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Evaluation of Galactomannan Enzyme Immunoassay and Quantitative Real-Time PCR for the Diagnosis of Invasive Pulmonary Aspergillosis in a Rat Model

  • Lin, Jian-Cong;Xing, Yan-Li;Xu, Wen-Ming;Li, Ming;Bo, Pang;Niu, Yuan-Yuan;Zhang, Chang-Ran
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1044-1050
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    • 2014
  • Since there is no consensus about the most reliable assays to detect invasive aspergillosis from samples obtained by minimally invasive or noninvasive methods, we compared the efficacy of an enzyme-linked immunosorbent assay (ELISA) for galactomannan (GM) detection and quantitative real-time PCR assay (qRT-PCR) for the diagnosis of invasive pulmonary aspergillosis. Neutropenic, male Sprague-Dawley rats (specific pathogen free; 8 weeks old; weight, $200{\pm}20g$) were immunosuppressed with cyclophosphamide and infected with Aspergillus fumigatus intratracheally. Tissue and whole blood samples were harvested on days 1, 3, 5, and 7 post-infection and examined with GM ELISA and qRT-PCR. The A. fumigatus DNA detection sequence was detected in the following number of samples from 12 immunosuppressed, infected rats examined on the scheduled days: day 1 (0/12), day 3 (0/12), day 5 (6/12), and day 7 (8/12) post-infection. The sensitivity and specificity of the qRT-PCR assay was 29.2% and 100%, respectively. Receiver operating characteristic curve (ROC) analysis indicated a Ct (cycle threshold) cut-off value of 15.35, and the area under the curve (AUC) was 0.627. The GM assay detected antigen in sera obtained on day 1 (5/12), day 3 (9/12), day 5 (12/12), and day 7 (12/12) post-infection, and thus had a sensitivity of 79.2% and a specificity of 100%. The ROC of the GM assay indicated that the optimal Ct cut-off value was 1.40 (AUC, 0.919). The GM assay was more sensitive than the qRT-PCR assay in diagnosing invasive pulmonary aspergillosis in rats.

Effect of Phlorotannins Isolated from the Ethyl Acetate Fraction of Ecklonia stolonifera on Peritoneal Macrophage Polarization (복강대식세포의 염증성 표현형에 대한 곰피(Ecklonia stolonifera) 유래 Phlorotannins의 효과)

  • Choi, Min-Woo;Choi, Jun-Hyeong;Kim, Hyeung-Rak;Kim, Jae-Il
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.4
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    • pp.439-446
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    • 2015
  • Inflammation is a protective response to infection or injury. However, prolonged inflammation can contribute to the pathogenesis of many diseases, such as cancer, diabetes, arthritis, atherosclerosis, and Alzheimer's disease. Recent studies have shown that activated macrophages, inflammatory effector cells, can react to tissue insults in a polarized manner, in which their phenotypes are polarized into two major subtypes, categorized as M1 or M2. Classical M1 activation involves the production of pro-inflammatory cytokines, such as interleukin (IL)-6 and tumor necrosis factor (TNF)-${\alpha}$, and free radicals, while M2 or alternative activation is an anti-inflammatory phenotype involved in homeostatic processes, such as wound healing, debris scavenging, and the dampening of inflammation via the production of very low levels of pro-inflammatory cytokines and high levels of anti-inflammatory mediators, including IL-10. As part of our ongoing effort to isolate anti-inflammatory compounds from seaweeds, we investigated the effects of phlorotannins isolated from the brown alga Ecklonia stolonifera on macrophage polarization. Mouse peritoneal macrophages were treated with various concentrations of the extracts, and real-time RT-PCR analyses were performed to examine the expression of polarization markers: IL-$1{\beta}$, IL-6, and TNF-${\alpha}$ for M1 and arginase-1, peroxisome proliferator-activated receptor (PPAR)-${\gamma}$, found inflammatory zone-1 (Fizz-1), chitinase 3-like 3 (Ym1), and$Kr{\ddot{u}}ppel$-like factor 4 (Klf-4) for M2. The pretreatment of cells with eckol, dieckol, and phlorofucofuroeckol-A (PFF-A), isolated from the ethyl acetate fraction of E. stolonifera ethanolic extract, potentiated the anti-inflammatory M2 phenotype of the macrophages. These results indicate that phlorotannins derived from E. stolonifera can be used to enrich macrophages with markers of the M2 anti-inflammatory state.

Effects of Glucuronic Acid Derivertives Isolated from Xylan an Antioxidative Defense System and Muscle Fatigue Recovery after Aerobic Exercise (Xylan으로부터 단리한 Glucuronic Acid의 유산소 운동 후 항산화 작총 및 근피로 회복효과)

  • 최향미;이수천;류승필;이인구;주길재;이순재
    • Journal of Nutrition and Health
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    • v.34 no.8
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    • pp.872-880
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    • 2001
  • The purpose of this study was to investigate the effects of glucuronic acid on antioxidative defense system and recovery of muscle fatigue in rat artier aerobic exercise. Sprague-Dawley male rats weighing 150 $\pm$ 10g were randomly assigned to one normal(N) group and three exercise training groups. Exercise training groups were classified into glucuronic acid free intubation group(T group), 250mg glucuronic acid/kg bw intubation group(TU group), and 500 mg glucuronic acid/kg bw intubation group(2TU group) according to glucuronic acid supplementation level. The glucuronic acids were administered to rats by oral intubation before exercise training. The experimental rats in exercise training groups(T, TU and 2TU) were exercised on glucuronic acid supplementation or rats in normal group were confined in cage for 4 weeks. And rats were sacrificed with an overdose of pentobarbital injection just after running. Liver xanthine oxidase(XOD) activities were not significantly different among four groups. The activity of superoxide dismutase(SOD) in T group was no significant difference from N group, but those of TU and 2TU groups were increased by 9% and 18%, respectively, compared with that of T group. Liver glutathione peroxidase(GSHpx) activites of T and TU groups showed a similar tendency to that of normal group, but increase 17% in 2TU group compared with normal group. The ratio of GSH/GSSG in liver of T group was lower than that of normal group, but those of TU and 2TU groups were a similar tendency to that of normal group. Contents of thiobarbituric acid reactive substance(TBARS) in T group was increased by 47%, compared with that of normal group but those of TU group and 2TU group were lower 27% and 35%, respectively, compared with that of T group. The contents of glycogen in soleus muscle significantly lower in all three trained exercise groups than that of normal group, but there were no significant differences among the trained exercise groups. Contents of hepatic glycogen in T group were decreased 27% compared with those of normal group while those of TU and 2TU groups were the same as normal group levels. The contents of serum lactic acid in T group were increased 240% of normal group, but hose of TU and 2TU groups were decreased 38%, 39%, respectively, by glucuronic acid supplementations, compared with that of T group. In conclusion, the effects of glucuronic acids in exercise training rats would appear to reduce peroxidation of tissue as an antioxidative defense mechanism and promote recovery of muscle fatigue.

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MicroRNA-155 Expression has Prognostic Value in Patients with Non-small Cell Lung Cancer and Digestive System Carcinomas

  • Xu, Tong-Peng;Zhu, Can-Hong;Zhang, Jian;Xia, Rui;Wu, Feng-Lei;Han, Liang;Shen, Hua;Liu, Ling-Xiang;Shu, Yong-Qian
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7085-7090
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    • 2013
  • Objective: Published data have shown that microRNAs (miRNAs) could play a potential role as diagnostic and prognostic indicators in cancers. Data for the predictive value of microRNA-155 are inconclusive. The aim of the present analysis was therefore to evaluate the role of miR-155 in prognosis for patients with a variety of carcinomas. Methods: Relevant studies were identified by searching PubMed and EMBASE. Data were extracted from studies comparing overall survival (OS), recurrence-free survival (RFS) or cancer-specific survival (CSS) in patients with carcinoma with higher miR-155 expression and those with lower levels. The pooled hazard ratios (HRs) and 95% confidence intervals (CIs) of miR-155 for clinical outcome were calculated. Results: A total of 15 studies were included. The pooled hazard ratio (HR) for OS of higher miR-155 expression in cancerous tissue was 1.89 (95% CI: 1.20-2.99, P=0.006), which could markedly predict poorer survival in general cancer. For RFS/CSS, elevated miR-155 was also associated with poor prognosis of cancer (HR=1.50, 95% CI: 1.10-2.05, P=0.01). On subgroup analysis, the pooled HR for OS in non-small cell lung cancer (NSCLC) was 2.09 (95% CI: 0.68-6.41, P > 0.05), but for RFS/CSS was 1.28 (95% CI: 1.05-1.55, P=0.015), with statistical significance; the pooled HRs for OS and RFS/CSS in digestive system neoplasms were 3.04 (95% CI: 1.48-6.24, P=0.003) and 2.61 (95% CI: 1.98-3.42, P<0.05), respectively. Conclusions: The results indicated that the miR-155 expression level plays a prognostic role in patients with cancer, especially NSCLCs and digestive system carcinomas.

Lipid Oxidation in Shellfish under the Different Conditions of Drying (패류의 건조조건에 따른 지질산화)

  • LEE Kang-Ho;CHO Tae-Yong;CHO Ho-Sung;LEE Jong-Ho;SHIM Ki-Hwan
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.2
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    • pp.143-148
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    • 1998
  • This study was carried out in order to investigate oxidative deterioration during dehydration at $40^{\circ}C,\;50^{\circ}C$ and $60^{\circ}C$ of sea mussel and baby clam. Moisture content was decreased with drying temperature and time. Sea mussel was dehydrated more rapidly than baby clam that had Harder muscle tissue. Both samples were not reached to Aw 0.62 in case of 10 hrs drying at $40^{\circ}C$, But it reached within 8 hrs in sea mussel and 10 hrs in baby clam at $50^{\circ}C$, respectively. Even if $60^{\circ}C$ could speed up drying, it caused to form more free fatty acid, peroxide, thiobarbituric acid and brown pigments. Lipophilic brown pigment was 10 times higher than hydrophilic and actively increased in all samples. fluorescence intensity was also increased with drying temperature and time. Particularly, it was higher sea mussel than baby clam more or less.

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Tactics and Pitfalls of MED(Micro Endoscopic Discectomy) System for Lumbar Disc - For Surgeons Who Wish to Attempt - (요추간반 탈출증에 있어 MED(Microendoscopic Discectomy) System을 이용한 수술의 효율성과 수기)

  • Hong, Hyun Jong;Oh, Seong Hoon;Bak, Kwang Hum;Kim, Jae Min;Kim, Choong Hyun;Kim, Young Soo;Ko, Yong;Oh, Suck Jun;Kim, Kwang Myung;Lee, Sang Gu;Kim, Nam Kyu
    • Journal of Korean Neurosurgical Society
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    • v.29 no.1
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    • pp.35-43
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    • 2000
  • Objective : Percutaneous lumbar approaches such as arthroscopic discectomy, laser discectomy, and nucleotome remain controversial and have technical limitations to free fragment disc, bony pathology and access to L5-S1, The purpose of this study was to determine efficacy of this new endoscopic system and to report techniques and tactics. Methods : From July 1997 to May 1998, we treated 40 consecutive patients(43 levels) with the MED system. Mean age was 32 years(range ; 18 to 62). There were 30 males and 10 females. All patients had sciatica with SLRT limitation. There were 23 patients with disc herniation at L4-5 and 14 patients at L5-S1. Three patients had 2 level disc herniations. There was one far lateral disc herniation at L4-5. Results : Using modified MacNab criteria, there were 37 excellent results and 3 good result. Most patients were discharged within 3-4 days except 2 patients with dural tearing. There were no other complications. Mean operation time was 1.5 hours(range : 40 minutes to 2.5 hours). Conclusion : The MED system is a reliable approach to lumbar disc herniations. This system combines the advantages of conventional open surgery and a minimally invasive technique. As tactics for the doctors who wish to attempt, "palpate" the lamina by first dilator, identification of interlaminar space by removal of overlying soft tissue and confirmation of the shoulder portion of nerve root before discectomy are important to this procedure. We conclude that lumbar disc herniations can be successfully treated with MED approach.

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The Effects of A High-Fat Diet on Pro- and Macro-Glycogen Accumulation and Mobilization During Exercise in Different Muscle Fiber Types and Tissues in Rats

  • Lee Jong-Sam;Eo Su-Ju;Cho In-Ho;Pyo Jae-Hwan;Kim Hyo-Sik;Lee Jang-Kyu;Kwon Young-Woo;Kim Chang-Keun
    • Nutritional Sciences
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    • v.8 no.3
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    • pp.181-188
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    • 2005
  • We investigated the effects of diet manipulation on pro- and macro-glycogen accumulation and mobilization during exercise in different kinds of muscle fiber and tissue. Thirty-two Sprague-Dawley rats were divided into groups representing one of two dietary conditions: high fat (HF, n=16) or standard chow (CHOW, n=16). Each dietary group was fm1her divided into control (REST, n=8) and exercise (EXE, n=8). After an eight-week dietary intervention period, the animals in EXE swam for 3 hours while the animals in REST remained at rest Skeletal muscle (soleus, red gastrocnemius and white gastrocnemius) and liver samples were then dissected out and used for analyses. 1here was no statistical difference in body weight between the animals in the HF and mow groups (p>.05). Three hours of exercise significantly increased plasma free fatty acid (FFA) concentration in the animals in the CHOW group but not in the animals in the HF group. Both citrate. synthase (CS) and $\beta$-hydroxyacyl dehydrogenase ($\beta$-HAD) activities in skeletal muscles were higher in the HF group than in the mow group. CS and $\beta$-HAD activities were also the highest in red gastrocnemius and the lowest in white gastrocnemius. At both time points (i.e., rest and immediately after exercise) intramuscular triglyceride (IMTG) and liver TG concentrations were significantly higher in the HF compared to the CHOW. IMTG and liver TG changed selectively in the CHOW. Except in white gastrocnemius muscle, there was no significant difference in total glycogen content between HF and mow at rest. Although exercise significantly lowered total glycogen content in all groups and tissues (p<.05), the degree of reduction was markedly greater in the mow than in the HF. Whereas changes in proglycogen concentration showed a trend similar to those of total glycogen, alterations in macroglycogen concentrations clearly differed from those of total glycogen. Specifically, the degree of reduction of macroglycogen following three hours of exercise was substantially greater in the CHOW than in the HF. These results suggest that metabolic alterations induced by a long-term high fat diet may be caused by macro-glycogen rather than pro-glycogen.

Patterns of care and treatment outcomes for primary thyroid lymphoma: a single institution study

  • Cha, Hyejung;Kim, Jun Won;Suh, Chang-Ok;Kim, Jin Seok;Cheong, June-Won;Lee, Jeongshim;Keum, Ki Chang;Lee, Chang Geol;Cho, Jaeho
    • Radiation Oncology Journal
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    • v.31 no.4
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    • pp.177-184
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    • 2013
  • Purpose: The aim of this study was to analyze the patterns of care and treatment outcomes in patients with primary thyroid lymphoma (PTL) in a single institution. Materials and Methods: Medical records of 29 patients with PTL treated between April 1994 and February 2012 were retrospectively reviewed. Diagnosis was confirmed by biopsy (n = 17) or thyroidectomy (n = 12). Treatment modality and outcome were analyzed according to lymphoma grade. Results: The median follow-up was 43.2 months (range, 3.8 to 220.8 months). The median age at diagnosis was 57 years (range, 21 to 83 years) and 24 (82.8%) patients were female. Twenty-five (86.2%) patients had PTL with stage IEA and IIEA. There were 8 (27.6%) patients with mucosa-associated lymphoid tissue (MALT) lymphoma and the remaining patients had high-grade lymphoma. Patients were treated with surgery (n = 2), chemotherapy (n = 7), radiotherapy (n = 3) alone, or a combination of these methods (n = 17). Treatment modalities evolved over time and a combination of modalities was preferred, especially for the treatment of high-grade lymphoma in recent years. There was no death or relapse among MALT lymphoma patients. Among high-grade lymphoma patients, 5-year overall survival (OS) and 5-year progression-free survival (PFS) were 75.6% and 73.9%, respectively. Complete remission after initial treatment was the only significant prognostic factor for OS (p = 0.037) and PFS (p = 0.003). Conclusion: Patients with PTL showed a favorable outcome, especially with MALT lymphoma. Radiotherapy alone for MALT lymphoma and chemotherapy followed by radiotherapy for high-grade lymphoma can be effective treatment options for PTL.

Effects of Mahwangbujaseshin-tang (Mahuangfuzixixintang) (麻黃附子細辛湯) on MIA-Induced Osteoarthritis Rats (마황부자세신탕(麻黃附子細辛湯)이 MIA로 유도된 골관절염 유발 Rat에 미치는 영향)

  • Lee, Hyung-Eun;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
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    • v.24 no.2
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    • pp.65-81
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    • 2014
  • Objectives This study was carried out to find out the anti-osteoarthritic effects of Mahwangbujaseshin- tang (Mahuangfuzixixintang ) on the monosodium iodoacetate (MIA)-induced osteoarthritis rats. Methods Osteoarthritis was induced by injecting MIA ($50{\mu}l$) into the knee joint of rats. Rats were divided into a 3 groups (n=7). The injection did not fit the normal group. A week later, after the injection of MIA, while control group took normal saline 2 ml, the extract of Mahwangbujaseshin-tang (Mahuangfuzixixintang ) (MBST) (200 mg/kg) was injected to treated group. After that, we examined hind paw weight bearing ability, functions of liver and kidney, serum TNF-$\alpha$, IL-$1{\beta}$, IL-6, $PGE_2$, $LTB_4$, TIMP-1, MMP-9 and hematology. Volume of cartilage was measured by micro CT arthrography. Injury of synovial tissue was measured by H & E, Safranin-O immunofluorescence. Results 1) DPPH and ABTS free radical scavenging activity of MBST was increased according to concentration of MBST and total phenolic contents were in high level. 2) In RAW 264.7 cells, ROS production was significantly decreased in MBST (at 10, $100{\mu}g/ml$) and NO was also decreased but meaningless in MBST (at $100{\mu}g/ml$). 3) In RAW 264.7 cells, IL-6 production was significantly decreased in MBST (at $100{\mu}g/ml$) and TNF-$\alpha$ and IL-$1{\beta}$ production were also decreased but meaningless in MBST (at $100{\mu}g/ml$). 4) In hind legs weight-bearing measurement, level of weight-bearing was increased. 5) Functions of liver and kidney were not affected. 6) TNF-$\alpha$, IL-$1{\beta}$, IL-6, $PGE_2$, $LTB_4$, MMP-9 and TIMP-1 production were significantly decreased. 7) In hematology, the levels of neutrophils, monocytes were significantly decreased and the levels of white blood cells, lymphocytes were also decreased but meaningless. 8) In micro CT-arthrography, cartilage volume was significantly increased. 9) Histopathologically, injury on cartilage and synovial membrane of MBST group was decreased. Conclusions Based on all results mentioned above, Mahwangbujaseshin-tang (Mahuangfuzixixintang) is believed to be meaningful for suppressing the progress of osteoarthritis. And it is related to inhibiting the activity of inflammatory cytokine and injury of volume in cartilage.

Effects of Differential Distribution of Microvessel Density, Possibly Regulated by miR-374a, on Breast Cancer Prognosis

  • Li, Jian-Yi;Zhang, Yang;Zhang, Wen-Hai;Jia, Shi;Kang, Ye;Tian, Rui
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.1715-1720
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    • 2013
  • Background: The discovery that microRNAs (miRNAs) regulate proliferation, invasion and metastasis provides a principal molecular basis of tumor heterogeneity. Microvessel distribution is an important characteristic of solid tumors, with significant hypoxia occurring in the center of tumors with low blood flow. The distribution of miR-374a in breast tumors was examined as a factor likely to be important in breast cancer progression. Methods: Breast tissue samples from 40 patients with breast cancer were classified into two groups: a highly invasive and metastatic group (HIMG) and a low-invasive and metastatic Group (LIMG). Samples were collected from the center and edge of each tumor. In each group, six specimens were examined by microRNA array, and the remaining 14 specimens were used for real-time RT-qPCR, Western blot and immunohistochemical analyses. Correlation analysis was performed for the miRNAs and target proteins. Follow-up was carried out during 28 months to 68 months after surgery, and survival data were analyzed. Results: In the LIMG, the relative content of miR-374a was lower in the center of the tumor than at its edge; in the HIMG, it was lower at the edge of the tumor, and miR-374a levels were lower in breast cancer tissues than in normal tissues. There was no difference between VEGF-A and VCAM-1 mRNA levels at the edge and center of the tumor; however, we observed a significant difference between VEGF-A and VCAM-1 protein expression levels in these two regions. There was a negative correlation between miR-374a and target protein levels. The microvessel density (MVD) was lower in the center of the tumor than at its edge in HIMG, but the LIMG vessels were uniformly distributed. There was a significant positive correlation between MVD and the number of lymph node metastases (Pearson correlation, r=0.912, P<0.01). The median follow-up time was 48.5 months. LIMG had higher rate of disease-free survival (100%, P=0.013) and longer median survival time (66 months) than HIMG, which had a lower rate of 75% and shorter median survival time (54 months). Conclusions: Our data demonstrated miR-374a to be differentially distributed in breast cancer; VEGF-A and VCAM-1 mRNA had coincident distribution, and the distribution of teh respective proteins was uneven and opposite to that for the miR-374a. These data might explain the differences in the distribution of MVD in breast cancer and variation in breast cancer prognosis.