP3 is a potential prognostic marker.

Untargeted metabolomics analysis by gas chromatography/time-of-flight mass spectrometry of human serum from methamphetamine abusers.

Methamphetamine (METH) abuse has become a global public health problem. However, the potential mechanisms involving METH-induced metabolic disorders have thus far remained poorly understood. Metabolomics can provide a clue for the cause of apparent changes and consequently be used to investigate the METH-induced dysregulation of metabolite expression and the mechanism of metabolic disorder mechanism. This laboratory investigation included 80 METH abusers and 80 healthy people. The serum metabolites were detected and analysed by gas chromatography/time-of-flight mass spectrometry. Raw data were processed with the software MS DIAL, which includes deconvolution, peak alignment and compound identification. The data matrix was processed by univariate and multivariate analyses for significant metabolite screening with the criteria of variable importance in projection values > 1, fold change > 1.5 and the t test (p value < 0.05). Significant differences in 16 metabolites (deoxycholic acid, cholic acid, hydroxylamine, etc.) in serum were found between the METH abuse group and the control group. Energy metabolic pathways and several amino acid metabolic pathways (alanine, aspartic acid and glutamate metabolism and tryptophan metabolism) were primarily involved. Further analysis indicated that the area under the receiver operating characteristic curve (AUC) was 0.998 for these 16 metabolites. Among the metabolites, three carbohydrates (d-ribose, cellobiose and maltotriose) had an AUC of 0.975, which were determined as potential markers of abuse. We observed metabolic disturbances in METH abusers, particularly perturbation in energy metabolism and amino acid metabolism, which can provide new insights into the search for biomarkers and the mechanisms underlying the adverse effects of METH on human health.

Endoscopic thyroidectomy using gasless axillary approach for low-risk papillary thyroid carcinoma.

To evaluate the application of endoscopic thyroidectomy using gasless axillary approach (ET-GA) for low-risk papillary thyroid carcinoma (PTC). Patients with T1N0M0 Ⅰ PTC undergoing unilateral thyroid lobectomy with central neck dissection in Taizhou Cancer Hospital during January 2019 to June 2021 were enrolled in the study, including 35 cases treated with ET-GA (ET-GA group) and 35 cases treated with conventional open thyroidectomy (COT group). The surgical treatment effect, cosmetic effect and the effect on neck function were compared between two groups. Neck function was evaluated by neck pain score, neck injury index and dysphagia index. Cosmetic effect was evaluated by cosmetic effect satisfaction score. In the ET-GA group, all unilateral thyroid lobectomy with central neck dissection were successfully completed, and no case was converted to open surgery. The number of central lymph nodes dissected in the ET-GA was not statistically different from that in the COT group (>0.05), but the operation time was longer than that of the COT group (<0.01). In the ET-GA group, 2 cases (5.7%) had transient vocal cord paralysis and 1 case (2.9%) had postoperative bleeding. In the COT group, 1 case (2.9%) had transient vocal cord paralysis, no postoperative bleeding. There was no significant difference in the complication rate between two groups (>0.05). At and postoperatively, there was no significant difference in neck pain score and neck injury index between two groups (both >0.05); dysphagia index was lower in ET-GA group, but the difference was not statistically significant (>0.05). The cosmetic effect satisfaction score of ET-GA group was higher than that in the COT group at postoperatively (4.3±0.6 vs.1.0, <0.01). ET-GA has the same efficacy and safety as conventional open thyroidectomy in the treatment of low-risk PTC, and it improves the satisfaction of postoperative cosmetics.

Advances in targeted therapy for anaplastic thyroid carcinoma.

Anaplastic thyroid carcinoma (ATC) is a highly malignant and aggressive thyroid malignancy with rapid onset and poor prognosis. There is no effective treatment for ATC yet. Molecular targeted therapy provides a new idea for ATC treatment. Tyrosine kinase inhibitor lenvatinib has potential in treating ATC patients with favorable efficacy in clinical trials. The effectiveness of the v-raf murine sarcoma viral oncogene homologue B1 () gene inhibitor dabrafenib in combination with trametinib for the treatment of positive ATC patients has been demonstrated in clinical trials. The has proposed dabrafenib in combination with trametinib as the preferred modality for the treatment of patients with positive ATC. The immune checkpoint inhibitor pembrolizumab can be applied to treat thyroid cancer with high tumor mutational load and may be considered as the preferred modality for the treatment of ATC patients with high programmed death ligand-1 expression. The mammalian target of rapamycin pathway inhibitors, peroxisome proliferators-activated receptor γ agonists, endothelial growth factor receptors-targeting monoclonal antibody cetuximab and novel vascular blocker fosbretabulin are still in the clinical research stage, which are expected to provide new directions for the development of novel targeted drugs. This article reviews the current research progress on targeted drugs for the treatment of ATC.

Opposite estrogen effects of estrone and 2-hydroxyestrone on MCF-7 sensitivity to the cytotoxic action of cell growth, oxidative stress and inflammation activity.

There are many kinds of estrogens, and endogenous estrogens produce a variety of estrogen metabolites with similar structure but with different physiological effects after metabolism in vivo. Studies have shown that estrone (E1) widely occurs in the environment and animal-derived food. Because of its estrogen effect, E1 can have adverse effects on the human body as an endocrine disruptor. In this study, we found that E1 and 2-hydroxyestrone (2-OH-E1), the hydroxylation metabolite of estrogen, have opposite proliferative effects on breast cancer cells (MCF-7) through cell proliferation experiments and comparison of their effects by molecular docking and detection of ROS, Ca2+, and cell pathway proteins. The effects of 2-methoxyestrone (2-MeO-E1) and 16α-hydroxyestrone (16α-OH-E1) on the biochemical and protein levels of MCF-7 were further studied to compare the effects of metabolic sites and modes on estrogen effects. Hydroxylation of E1 at the C2 site weakened the estrogen effect, down-regulated the expression of the mammalian target of rapamycin (mTOR) and protein kinase B (Akt) pathway proteins, inhibited the proliferation of cancer cells, and enhanced anti-oxidative stress and anti-inflammation. Methoxylation at the C2 position also inhibited the expression of inflammatory and oxidative stress pathway proteins but did not greatly affect the estrogen effects. However, hydroxylation on C16 had no significant effect on the biological effects of estrogen. Therefore, the structural changes of estrogen on C2 are important reasons for the different physiological effects of estrogen and its metabolites. Thus, by regulating the gene Cytochrome P450 1B1(CYP1B1), which affects the hydroxylation metabolism of estrogen, and promoting the hydroxylation of estrone at the C2 position, the estrogen effect of estrone can be effectively reduced, thus reducing the harm its poses in food and the environment.

Unreduced Male Gamete Formation in Cymbidium and Its Use for Developing Sexual Polyploid Cultivars.

Polyploidy plays an important role in crop improvement. Polyploid plants, particularly those produced through unreduced gametes (2n gametes), show increased organ size, improved buffering capacity for deleterious mutations, and enhanced heterozygosity and heterosis. Induced polyploidy has been widely used for improving floriculture crops, however, there are few reported sexual polyploid plants in the floriculture industry. This study evaluated nine cultivars of Cymbidium Swartz and discovered that 2n male gametes occurred in this important orchid. Depending on cultivars, 2n male gamete formation frequencies varied from 0.15 to 4.03%. Interspecific hybrids generally produced more 2n male gametes than traditional cultivars. To generate sexual polyploid plants, seven pairs of crosses were made, which produced five triploid and two tetraploid hybrids. Two triploid hybrids were evaluated for in vitro regeneration and growth characteristics. Compared to the diploid parents, the triploids were more easily regenerated through rhizomes or protocorms, and regenerated plants had improved survival rates after transplanting to the greenhouse. Furthermore, the sexual polyploid plants had more compact growth style, produced fragrant flowers, and demonstrated heterosis in plant growth. Through this study, a reliable protocol for selection of appropriate parents for 2n gamete production, ploidy level evaluation, in vitro culture of polyploid progenies, and development of new polyploid cultivars was established. Our study with Cymbidium suggests that the use of 2n gametes is a viable approach for improving floriculture crops.

[Onset time and efficacy of sublingual immunotherapy with Dermatophagoides farinae drops in children with allergic rhinitis].

OBJECTIVE: To investigate the onset time and efficacy of sublingual immunotherapy (SLIT) with Dermatophagoides farinae drops in children with house dust mites (HDM)-induced allergic rhinitis (AR). METHODS: One hundred and forty three children with perennial moderate to severe HDM-induced AR were treated by SLIT with standardized Dermatophagoides farinae extract. One hundred children who finally completed two years treatment were divided into two groups according to the age: younger children group (aged 4-8 years, n = 52) and older children group (aged 9-14 years, n = 48). Respectively, Each children was assessed before and after 1st, 2nd, 3rd, 6th, 12th, 24th months of the treatment. Total nasal symptom score (TNSS), total medication score (TMS) and visual analogue scale (VAS) were evaluated at each visit. All clinical data were analyzed retrospectively with the SPSS 19.0 software. RESULTS: TNSS, TMS and VAS of two groups decreased significantly after three months of the treatment compared with before (younger children group: Z value was -3.843, -3.534, -3.940, older children group: Z value was -3.938, -3.405, -3.953, all P < 0.05). TNSS and VAS of younger children group decreased significantly after two months of the treatment compared with before (6.4 ± 1.6, 5.3 ± 1.4 vs 8.6 ± 1.2, 7.9 ± 1.6, Z value was -3.843, -3.940, both P < 0.05). Five children (5%) experienced local adverse events and 2 children (2%) experienced mild systemic adverse events. No severe adverse events happened during the treatment. CONCLUSIONS: SLIT with Dermatophagoides farinae drops is an efficient and safe treatment for children with HDM-induced AR. Its onset of action can be observed as early as 3 months after treatment.

Health risk assessment of odors emitted from urban wastewater pump stations in Tianjin, China.

In order to evaluate the potential adverse health effects of odor emissions from wastewater pump stations (WWPSs) to human, a health risk assessment was performed to study the odors emitted from an urban WWPS in a residential area, Tianjin (in North China). First, 15 types of volatile organic compounds in the WWPS were collected and analyzed using gas chromatography-mass spectrometry. Next, Monte Carlo probabilistic modeling was applied to evaluate the potential health effects of four odors (chlorobenzene, dichloromethane, hydrogen sulfide, and carbon disulfide), which had higher concentrations. The results revealed that the 95th percentile of the total non-carcinogenic risk was approximately 1.73, which poses a threat to human health. In addition, hydrogen sulfide had the highest non-carcinogenic risk value of the four; the hazard quotient of hydrogen sulfide was estimated to be 1.60 at the 95th percentile, higher than the upper confidence limit (1.0). The 95th percentile of the carcinogenic risk was approximately 5.47E-08, much lower than the maximum acceptable level (1.0E-06). Finally, the influence of the input variables on the output was evaluated using sensitivity analysis, and contaminant concentration, reference concentration, and inhalation unit risk were the most influential variables.

[Clinical characteristics of benign paroxysmal positional vertigo secondary to sudden deafness].

OBJECTIVE: To retrospectively analyze the clinical characteristics of the benign paroxysmal positional vertigo (BPPV) secondary to the sudden deafness (SD) and to explore pathogenetic mechanism. METHOD: One hundred and seventy-eight cases of the SD in our department were retrospectively analyzed. They were all treated under the guidance of clinical guidelines. RESULT: (1) In all these patient's with SD, there were 31 cases with BPPV secondary to the SD. There were 26 cases of BPPV of posterior semicircular canal and 5 cases of BPPV of lateral BPPV semicircular canal. All patients with BPPV were diagnosed as the same ears as the SD, including 16 cases on left sides and 15 on right sides. (2) The interval between the onset of SD and BPPV was less than one week in 27 cases, between one week and one month in 3 cases, and between one and three months in 1 case. (3) All patients with BPPV secondary to the SD were cured with Epley maneuver or Barbecue roll maneuver. CONCLUSION: The occurrence of BPPV may follow SD, and the major of BPPV secondary to the SD occurs in the posterior semicircular canal. The canalith repositioning is an effective therapy to the secondary BPPV.

Room temperature phosphorescence sensor for Hg2+ based on Mn-doped ZnS quantum dots.

Mercury pollution is one of the most serious concerns to human health and the environment. The development of highly sensitive and selective sensors to detect toxic mercury ions has been the focus of scientific research. In this study, L-cysteine-capped Mn-doped ZnS quantum dots (QDs) have been synthesized and used for the room temperature phosphorescence detection of Hg2+. The phosphorescence of the Mn-doped ZnS QDs could be selectively quenched in the presence of Hg2+. Under optimal conditions, good linear correlations were obtained over the concentration range from 2.0 x 10(-8) mol/L to 4.5 x 10(-6) mol/L with a detection limit of 3.8 x 10(-9) mol/L. Additionally, the long lifetime of the phosphorescence of the Mn-doped ZnS QDs can avoid the interference of the autofluorescence and scattering light of the background, which facilitates their application in real samples. The possible quenching mechanism was examined by UV-vis absorption spectra and Rayleigh scattering spectra.

Preparation of whey protein hydrolysates using a single- and two-stage enzymatic membrane reactor and their immunological and antioxidant properties: characterization by multivariate data analysis.

An initial 5% (w/v), followed thereafter with replacement aliquots of 3% (w/v), whey protein isolate (WPI) (ca. 86.98% Kjeldahl N x 6.38), was hydrolyzed using Protease N Amano G (IUB 3.4.24.28, Bacillus subtilis) in an enzymatic membrane reactor (EMR) fitted with either a 10 or 3 kDa nominal molecular weight cutoff (NMWCO) tangential flow filter (TFF) membrane. The hydrolysates were desalted by adsorption onto a styrene-based macroporous adsorption resin (MAR) and washed with deionized water to remove the alkali, and the peptides were desorbed with 25, 50, and 95% (v/v) ethyl alcohol. The desalted hydrolysates were analyzed for antibody binding, free radical scavenging, and molecular mass analysis as well as total and free amino acids (FAA). For the first time a quantity called IC50, the concentration of peptides causing 50% inhibition of the available antibody, is introduced to quantify inhibition enzyme-linked immunosorbent assay (ELISA) properties. Principal component analysis (PCA) was used for data reduction. The hydrolysate molecular mass provided the most prominent influence (PC1 = 57.35%), followed by inhibition ELISA (PC2 = 18.90%) and the antioxidant properties (PC3 = 10.43%). Ash was significantly reduced in the desalted fractions; the protein adsorption recoveries were high, whereas desorption with alcohol was prominently influenced by the hydrophobic/ hydrophilic amino acid balance. After hydrolysis, some hydrolysates showed increased ELISA reactivity compared with the native WPI.

[Separation and identification of volatile flavors of Jinhua ham by gas chromatography-mass spectrometry coupled with head space solid phase microextraction].

Head space solid phase microextraction (SPME) technique was employed to extract volatile flavor compounds from the Chinese traditional food, Jinhua ham. Volatile flavor components of Jinhua ham were isolated and identified successfully by capillary gas chromatography-mass spectrometry (GC-MS). A 75 microm carboxen/poly (dimethysiloxane) (CAR-PDMS) fiber was selected to be the optimum fiber among four different fibers. Optimum adsorption time was 40 min at 60 degrees C. Aldehydes, carboxylic acids and acetones were the major flavor compounds of Jinhua ham.