The improvement of postoperative blood pressure and associated factors in patients with hormone-negative adrenal adenoma and hypertension.

OBJECTIVE: To investigate the effect of adrenal surgery on blood pressure (BP) improvements in patients with hormone-negative adrenal adenoma (HNA) concomitant with hypertension and analyze associated prognostic factors. METHODS: We retrospectively reviewed the clinical data of patients with HNA and hypertension and patients with aldosterone-producing adenoma (APA) and hypertension who underwent adrenal surgery at our center between 2019 and 2022. Hypertension outcomes were evaluated in all patients and subjects were divided into three groups according to follow-up BP and the administration of anti-hypertensive agents: a clinical curation group, an improvement group, and a no-improvement group. Logistic regression analysis was performed to predict factors associated with clinical curation in patients with HNA post-surgery. RESULTS: Of the 182 patients with HNA, clinical curation was achieved in 58 patients (31.9%), improvement in 72 (39.5%), and no improvement in 52 (28.6%). The clinical curation, improvement and no improvement rates in patients with APA were 64.8% (n = 118), 15.9% (n = 29), and 19.2% (n = 35). Multivariate logistic regression analysis indicated that a duration of hypertension ≤6 years and a plasma aldosterone level >160 pg/ml were both independent factors for the clinical curation of hypertension in patients with HNA after adrenal surgery. CONCLUSION: Adrenal surgery can cure or improve hypertension in most patients with HNA, especially in a short duration of hypertension and high plasma levels of aldosterone.

High-power green-light laser endoscopic submucosal dissection for non-muscle-invasive bladder cancer: A technical improvement and its initial application.

Background: The technique of laser en bloc resection of bladder tumor (ERBT) has been a valuable alternative technique to transurethral resection of bladder tumor (TURBT). However, the combination of laser ERBT and endoscopic submucosal dissection (ESD) technique has not been well studied. Here, a novel technique integrating a high-power green-light laser with ESD was presented. This study aimed to evaluate the safety and efficacy of high-power green-light laser endoscopic submucosal dissection (HPL-ESD) for the treatment of primary non-muscle-invasive bladder cancer (NMIBC). Materials and Methods: From January 2015 to December 2018, a total of 56 patients with NMIBC underwent HPL-ESD. All tumors were transurethral en bloc resected in the ESD technique. Perioperative clinical data were retrospectively collected and analyzed. Results: All operations were safely performed by the technique of HPL-ESD without blood transfusion. The mean tumor diameter was 2.04 ± 0.65 cm, ranging from 0.5 to 3.5 cm. The mean operative time was 28.39 ± 16.04 min. The average serum hemoglobin decrease was 0.88 ± 0.54 g/dL. The mean postoperative catheterization time was 2.88 ± 0.94 days. The pathologic stages included pTa (32 cases), and pT1 (24 cases). Double-J stent indwelling was not performed for four patients whose tumors were adjacent to the ureteral orifice and no postoperative hydronephrosis was observed. Only one case of ectopic bladder tumor recurred due to irregular bladder irrigation during the 36-month follow-up. Conclusion: HPL-ESD is a safe and effective alternative for the treatment of primary NMIBCs, especially for tumors adjacent to the ureteral orifice.

Identification of tumor antigens and immune landscapes for bladder urothelial carcinoma mRNA vaccine.

Background: Bladder urothelial carcinoma (BLCA) is associated with high mortality and recurrence. Although mRNA-based vaccines are promising treatment strategies for combating multiple solid cancers, their efficacy against BLCA remains unclear. We aimed to identify potential effective antigens of BLCA for the development of mRNA-based vaccines and screen for immune clusters to select appropriate candidates for vaccination. Methods: Gene expression microarray data and clinical information were retrieved from The Cancer Genome Atlas and GSE32894, respectively. The mRNA splicing patterns were obtained from the SpliceSeq portal. The cBioPortal for Cancer Genomics was used to visualize genetic alteration profiles. Furthermore, nonsense-mediated mRNA decay (NMD) analysis, correlation analysis, consensus clustering analysis, immune cell infiltration analysis, and weighted co-expression network analysis were conducted. Results: Six upregulated and mutated tumor antigens related to NMD, and infiltration of APCs were identified in patients with BLCA, including HP1BP3, OSBPL9, SSH3, ZCCHC8, FANCI, and EIF4A2. The patients were subdivided into two immune clusters (IC1 and IC2) with distinct clinical, cellular and molecular features. Patients in IC1 represented immunologically 'hot' phenotypes, whereas those in IC2 represented immunologically 'cold' phenotypes. Moreover, the survival rate was better in IC2 than in IC1, and the immune landscape of BLCA indicated significant inter-patient heterogeneity. Finally, CALD1, TGFB3, and ANXA6 were identified as key genes of BLCA through WGCNA analysis, and their mRNA expression levels were measured using qRT-PCR. Conclusion: HP1BP3, OSBPL9, SSH3, ZCCHC8, FANCI, and EIF4A2 were identified as potential antigens for developing mRNA-based vaccines against BLCA, and patients in IC2 might benefit more from vaccination.

Nitrogen removal performance and rapid start-up of anammox process in an electrolytic sequencing batch reactor (ESBR).

In this study, the electrolytic sequencing batch reactor (ESBR) with different current densities was constructed to investigate the nitrogen removal performance and rapid start-up of anaerobic ammonia oxidation (anammox) process. The changes of total nitrogen removal rate (TNRR), specific anammox activity (SAA) and nitrogen concentration under different current densities were analyzed, and then the effect of the optimal current density on the start-up of anammox in ESBR was explored. The results showed that ammonium nitrogen removal efficiency (92.7%), nitrite nitrogen removal efficiency (15.5%) and total nitrogen removal efficiency (28.1%) were obtained with the TNRR and SAA were 0.0118 g N L-1 d-1 and 0.0050 g N (g Vss d)-1, respectively under the optimal conditions (i.e., current density = 0.10 mA cm-2, temperature = 36 °C and pH = 7.6). In addition, the stoichiometric ratio indicated that anammox was initiated successfully for 91 days in ESBR with the current density of 0.10 mA cm-2, which was shortened by 10 days compared with the conventional SBR without current density. These results suggest that an array of rapid start-up processes of anammox can be developed through applying current density to stimulate the activity of anammox bacteria (AnAOB).

Genome-Wide Association and Expression Analysis Revealed the Candidate Variants and Molecular Underpinnings of Cold-Stress Response in Large Yellow Croaker.

Large yellow croaker (Larimichthys crocea) is one of the most economically important fish in China. Recently, global climate change has caused more and more intense and extreme low temperature weathers, resulting in huge losses to the large yellow croaker industry. Therefore, it is essential to understand the mechanisms of low-temperature tolerance in large yellow croaker. Here, we conducted an integrative analysis of genome-wide association study (GWAS) and transcriptome analysis to identify candidate variants and reveal the molecular underpinning of cold-stress response in large yellow croaker. A total of 8 significant single nucleotide polymorphisms (SNPs) loci on 6 chromosomes were identified in the GWAS analysis, and 5764 (gill) and 3588 (liver) differentially expressed genes (DEGs) were detected in cold-stressed large yellow croaker, respectively. Further comparative and functional analysis of the candidate genes and DEGs highlighted the importance of pathways/genes related to immune response, cellular stress response, lipid transport, and metabolism in the cold-stress response of large yellow croaker. Our results provide insights into the cold tolerance of large yellow croaker and contribute to genomic-based selection for low-temperature-resistant large yellow croaker.

Transcriptome Analysis Reveals Molecular Underpinnings of Common Carp (Cyprinus carpio) Under Hypoxia Stress.

As an essential environmental factor that affects the economic benefits of aquaculture, hypoxia is one of the urgent problems to be solved in the aquaculture fish breeding industry. Common carp (Cyprinus carpio) is a critical economic fish in China, and at present, there are many breeding strains of common carp with different character advantages in China, including Hebao red carp (C. carpio var wuyuanesis) and Songpu mirror carp (C. carpio var specularis). Even if the environmental adaptation of common carp is generally strong, the genetic background of hypoxia tolerance in different strains of common carp is unclear yet. This study tested the hypoxia tolerance of Songpu minor carp, Hebao red carp, and their hybrid F1 population by an acute hypoxia treatment. Muscle and liver tissues were used for transcriptome sequencing analysis to identify the key factors for hypoxia tolerance and explore the potential genetic mechanism for breeding high hypoxia tolerance in common carp. The comparative transcriptomic analysis revealed abundant hypoxia response-related genes and their differential regulation mechanism in these two tissues of different common carp strains under acute hypoxia, including immune response, cellular stress response, HIFs (hypoxia-inducible factors), MAP kinase, iron ion binding, and heme binding. Our findings will facilitate future investigation on the hypoxia response mechanism and provide a solid theoretical basis for breeding projects in common carp.

Construction of a Lactate-Related Prognostic Signature for Predicting Prognosis, Tumor Microenvironment, and Immune Response in Kidney Renal Clear Cell Carcinoma.

Kidney renal clear cell carcinoma (KIRC) is one of the most prevalent primary malignancies with high heterogeneity in the urological system. Growing evidence implies that lactate is a significant carbon source for cell metabolism and plays a vital role in tumor development, maintenance, and therapeutic response. However, the global influence of lactate-related genes (LRGs) on prognostic significance, tumor microenvironment characteristics, and therapeutic response has not been comprehensively elucidated in patients with KIRC. In the present study, we collected RNA sequencing and clinical data of KIRC from The Cancer Genome Atlas (TCGA), E-MTAB-1980, and GSE22541 cohorts. Unsupervised clustering of 17 differentially expressed LRG profiles divided the samples into three clusters with distinct immune characteristics. Three genes (FBP1, HADH, and TYMP) were then identified to construct a lactate-related prognostic signature (LRPS) using the least absolute shrinkage and selection operator (LASSO) and Cox regression analyses. The novel signature exhibited excellent robustness and predictive ability for the overall survival of patients. In addition, the constructed nomogram based on the LRPS-based risk scores and clinical factors (age, gender, tumor grade, and stage) showed a robust predictive performance. Furthermore, patients classified by risk scores had distinguishable immune status, tumor mutation burden, response to immunotherapy, and sensitivity to drugs. In conclusion, we developed an LRPS for KIRC that was closely related to the immune landscape and therapeutic response. This LRPS may guide clinicians to make more precise and personalized treatment decisions for KIRC patients.

Efficient degradation of naproxen in a three dimensional biofilm electrode magnetism reactor (3DBEMR): Removal performance and microbial community.

A three-dimensional biofilm electrode magnetism reactor (3DBEMR) was constructed to removal naproxen (NPX). This study evaluated 3DBEMR performance in removal of refractory NPX, while also discussing the effect of the electro-magnetic superposition on microbial community by high throughput sequencing. Results indicated that 3DBEMR's average removal rate for NPX stood at 88.36%, representing an increase by 75.24%, 65.03% and 12.36%, respectively, compared to 3DBR (Three-Dimensional Biofilm Reactor), 3DBMR (Three-Dimensional Biofilm Magnetism Reactor) and 3DBER (Three-Dimensional Biofilm Electrode Reactor). This was attributed to the influence of electro-magnetic adsorption, electro-oxidaton/catalysis, and electro-magnetic biodegradation. Another major contributing factor to NPX removal was the presence in 3DBEMR of high-abundance genera such as Rhodobacter, Porphyrobacter, Methyloversatilis, Sphingopyxis,Bosea, Singulisphaera, Sphingomonas. Therefore, the 3DBEMR was successfully demonstrated to be a flexible and effective technique in NPX degradation, which would help to better understand the effect of superposition of electric and magnetic fields on microbial community.

Electro/magnetic superposition effects on diclofenac degradation: Removal performance, kinetics, community structure and synergistic mechanism.

Electric and magnetic fields characterized by high efficiency, low consumption and environment-friendly performance have recently generated interest as a possible measure to enhance the performance of the biological treatment process used to remove refractory organics. Few studies have been carried out to-date regarding the simultaneous application of electric and magnetic fields on biofilm process to degrade diclofenac. In this study, 3DEM-BAF was designed to evaluate the electrio-magnetic superposition effect on diclofenac removal performance, kinetics, community structure and synergistic mechanism. The results show that 3DEM-BAF could significantly increase the average removal rate of diclofenac by 65.30 %, 57.46 %, 9.48 % as compared with that of BAF, 3DM-BAF, 3DE-BAF, respectively. The diclofenac degradation kinetic constants and dehydrogenase activity of 3DEM-BAF were almost 6.72 and 2.53 times higher than those of BAF. Microorganisms of 3DEM-BAF in the Methylophilus and Methyloversatilis genera were distinctively enriched, which was attributed to the screening function of electric field and propagation effect of magnetic field. Moreover, three processes were found to contribute to diclofenac degradation, namely electro-magnetic-adsorption, electro-chemical oxidation and electro-magnetic-biodegradation. Thus, the simultaneous application of electric and magnetic fields on biofilm process was demonstrated to be a promising technique as well as a viable alternative in diclofenac degradation enhancement.

Production of an electro-biological particle electrode (EBPE) from lithium slag and its removal performance to salicylic acid in a three-dimensional electrocatalytic biological coupling reactor (3D-EBCR).

Electro-biological particle electrode (EBPE) prepared by lithium slag was used to remove salicylic acid in a three-dimensional electrocatalytic biological coupling reactor (3D-EBCR). The physical and chemical properties of the EBPE, the removal performance of salicylic acid and the degradation mechanism were studied. Results revealed as follows: (1) the EBPE prepared by lithium slag contained effective catalytic components including Fe2O3, SnO2, ZnO, MnO, Rb2O and TiO2, with stable structure and good adsorption performance; (2) the 3D-EBCR with EBPE had strong adaptability to the current intensity in the range of 0.25-0.40 A, and the removal rates of COD and salicylic acid were maintained above 87.1% and 85.2% respectively; (3) salicylic acid was removed through the synergistic action of adsorption, electrochemical oxidation and biological action.

Electrocatalysis degradation of tetracycline in a three-dimensional aeration electrocatalysis reactor (3D-AER) with a flotation-tailings particle electrode (FPE): Physicochemical properties, influencing factors and the degradation mechanism.

Novel particle electrodes, i.e. flotation tailings particle electrode (FPE), were prepared using flotation tailings, garden soil, and soluble starch with a mass ratio of 16:3:1, and then used in tetracycline wastewater treatment. The physicochemical properties of FPE were systematically characterized using SEM, XRD, FT-IR and XRF. Tetracycline adsorption and its adsorption mechanism onto FPE was explored for the first time. Parameters affecting FPE's degradation efficiency and energy consumption such as current density, electrolysis time, initial concentration, initial pH and aeration rate were examined. The electrocatalytic degradation of tetracycline shows that the degradation of tetracycline meets the pseudo-first-order kinetics. Moreover, the numbers of •OH produced on the surfaces of the cathode, anode and particle electrode were compared. Results showed that the adsorption-saturated FPE can be regenerated by electrochemical action to induce further absorption and form in-situ electrocatalysis. In order to find out the transformation products in water and degradation pathways of Tetracycline, UHPLC method was used to obtain the degradation pathways for Tetracycline. So, this work could provide a fabrication of high-efficiency and low-cost electrocatalytic for removal of pharmaceuticals pollutants from waste water as well as deeper insight into electrocatalytic mechanism, transformation products, and degradation pathways of Tetracycline in water.

Crossed-fused renal ectopia with renal calculi: Two case reports and a review of the literature.

RATIONALE: Crossed renal ectopia (CRE) is a rare congenital anomaly that is frequently associated with gastrointestinal, cardiovascular, genital and bone malformations. To the best of our knowledge, only 35 cases of crossed renal ectopia involving calculi and 30 cases of CRE associated with renal carcinoma have been reported to date. PATIENT CONCERNS: Here, we present 2 cases of crossed renal ectopia. A 59-year-old woman with diabetes presented to our hospital with abdominal pain. The second patient was a 24-year-old woman who complained with abdominal pain with a duration of 1 day. DIAGNOSES: On the basis of abdominal ultrasonography, we suspected a solitary kidney both in the two patients. Combined with retrograde pyelography and 3D computed tomography, case 1 was diagnosed as an S-shaped right-to-left crossed-fused ectopic kidney with many stones in the left (normal) renal pelvis and case 2 was confirmed to have lump right-to-left crossed-fused renal ectopia with two 3-mm stones in the renal pelvis of the 2 kidneys. INTERVENTIONS: Case 1 underwent percutaneous nephrolithotomy while case 2 refused to undergo surgery and underwent conservative treatment for pain relief. OUTCOMES: Two patients have been followed up and have no stones recurrence. LESSONS: Crossed fused renal ectopia is easily misdiagnosed as a solitary kidney. CRE is so rare that the recognition of the disease needs to be improved and effective treatment should be taken timely. According to the two cases and literature review, minimally invasive surgery has become increasingly common to treat CRE with stones and carcinoma.

Magnetic multiwalled carbon nanotubes with controlled release of epirubicin: an intravesical instillation system for bladder cancer.

BACKGROUND: Traditional intravesical instillation treatment in bladder cancer has limited efficacy, which results in a high frequency of recurrence. PURPOSE: The aim of this study was to report on an epirubicin (EPI)-loaded magnetic multi-walled carbon nanotube (mMWCNTs-EPI) system for intravesical instillation in place of the current formulation. METHODS: The mMWCNTs-EPI system was formulated with carboxylated MWCNTs, Fe3O4 magnetic nanoparticles, and EPI. Features and antitumor activity of the system were investigated. RESULTS: Under the effect of external magnets, the mMWCNTs-EPI system showed sustained release and prolonged retention behavior and better antitumor activity than free EPI. The mMWCNTs-EPI system had higher efficiency in enhancing cytotoxicity and inhibiting proliferation in vitro and in vivo than free EPI. Our studies also revealed the atoxic nature of mMWCNTs. CONCLUSION: These findings suggested that mMWCNTs are effective intravesical instillation agents with great potential for clinical application.

Removal of diclofenac by three-dimensional electro-Fenton-persulfate (3D electro-Fenton-PS).

Diclofenac (DIC) is a new type of contaminant that has been widely detected in the water environment, posing threats to the ecological environment and human health. However, the conventional wastewater treatment process has a very limited ability to reduce DIC. In this research, persulfate is added to electro-Fenton with the three-dimensional particle electrode (TDE) process whose particle electrodes were formed from manganese slag with loaded active substance (Cu: Fe = 1:1) to construct a three-dimensional electro-Fenton-persulfate (3D electro-Fenton-PS) process to investigate the removal rate of DIC under the optimum working conditions. The effects of different persulfate addition, activator addition and different activators on the removal rate of DIC were researched, respectively. The removal rate of DIC reached 96.3% when the persulfate and the Fe0 addition were 1.50 mM and 3.00 mM, respectively. The results showed that and OH existed simultaneously in the reaction system, and the removal of DIC was the result of the two free radicals. Moreover, degradation pathways and mechanism of DIC were also discussed. The study may provide a new theoretical basis and technical support for the treatment of DIC in municipal wastewater.

In-situ ion exchange electrocatalysis biological coupling (i-IEEBC) for simultaneously enhanced degradation of organic pollutants and heavy metals in electroplating wastewater.

The goal of this research was to develop a new process for simultaneously removing organics and heavy metals of electroplating wastewater by in-situ ion exchange electrocatalysis biological coupling (i-IEEBC). The study evaluated the removal efficiency of coexisting refractory organics and heavy metal ions, and investigated the effects of current density (CD) on the removal performance of the i-IEEBC method. The results indicated the i-IEEBC reactor exhibited higher average removal rates of COD, TOC, Cr and Cu ions, i.e. 87.23%, 80.42%, 91.25%, and 95.97% in that order, which represented an increase by 32.59%, 40.10%, 31.86%, and 33.82%, respectively, compared with BAF. The optimum CD for simultaneously removing organics and heavy metals of electroplating wastewater in i-IEEBC was 0.40 mA/(cm)2. The change of biodegradability and the presence of short chain organic compounds also indirectly confirmed the excellent removal organic pollutants performance of i-IEEBC at the optimum CD. In addition, the composition and construction of CER before and after the application, under the optimum CD, SEM, EDS and FT-IR spectroscopy also showed that the cation exchange properties of CER improved the catalytic lifetime of the particle electrodes. This research provides a highly efficient new alternative to electroplating wastewater treatment technology.

Preparation of particle electrodes from manganese slag and its degradation performance for salicylic acid in the three-dimensional electrode reactor (TDE).

Salicylic acid (SA) is a class of trace pollutants widely presented in the environment belonged to pharmaceuticals and personal care products. It is difficult to remove SA by the traditional treatment processes because of its toxicity. In this paper, the degradation of SA by Mn-loaded Cu/Fe particle electrodes was studied. Firstly, the particle electrodes were prepared by impregnation-roasting method and then characterized by SEM, XRF and XRD. The diffraction peaks of Fe2O3 and CuO in the XRD patterns of the particle electrodes which had the dense spherical particles were significantly increased and the content of CuO and Fe2O3 increased by 1.9% and 3.6% respectively. Secondly, single factor experiments were carried out under conditions of cell voltage, electrolyte concentration, pH, HRT, inter-electrode distance and initial pollutant concentration. Under the optimum conditions of all the factors, the degradation rate of SA reached 76.9%. Then, HPLC and GCMS were employed to deduce the degradation pathways of SA by the TDE with Mn-particle electrodes (Mn-PETDE). Under the action of •OH, SA underwent decarboxylation and substitution reactions and then mineralized after the ring-opening reaction. All results demonstrated that this Mn-PETDE was effective for degradation of SA.

Performance and microbial community of an electric biological integration reactor (EBIR) for treatment of wastewater containing ibuprofen.

Electric biological integration reactor (EBIR) was designed and built for the treatment of wastewater containing ibuprofen. This study evaluates the removal performance of EBIR by comparison with biological aerated filter (BAF), while also discussing the optimal operational parameters of EBIR within the context of the response surface methodology. The results indicate that EBIR exhibits higher average removal rates of ibuprofen, chemical oxygen demand (COD) and NH4+-N, i.e. 93.48%, 86.72% and 85.19%, representing an increase by 61.59%, 14.57% and 10.49%, respectively, compared with BAF. The optimal conditions for EBIR were 12.73 A/m2 current density (CD), 3.5 h hydraulic retention time and 0.08 mg/L influent ibuprofen concentration. In addition, microbial community structures were detected using an Illumina Miseq PE300 system, which were different at the phylum, class, and genus levels between EBIR and BAF. The microbial communities of EBIR, including mainly Trichococcus, Aeromonas, Saprospiraceae_uncultured, Thiobacillus, Aeromonas Flavobacterium, Sphingopyxis, Candidate_division_TM7_norank, Acinetobacter and physicochemical properties indirectly confirmed the excellent removal performance at 12.73 A/m2 CD.

Anatomic study on mental canal and incisive nerve canal in interforaminal region in Chinese population.

PURPOSE: This study was aimed to detect the positions of mental canal and incisive nerve canal as well as the prolongation of mandibular canal in interforaminal region in Chinese population to supply the reference data of the surgical safe zone in chin for clinicians. MATERIALS AND METHODS: A total of 80 formalin-fixed semi-mandibles of Chinese adult cadavers were dissected, the positions and courses of mental canal and incisive nerve canal as well as the prolongation of mandibular canal in interforaminal region were measured. RESULTS: The mental foramina were present in all cases (100 %), and most of them were located below 2nd premolar (58.75 %). Accessory mental foramina were observed in 5 %. The anterior end of mandibular canal, extending along the course of 7.37 ± 1.10 mm above the lower border of mandible to interforaminal region about 3.54 ± 0.70 mm medial to the mental foramen, most often ended below between the two premolars (73.75 %), where it continued as the incisive nerve canal (100 %) and the mental canal (96.25 %). Mental canal, with the wall formed by compact bone, being 2.60 ± 0.60 mm in diameter and 4.01 ± 1.20 mm in length, opened into mental foramen. Incisive nerve canal, with the wall formed by thin compact bone and/or partly or completely by spongy bone, being 1.76 ± 0.27 mm in diameter and 24.87 ± 2.23 mm in length, extended to the incisor region along the course of 9.53 ± 1.43 mm above the lower border of mandible, and most often ended below the lateral incisor (70.00 %). CONCLUSION: This research recommended for chin operations in Chinese population: the surgical safe zone could be set in the region about over 4 mm anterior to the mental foramen, and over 12 mm above inferior border of mandible for anterior alveolar surgery, or within 9 mm above inferior border of mandible for genioplasty.

CCL21/CCR7 axis activating chemotaxis accompanied with epithelial-mesenchymal transition in human breast carcinoma.

Secondary lymphoid tissue chemokine (SLC/CCL21) and its receptor CCR7 have been implicated in lymph node metastasis, whereas the mechanism of which remains unclear. Epithelial-mesenchymal transition (EMT) plays an important role in invasion and migration of cancer cells. We presumed that CCL21/CCR7 axis activates EMT process to induce cancer cell invasion and metastasis. Firstly, the expressions of CCR7 and EMT markers were examined by immunohistochemical staining in the primary breast carcinoma tissues from 60 patients who underwent radical mastectomy. Then, we investigated whether CCL21/CCR7 induces EMT process during mediating cancer cell invasion or migration in vitro. By immunohistolochemistry, high expressions of CCR7, Slug and N-cadherin were seen in 60, 65, and 76.67 % of tumors, respectively, and significantly associated with lymph node metastases as well as clinical pathological stage. Furthermore, the CCR7 expression was significantly correlated to Slug and N-cadherin. In vitro, stimulating breast cancer cell lines 1428, MCF-7 and MDA-MB-231 with CCL21, the invasion and migration of tumor cells were promoted, and simultaneously, EMT phenotype of tumor cells was enhanced, including down-regulation of E-cadherin, up-regulation of Slug, Vimentin and N-cadherin at both protein and mRNA levels. Inversely, knockdown of CCR7 by shRNA suppressed tumor cell invasion, migration and EMT phenotype induced by CCL21. These results indicated that CCL21/CCR7 axis could activate EMT process during chemotaxis of breast carcinoma cells.

An effective method of isolating endothelial cells from intact rat aorta.

Rat aorta has been widely utilized in studies on cardiovascular diseases. Due to its small size, the endothelial cells are difficult to isolate. Here, we developed a modified method to obtain the endothelial cells from the rat aorta. The thoracic aorta of Wistar rat was harvested with its inside was turned out to expose the tunica intima. With its both ends ligated and cultured in ECM in T25 flask, a small amount of cells grew out from the aorta on day 4 and manifested the characteristic cobble stone-like monolayer on day 9-10. The cells were identified to be endothelial cells using reverse transcription (RT)-polymerase chain reaction (PCR), immunocytochemical staining for Von Willebrand Factor (vWF), Dil-Ac-LDL uptake assay, and capillary tube formation assay. This method serves as an easy way to get a large amount of endothelial cells from small-sized vessels, which, therefore, will greatly benefit the researches of vascular disease.