MED27 promotes malignant behavior of cells by affecting Sp1 in breast cancer.

OBJECTIVE: The aim of this study was to investigate the expression of mediator complex subunit 27 (MED27) in breast cancer (BC) and explore its effects on the proliferation and apoptosis of BC cells. PATIENTS AND METHODS: The expression of MED27 in 60 BC tissues and para-cancer tissues was detected. Based on the significantly high expression level of MED27 in tumors, the tumor samples were divided into high-expression group and low-expression group according to the median standard, with 30 samples in each group. Then, the association between MED27 expression and clinicopathological features of patients was analyzed. The correlation between MED27 expression and survival time of patients was estimated using the Kaplan-Meier method. Next, the expression level of MED27 in cells was also measured using qRT-PCR assay. In vitro study, si-MED27 was designed to interfere with the expression of MED27 in MDA-MB-231 cells. To further explore the mechanism of MED27 in BC, the expression level of SP1 in cells was examined after different treatments. RESULTS: In quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay, MED27 was found to be highly expressed in both BC tissues and cells. Then, the relationship between MED27 expression and clinical pathological data was statistically analyzed, and it was found that MED27 expression was correlated with tumor size and grade. In the 60-month follow-up and Kaplan-Meier analysis, patients with high expression of MED27 had a poor prognosis. In in vitro study, MED27 expression in cells was down-regulated by transfection with si-MED27. Western blot (WB) analysis suggested that si-MED27 could effectively reduce the protein expression level of MED27 in cells, and the specificity protein 1 (Sp1) expression was also limited. In CCK-8, clone formation and flow cytometry experiments, the proliferation of cells with low MED27/Sp1 expression was suppressed, while cell apoptosis was promoted. CONCLUSIONS: MED27 acted as an oncogene in BC. By affecting Sp1, MED27 could be a new therapeutic target for the treatment of BC.

Utilizing integrating network pharmacological approaches to investigate the potential mechanism of Ma Xing Shi Gan Decoction in treating COVID-19.

Beginning in December 2019, coronavirus disease 2019 (COVID-19), due to 2019-nCoV infection, emerged in Wuhan and spread rapidly throughout China and even worldwide. Employing combined therapy of modern medicine and traditional Chinese medicine has been proposed, in which Ma Xing Shi Gan Decoction (MXSGD) was recommended as a basic prescription and applied widely in the clinical treatment of COVID-19. We investigated the underlying mechanism of MXSGD in treating COVID-19 utilizing the approaches of integrating network pharmacology. A total of 97 active ingredients of MXSGD were screened out, and 169 targets were predicted. The protein-protein interaction network exhibited hub targets of MXSGD, such as Heat shock protein 90, RAC-alpha serine/threonine-protein kinase, Transcription factor AP-1, Mitogen-activated protein kinase 1, Cellular tumor antigen p53, Vascular endothelial growth factor A, and Tumour necrosis factor. Gene Ontology functional enrichment analysis demonstrated that the biological processes altered within the body after taking MXSGD were closely related to the regulation of such processes as the acute inflammatory response, chemokine production, vascular permeability, response to oxygen radicals, oxidative stress-induced apoptosis, T cell differentiation involved in the immune response, immunoglobulin secretion, and extracellular matrix disassembly. KEGG enrichment analysis indicated that the targets of MXSGD were significantly enriched in inflammation-related pathways, immunomodulation-related pathways, and viral infection-related pathways. The therapeutic mechanisms of MXSGD on COVID-19 may primarily involve the following effects: reducing inflammation, suppressing cytokine storm, protecting the pulmonary alveolar-capillary barrier, alleviating pulmonary edema, regulating the immune response, and decreasing fever.

[The significance of eosinophils in the correlation of upper and lower airway inflammation in patients with chronic rhinitis].

Objective: To explore the predictor of lower airway inflammation among the index of nasal inflammation by investigating the expression and association of eosinophils (EOS) in the upper-lower airways and blood of patients with chronic rhinitis. Methods: A total of 162 patients with allergic rhinitis (AR), 117 patients with non-allergic rhinitis (NAR) and 104 controls were enrolled from June 2010 to December 2013 from General Hospital of Eastern Theater Command, People's Liberation Army. All subjects were required detailed medical history collection and nasal resistance measurement. Skin prick test (SPT), blood total immunoglobin E (tIgE) and blood EOS, nasal lavage and induced sputum EOS, nasal provocation and bronchial provocation test (NPT, BPT), nasal and forced exhaled nitric oxide (NNO, FeNO) were performed in all patients. One-way analysis of variance was used for comparison between groups. LSD t test or Mann-Whitney U test was used for comparison between the two groups. Pearson or Spearman related parameter test was used for correlation analysis. Results: The nasal lavage EOS, NNO, induced sputum EOS, FeNO, blood EOS and tIgE were higher in the AR group than that in the NAR group (3.70[1.20, 14.23]/200 HP vs 1.40[0.20, 3.40]/200 HP, 673.50[466.80, 936.00] ppb vs 455.80[248.10, 705.60] ppb, 2.97[0.00, 10.63]% vs 1.00[0.23, 2.00]%, (49.28±26.37)ppb vs (34.07±19.11)ppb, 4.00[2.00, 7.00]% vs 2.00[1.00, 5.00]%, 208.01[61.70, 387.50] IU/ml vs 43.30[19.00, 122.00] IU/ml, F or χ(2) value was 11.442, 19.440, 70.727, 69.449, 47.453, 46.525, respectively, all P<0.05). But there was no significant difference in nasal resistance, NPT and BPT between the two groups. Nasal lavage EOS in AR group and NAR group was correlated with induced sputum EOS, FeNO, tIgE and blood EOS (r value of AR group was 0.448, 0.202, 0.159, 0.321, r value of NAR group was 0.442, 0.268, 0.268, 0.334, respectively, all P<0.05), but not with BPT. After adjustment for gender, age, height and weight, nasal EOS was positively correlated with sputum EOS. Multiple linear regression analysis showed that nasal EOS, blood EOS and SPT were factors affecting sputum EOS levels. The optimal threshold for nasal EOS to determine induced sputum EOS was 3.30/200 HP by (receiver operating characteristic,ROC) analysis. Conclusion: The nasal EOS is correlated with multiple lower airway and systemic inflammatory markers, and is a risk factor for the induced sputum EOS, which can be used as an inflammation biomarker to predict the lower air inflammation.

[Different care models and symptom progression of Alzheimer's disease in China].

Objective: To compare the differences of cognitive function, daily living ability and neuropsychiatric symptoms in patients with sporadic type of Alzheimer's disease (AD) under different care modes, and find the most favorable care mode for delaying the progress of disease. Methods: One hundred and twenty cases of AD patients were divided into three groups: Spouse Care Group, Adult Child Care Group and Nursing Home Group. Medical history collection and scale evaluation were carried out by trained specialists on 3 groups of patients and caregivers. Assessment included socio-demographic data, including name, gender, age, course of the disease, the year of education and the way of care, Mini mental state examination (MMSE), Activity of Daily Living (ADL), Alzheimer's Disease Assessment Scale-Cognitive section (ADAS-Cog), Neuropsychiatric Inventory (NPI), and the Relevant Outcome Scale for Alzheimer's disease (ROSA). All the evaluations were completed upon enrollment. The differences in cognitive function, daily living ability and neuropsychiatric symptoms were compared among the three groups. Results: There was no significant difference in age, gender, education duration and course of disease between the three groups (P>0.05). The MMSE average scores of Spouse Care Group, Adult Child Care Group and Nursing Home Group were 19±7, 15±6, 13±7 respectively. The ADAS-Cog median scores of Spouse Care Group, Adult Child Care Group and Nursing Home Group were 17.32(9.78, 26.50), 30.00(16.10, 38.55), 33.15 (16.28, 50.68). The NPI median total scores of Spouse Care Group, Adult Child Care Group and Nursing Home Group were 5.00(1.00, 13.00), 9.00(4.00, 20.00), 19.50(8.50, 28.50) respectively. The ADL average scores of Spouse Care Group, Adult Child Care Group and Nursing Home Group were 21±9, 25±9, 35±11. The difference of MMSE, ADAS-Cog, ADL and NPI was statistically significant among the three groups (P<0.05). No significant difference was found in care burden among the three groups (P>0.05). Conclusions: The AD patients with spouse care tend to suffer from mild diseases severitys, no matter in terms of cognitive function, daily living ability or neuropsychiatric symptoms. Close, familiar and comprehensive care plays an important role in delaying the progress of AD.

[Different phenotypes in patients with chronic sinusitis with bilateral nasal polyps].

Objective: To investigate the clinical features of patients with chronic sinusitis with bilateral nasal polyps. Method: The data of 134 patients with chronic sinusitis with bilateral nasal polyps were analyzed, retrospectively. Based on the results of the histopathological examination, patients were divided into eosinophilic chronic sinusitis with bilateral nasal polyps(ECRSwBNP) group and noneosinophilic chronic sinusitis with bilateral nasal polyps(nonECRSwBNP) group. The clinical data including medical history collection, skin prick test, nasal secretion examination, serum total IgE level, peripheral blood test, nasal endoscopy, and sinus CT examination were analyzed and compared. Result: The rate of concomitant with asthma(19.0%, 0), atopy(26.6%, 9.1%), NP recurrence rate(54.4%, 30.9%), the ratio of EOS in peripheral blood(4.8%, 1.9%), the ratio of EOS in nasal secretions(10.0%, 2.0%) and total serum IgE levels(82.15 IU/L, 25.80 IU/L) were significantly(P<0.05 ) different between two groups. In the sinus CT score, posterior ethmoid sinus(2.0, 1.0), total ethmoid sinus(5.0, 4.0), sphenoid sinus(3.0, 2.0), Olfactory cleft(OC)(4.0, 2.0), the ratio of thee thmoid sinus/maxillary sinus(2.0, 1.0) was statistically significant(P<0.05), while the rest were not statistically significant. The OC score, the ratio of EOS in peripheral blood, the ratio of the posterior ethmoid sinus/ anterior ethmoid sinus, and the ratio of the ethmoid sinus/maxillary sinus can be used for subtype in CRSwBNP patients (AUC> 0.5), while the OC score under the ROC curve is the largest area with the highest diagnostic value. When the OC score is 3 or higher, the predicted value specificity is 80.9% and the sensitivity is 52.1%. Conclusion: Patients with chronic sinusitis with bilateral nasal polyps often comorbid asthma and atopy, while these patients are trend to recurrence. The patients with ECRSwBNP comorbid asthma mainly characterized by eosinophil infiltration in tissues are more likely to recurrence. Targeted treatment based on phenotype should be considered in the clinic.

[The role of NF-κB signaling pathway in laryngeal leukoplakia recurrent with laryngeal reflux].

Objective: To study the mechanism of vocal mucosal barrier damage mediated by NF-κB and NF-κB-regulated signaling pathway via probing the expression of inflammatory factors and essential proteins for node of NF-κB signaling pathway. Methods: The patients suffering from vocal leukoplakia accompanied with larygopharyngeal reflux(LPR) were treated with oral administration of proton pump inhibitor(PPI). Mucosal specimens of vocal cord were collected from all patients before PPI treatment. And the mucosal specimens of vocal cord were collected from the patients with suspected recurrence at 8 weeks after PPI treatment. HE staining was used to observe the histopathological changes of the mucosa. ELISA was utilized to detect the levels of inflammatory factors including tumor necrosis factor(TNF)-α, interleukin(IL)-1 and IL-6. Western blot was used to detect the expression of p-p65, p-IKK and p-IκB. Immunofluorescence method was adopted to detect the entrance of p65 to cell nucleus.Data was analyzed by SPSS 23.0 software. Results: In PPI untreated group, the expressions of IL-1ß, TNF-α and IL-6 in the specimens of 8 weeks after operation were not different significantly from those obtained during operation.But in the PPI-treated group, the expressions were down-regulated.The expression of p-p65 in the middle and high grade heterogenous hyperplasia group was higher than that of low level heterogenous hyperplasia group.The difference of p65 and p-p65 expression between 8 weeks after surgery and surgery in PPI-untreated group was statistically insignificant (P>0.05). The difference of p65 expression between PPI-treated group and PPI pre-treatment group was statistically insignificant (P>0.05). The expression of p-p65 in the PPI-treated group was lower than that of the PPI pre-treatment group (P<0.05). The expressions of IL-1ß, TNF-α and IL-6 were positively related with that of NF-κB-p65. Immun of luorescence method revealed the entrance of p65 to cell nucleus in PPI pre-treatment group, which meant that NF-κB was activated. In the PPI-treated group, few activated p65 could be observed in the cell nucleu. Conclusion: The possible mechanism of vocal mucosal barrier damage in vocal leukoplakia accompanied with LPR maybe the vocal mucosal inflammation mediated by NF-κB and NF-κB-regulated signaling pathway activated with refluxed materials.

Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) promotes EGF receptor signaling of oral squamous cell carcinoma metastasis via the complex N-glycosylation.

Aberrant protein glycosylation could be a distinct surface-marker of cancer cells that influences cancer progression and metastasis because glycosylation can regulate membrane protein folding which alters receptor activation and changes epitope exposure for antibody (Ab) recognition. Carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6), a glycophosphoinositol-anchored protein, is a heavily glycosylated tumor antigen. However, the clinical significance and biological effect of CEACAM6 glycosylation has not been addressed in cancers. We recently developed an anti-CEACAM6 Ab (TMU) from an immune llama library which can be engineered to a single-domain (sd)Ab or a heavy-chain (HC)Ab. The TMU HCAb specifically recognized glycosylated CEACAM6 compared to the conventional antibodies. Using the TMU HCAb, we found that glycosylated CEACAM6 was a tumor marker associated with recurrence in early-stage OSCC (oral squamous cell carcinoma) patients. CEACAM6 promoted OSCC cell invasion, migration, cytoskeletal rearrangement, and metastasis via interaction with epidermal growth factor (EGF) receptor (EGFR) and enhancing EGFR activation, clustering and intracellular signaling cascades. These functions were modulated by N-acetylglucosaminyltransferase 5 (MGAT5) which mediated N-glycosylation at Asn256 (N256) of CEACAM6. Finally, the TMU sdAb and HCAb treatment inhibited the migration, invasion and EGF-induced signaling in CEACAM6-overexpressing cells. In conclusion, the complex N-glycosylation of CEACAM6 is critical for EGFR signaling of OSCC invasion and metastasis. Targeting glycosylated CEACAM6 with the TMU sdAb or TMU HCAb could be a feasible therapy for OSCC.

Analysis on the local structures for 3d1 impurities (Ti3+ and V4+ ) in KTiPO4.

Making use of the perturbation formulae for 3d1 ions (Ti3+ and V4+ ) under orthorhombically compressed octahedra, the spin Hamiltonian parameters (g factors: gx , gy , gz and hyperfine structure constants: Ax , Ay , Az ) and local structures of the 3d1 impurity centres C1 , C2 , and C3 in KTiOPO4 crystals are theoretically analyzed in a consistent way. The remarkable local distortions (i.e., the relative axial compression ratios 11.2%, 7.0%, and 5.5% along Z axis and the relative planar bond length variation ratios 15.9%, 7.0%, and 6.0%) are obtained for the [Ti2O6 ]9- cluster on Ti2 site and [VO6 ]8- clusters on Ti1 and Ti2 sites, respectively, in view of the Jahn-Teller effect. The above local orthorhombic distortion parameters in the impurity centres are found to be more significant than the host Ti1 and Ti2 sites in pure KTiOPO4 . The sequences (C1  > C2  > C3 ) of the local orthorhombic distortion parameters ρ and τ are in accordance with those of the axial and perpendicular anisotropies Δg and δg of g factors, respectively.

[Efficacy and tolerance of Memantine monotherapy and combination therapy with Reinhartdt And Sea Cucumber Capsule on agitation in moderate to severe Alzheimer disease].

Objective: To explore the efficacy and tolerance of Memantine combined Reinhartdt And Sea Cucumber Capsule (R.S.C) on treating agitation in patients with moderate-severe Alzheimer disease (AD). Methods: One hundred and fifty-eight moderate-sever AD patients from Sep.2013 to Sep.2014 in Zhejiang Provincial People's Hospital were randomly divided into two groups: group of Memantine combined R. S.C and group of single Memantine. Then Mini-Mental Sate Examination (MMSE) and Neuropsychiatric Inventory (NPI) were used to evaluate cognition symptom, behavioral and psychological symptoms of dementia (BPSD) and agitation symptom at the baseline and the end of 24 weeks.The Treatment Emergent Symptom Scale (TESS) was used to assess adverse reaction and tolerance.At last, the data was analyzed by chi-square test, t-test and covariance test. Result: At the terminal of experience, the total NPI scores and agitation factor decreased markedly in both of the two groups (P<0.05). Among the patients who had agitation symptom at the baseline, the total NPI scores and agitation factor (18±5, 3.7±2.6) in group of Memantine combined R. S.C were notably lower than those in the group of single Memantine (21±6, 5.3±2.5) (P<0.05). The incidence of adverse reaction between the two groups had no significant difference (combined treatment group was 27.7%, single treatment group was 23.2%). One patient dropped out because of skin allergy, and most adverse reactions were tolerant. Conclusions: Both two groups are effective in agitation and BPSD, and Memantine combined R. S.C is better than single treatment.R.S.C dose not aggravate adverse reaction and can be well tolerated.

Targeting palmitoyl acyltransferase ZDHHC21 improves gut epithelial barrier dysfunction resulting from burn-induced systemic inflammation.

Clinical studies in burn patients demonstrate a close association between leaky guts and increased incidence or severity of sepsis and other complications. Severe thermal injury triggers intestinal inflammation that contributes to intestinal epithelial hyperpermeability, which exacerbates systemic response leading to multiple organ failure and sepsis. In this study, we identified a significant function of a particular palmitoyl acyltransferase, zinc finger DHHC domain-containing protein-21 (ZDHHC21), in mediating signaling events required for gut hyperpermeability induced by inflammation. Using quantitative PCR, we show that ZDHHC21 mRNA production was enhanced twofold when intestinal epithelial cells were treated with TNF-α-IFN-γ in vitro. In addition, pharmacological targeting of palmitoyl acyltransferases with 2-bromopalmitate (2-BP) showed significant improvement in TNF-α-IFN-γ-mediated epithelial barrier dysfunction by using electric cell-substrate impedance-sensing assays, as well as FITC-labeled dextran permeability assays. Using acyl-biotin exchange assay and click chemistry, we show that TNF-α-IFN-γ treatment of intestinal epithelial cells results in enhanced detection of total palmitoylated proteins and this response is inhibited by 2-BP. Using ZDHHC21-deficient mice or wild-type mice treated with 2-BP, we showed that mice with impaired ZDHHC21 expression or pharmacological inhibition resulted in attenuated intestinal barrier dysfunction caused by thermal injury. Moreover, hematoxylin and eosin staining of the small intestine, as well as transmission electron microscopy, showed that mice with genetic interruption of ZDHHC21 had attenuated villus structure disorganization associated with thermal injury-induced intestinal barrier damage. Taken together, these results suggest an important role of ZDHHC21 in mediating gut hyperpermeability resulting from thermal injury.NEW & NOTEWORTHY Increased mucosal permeability in the gut is one of the major complications following severe burn. Here we report the novel finding that zinc finger DHHC domain-containing protein-21 (ZDHHC21) mediates gut epithelial hyperpermeability resulting from an experimental model of thermal injury. The hyperpermeability response was significantly attenuated with a pharmacological inhibitor of palmitoyl acyltransferases and in mice with genetic ablation of ZDHHC21. These findings suggest that ZDHHC21 may serve as a novel therapeutic target for treating burn-induced intestinal barrier dysfunction.

[The standardization of inflammation detecting methods in upper and lower airways].

Objective:To investigate the standardization of inflammation detecting methods in upper and lower airways. Method:After a five year cooperation with Guangzhou Institute of Respiratory Diseases on inflammatory diseases of airways, we have found a series of evaluation methodology and normative values in upper and lower airways (NO), airway hyper reactivity and cytology (Eos). Result:The normative range of nasal and pulmonary NO is 400-900 ppb and 5-25 ppb respectively. The nasal resistance increased ≥100% and FEV1 fell ≥20% when compared with their respective baselines both illustrating a positive result. The positive value of nasal and pulmonary Eos are ≥2.00/HP and ≥2.5% respectively. Conclusion:The standardization of evaluation methods for upper and lower airway inflammation provides the methodology and research basis for follow-up studies of upper and lower airways.

Interleukin-1ß Mediates ß-Catenin-Driven Downregulation of Claudin-3 and Barrier Dysfunction in Caco2 Cells.

BACKGROUND: IL-1ß is a cytokine involved in mediating epithelial barrier dysfunction in the gut. It is known that IL-1ß mediates activation of non-muscle myosin light chain kinase in epithelial cells, but the precise mechanism by which epithelial barrier dysfunction is induced by IL-1ß is not understood. METHODS AND RESULTS: Using a Caco2 cell model, we show that the expression of the tight junction protein, claudin-3, is transcriptionally downregulated by IL-1ß treatment. In addition, after assessing protein and mRNA expression, and protein localization, we show that inhibition of nmMLCK rescues IL-1ß-mediated decrease in claudin-3 expression as well as junction protein redistribution. Using chromatin immunoprecipitation assays, we also show that ß-catenin targeting of the claudin-3 promoter occurs as a consequence of IL-1ß-mediated epithelial barrier dysfunction, and inhibition of nmMLCK interferes with this interaction. CONCLUSIONS: Taken together, these data represent the first line of evidence demonstrating nmMLCK regulation of claudin-3 expression in response to IL-1ß-treated epithelial cells.

The integrated spintronic functionalities of an individual high-spin state spin-crossover molecule between graphene nanoribbon electrodes.

The spin-polarized transport properties of a high-spin-state spin-crossover molecular junction with zigzag-edge graphene nanoribbon electrodes have been studied using density functional theory combined with the nonequilibrium Green's-function formalism. The molecular junction presents integrated spintronic functionalities such as negative differential resistance behavior, spin filter and the spin rectifying effect, associated with the giant magnetoresistance effect by tuning the external magnetic field. Furthermore, the transport properties are almost unaffected by the electrode temperature. The microscopic mechanism of these functionalities is discussed. These results represent a step toward multifunctional molecular spintronic devices on the level of the individual spin-crossover molecule.

A novel missense mutation nt737T>G of JK gene with Jk(a-b-) phenotype in Chinese blood donors.

OBJECTIVES: The aim of this study was to investigate the molecular mechanism of the JK-null phenotype in the Chinese population. BACKGROUND: The Jk(a-b-) phenotype is vanishingly rare and the molecular basis differs between ethnic groups. The information regarding the molecular basis of JK-null alleles in the Chinese population is limited. MATERIALS AND METHODS: Three unrelated Jk(a-b-) phenotype donors were selected from 52 260 randomly blood samples through the urea lysis test and serological analysis. The JK gene-coding regions were amplified by the polymerase chain reaction and the products were sequenced directly. RESULTS: Sequencing results revealed that one sample of JK(*) B alleles carried the well-known Polynesian Jk(a-b-) mutation IVS5-1g>a. Another null allele, also on the JK(*) B background, presented with two heterozygous missense mutation, including nt222C>A(Asn74Lys) in exon 5 and nt896G>A(Gly299Glu) in exon 9. The third null allele carried two heterozygous missense mutations, nt222C>A and a novel allele nt737T>G(Leu246Arg) in exon 8. The family investigation revealed that the proband was JK(*) A(737T>G)/JK(*) B(222C>A). CONCLUSION: The Jk(a-b-) phenotype in the Chinese population shows several different molecular mechanisms. A novel missense mutation nt737T>G of JK gene was found as associated with Jk(a-b-) phenotype.

Protein tyrosine kinase 6 mediates TNFα-induced endothelial barrier dysfunction.

A key event in the progression of systemic inflammation resulting from severe trauma or shock involves microvascular hyperpermeability, which leads to excessive plasma fluid and proteins accumulating in extravascular space resulting in tissue edema. The precise molecular mechanism of the hyperpermeability response is not completely understood. Protein tyrosine kinase 6 (PTK6, also known as breast tumor kinase BRK) is a non-receptor tyrosine kinase related to Src-family proteins. Although it has also been shown that PTK6 participates in regulating epithelial barrier function, the role of PTK6 in endothelial barrier function has not been reported. In this study, we hypothesized that PTK6 is (1) expressed in vascular endothelial cells, and (2) contributes to vascular endothelial hyperpermeability in response to TNFα. Results showed that PTK6 was detected in mouse endothelial cells at the level of protein and mRNA. In addition, PTK6 knockdown attenuated TNFα induced decrease in endothelial barrier function as measured by electric cell-substrate impedance sensing (ECIS) and in vitro transwell albumin-flux assays. Furthermore, we showed that TNFα treatment of endothelial cells increased active PTK6 association with p120-catenin at endothelial cell-cell junctions. Further analysis using immunocytochemistry and immunoprecipitation demonstrated that PTK6 knockdown attenuated TNFα induced VE-cadherin internalization as well as promoting its association with p120-catenin. Our study demonstrates a novel role of PTK6 in mediating endothelial barrier dysfunction.

A microwave applicator for uniform irradiation by circularly polarized waves in an anechoic chamber.

Microwave applicators are widely employed for materials heating in scientific research and industrial applications, such as food processing, wood drying, ceramic sintering, chemical synthesis, waste treatment, and insect control. For the majority of microwave applicators, materials are heated in the standing waves of a resonant cavity, which can be highly efficient in energy consumption, but often lacks the field uniformity and controllability required for a scientific study. Here, we report a microwave applicator for rapid heating of small samples by highly uniform irradiation. It features an anechoic chamber, a 24-GHz microwave source, and a linear-to-circular polarization converter. With a rather low energy efficiency, such an applicator functions mainly as a research tool. This paper discusses the significance of its special features and describes the structure, in situ diagnostic tools, calculated and measured field patterns, and a preliminary heating test of the overall system.

MCT-1 expression and PTEN deficiency synergistically promote neoplastic multinucleation through the Src/p190B signaling activation.

Multinucleation is associated with malignant neoplasms; however, the molecular mechanism underlying the nuclear abnormality remains unclear. Loss or mutation of PTEN promotes the development of malignant tumors. We now demonstrate that increased expression of the oncogene MCT-1 (multiple copies in T-cell malignancy 1) antagonizes PTEN gene presentation, PTEN protein stability and PTEN functional activity, thereby further promoting phosphoinositide 3 kinase/AKT signaling, survival rate and malignancies of the PTEN-deficient cells. In the PTEN-null cancer cells, MCT-1 interacts with p190B and Src in vivo, supporting that they are in proximity of the signaling complexes. MCT-1 overexpression and PTEN loss synergistically augments the Src/p190B signaling function that leads to inhibition of RhoA activity. Under such a condition, the incidence of mitotic catastrophes including spindle multipolarity and cytokinesis failure is enhanced, driving an Src/p190B/RhoA-dependent neoplastic multinucleation. Targeting MCT-1 by the short hairpin RNA markedly represses the Src/p190B function, improves nuclear structures and suppresses xenograft tumorigenicity of the PTEN-null breast cancer cells. Consistent with the oncogenic effects in vitro, clinical evidence has confirmed that MCT-1 gene stimulation is correlated with p190B gene promotion and PTEN gene suppression in human breast cancer. Accordingly, MCT-1 gene induction is recognized as a potential biomarker of breast tumor development. Abrogating MCT-1 function may be a promising stratagem for management of breast cancer involving Src hyperactivation and/or PTEN dysfunction.

Cardiac rehabilitation in a pediatric patient with heart retransplantation. A single case study.

BACKGROUND: Cardiac rehabilitation (CR) after heart transplantation is known to benefit physical capacity in adults, but the advantages of CR on pediatric patients with heart retransplantation remain undetermined. AIM: The purpose of the present study was to report the effect of structured CR for a boy receiving heart transplantations twice. DESIGN: Single case study. SETTING: Inpatient and outpatient rehabilitation department. POPULATION: A pediatric patient underwent heart transplantation due to dilated cardiomyopathy at 13.6 year-old and retransplantation owing to severe cardiac allograft vasculopathy at 16.2 year-old. METHODS: CR was arranged after both transplantations. Bicycle or treadmill exercises were conducted three times weekly with the intensity adjusted to the ventilatory threshold. Serial cardiopulmonary exercise tests were performed to evaluate the sequential cardiorespiratory function changes using the peak oxygen uptake (VO2peak) as the primary outcome. RESULTS: The patient had undergone 10 times of exercise tests during rehabilitation. The VO2peak increased from 12.27 to 15.63 mL·kg-1·min-1 within 6 months after the primary transplantation. However, the VO2peak dropped intensively after a rejection episode and failed to improve since the development of cardiac allograft vasculopathy. Following retransplantation, the VO2peak appeared worse initially but increased gradually with rehabilitation. One year subsequent to retransplantation, the VO2peak reached 17.7 mL·kg-1·min-1 with a 7.22 mL·kg-1·min-1 improvement compared with his baseline value. CONCLUSION: Structured CR improves aerobic capacity of a pediatric patient with heart retransplantation. CLINICAL REHABILITATION IMPACT: CR is safe and beneficial for pediatrics with heart retransplantation. Cardiopulmonary exercise testing can be considered as an adjuvant tool for detecting rejection or cardiac allograft vasculopathy in pediatric heart transplantation recipients.