MiR-124-3p targeting PDE4B attenuates LPS-induced ALI through the TLR4/NF-κB signaling pathway.

BACKGROUND: A large number of studies have found that microRNAs (miRNAs) and phosphodiesterase 4 (PDE4) are crucial regulators of inflammatory responses in acute lung injury (ALI). OBJECTIVE: This study will explore the protective effect of miR-124-3p on ALI and its related mechanism. METHODS: The ALI mouse model was established by intratracheal administration of lipopolysaccharide (LPS) and evaluated by haematoxylin and eosin (HE) staining, lung injury score, inflammation factors, polymorphonuclear leukocyte (PMN) count, total protein and lung wet weight/dry weight (W/D) ratio. MiR-124-3p was overexpressed in vivo by intratracheal administration of miR-agomir, and PDE4B was expressed at low level in vivo by intratracheal administration of a PDE4B inhibitor. The mRNA expression level was detected by qRT-PCR, and the protein expression level was detected by Western blot. The relationship between miR-124-3p and PDE4B was detected by dual-luciferase activity assay. RESULTS: We found that miR-124-3p was downregulated in LPS-induced ALI. Overexpression of miR-124-3p alleviated lung injury by inhibiting the Toll-like receptor 4 (TLR4)/nuclear factor-kappa B (NF-κB) signaling pathway. Furthermore, we confirmed that miR-124-3p suppressed the TLR4/NF-κB signaling pathway by directly targeting PDE4B. CONCLUSION: miR-124-3p targeting PDE4B had a protective effect on LPS-induced ALI by inhibiting the TLR4/NF-κB signaling pathway.

Butter-Derived Ruminant Trans Fatty Acids Do Not Alleviate Atherosclerotic Lesions in High-Fat Diet-Fed ApoE-/- Mice.

Atherosclerosis (AS) is the most common cardiovascular disease (CVD). Currently, it is widely believed that R-TFA and I-TFA may cause different biological effects. In the present study, we aim to elucidate the effect of mixed R-TFA derived from butter on the development of AS in high-fat diet-fed ApoE-/- mice and find the possible mechanism. It was shown that butter-derived R-TFA promoted dyslipidemia, reduced thoracic and abdominal aorta diameters, and induced aortic lipid deposition and atherosclerotic lesions in high-fat diet-fed ApoE-/- mice. Meanwhile, butter-derived R-TFA affected the serum lipid profile of high-fat diet-fed ApoE-/- mice and the lipid metabolism of human umbilical vein endothelial cells (HUVECs). Through lipidomic techniques, we found that butter-derived R-TFA had a significant effect on the glycerophospholipid metabolic pathway. In conclusion, our results demonstrated that butter-derived R-TFA does not alleviate but promotes atherosclerotic lesions in high-fat diet-fed ApoE-/- mice and that the glycerophospholipid metabolic pathway plays a major role in this pro-atherosclerotic effect.

[An experimental research of hyperthermic preconditioning on the spinal cord injury].

OBJECTIVE: To assess the effects of hyperthermic preconditioning on spinal cord injury(SCI) in rats. METHODS: A total of 96 SD rats were divided into control(A), trauma(B) and hyperthermic preconditioning (C) group. SCI models were made by static compression. The animals were decapitated at 6 h, 12 h, 1 d, 2 d, 3 d, 1 w, 2 w and 3 w after injury. With in situ hybridization and immunohistochemistry, the expression of HSP70 after spinal cord injury was studied at the transcript level and translation level respectively. Neurological outcome was evaluated by the combined behavioral score (CBS). RESULTS: Neurological outcome in Group C was significantly higher than that in Group B (P < 0.05). In-situ hybridization and Immunohistochemistry analysis showed that expression of HSP70 in spinal cord elevated after hyperthermic preconditioning; furthermore, the level of HSP70 in Group C was significantly higher than that in Group B (P < 0.05). CONCLUSION: Hyperthermic preconditioning might improve neurological outcome after spinal cord injury in rats, and the protective mechanism in this connection might involve the induction of HSP70 synthesis in spinal cord.

[A study of correlation between the blood supply types of PHcc on spiral CT and the level of VEGF expression in PHcc].

OBJECTIVE: To study the correlation between the blood supply types of primary hepatocellar carcinoma (PHcc) on Spiral CT and the level of vascular endothelial growth factor (VEGF) expression in PHcc for improvement in treatment strategies and prognostication. METHODS: Forty-five cases of PHcc identified by operation and pathology were investigated. Immunohistochemistry staining in SP was performed. The relationships between blood supply types of PHcc on Spiral CT during dual-phase scanning and the expression levels of VEGF in well-differentiated, moderately differentiated and poorly differentiated PHcc were analyzed. RESULTS: There were four blood supply types of PHcc on Spiral CT. Both the strong positive staining and the positive staining were most frequently seen in the hepatic artery blood supply type, and then were frequently in the double blood supply type, i. e. the hepatic arterial supply combining with the portal blood supply type. The positive staining results were rarely seen in the portal blood supply type and poor blood supply type (P<0.001). And with the escalation of the Edmonson- Steiner histological grades, the VEGF expression levels were shown to increase correspondingly (P < 0.05). CONCLUSION: We can ascertain the blood supply type of PHcc and infer the VEGF expression levels that reflect the PHcc angiogenesis condition and the histological grades by means of Spiral CT during dual- phase scanning. Hence this method is useful for the selection of PHcc treatment plans, including anti-angiogenesis and evaluating the prognosis.

Neuroprotective effect of exogenous vascular endothelial growth factor on rat spinal cord neurons in vitro hypoxia.

BACKGROUND: Vascular endothelial growth factor (VEGF) is well known as a hypoxia-induced protein. That it markedly increased expression of VEGF and improvement of rat motor function after spinal cord injury suggested that VEGF could play a neuroprotective role in ischaemic tolerance. This study investigated whether vascular endothelial growth factor has direct neuroprotective effects on rat spinal cord neurons. METHODS: We employed primary cultures of embryonic rat spinal cord neurons, then administrated different concentrations of VEGF164 in the culture medium before hypoxia when the number of neurons was counted and the cell viability was detected by MTT. The neuronal apoptosis and expression of VEGF and its receptor genes were evaluated by terminal deoxynucleotidyl transferase mediated dUTP nick-end labelling (TUNEL) and immunohistochemistry. The VEGFR2/FLK-1 inhibitor, SU1498, was used to confirm whether the neuroprotective effect of VEGF was mediated through VEGFR2/Flk-1 receptors. RESULT: In hypoxic conditions, the number and viability of neurons decreased progressively, while the number of TUNEL-positive cells increased along with the prolongation of hypoxic exposure. When the concentration of VEGF in cell culture medium reached 25 ng/ml, the cell viability increased 11% and neuronal apoptosis reduced to half, this effect was dose dependent and led to an approximately 25% increase in cell viability and about threefold decrease in TUNEL-positive cells at a maximally effective concentration of 100 ng/ml. In normal conditions, VEGF/Flk-1 but not VEGF/Flt-1 gene expressed at a low level: after hypoxia, the expression of VEGF/Flk-1, but not VEGF/Flt-1 was significantly increased. The protective effect of VEGF was blocked by the VEGFR2/Flk-1 receptor tyrosine kinase inhibitor, SU1498. CONCLUSIONS: VEGF has direct neuroprotective effects on rat spinal cord neurons, which may be mediated in vitro through VEGFR2/Flk-1 receptors.

Tumor micrometastases in mesorectal lymph nodes and their clinical significance in patients with rectal caner.

AIM: To investigate the number, size, and status of lymph nodes within the mesorectum and to explore the prognostic significance of lymph node micrometastases in patients with rectal cancer. METHODS: Thirty-one patients with rectal cancer undergone total mesorectal excision between October 2001 and October 2002 were included. Mesorectal nodes retrieved from the resected specimens were detected with a combination of haematoxylin and eosin (HE) staining and immunohistochemistry (IHC). The relations between lymph node metastases, micrometastases and postoperative recurrence were analyzed. RESULTS: A total of 548 lymph nodes were harvested, with 17.7+/-8.2 nodes per case. The average number of metastatic nodes in HE-positive patients and micrometastatic nodes in IHC-positive patients was 5.2+/-5.1 per case and 2.2+/-1.3 per case, respectively. The mean size of all nodes and metastatic nodes was 4.1+/-1.8 mm and 5.2+/-1.7 mm in diameter, respectively. The mean size of micrometastatic nodes was 3.9+/-1.4 mm in diameter. The size of the majority of mesorectal nodes (66.8%), metastatic nodes (52.6%), and micrometastatic nodes (79.5%) was less than 5 mm in diameter. During a median follow-up period of 24.6+/-4.7 mo, 5 patients (16.7%) had recurrence, of them 2 died and 3 survived. Another case died of tumor unrelated cause and was excluded. All 5 recurrent cases had 3 or more nodes involved, and one of them developed only lymph node micrometastases. The mean number of both metastatic and micrometastatic nodes per case differed significantly between the recurrent and non-recurrent groups (P<0.01 and P = 0.01, respectively). CONCLUSION: The majority of lymph nodes, metastatic, and micrometastatic lymph nodes within the mesorectum are smaller than 5 mm in diameter. The nodal status and the number of lymph nodes involved with tumor metastases and micrometastases are related to the rapid postoperative recurrence.

[The effects of anti-inflammatory and anti-asthmatic agents on CD34+ hematopoietic cells in bone marrow of asthmatic mice].

OBJECTIVE: To observe the effects of glucocorticoids and cysteinyl leukotrienes 1 receptor antagonist on CD(34)(+) hematopoietic cells, and to study the rationality of a bone marrow-targeting anti-inflammatory strategy. METHODS: Twenty-four BALB/c mice were sensitized and challenged by 1% ovalbumin (OVA) to establish the asthmatic model. Asthmatic mice were challenged by 1% OVA and divided into 4 groups: fed by sterile saline (group A), prednisone (group B), montelukast (group C) and prednisone plus montelukast (group D) respectively for two consecutive weeks. The mice were killed at 24 h after the last challenge, then bronchoalveolar lavage fluid (BALF), peripheral blood and bone marrow were prepared. Eosinophils in peripheral blood and BALF, nucleate cells in BALF, peripheral blood and bone marrow were counted. The percentage of CD(34)(+) cells, CD(4)(+), CD(8)(+) T lymphocyte to nucleate cells in peripheral blood and bone marrow were counted by flow cytometry. Immunocytochemistry and in situ hybridization were employed to detect the hematopoietic cells expression of CD(34)(+) and IL-5Ralpha mRNA in bone marrow (CD(34)(+) IL-5Ralpha mRNA(+) cells). RESULTS: The number of EOS in BALF and peripheral blood and the number of CD(34)(+) cells in peripheral blood and bone marrow in group A were [(18.3 +/- 1.3) x 10(5)/L], [(2.5 +/- 0.4) x 10(8)/L], [(9.6 +/- 5.1) x 10(7)/L] and [(7.7 +/- 3.2) x 10(7)/femur] respectively, compared with the corresponding indices in group B [(4.6 +/- 1.7) x 10(5)/L, (1.5 +/- 0.3) x 10(8)/L, (3.9 +/- 2.1) x 10(7)/L, (3.3 +/- 1.8) x 10(7)/femur] and group D [(3.7 +/- 1.4) x 10(5)/L, (1.7 +/- 0.3) x 10(8)/L, (4.1 +/- 1.8) x 10(7)/L, (2.2 +/- 0.7) x 10(7)/femur]; the differences all were significant (all P < 0.01). The number of bone marrow CD(34)(+) IL-5Ralpha mRNA(+) in group B and D were (23 +/- 7)% and (21 +/- 4)%, as compared with the corresponding index in group A [(37 +/- 4)%], the differences were significant (P < 0.01); the number of eosinophils in BALF in group C was (12.2 +/- 1.1) x 10(5)/L, as compared with the corresponding index in group A [(18.3 +/- 1.3) x 10(5)/L], the difference was significant (P < 0.05). CONCLUSIONS: Prednisone probably inhibits the proliferation, differentiation and emigration of CD(34)(+) cells in the bone marrow of asthmatic mice, and inhibits eosinophilopoiesis in bone marrow, eosinophil migration into peripheral blood and recruitment to the airways. Montelukast may suppress eosinophil infiltrating into lungs of asthmatic mice, but it does not inhibit the proliferation and emigration of CD(34)(+) cells and does not show apparent synergistic effect with prednisone.

Distribution and patterns of lymph nodes metastases and micrometastases in the mesorectum of rectal cancer.

BACKGROUND AND OBJECTIVES: Facts buried in the mesorectum remain to be unveiled. This study investigated the number, size, and detailed distribution of lymph nodes metastases and micrometastases within the mesorectum of rectal cancer. METHODS: Thirty-one patients who underwent total mesorectal excision (TME) were treated with lymph node revealing solution to retrieve lymph nodes, which were submitted to hematoxylin and eosin (HE) examination and immunohistochemical (IHC) staining. RESULTS: The mean number of mesorectal nodes per case was 17.7. The mean size of metastatic, micrometastatic, and isolated tumor cells (ITC) harbored nodes was 5.2 mm, 4.5 mm, and 3.3 mm, respectively. Most of the metastatic (92.1%), micrometastatic and ITC-involved nodes (69.2%) were located along the superior rectal artery (SRA). Posterior-wall located tumor might spread along both sides of the mesorectum simultaneously (P = 0.34), while lateral-wall located tumor spread preferably to ipsolateral side versus contralateral side (P = 0.012). CONCLUSION: Most of the metastases and micrometastases positive lymph nodes were smaller than 5 mm and distributed along the SRA. The patterns of lymph nodes spread were related to the circumferential situation of tumor in the rectal wall. Surgical excision of the rectal cancer should completely remove the whole mesorectum, especially to avoid any damage of the mesorectum on tumor side.