Comparative pharmacokinetics of six components in normal and rheumatoid arthritis rats after intragastrical administration of Qianghuo Shengshi Decoction granules by LC-MS/MS.

Objective: To investigate the plasma pharmacokinetics of six representative components (nodakenin, osthole, 5-O-methylvisammioside, ferulic acid, liquiritigenin, and liquiritin), which were the ingredients of Qianghuo Shengshi Decoction (QSD) granules, in normal and rheumatoid arthritis (RA) rats administrated QSD granules intragastrically. Methods: A rapid and accurate ultra-high performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of six components in plasma, and it showed a good specificity, linearity, intra-day and inter-day precision, intra-day and inter-day accuracy, extraction recovery, stability, and the less matrix effect. Results: The validated LC-MS/MS method was successfully used to compare the plasma pharmacokinetics of six ingredients between normal and RA rats after intragastrical administration of QSD granules and differences in the pharmacokinetics were found in two types of rats. The absorption rate in the RA rats was lower for nodakenin, osthole, 5-O-methylvisammioside, liquiritigenin and liquiritin than in the normal group, while the absorption rate of ferulic acid remained constant in two groups. In comparison with the normal rats, the exposure concentration of nodakenin was higher and that of other five components except for nodakenin was lower under pathological conditions. Additionally, the absorptive amount of nodakenin, osthole, 5-O-methylvisammioside and liquiritin was increased and that of ferulic acid and liquiritigenin was reduced in the RA rats than in the normal rats. Compared with the normal rats, the retention time of nodakenin, ferulic acid and liquiritin was reduced in vivo, whereas the retention time of osthole, 5-O-methylvisammioside and liquiritigenin was raised in the body for the RA rats. In contrast to the normal rats, the data demonstrated an increase in the elimination velocity of nodakenin and a decrease in the elimination velocity of the other five components except for nodakenin in the pathological state. Conclusion: This study showed that the pharmacokinetic behavior of the six components, nodakenin, osthole, 5-O-methylvisammioside, ferulic acid, liquiritigenin, and liquiritin, is different in vivo between normal and pathological states of rats, and this research provided the necessary experimental data to explain the pharmacokinetics of QSD granules in both normal and pathological states and provide some references for its clinical application at some level.

The R2R3 MYB gene TaMYB305 positively regulates anther and pollen development in thermo-sensitive male-sterility wheat with Aegilops kotschyi cytoplasm.

MAIN CONCLUSION: The loss of TaMYB305 function down-regulated the expression of jasmonic acid synthesis pathway genes, which may disturb the jasmonic acid synthesis, resulting in abnormal pollen development and reduced fertility. The MYB family, as one of the largest transcription factor families found in plants, regulates plant development, especially the development of anthers. Therefore, it is important to identify potential MYB transcription factors associated with pollen development and to study its role in pollen development. Here, the transcripts of an R2R3 MYB gene TaMYB305 from KTM3315A, a thermo-sensitive cytoplasmic male-sterility line with Aegilops kotschyi cytoplasm (K-TCMS) wheat, was isolated. Quantitative real-time PCR (qRT-PCR) and promoter activity analysis revealed that TaMYB305 was primarily expressed in anthers. The TaMYB305 protein was localized in the nucleus, as determined by subcellular localization analysis. Our data demonstrated that silencing of TaMYB305 was related to abnormal development of stamen, including anther indehiscence and pollen abortion in KAM3315A plants. In addition, TaMYB305-silenced plants exhibited alterations in the transcriptional levels of genes involved in the synthesis of jasmonic acid (JA), indicating that TaMYB305 may regulate the expression of genes related to JA synthesis and play an important role during anther and pollen development of KTM3315A. These results provide novel insight into the function and molecular mechanism of R2R3-MYB genes in pollen development.

Smart Responsive and Controlled-Release Hydrogels for Chronic Wound Treatment.

Chronic wounds are a major health challenge that require new treatment strategies. Hydrogels are promising drug delivery systems for chronic wound healing because of their biocompatibility, hydration, and flexibility. However, conventional hydrogels cannot adapt to the dynamic and complex wound environment, which involves low pH, high levels of reactive oxygen species, and specific enzyme expression. Therefore, smart responsive hydrogels that can sense and respond to these stimuli are needed. Crucially, smart responsive hydrogels can modulate drug release and eliminate pathological factors by changing their properties or structures in response to internal or external stimuli, such as pH, enzymes, light, and electricity. These stimuli can also be used to trigger antibacterial responses, angiogenesis, and cell proliferation to enhance wound healing. In this review, we introduce the synthesis and principles of smart responsive hydrogels, describe their design and applications for chronic wound healing, and discuss their future development directions. We hope that this review will inspire the development of smart responsive hydrogels for chronic wound healing.

[The predicting role of postoperative changes in self-reported symptoms in patients with recurrence eosinophilic chronic sinusitis with nasal polyps].

Objective:To investigate whether changes in postoperative symptoms and signs in patients can predict the recurrence of ECRS after nasal endoscopic sinus surgery. Methods:A total of 70 adult patients with ECRS were enrolled for ESS surgery from June 2020 to March 2022 in a single center. There were 50 males and 20 females, with an average age of （46.9±14.5） years. Follow-up after ESS was at least 52 weeks. Patients undergo peripheral blood tests, CT of the sinuses, olfactory T&T test, visual analogue scale of symptoms（VAS）, and endoscopic scoring. Results:VAS scores and endoscopic scores were analyzed at preoperative and 6th week, 12th week, 24th week and 52th week postoperative. After 12th week postoperatively, there was a clear correlation between symptom scores and endoscopic scores. Moreover, olfactory disorder and nasal discharge were the two most obvious symptoms. There were differences in the expression of multiple preoperative clinical inflammatory indicators between the symptom-controled group and the symptom-uncontrolled group（previous surgical history, concomitant asthma, nasal smear eosinophil, serum EOS%, total IgE, CT score, olfactory score, and symptom score, all with P<0.05）, while there was no difference in baseline endoscopic score（P>0.05）. At 12th week postoperative, the two groups of patients showed significant differences in both symptom scores and endoscopic scores. The symptoms and endoscopic score at the 12th week point of follow-up were used as predictive indicators for recurrence, with sensitivity and specificity of 62.5% and 83.3%, respectively. Conclusion:The changes in postoperative symptom score and endoscopic score in ECRSwNP patients indicated that the recurred ECRS. In the symptom-uncontrolled group, symptomatic and endoscopic scores showed consistent increased scores; In the symptom-controlled group, conflicting results between increased endoscopic scores and stable symptoms suggest that the presence of asymptomatic recurrence must be considered. The changes in symptoms and signs at the 12th week point of follow-up can serve as clinical indicators for preventing disease recurrence.

A geomagnetic vector compensation method compatible with nonlinear interferences based on back propagation network and 3D Helmholtz coil.

Magnetic interferential compensation plays a vital role in geomagnetic vector measurement applications. Traditional compensation accounts for only the permanent interferences, induced field interferences, and eddy-current interferences. However, nonlinear magnetic interferences are found, which also have a great impact on measurement, and it cannot be fully characterized by a linear compensation model. This paper proposes a new compensation method based on a back propagation neural network, which can reduce the influence of the linear model on compensation accuracy due to its good nonlinear mapping capabilities. The high-quality network training requires representative datasets, yet it is a common problem in the engineering field. To provide adequate data, this paper adopts a 3D Helmholtz coil to restore the magnetic signal of a geomagnetic vector measurement system. A 3D Helmholtz coil is more flexible and practical than the geomagnetic vector measurement system itself when generating abundant data under different postures and applications. Simulations and experiments are both conducted to prove the superiority of the proposed method. According to the experiment, the proposed method can reduce the root mean square errors of north, east, and vertical components and the total intensity from 73.25, 68.54, 70.45, and 101.77 nT to 23.35, 23.58, 27.42, and 29.72 nT, respectively, compared with the traditional method.

Cancer-associated fibroblasts suppress ferroptosis and induce gemcitabine resistance in pancreatic cancer cells by secreting exosome-derived ACSL4-targeting miRNAs.

BACKGROUND: Pancreatic cancer continues to be one of the world's most lethal cancers. Chemotherapy resistance in patients with advanced pancreatic cancer often accompany with dismal prognosis, highlighting the need to investigate mechanisms of drug resistance and develop therapies to overcome chemoresistance. METHODS: This research was filed with the Chinese Clinical Trial Registry (ChiCTR2200061320). In order to isolate primary normal fibroblasts (NFs) and cancer-associated fibroblasts (CAFs) samples of pancreatic ductal adenocarcinoma (PDAC) and paracancerous pancreatic tissue from individuals diagnosed with PDAC were obtained. The exosomes were obtained using ultracentrifugation, and their characteristics were determined by Western blotting, nanoparticle tracking analysis, and transmission electron microscopy. CAF-derived miRNAs were analyzed by RT-qPCR and high-throughput sequencing. Gemcitabine (GEM) was employed to promote ferroptosis, and ferroptosis levels were determined by monitoring lipid reactive oxygen species (ROS), cell survival, and intracellular Fe2+ concentrations. To assess in vivo tumor response to GEM therapy, a xenograft tumor mouse model was utilized. RESULTS: Exosomes derived from CAFs in PDAC did not exhibit innate GEM resistance. CAFs promoted chemoresistance in PDAC cells following GEM treatment by secreting exosomes, and maintaining signaling communication with cancer cells. Mechanistically, miR-3173-5p derived from CAF exosomes sponged ACSL4 and inhibited ferroptosis after uptake by cancer cells. CONCLUSION: This work demonstrates a novel mode of acquired chemoresistance in PDAC and identifies the miR-3173-5p/ACSL4 pathway as a promising treatment target for GEM-resistant pancreatic cancer.

Identification of Perna viridis contaminated with diarrhetic shellfish poisoning toxins in vitro using NIRS and a discriminative non-negative representation-based classifier.

Diarrhetic shellfish poisoning (DSP) toxins are one of the most widespread marine biotoxins that affect aquaculture and human health, and their detection has become crucial. In this study, near-infrared reflectance spectroscopy (NIRS) with non-destructive characteristics was used to identify DSP toxins in Perna viridis. The spectral data of the DSP toxin-contaminated and non-contaminated Perna viridis samples were acquired in the 950-1700 nm range. To solve the discrimination of spectra with crossover and overlapping, a discriminative non-negative representation-based classifier (DNRC) has been proposed. Compared with collaborative and non-negative representation-based classifiers, the DNRC model exhibited better performance in detecting DSP toxins, with a classification accuracy of 99.44 %. For a relatively small-scale sample dataset in practical applications, the performance of the DNRC model was compared with those of classical models. The DNRC model achieved the best results for both identification accuracy and F-measure, and its detection performance did not significantly decrease with decreasing sample size. The experimental results validated that a combination of NIRS and the DNRC model can facilitate rapid, convenient, and non-destructive detection of DSP toxins in Perna viridis.

FBG-Based Sensitivity Structure Based on Flexure Hinge and Its Application for Pipeline Pressure Detection.

With the widespread application of pipelines in engineering, more and more accidents occur because of pipeline leakage. Therefore, it is particularly important to continuously monitor the pipeline pressure. In this study, a non-intrusive and high-sensitivity structure based on FBG (Fiber Bragg grating) sensor is proposed. Firstly, the basic sensing theory of FBG and the state of a pipeline wall under inner pressure are analyzed. Then, structural sensitivity is deduced based on the flexure hinge and mechanical lever. Subsequently, finite element simulation for the whole sensitization structure is carried out, and optimal parameters are determined to obtain the maximum sensitivity. Finally, laboratory experiments are conducted to verify the function of the designed sensitivity structure. The experimental results show a good agreement with the simulation results. In the experiment, it can be found that the designed structure has a strain sensitivity of 9.59 pm/µÎµ, which is 11.51 times the pipeline surface strain. Besides, the structure is convenient to operate and has a good applied prospect for the engineering practice.

TRPM2 promotes pancreatic cancer by PKC/MAPK pathway.

The mechanism of pancreatic cancer (PA) is not fully understanded. In our last report, TRPM2 plays a promising role in pancreatic cancer. However, the mechanism of TRPM2 is still unknown in this dismal disease. This study was designed to investigate the role and mechanism of TRPM2 in pancreatic cancer. TRPM2 overexpressed and siRNA plasmid were created and transfected with pancreatic cancer cell line (BxPC-3) to construct the cell model. We employed CCK-8, Transwell, scratch wound, and nude mice tumor-bearing model to investigate the role of TRPM2 in pancreatic cancer. Besides, we collected the clinical data, tumor tissue sample (TT) and para-tumor sample (TP) from the pancreatic cancer patients treated in our hospital. We analyzed the mechanism of TRPM2 in pancreatic cancer by transcriptome analysis, western blot, and PCR. We blocked the downstream PKC/MEK pathway of TRPM2 to investigate the mechanism of TRPM2 in pancreatic cancer by CCK8, scratch wound healing, and transwell assays. Overexpressed TRPM2 could promote pancreatic cancer in proliferation, migration, and invasion ability in no matter the cell model or nude mice tumor-bearing model. TRPM2 level is highly negative correlated to the overall survival and progression-free survival time in PA patients, however, it is significantly increased in PA tissue as the tumor stage increases. The transcriptome analysis, GSEA analysis, western-blot, and PCR results indicate TRPM2 is highly correlated with PKC/MAPK pathways. The experiments of PKC/MEK inhibitors added to TRPM2 overexpressed BxPC-3 cell showed that significant inhibition of PA cells happened in CCK8, transwell, and wound-healing assay. TRPM2 may directly activate PKCα by calcium or indirectly activate PKCε and PKCδ by increased DAG in PA, which promote PA by downstream MAPK/MEK pathway activation.

Use of Nasal Nitric Oxide in the Diagnosis of Allergic Rhinitis and Nonallergic Rhinitis in Patients with and without Sinus Inflammation.

BACKGROUND: Nasal nitric oxide (nNO) has been evaluated in patients with chronic rhinosinusitis with and without nasal polyps. However, nNO levels in patients with allergic rhinitis (AR) and nonallergic rhinitis (NAR) have shown conflicting results in previous studies. OBJECTIVE: To evaluate the value of nNO in diagnosing AR and NAR and the impact of absence or presence of sinus inflammation (SI). METHODS: A total of 173 consecutive patients scanned with high-resolution computed tomography (CT) and 46 normal controls (NCs) were included in our study. Patients were evaluated according to their medical history, nasal symptoms, endoscopic examinations, and skin prick test results. On the basis of CT scans (Lund-Mackay score >2), all patients were subgrouped as AR with SI (ARwSI) and AR without SI (ARsSI), or NAR with SI (NARwSI) and NAR without SI (NARsSI). nNO levels were measured with the NIOX, and eosinophils in nasal smears were evaluated simultaneously. Receiver-operating characteristic analysis was performed for differential diagnosis of AR, NAR, and subgroups. RESULTS: Ninety-four patients were diagnosed with AR and 79 patients with NAR. The levels of nNO were significantly higher in patients with AR and in NCs compared with patients with NAR (939 ± 335 in AR and 813 ± 272 in NCs vs 670 ± 188 in NAR; P < .001 for both), and significantly higher in patients with AR compared with NCs (P < .05). On the basis of sinus CT scans, 49% patients with AR (46 of 94) were defined as ARwSI and 51% patients with NAR (40 of 79) were defined as NARwSI. Patients with ARsSI showed the highest nNO levels (1180 ± 289) compared with other subgroups (P < .001), and patients with NARwSI showed the lowest nNO levels (522 ± 120) compared with other subgroups (P < .001). nNO could be used to discriminate AR, NAR, and subgroups with acceptable sensitivity and specificity. Moreover, patients with nasal smear eosinophilia had lower nNO levels than did patients with NAR (P < .05). CONCLUSIONS: nNO levels are different between patients with ARwSI/ARsSI and NARwSI/NARsSI, and may be used to discriminate these phenotypes.

Identification of heat damage in imported soybeans based on hyperspectral imaging technology.

BACKGROUND: Imported soybeans are prone to heat damage due to high storage temperatures or poor ventilation during transportation. Heat damage directly degrades the quality of the produce and greatly reduces the edible value of soybeans. Rapid and nondestructive techniques for assessing the quality of imported soybeans are in demand. Hyperspectral imaging (HSI) technology was used to distinguish sound soybeans from heat-damaged soybeans. RESULTS: For testing the effectiveness of preprocessing methods in enhancing model performance, five different preprocessing methods were implemented to original spectra. To solve problems related to accuracy, efficiency, and model interpretability caused by high-dimensional HSI data, three waveband selection algorithms - dependency measure (DM-NRS), mutual information (MI-NRS) and variable precision (VP-NRS) - based on neighborhood rough set (NRS) theory were proposed to identify the waveband subsets with optimal distinguishing ability. The effectiveness of preprocessing methods and waveband selection algorithms was validated by establishing two kinds of models: extreme learning machine (ELM) and random forest (RF) models. In addition to the classification accuracy, the robustness of the waveband selection algorithms was studied. The results demonstrated that the Savitzky-Golay (SG) smoothing preprocessing method combined with the MI-NRS waveband selection algorithm and the ELM classifier achieved the best classification and robustness results. Classification accuracy reached 99.98% when using only two optimal wavebands, and reached 100% when using more than four optimal wavebands. CONCLUSION: The results prove that the HSI technology is an accurate, effective, and nondestructive technique for classifying sound and heat-damaged soybeans. © 2019 Society of Chemical Industry.

Relationship between cFGF23/Klotho ratio and phosphate levels in patients with chronic kidney disease.

PURPOSE: To characterize the relationship between the cFGF23/Klotho ratio and phosphate level in patients with chronic kidney disease (CKD). METHODS: A total of 152 patients with CKD stage 3-5 (CKD stage 3: n = 74; CKD stage 4: n = 60; CKD stage 5: n = 18) were included in the study. Thirty healthy volunteers served as controls. Intact-FGF23, cFGF23, Klotho, serum calcium, serum phosphate, and serum creatinine were measured, and estimated glomerular filtration rate (eGFR) was calculated. The Kruskal-Wallis H test was used for comparison between groups, and the Spearman test was used for correlation analysis. RESULTS: In CKD stage 3-5, creatinine and iFGF23 levels, as well as the cFGF23/Klotho ratio, were higher (P < 0.01), phosphate levels were higher (P < 0.05), and Klotho levels were lower (P < 0.01), compared with controls. C-terminal-FGF23 levels were higher in CKD phase 4-5 (P < 0.05). In CKD stage 4-5, creatinine, iFGF23, and phosphate levels, as well as the cFGF23/Klotho ratio, were higher (P < 0.01), cFGF23 levels were higher (P < 0.05), and Klotho levels were lower (P < 0.05), compared with CKD stage 3. In CKD stage 5, creatinine and cFGF23 levels, as well as the cFGF23/Klotho ratio, were higher (P < 0.01), phosphate and iFGF23 levels were higher (P < 0.05), and Klotho levels were lower (P < 0.01), compared with CKD stage 4. Phosphate was positively correlated with the cFGF23/Klotho ratio (r = 0.235, P < 0.01). CONCLUSIONS: EGFR reduction was associated with an increased cFGF23/Klotho ratio, and the cFGF23/Klotho ratio was positively correlated with phosphate. This suggests that the phosphate level can be controlled by modifying the cFGF23/Klotho ratio.

TRPM2 promotes the proliferation and invasion of pancreatic ductal adenocarcinoma.

The aim of the present study was to investigate transient receptor potential cation channel subfamily M member 2 (TRPM2), a promising therapeutic target and biomarker for pancreatic ductal adenocarcinoma (PDAC) prognosis, in addition to determining its effects regarding tumor progression and invasion. PDAC is a fatal disease with a poor prognosis, and its associated pathogenic molecular mechanisms remain to be determined. In the present study, combined analysis using genomic and transcriptomic data from two PDAC studies was performed to discover a survival­associated biomarker of PDAC. Survival analysis for genes mutated in ≥10 patients was performed using a Kaplan­Meier curve and tested for significance using a log­rank test. Furthermore, gene­expression correlation analysis was performed to determine the genes with the strongest correlations to TRPM2. In addition, a Cell Counting Kit­8 assay, a scratch wound­healing assay and a Transwell assay were performed in the present study to investigate the proliferative, invasive and metastatic ability of PANC­1 cells in TRPM2­overexpressing and downregulated groups. The mutated TRPM2 gene had a strong negative correlation with patient survival probability compared with the normal control group (P=1.06x10­4). Expression of TRPM2 was strongly correlated with expression of probable phospholipid­transporting ATPase IM, γ­parvin, tudor domain containing 9, Toll­like receptor 7 and Scm­like with four MBT domains protein 2 according to the criterion of a correlation coefficient >0.5. Furthermore, the results of the present study demonstrated that the TRPM2 overexpression in a PDAC cell line (PANC­1) promoted cell proliferation, invasion and metastatic ability. TRPM2 represents a potential therapeutic target and prognostic marker for patients with PDAC. TRPM2 regulates cell proliferation, invasion and migration; however, the underlying mechanism requires further investigation in future studies.

Bioinformatics analysis to screen key genes implicated in the differentiation of induced pluripotent stem cells to hepatocytes.

Due to the lack of potential organs, hepatocellular transplantation has been considered for treating end-stage liver disease. Induced pluripotent stem cells (iPSCs) are reverted from somatic cells and are able to differentiate into hepatocytes. The present study aimed to investigate the mechanisms underlying iPSC differentiation to hepatocytes. GSE66076 was downloaded from the Gene Expression Omnibus; this database includes data from 3 undifferentiated (T0), 3 definitive endoderm (T5), and 3 early hepatocyte (T24) samples across hepatic­directed differentiation of iPSCs. Differentially expressed genes (DEGs) between T0 and T5 or T24 samples were identified using the linear models for microarray data package in Bioconductor, and enrichment analyses were performed. Using the weighted correlation network analysis package in R, clusters were identified for the merged DEGs. Cytoscape was used to construct protein­protein interaction (PPI) networks for DEGs identified to belong to significant clusters. Using the ReactomeFI plugin in Cytoscape, functional interaction (FI) networks were constructed for the common genes. A total of 433 and 1,342 DEGs were identified in the T5 and T24 samples respectively, compared with the T0 samples. Blue and turquoise clusters were identified as significant gene clusters. In the PPI network for DEGs in the blue cluster, the key node fibroblast growth factor 2 (FGF2) could interact with bone morphogenetic protein 2 (BMP2). Cyclin­dependent kinase 1 (CDK1) was demonstrated to have the highest degree (degree=71) in the PPI network for DEGs in the turquoise cluster. Enrichment analysis for the common genes, including hepatocyte nuclear factor 4α (HNF4A) and epidermal growth factor (EGF), in the FI network indicated that EGF and FGF2 were enriched in the Ras and Rap1 signaling pathways. The present results suggest that FGF2, BMP2, CDK1, HNF4A and EGF may participate in the differentiation of iPSCs into hepatocytes.

A partially penalty immersed Crouzeix-Raviart finite element method for interface problems.

The elliptic equations with discontinuous coefficients are often used to describe the problems of the multiple materials or fluids with different densities or conductivities or diffusivities. In this paper we develop a partially penalty immersed finite element (PIFE) method on triangular grids for anisotropic flow models, in which the diffusion coefficient is a piecewise definite-positive matrix. The standard linear Crouzeix-Raviart type finite element space is used on non-interface elements and the piecewise linear Crouzeix-Raviart type immersed finite element (IFE) space is constructed on interface elements. The piecewise linear functions satisfying the interface jump conditions are uniquely determined by the integral averages on the edges as degrees of freedom. The PIFE scheme is given based on the symmetric, nonsymmetric or incomplete interior penalty discontinuous Galerkin formulation. The solvability of the method is proved and the optimal error estimates in the energy norm are obtained. Numerical experiments are presented to confirm our theoretical analysis and show that the newly developed PIFE method has optimal-order convergence in the [Formula: see text] norm as well. In addition, numerical examples also indicate that this method is valid for both the isotropic and the anisotropic elliptic interface problems.

Sensitization patterns and minimum screening panels for aeroallergens in self-reported allergic rhinitis in China.

Optimization of skin prick test (SPT) panels, especially in view of significant differences in sensitizations patterns within different geographical areas, is an unmet need within China. Our aim was to assess the patterns and clinical relevance of aeroallergen sensitizations in allergic rhinitis (AR) and define the minimal battery of SPT allergens, classified according to the Köppen-Geiger climate map. Overall, 7148 subjects with self-reported AR completed a standard questionnaire and were assessed for sensitization to relevant allergens by SPT. 6340 (88.7%) patients had at least one positive skin prick reaction, and demonstrated unique sensitization patterns by stratification with age, gender, and geographic region. Sensitization to house dust mites (HDM) was highest in south China, whereas the three most prevalent aeroallergens were mugwort, ragweed and dandelion pollen in north-west China. Higher sensitization rates and multiple sensitizations were associated with AR comorbidities. Eight allergens (Der f, mugwort, Blatella, hazel, goosefoot, Penicillium notatum, animal dander and Der p) allowed identification >96% of sensitized subjects in central China. Differences in optimal panels were observed between regions, with five to six allergens being sufficient for north-east, north-west and south China. These SPT panels may provide a cost-effective tool for screening sensitized patients in China.

Ring finger protein 125, as a potential highly aggressive and unfavorable prognostic biomarker, promotes the invasion and metastasis of human gallbladder cancers via activating the TGF- ß1-SMAD3-ID1 signaling pathway.

Human gallbladder cancer (GBC) is a lethal aggressive malignant neoplasm. Identification of potential molecular biomarkers and development of targeted therapeutics for GBC patients is very necessary. In this study, we firstly investigated the correlation between ring finger protein 125 (RNF125) expression and the metastasis and prognosis of GBC, and the underlying molecular mechanism. RNF125 expression in a cohort of GBC tissues was examined; its correlation with clinicopathological and prognostic factors of GBC patients was analyzed. Moreover, the metastasis-related difference expressed genes in highly and lowly aggressive GBC cell lines were identified; and the influence of RNF125 knockdown on the metastatic phenotypes and characteristic EMT markers in highly aggressive GBC NOZ cells was detected. Furthermore, the underlying molecular mechanism of RNF125 effect was explored. The results showed that RNF125 was highly expressed in GBC tissues and related with aggressive characteristics such as Nevin stage (P = 0.041) etc. and unfavorable prognosis of GBC patients (P = 0.023, log-rank test). And, RNF125 was proved to a positive metastasis-related gene in vitro. RNF125 knockdown inhibited the invasion and migration, enhanced the adhesion, upregulated E-cadherin and ß-catenin expression, and downregulated vimentin and N-cadherin expression (all P < 0.001) of NOZ cells in vitro. RNF125 promoting effect on GBC tumor progression was identified to relate with the activation of TGF-ß1-SMAD3-ID1 signaling pathway. These findings firstly confirm that high RNF125 expression is related with aggressive characteristics and unfavorable prognosis of GBC patients; RNF125 promotes the invasion and metastasis of human GBCs via activating the TGF-ß1-SMAD3-ID1 signaling pathway.

Establishment and characterization of a novel highly aggressive gallbladder cancer cell line, TJ-GBC2.

BACKGROUND: Human gallbladder cancer (GBC) is an aggressive malignant neoplasm with a poor prognosis. The development of ideal tools for example tumor cell lines for investigating biological behavior, metastatic mechanism and potential treatment in GBCs is essential. In present study, we established and characterized a GBC cell line derived from primary tumor. METHODS: Primary culture method was used to establish this cell line from a primary GBC. Light and electron microscopes, flow cytometry, chromosome analysis, heterotransplantation and immunohistochemistry were used to characterize the epidemic tumor characteristics and phenotypes of this cell line. RESULTS: A novel GBC cell line, named TJ-GBC2, was successfully established from primary GBC. This cell line had characteristic epithelial tumor morphology and phenotypes in consistent with primary GBC, such as polygon and irregular cell shape, increased CA19-9 and AFP levels, and positive expression of CK7, CK8, CK19 and E-cadherin with negative vimentin. Moreover, about 25% of the cells were in the S-G2/M phase; abnormity in structure and number of chromosome with a peak number of 90-105 and 80% hypertetraploid were observed. Furthermore, this cell line had higher invasion and highest migration abilities compared to other GBC cell lines; and metastatic-related marker MMP9 and nm23 were positively expressed. CONCLUSIONS: A novel highly aggressive GBC cell line TJ-GBC2 was successfully established from primary GBC. TJ-GBC2 cell line may be efficient tool for further investigating the biological behaviors, metastatic mechanism and potential targeted therapy of human GBC.

A potential small-molecule synthetic antilymphangiogenic agent norcantharidin inhibits tumor growth and lymphangiogenesis of human colonic adenocarcinomas through blocking VEGF-A,-C,-D/VEGFR-2,-3 "multi-points priming" mechanisms in vitro and in vivo.

BACKGROUND: Tumor lymphangiogenesis plays an important role in promoting growth and metastasis of tumors, but no antilymphangiogenic agent is used clinically. Based on the effect of norcantharidin (NCTD) on lymphangiogenesis of human lymphatic endothelial cells (LECs), we firstly investigated the antilymphangiogenic activity of NCTD as a tumor lymphangiogenic inhibitor for human colonic adenocarcinomas (HCACs). METHODS: In vivo and in vitro experiments to determine the effects of NCTD on tumor growth and lymphangiogenesis of the in-situ colonic xenografts in nude mice, and lymphatic tube formation of the three-dimensional (3-D) of the co-culture system of HCAC HT-29 cells and LECs were done. Proliferation, apoptosis, migration, invasion, Ki-67, Bcl-2 and cell cycle of LECs and the co-culture system in vitro were respectively determined. Streparidin-peroxidase staining, SABC, western blotting and RT-PCR were respectively used to examine the expression of LYVE-1, D2-40, CK20 (including their LMVD), and VEGF-A, VEGF-C, VEGF-D, VEGFR-2 and VEGFR-3 in vitro and in vivo. RESULTS: NCTD inhibited tumor growth and lymphangiogenesis of the in-situ colonic xenografts in vivo, and these observations were confirmed by facts that lymphatic tube formation, proliferation, apoptosis, migration, invasion, S-phase cell cycle, and Ki-67 and Bcl-2 expression in vitro, and LYVE-1, D2-40, CK20 expression and their LMVD in vitro and in vivo were inhibited and affected. Furthermore, the expression of VEGF-A, VEGF-C, VEGF-D, VEGFR-2 and VEGFR-3 at protein/mRNA levels in the process of lymphatic tube formation in vitro and tumor lymphangiogenesis in vivo was downregulated; NCTD in combination with mF4-31C1 or Sorafenib enhanced these effects. CONCLUSIONS: NCTD inhibits tumor growth and lymphangiogenesis of HCACs through "multi-points priming" mechanisms i.e. affecting related malignant phenotypes, inhibiting Ki-67 and Bcl-2 expression, inducing S-phase cell cycle arrest, and directly or indirectly downregulating VEGF-A,-C,-D/VEGFR-2,-3 signaling pathways. The present finding strongly suggests that NCTD could serve as a potential antilymphangiogenic agent for tumor lymphangiogenesis and is of importance to explore NCTD is used for antitumor metastatic comprehensive therapy for HCACs.

Frequency, suppressive capacity, recruitment and induction mechanisms of regulatory T cells in sinonasal squamous cell carcinoma and nasal inverted papilloma.

BACKGROUND: Sinonasal squamous cell carcinoma (SSCC) and nasal inverted papilloma (NIP) represent the predominant type of malignant and benign tumors in sinonasal tract, respectively. CD4+ CD25+ Foxp3+ natural regulatory T (Treg) cells might play critical role(s) in the suppression of anti-tumor immune response and thus shed light on tumor progression from benign to malignant. OBJECTIVE: This study aimed to evaluate the frequency and suppressive capacity of Treg cells in SSCC compared to NIP and further to explore the underlying mechanisms. PATIENTS AND METHODS: Frequencies of Treg, Th1 and Th2 cells were evaluated by flow cytometry in tissue homogenate and peripheral blood from 31 SSCC patients, 32 NIP patients and 35 normal controls. Treg cells were tested for regulatory function by co-culture with effector T cells. CCR4 and its ligands, CCL22 and CCL17, were analyzed by flow cytometry and Luminex, respectively. The chemoattractant properties of CCR4/CCL22 and CCR4/CCL17 for Treg cells were assessed using the Boyden chamber technique, to elucidate the potential mechanisms of Treg recruitment in tumor microenvironment. Treg cells induction via TGF-ß was assessed with transwells after local CD4+ Foxp3+ T cells were assessed by immunohistochemistry and TGF-ß concentration was measured by Luminex. RESULTS: Tumor-infiltrating Treg cells increased significantly from normal to NIP to SSCC (P ≤ 0.001 for normal vs. NIP and P = 0.004 for NIP vs. SSCC). Significantly elevated frequency and enhanced suppression capacity of circulating Treg cells in SSCC were detected compared to NIP and healthy controls, concomitant with Th1 decrease and Th2 increase. Apparently increased CCL22 attracted CCR4-expressing Treg cells to tumor microenvironment in SSCC, compared to NIP. SSCC produced significantly more TGF-ß than NIP and thus possessed greater potential for Treg cell induction. CONCLUSION: Frequency and suppressive capacity of Treg cells enhanced with progression of malignancy from NIP to SSCC. Circulating Treg cells were recruited to tumor tissue via CCR4/CCL22 signalling, whereas tumor-synthesised TGF-ß contributed to induction of peripheral Treg cells.