LncRNA NEAT1-associated aerobic glycolysis blunts tumor immunosurveillance by T cells in prostate cancer.

Long noncoding RNA (lncRNA) nuclear enriched abundant transcript 1 (NEAT1) is nuclear-located and transcribed from chromatin 11. To date, little is known about the cellular functions and regulatory mechanisms of NEAT1 in prostate cancer (PCa). In this study, whole-genome RNA sequencing data were downloaded from TCGA and GEO databases. Biological information was used to analyze the different expressions of NEAT1. In situ hybridization (ISH) was performed to detect the expression of NEAT1 in PCa and paracarcinoma clinical samples. Then, NEAT1 was knocked down in PC3 cells through lentiviral infection with a plasmid construct. Bioinformatics and integrative analytical approaches were utilized to identify the relationships of NEAT1 with specific cancer-related gene sets. Cell proliferation assay and colony formation assay were performed to evaluate the cell proliferative ability. Glycolysis stress test, metabolism assay, and infiltrating T-cell function analysis were implemented to assess the changes in metabolism and immune microenvironment of PCa. We found that the expression of NEAT1 was higher in PCa than in non-neoplastic tissues. The cell proliferative capability of PCa cells was significantly reduced in the NEAT1 knockdown group. PCR array and bioinformatics analysis revealed that the enrichment of acidic substance-related gene sets was associated with NEAT1 expression. NEAT1 depletion inhibited PCa cell aerobic glycolysis accompanied by the reduction of lactate levels in the medium. Further, we found that lactate dehydrogenase A (LDHA) expression was positively regulated by NEAT1. At last, co-culture systems indicated that NEAT1 or LDHA knockdown promoted the secretion of CD8+ T-lymphocyte factors, including TNF-α, IFN-γ, and Granzyme B, and enhanced the antitumor effects.

Serum total indoxyl sulfate levels and all-cause and cardiovascular mortality in maintenance hemodialysis patients: a prospective cohort study.

BACKGROUND: The association between serum total indoxyl sulfate (tIS), and cardiovascular disease (CVD) and all-cause mortality is a matter of debate. In the current study we sought to determine the association, if any, between serum tIS, and all-cause and CVD-associated mortality in patients on maintenance hemodialysis (MHD). METHODS: A prospective cohort study was conducted involving 500 MHD patients at Dalian Municipal Central Hospital from 31 December 2014 to 31 December 2020. Serum tIS levels were measured at baseline and classified as high (≥44.16 ng/ml) or low (< 44.16 ng/ml) according to the "X-tile" program. Besides, the associations between continuous serum tIS and outcomes were also explored. Predictors were tested for colinearity using variance inflation factor analysis. Hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated using Cox proportional hazards regression models. Restricted cubic spline model was performed to assess dose-response relationships between tIS concentration and all-cause and CVD mortality. RESULTS: During a 58-month median follow-up period, 224 deaths (132 CVD deaths) were documented. After adjustment for potential confounders, the serum tIS level was positively associated with all-cause mortality (HR = 1.02, 95% = 1.01-1.03); however, we did not detect a significant association when tIS was a dichotomous variable. Compared with the MHD population with a serum tIS level < 44.16 ng/ml, the adjusted HR for CVD mortality among those with a serum tIS level ≥ 44.16 ng/ml was 1.76 (95% = 1.10-2.82). Furthermore, we also noted the same association when the serum tIS level was a continuous variable. CONCLUSION: The serum tIS level was associated with higher risk of all-cause and CVD mortality among MHD patients. Further prospective large-scale studies are required to confirm this finding.

Bone marrow mesenchymal stem cells inhibit cardiac hypertrophy by enhancing FoxO1 transcription.

Bone marrow-derived mesenchymal stem cells (BMSCs) have therapeutic potential for certain heart diseases. Previous studies have shown that stem cells inhibit cardiac hypertrophy; however, it is necessary to explore the mechanisms underlying this effect. This study aimed to investigate the possible mechanism underlying the inhibitory effect of BMSCs on cardiomyocyte hypertrophy. We induced cardiomyocyte hypertrophy in cultured rat cells through isoproterenol (ISO) treatment with or without BMSC coculture. A microarray was performed to analyze messenger RNA expression in response to ISO treatment and BMSC coculture. Pathway enrichment analysis showed that the expression of differential genes was closely related to the 5'-adenosine monophosphate-activated protein kinase (AMPK) signaling pathway and that the expression of forkhead box O 1 (FoxO1) was significantly increased in the presence of BMSCs. Furthermore, we determined the expression levels of p-AMPK/AMPK and p-FoxO1/FoxO1 by western blot analysis. The expression of p-AMPK/AMPK was upregulated, whereas that of p-FoxO1/FoxO1 was downregulated upon coculturing with BMSCs. The AMPK-specific antagonist Compound C inhibited the downregulation of p-FoxO1/FoxO1 induced by the BMSC coculture. Furthermore, treatment with the specific FoxO1 antagonist AS1842856 reduced the inhibitory effects of BMSCs on cardiomyocyte hypertrophy in vivo and in vitro. Our present study demonstrates the inhibition of cardiomyocyte hypertrophy by BMSCs, which occurs partly through the AMPK-FoxO1 signaling pathway.

Pulmonary endothelium-derived PD-L1 induced by the H9N2 avian influenza virus inhibits the immune response of T cells.

BACKGROUND: The PD-1/PD-L1 pathway is an inhibitory signaling pathway that maintains the balance between the immune response and immunotolerance, and its overactivation in cancer and viral infections inhibits T cell function. The target cells of various viruses, microvascular endothelial cells (MECs) have been shown to be key regulatory points in immune regulation and virion diffusion in vivo during infection with multiple influenza virus subtypes. Furthermore, avian influenza virus (AIV) infection can induce immunosuppression by causing imbalances in immune responses and immune organ damage. Thus, the aim of this study was to investigate whether the H9N2 virus inhibited the immune function of T cells that migrated across MECs by upregulating PD-L1 expression on MECs. METHODS: The susceptibility of rat pulmonary microvascular endothelial cells (RPMECs) to the H9N2 virus was evaluated by a plaque-forming assay and immunofluorescence staining. Then, we quantified the mRNA and protein levels of PD-L1 in RPMECs induced by H9N2 virus infection using quantitative real-time PCR and flow cytometry. The interaction between the activated T cells and RPMECs infected with the H9N2 virus was revealed using a coculture system. The effect of endothelial-derived PD-L1 on T cell function was investigated by using ELISA and flow cytometry with or without a PD-L1-specific antibody. RESULTS: Surface staining and the plaque-forming assay showed that the H9N2 virus infected and replicated in RPMECs. Both the PD-L1 mRNA level and PD-L1 protein level were upregulated in RPMECs infected with the H9N2 virus. H9N2 virus-induced PD-L1 expression significantly reduced the secretions of IL-2, IFN-γ and granzyme B and perforin expression in T cells. The above data were significantly increased after treatment with an anti-PD-L1 antibody, confirming the above mentioned findings. In addition, the induction of PD-L1 expression decreased the proliferative capacity of the cocultured T cells but did not affect the apoptosis rate of T cells. CONCLUSIONS: Taken together, the results suggest that the H9N2 virus is able to inhibit the T cell immune response by upregulating PD-L1 expression in pulmonary microvascular endothelial cells.

Synchronous vasomotion of skin microvessels at acupoints on the twelve main meridians of beagle dogs.

BACKGROUND/PURPOSE: Synchronous microvascular vasomotion was detected at acupoints in our previous human study. This present study aimed to characterize the skin microvascular vasomotion at acupoints on the twelve meridians of beagle dogs. MATERIALS AND METHODS: Two acupoints were selected on each meridian and exactly located at the rosy red spots by an electrochemical color-appearing method, where the electrical resistance was measured. The skin blood flow at acupoints was recorded by laser Doppler flowmetry (LDF), and microvascular vasomotion was analyzed according to LDF waveforms. RESULTS: The skin electrical resistance at acupoints was significantly lower than that at control non-acupoints. The LDF waveforms at acupoints were sinusoidal, which showed the synchronization of the microvascular vasomotion. The spectral analysis revealed that the vasomotion frequencies at acupoints on the same meridian were identical but not among different meridians, and the frequencies on the twelve main meridians displayed a constant order. CONCLUSION: The skin microvascular vasomotion is synchronous at acupoints of beagle dogs and has a specific frequency along the meridian, and the electrochemical color-appearing method is a feasible strategy for the precise and visual location of acupoints. The study provides evidence for the universality of synchronous vasomotion of skin microvessels at acupoints.

Effect of Andrographolide on Gene Expression Profile and Intracellular Calcium in Primary Rat Myocardium Microvascular Endothelial Cells.

Andrographolide (ANDRO) is a diterpene lactone compound with extensive biological effects, such as antibacterial, antitumor and treatment of cardiovascular diseases. Until now, studies on the pharmacological functions of ANDRO are still in progress. However, little is known about the gene expression profile and calcium response of endothelial cells to ANDRO. In this study, we used a microarray technology to investigate the gene expression responses in primary rat myocardium microvascular endothelial cells treated with 10 µg/mL ANDRO. The expression of caveolin-1 and 1-phosphatidylinositol 4, 5-bisphosphate phosphodiesterase δ3 was verified by RT-PCR and western blot. In addition, we investigated the effect of ANDRO on intracellular calcium induced by exogenous adenosine triphosphate and on inflammatory response induced by lipopolysaccharide. Results showed that ANDRO treatment induced an abundance of differential expressed genes, exhibiting a multitarget regulatory effect. ANDRO significantly decreased caveolin-1 and phosphodiesterase δ3 expression, lipopolysaccharide-induced IL-6 and TNF-α levels and expression of several chemokine genes, which are associated with reducing inflammation response and decreasing calcium release without affecting normal endothelia cell function, suggesting that ANDRO may be a potential candidate to treat cardiovascular diseases with less toxicity.

Effects of aqueous extracts of Taraxacum Officinale on expression of tumor necrosis factor-alpha and intracellular adhesion molecule 1 in LPS-stimulated RMMVECs.

BACKGROUND: Mastitis gives rise to big financial burden to farm industry (mainly dairy production) and public health. Its incidence is currently high and therefore, highly effective treatments for therapy, especially with natural products are required. Taraxacum officinale has been reported to use for anti-inflammation. However, its effect on endothelium during mastitis has not been reported. METHODS: We firstly established inflammation experimental model of rat mammary microvascular endothelial cells (RMMVECs). We evaluated the effects of dandelion leaf aqueous extracts (DAE) on LPS-induced production of inflammatory mediators in RMMVECs by enzyme-linked immunosorbent assay and Western blot. We treated RMMVECs with 1 µg/ml LPS for 4 h and then incubated with 10, 100 and 200 µg/mL DAE for 4, 8, 12 and 24 h. The expression (mRNA and protein level) of targets (tumor necrosis factor-alpha (TNF- α) and Intracellular Adhesion Molecule 1 (ICAM1) was analyzed by employing real-time PCR and Western blots. The in vivo anti-inflammatory effect of DAE on mastitis within an Staphylococcus aureus-induced mouse model was also determined. RESULTS: The obtained results showed that dandelion extracts at the concentration of 100 and 200 µg/mL could significantly inhibit both TNF-α and ICAM-1 expression in all time points checked while 10 µg/mL of dandelion only suppress both expression at 8 and 12 h post-treatment. The in vivo tests showed that the DAE inhibited the expression of TNF-α and ICAM-1 in a time-dependent manner. CONCLUSIONS: All results suggest that the endothelium may use as as a possible target of dandelion for anti-inflammation.

[Pulmonary capillary hemangiomatosis: a case report and literature review].

We reported a case of pulmonary capillary hemangiomatosis (PCH) and introduced its diagnosis, differential diagnosis, pathogenesis and development of treatment based on the review of Dana Point 2008 Classification of Pulmonary Hypertensiona and current literatures. A 43-year-old female presented progressive dyspnea, elevated pulmonary arterial pressures and CT pulmonary angiography (CTPA) imaging of main pulmonary arterial enlargement and wide spread ill-defined centrilobular nodules of ground-glass opacity. Her histologic features were proliferation of capillary channels within alveolar walls as well as muscularization of arterioles and medial hypertrophy of muscular pulmonary arteries. The treatment with diuretics and warfarin was used promptly, but unfortunately was ineffective. The patient died three months after diagnosis. PCH is a very rare vascular disease with poor prognosis. The diagnosis of PCH rests on the integration of clinical and radiographic information with pathologic features, however pathology is the most reliable means. Because clinical symptoms, imaging and histological features of pulmonary veno-occlusive disease (PVOD) and PCH broadly overlap, differential diagnosis should be made carefully. Among the various pathologic features proliferation of capillaries within alveolar walls is the key point for diagnosing PCH, which is also the most critical criteria for differentiating PCH from PVOD. So far the only definitive treatment for PCH is lung transplantation, without which the patient will die several months after diagnosis.

Synergistic Copper-Aminocatalysis for Direct Tertiary α-Alkylation of Ketones with Electron-Deficient Alkanes.

In this study, a novel approach for the tertiary α-alkylation of ketones using alkanes with electron-deficient C─H bonds is presented, employing a synergistic catalytic system combining inexpensive copper salts with aminocatalysis. This methodology addresses the limitations of traditional alkylation methods, such as the need for strong metallic bases, regioselectivity issues, and the risk of over alkylation, by providing a high reactivity and chemoselectivity without the necessity for pre-functionalized substrates. The dual catalytic strategy enables the direct functionalization of C(sp3)─H bonds, demonstrating remarkable selectivity in the presence of conventional C(sp3)─H bonds that are adjacent to heteroatoms or π systems, which are typically susceptible to single-electron transfer processes. The findings contribute to the advancement of alkylation techniques, offering a practical and efficient route for the construction of C(sp3)─C(sp3) bonds, and paving the way for further developments in the synthesis of complex organic molecules.

Capillary blood flow in patients with dysmenorrhea treated with acupuncture.

OBJECTIVE: To observe capillary blood flow at acupoints during acupuncture treatment of primary dysmenorrhea and gain new insights into its analgesic mechanism. METHODS: Patients with primary dysmenorrhea were enrolled and randomly assigned to a treatment or control group. Subjects' symptoms were differentiated into various Traditional Chinese Medicine (TCM) syndromes and treated for 10 sessions with puncturing acupuncture or self-pressing right-hand Hegu (LI 4), adding other acupoints based on syndrome. Laser speckle was used to compare the change in the vasomotor amplitude and perfusion of the capillaries in Hegu (LI 4) before and during the treatment. Each subject was required to finish the period pain symptoms observation form, verbal rating scales, numerical rating scale, pain rating index, face rating scale, Zung self-rating depression scale, Zung self-rating anxiety scale, and numerical rating scale before and after treatments. RESULTS: After 10 sessions, the symptom scores, pain index (PI), and visual analog scale (VAS) decreased significantly in treatment group. The volume of blood flow in Hegu (LI 4) declined slightly. No significant evidence supported that needling caused capillary contraction, but the capillary vasomotor amplitude at Hegu (LI 4) increased remarkably. CONCLUSION: Acupuncture can increase the capillary blood flow, thus promoting the flow of Qi and blood in terms ofTCM theory, which facilitates pain relief.

[Retrospective analysis of 11 cases of respiratory amyloidosis].

OBJECTIVE: To investigate the clinical manifestation, diagnosis and treatment of respiratory amyloidosis. METHODS: Data of 11 patients with respiratory amyloidosis diagnosed by biopsy in Peking University First Hospital from January 2002 to January 2012 were analyzed, and the related literatures were reviewed. RESULTS: In the last decade, 250 of 389 402 hospitalized patients were pathologically diagnosed as having amyloidosis, and 11 cases were pathologically confirmed to be respiratory amyloidosis. In these 11 patients, 4 cases were with serum amyloid A (AA) amyloidosis and 7 with light-chain (AL) amyloidosis. The main clinical manifestations included hoarseness, cough and dyspnea. In 4 cases with AA type unilateral larynx was involved and there was no recurrence after surgical resection. Of 7 cases with AL type, 2 cases had involvement of bilateral larynxes and both relapsed after surgery. Diffuse involvement of trachea and bronchi was found in 4 cases, and the chest CT scans showed diffuse thickening and local calcification of the airway wall, bronchial stenosis and nodules protruding into the lumen. Bronchoscopy showed airway mucosal hypertrophy, hyperemia, edema and bronchial stenosis. Lung involvement was found in 3 cases, 2 of which presented with diffuse pulmonary interstitial infiltrates, and another case presented with solitary pulmonary mass and extrapulmonary lesions. Of the 7 cases with AL type, 3 cases were treated by chemotherapy and/or radiotherapy, 3 received surgery, 2 underwent autologous hematopoietic stem cell transplantation, and 2 underwent bronchoscopic interventional therapy. Within 3 years of follow-up, 4 patients were alive, 2 dead and 1 lost to follow up. CONCLUSIONS: Respiratory amyloidosis, which can be divided into AA and AL types, is clinically rare. Patients with AA type usually present with local lesions, which can be cured by surgery, while patients with AL type often present with diffuse lesions and require integrated therapies including surgery, interventional treatment, chemotherapy, radiotherapy, and autologous hematopoietic stem cell transplantation.

Mechanism Study on Chinese Medicine in Treatment of Nodular Goiter.

Nodular goiter has become increasingly prevalent in recent years. Clinically, there has been a burgeoning interest in nodular goiter due to the risk of progression to thyroid cancer. This review aims to provide a comprehensive summary of the mechanisms underlying the therapeutic effect of Chinese medicine (CM) in nodular goiter. Articles were systematically retrieved from databases, including PubMed, Web of Science and China National Knowledge Infrastructure. New evidence showed that CM exhibited multi-pathway and multi-target characteristics in the treatment of nodular goiter, involving hypothalamus-pituitary-thyroid axis, oxidative stress, blood rheology, cell proliferation, apoptosis, and autophagy, especially inhibition of cell proliferation and promotion of cell apoptosis, involving multiple signal pathways and a variety of cytokines. This review provides a scientific basis for the therapeutic use of CM against nodular goiter. Nonetheless, future studies are warranted to identify more regulatory genes and pathways to provide new approaches for the treatment of nodular goiter.

Observation of Microvascular Perfusion in the Hegu (LI4) Acupoint Area after Deqi Acupuncture at Quchi (LI11) Acupoint Using Speckle Laser Blood Flow Scanning Technology.

The aim of this study was to investigate the traditional meridian theory using speckle laser blood flow scanning technology to observe microcirculation of the Hegu acupoint area after acupuncture stimulation on distant points. An observational study was conducted to observe the microvascular perfusion of Hegu (LI4) and control points after acupuncturing Quchi (LI11). Thirty healthy volunteers (mean age 31.6 ± 8.7 years) received deqi acupuncture on Quchi (LI11, right side), and simultaneously changes in microvascular perfusion of Sanjian (LI3), Hegu (LI4), Yangxi (LI5), and two control points were observed before, during, and after needling using a MOOR speckle laser. The results showed that the changes in microvascular perfusion of the observed points are not regular. After correction, the experiment showed that the blood perfusion on 3 meridian acupoints was increased while the perfusion on 2 control points was decreased following acupuncture stimulation, the changes at Hegu (LI4) being the statistically most significant ones. Deqi acupuncture can help in regulating the body's blood flow, with a certain degree of meridian specificity.

[Retrospective analysis of relationship between radiological features and prognoses of pneumocystis pneumonia in non-AIDS immunocompromised patients].

OBJECTIVE: To elucidate the relationship between the radiological features and prognoses of pneumocystis pneumonia (PCP) in non-acquired immunodeficiency syndrome (non-AIDS) immunocompromised patients. METHODS: Retrospective analyses were performed for the radiological features and prognoses of 36 non-AIDS immunocompromised PCP patients hospitalized at Peking University First Hospital from April 2006 to April 2010. There were 24 males and 12 females with an average age of (51.7 ± 18.9) years. RESULTS: All patients underwent chest photographic examinations and chest computed tomography (CT) was performed on 29 of them. The overall case fatality rate (CFR) of 36 patients was 33.3% (12/36). At the start of anti-PCP treatment, there were normal chest radiography (n = 7), bilateral diffuse infiltration (n = 21), bilateral diffuse consolidation (n = 8), aerothorax, mediastinal emphysema or pneumohypoderma (n = 6) and numbers of deceased patients (n = 0, 8, 5 and 5 respectively). The CFRs of the patients with normal chest radiography and aerothorax, mediastinal emphysema or pneumohypoderma were both statistically significant as compared with the overall rate (P < 0.05). At the start of anti-PCP treatment, there were bilateral diffuse ground glass opacities (GGO) (n = 29), "mosaic sign" (n = 19), air bronchogram (n = 19), pulmonary consolidation (n = 17), bilateral pleural effusion (n = 7), thickened lobular septa (n = 6), cystic lesions (n = 5), aerothorax, mediastinal emphysema or pneumohypoderma (n = 4) and numbers of deceased patients (n = 8, 5, 5, 5, 2, 2, 2 and 4 respectively). The CFRs of the patients with aerothorax, mediastinal emphysema or pneumohypoderma were statistically significant as compared with the overall rate (P < 0.05). CONCLUSIONS: The major radiological features of PCP in non-AIDS immunocompromised patients are bilateral diffuse infiltration. And bilateral diffuse GGO on chest CT scans are usually accompanied with "mosaic" sign, air bronchogram or pulmonary consolidation. The prognoses are excellent for the patients with normal chest radiograph and poor for those with aerothorax, mediastinal emphysema or pneumohypoderma.

[Study on the chemical constituents of Lonicera similes].

OBJECTIVE: To study the chemical constituents of flower buds of Lonicera similes. METHODS: The chemical constituents were isolated and purified with macroporous resin, silica gel column chromatography, gel chromatography, ect. The structures were elucidated according to the references combined with the application of spectrum techniques such as 1H-NMR, 13C-NMR ect. RESULTS: Nine compounds were isolated and their structures were identified as 5-hydroxyl-7,4'-dimethoxyflavone (I), rutin (II), quercetin (III), amentoflavone (IV), aesculetin (V), beta-sitosterol (VI), 5-hydroxyl-7,3', 4'-trimethoxyflavone (VIII), oleanolic acid (VIII), beta-daucosterin (IX). CONCLUSION: Among the nine isolated compounds, compounds I, II, III, IV, V, are flavonoids. Compounds I , IV, V, VII, VIII, IX are isolated from this plant for the first time.

Microvascular endothelial cells play potential immunoregulatory roles in the immune response to foot-and-mouth disease vaccines.

Microvascular endothelial cells (MVECs) have been documented to have important immunoregulatory effects. Exploring their roles in the immune response to foot-and-mouth disease (FMD) vaccines would help to improve their efficacy. In this study, the effects of FMD vaccine 146s antigens on gene expression profiles of rat intestinal mucosal and myocardial MVECs were analysed using microarray, and their effects on transendothelial migration (TEM) of peripheral blood mononuclear cells (PBMC) were investigated by the Transwell migration assay. Both kinds of MVECs displayed significant responses to 146s antigens, and 252 and 67 genes were differentially expressed in rat intestinal mucosal and myocardial MVECs, respectively. Despite different altered gene expression patterns, many immune-associated genes were involved in both kinds of MVECs. The gene expression changes by microarray were confirmed by real-time reverse transcription-polymerase chain reaction. Transwell migration analysis indicated that the TEM of PBMC was increased by 146s antigens, which could be partially inhibited by blocking vascular cell adhesion molecule 1 in MVECs. This study suggests that MVECs play potential immunoregulatory roles in the immune response to FMD vaccines, one of which is influencing the TEM of immune cells.

Establishment and characterization of a rat intestinal microvascular endothelial cell line.

Intestinal microvascular endothelial cell (IMVEC) is a fundamental and essential component of gut-vascular barrier which is closely associated with intestinal disorders However, there is still a lack of established intestinal microvascular endothelial cell line. In the present study, a newly established rat intestinal microvascular endothelial cell line termed RIMVEC-11 was described and characterized which has been stably cultured for more than 90 passages so far. RIMVEC-11 was characterized by endothelial features with the cobblestone morphology under light microscopy, the Weibel-Palade body and rich vesicles in the cytoplasm on the ultrastructural level, and positive endothelial specific markers CD31 and von Willebrand factor by immunocytochemistry analysis. Meanwhile, RIMVEC-11 maintained the fundamental physiological function of the microvascular endothelial cells. Tube formation assay confirmed that RIMVEC-11 retained the potential for capillaries formation. Scratch assay confirmed the endothelial cell migration potential of RIMVEC-11. Thus, a novel IMVEC cell line RIMVEC-11 was established, which could be used as a promising model for the gut-vascular barrier research.

Key genes and proteins involved in CTCM-reducing microvascular endothelial cell permeability induced by SLT-IIv using gene chips and DIGE.

An Affymetrix mouse genome array and differential in-gel electrophoresis (DIGE) techniques were used to investigate the pharmacological mechanisms of a mixture of herbs, designated CTCM, a compound of traditional Chinese medicine, for the treatment of increased permeability in mouse intestinal microvascular endothelial cells (MIMECs) induced by the Shiga-like toxin type II variant (SLT-IIv). MIMECs were challenged with 10microg/ml SLT-IIv for 12h and then treated with CTCM at a concentration of 200microg/ml for 12h. Total RNA and proteins from each treatment group were extracted from cultured MIMECs for analysis by the Affymetrix GeneChip Mouse Genome 430 2.0 microarray and DIGE. The results obtained demonstrated that there were one genes downregulated and one genes upregulated, one protein downregulated and four proteins upregulated in the SLT-IIv group compared to the control group. In the CTCM group, four genes were upregulated, three genes were downregulated, a single protein was downregulated and a single protein was upregulated when compared to the control group. When the CTCM-treated group was compared to the SLT-IIv group, expression of one gene was found to be increased, and all other genes were decreased, with five proteins downregulated. Analysis of the data suggested that CTCM specifically and effectively reduced microvascular endothelial cell permeability to SLT-IIv in the treatment of pig edema disease. In the CTCM-treated group, hspa9 expression was increased in both gene chip and DIGE analysis, so it may be a key protein in reducing cell permeability and utilized in medical treatments.

IFN-γ induces IFN-α and IFN-ß expressions in cultured rat intestinal mucosa microvascular endothelial cells.

Although researchers have recently begun to pay more attention to the immunological characteristics of microvascular endothelial cells (MVECs), there are no reports on whether activation of MVECs by interferon-γ (IFN-γ) exerts any influence on the expressions of IFN-α/ß. In the present study, we examined the influence of IFN-γ on the expressions of IFN-α/ß in rat intestinal mucous MVECs (RIMMVECs). Different concentrations of IFN-γ were used to stimulate cultured RIMMVECs in vitro, and the cells and cell supernatants were collected at different time intervals. The influence of IFN-γ on the expressions of IFN-α/ß in the RIMMVECs was examined at the mRNA and protein levels by real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA), respectively. The results indicated that IFN-γ was able to activate RIMMVECs, thereby leading to upregulated expressions of IFN-α/ß. The real-time quantitative PCR analyses indicated that the IFN-α/ß mRNA expression levels in RIMMVECs achieved their peak values after stimulation with IFN-γ at 20 ng/mL for 6 h and were increased by 14.88- and 3.82-fold, respectively, when compared with the levels in negative control cells. The ELISA analyses revealed that the IFN-α/ß protein expression levels achieved their peak values after stimulation with IFN-γ at 40 ng/mL. The expression of IFN-α protein achieved its peak value at 12 h, while the expression of IFN-ß protein achieved its peak value after 6 h. The present results suggest that the expression and secretion of IFNs may participate in the immunologic barrier function of MVECs.

A treatment planning comparison between proton beam therapy and intensity-modulated x-ray therapy for recurrent nasopharyngeal carcinoma.

BACKGROUND AND PURPOSE: A comparative treatment planning study was undertaken between proton beam therapy (PBT) and intensity-modulated x-ray therapy (IMXT) for patients with recurrent nasopharyngeal carcinoma (NPC), to assess the potential benefits and limitations of the two treatment modalities. MATERIALS AND METHODS: We retrospectively analyzed seven patients with recurrent NPC treated at our proton center with PBT from 2004 to 2007. Eclipse proton treatment planning system and Eclipse inverse treatment planning system for IMXT were employed. For each patient, the IMXT plan was optimized with nine evenly spaced coplanar fields. Three coplanar beams and passive scattering mode were used for PBT. The dose prescription in cobalt Gray equivalent (CGE) for gross tumor volume (GTV) was 66CGE and for planning target volume (PTV), 62.7CGE. Dose-volume histograms (DVH) were used to evaluate the difference in dosimetric distributions for the target volume and the organs at risk (OARs). RESULTS: Optimal target volume coverage and similar target conformation were achieved in both PBT and IMXT. Median conformity index was 0.72 and 0.75 (p=0.15) and median inhomogeneity coefficient was 0.14 and 0.10 (p=0.08) for PBT and IMXT, respectively. Dose to OARs was significantly lower in PBT plans than IMXT. Median maximal dose to the brainstem was 27.89CGE(cobalt Gray equivalent) and 42.45Gy (p< 0.01), and the dose to 5% of the brainstem(D5) was 12.83CGE and 19.47Gy (p< 0.001 ), for PBT and IMXT, respectively. Median maximal dose to the spinal cord was 8.38CGE and 22.91Gy (p< 0.004), and the dose to 5% of the spinal cord was 2.18CGE and 13.62Gy (p< 0.001), for PBT and IMXT, respectively. CONCLUSIONS: The use of PBT, when compared with IMXT, resulted in similar levels of tumor conformation. PBT, however, exposed the OARs to a significantly lower dose, effectively sparing the brainstem, spinal cord, optic nerve and chiasm, temporal lobes and parotid glands. The superior dose distributions possible with PBT should translate into reduced morbidity and improved quality of life.