Previous studies have demonstrated that exocytotic release from large dense cored vesicle(LDV)at structurally non-specialized areas within axon terminal of the medullary dorsal horn and it has been speculated that non-synaptic exocytosis from LDV may be a probable mechanism for neuropeptide release. This study provides the evidence that SP containing LDV can release their contents by exocytosis at non-synaptic sites of axon terminal within superficial dorsal horn of medulla oblongata in the rat with ultrastructural immunohistochemistry staining by an unilateral deafferentation. The results of this study confirmed previous hypothesis that SP and perhaps other peptides containing LDV release their contents by exocytosis at non-specialized sites and the significance of such release is discussed.

The ultrastructural localization of substance P (SP) immunoreactivity, especially the morphology, number and distribution of positive large granular vesicles (LGV) in SP axon terminals of the trigeminal subnucleus caudalis of the rat were studied by electron microscopic immunocytochemistry. This study revealed that SP immunoreactivity was mostly located in axon terminals and unmyelinated fibers. SP axon terminals contained both clear round vesicles and LGV. SP immunoreactivity was found in LGV, and on the surface of clear round vesicles and outer membrane of mitochondria. Positive LGV were spherical or oval in shape (60～120nm in diameter). The number of LGv was mostly 2～3 in a SP axon terminal. LGV often apposed to the axolemma or scattered in the centre of terminal. LGV were far from the presynaptic sites of the SP terminals which formed synapses. The number of LGV closed to the terminal membrane was significantly (P

Objective To observe the anti-tumor action of Chang'aishu and its influence on the expression of the Ki67 and PCNA. Methods Forty Balb/c mice with CT-26 colorectal cancer were randomly divided into four groups, including the control group (normal saline, 0.6 mL), the chemotherapy group (Xeloda 205.5 mg/kg), the high dose group (Chang'aishu 51.38 g/kg) and the low dose group (Chang'aishu 17.13 g/kg). Each group of mice was treated with intragastric administration every two days. After 15 days, the anti-tumor rate was calculated and the expression of Ki67 and PCNA were analyzed by immunohistochemistry. Results Compared with the control group, inhibitive rate of the chemotherapy group, the high dose group and the low dose group was 43.35%, 29.48% and 13.30%, respectively. Compared with the control group, the treatment group showed statistical significant difference in Ki67 and PCNA (P<0.05, P<0.01). Conclusion Chang'aishu had inhibition effect on the growth of colorectal cancer, which may be related to down-regulating the expression of Ki67 and PCNA.

PURPOSE: The present study was designed to determine whether rapamycin could inhibit transforming growth factor beta1 (TGF-beta1)-induced fibrogenesis in primary lung fibroblasts, and whether the effect of inhibition would occur through the mammalian target of rapamycin (mTOR) and its downstream p70S6K pathway. MATERIALS AND METHODS: Primary normal human lung fibroblasts were obtained from histological normal lung tissue of 3 patients with primary spontaneous pneumothorax. Growth arrested, synchronized fibroblasts were treated with TGF-beta1 (10 ng/mL) and different concentrations of rapamycin (0.01, 0.1, 1, 10 ng/mL) for 24 h. We assessed m-TOR, p-mTOR, S6K1, p-S6K1 by Western blot analysis, detected type III collagen and fibronectin secreting by ELISA assay, and determined type III collagen and fibronectin mRNA levels by real-time PCR assay. RESULTS: Rapamycin significantly reduced TGF-beta1-induced type III collagen and fibronectin levels, as well as type III collagen and fibronectin mRNA levels. Furthermore, we also found that TGF-beta1-induced mTOR and p70S6K phosphorylation were significantly down-regulated by rapamycin. The mTOR/p70S6K pathway was activated through the TGF-beta1-mediated fibrogenic response in primary human lung fibroblasts. CONCLUSION: These results indicate that rapamycin effectively suppresses TGF-beta1-induced type III collagen and fibronectin levels in primary human lung fibroblasts partly through the mTOR/p70S6K pathway. Rapamycin has a potential value in the treatment of pulmonary fibrosis.
Cells, Cultured ; Collagen Type III/metabolism ; Fibroblasts/*drug effects/metabolism/physiology ; Fibronectins/metabolism ; Humans ; Lung/cytology/drug effects ; Pulmonary Fibrosis/drug therapy ; Signal Transduction/drug effects ; Sirolimus/*pharmacology ; TOR Serine-Threonine Kinases/metabolism/physiology ; Transforming Growth Factor beta1/*antagonists & inhibitors/physiology

OBJECTIVETo investigate the effects of sevoflurane on cytosolic phospholipase A₂ (C-PLA₂) and clara cell secretory protein (CCSP) in lung tissues of rabbits with one-lung ventilation (OLV)-induced lung injuries.

METHODSThirty-six healthy Japanese white rabbits were randomized into sham-operated group, OLV group, and OLV plus sevoflurane group subdivided into 4 subgroups with sevoflurane concentrations of 1%, 2%, 3% and 4%. CCSP and C-PLA₂ mRNA and protein expressions in rabbit lung tissues were detected by Western blotting and real-time PCR, and the content of arachidonic acid (AA) was measured using ELISA. The severities of the lung injury were evaluated according to lung wet/dry weight (W/D) ratio and histological scores.

RESULTSIn the OLV group and OLV+ sevoflurane groups, pulmonary CCSP expressions were significantly lower, while C-PLA₂ expression, lung W/D ratios and lung histological scores were significantly higher than those in the sham-operated group (P<0.05). Compared with OLV group, the OLV+sevoflurane groups showed significantly increased expressions of CCSP and reduced C-PLA₂ expression, lung W/D ratios and histological scores (P<0.05). In the 4 OLV+sevoflurane groups, CCSP expressions underwent no significant changes as sevoflurane concentration increased, but C-PLA₂ expressions, lung W/D ratios and histological scores all decreased gradually as the concentrations of sevoflurane increased (P<0.05).

CONCLUSIONOLV can result in down-regulated CCSP expressions and up-regulated C-PLA₂ expressions in rabbit lung tissues. Sevoflurane can protect against OLV-induced acute lung injury possibly by inhibiting C-PLA₂ expression via up-regulation of CCSP expressions or through other mechanisms resulting in down-regulated expression of C-PLA₂.

Animals ; Female ; Lung ; metabolism ; pathology ; Male ; Methyl Ethers ; pharmacology ; One-Lung Ventilation ; adverse effects ; Phospholipases A2 ; metabolism ; Rabbits ; Uteroglobin ; metabolism ; Ventilator-Induced Lung Injury ; metabolism

OBJECTIVETo explore the protective mechanisms of sevoflurane against acute lung injury (ALI) induced by one-lung ventilation (OLV) in view of cyclooxygenase-2 (COX2) and 5-lipoxygenase (5-LOX) pathways.

METHODEighteen healthy Japanese white rabbits were randomized into sham-operated group (S group), OLV group (O group) and OLV + sevoflurane group (OS group). COX2 and 5-LOX protein and mRNA expressions in the lungs were detected by Western blotting and real-time PCR, respectively. Prostaglandin I2 (PGI2), thromboxane A2 (TXA2) and leukotrienes B2 (LTB2) in the lung tissues were quantified with ELISA. Histological scores and lung wet/dry weight (W/D) ratios were determined for lung injury assessment.

RESULTSCOX2 and 5-LOX protein and mRNA expressions and the contents of LTB2, TXA2 and PGI2 in the lungs, lung W/D ratio and histological scores were significantly higher while PGI2/TXA2 ratio was significantly lower in O group and OS group than in S group (P<0.05). Compared with those in O group, COX2 and 5-LOX expressions, pulmonary contents of LTB2, TXA2 and PGI2, and lung W/D ratio all decreased significantly but PGI2/TXA2 ratio was significantly elevated in OS group (P<0.05).

CONCLUSIONOLV may activate COX2 and 5-LOX pathways to result in increased production of arachidonic acid metabolites. Sevoflurane protects against OLV-induced ALI probably by reducing AA metabolites and regulating PGI2/TXA2 ratio through inhibitions of COX2 and 5-LOX pathways.

Acute Lung Injury ; etiology ; metabolism ; Animals ; Arachidonate 5-Lipoxygenase ; metabolism ; Cyclooxygenase 2 ; metabolism ; Lung ; drug effects ; metabolism ; Methyl Ethers ; adverse effects ; One-Lung Ventilation ; adverse effects ; RNA, Messenger ; genetics ; Rabbits

OBJECTIVE：To establish a method for online detection of antioxidant active components in Glycyrrhiza uroalensis decoction pieces ，and to identify it. METHODS ：The free radical scavenging rate of 1，1-diphenyl-2-trinitrobenzene hydrazine （DPPH）was determined to evaluate the antioxidant activity of G. uralensis decoction pieces. HPLC-UV-DPPH method was used to screen the anti oxidant active components of G. uralensis decoction pieces. HPLC-TOF/MS was used to obtain mass spectrum data and Qualitive Analyst B 06.00 Build 6.0.633.0 software was used to analyze data. Through contrast analysis of UV absorption spectrum，online chromatogram ，mass spectrum information of G. uralensis and the retention time of each compound ，accurate molecular weight ，antioxidant active components were identified by referring to relevant literature. Validation test was also conducted. RESULTS ：DPPH free radical scavenging rate in 8 batches of G. uralensis decoction pieces ranged 55.71％-60.17％. Seven antioxidative active compounds ，including avolomotor ，8-isopentenyl naringin ，yellow lupulin weitone ，isoflavone B ，3′， 4′-dimethoxy3-hydroxy-6-methyl flavone ，glycyrrhizin E and glycyrrhizin H ，could be screened from G. uralensis decoction pieces. After validation ，the peak area of inverted peak generated by online reaction was positively correlated with DPPH free radical scavenging rate. CONCLUSIONS ：Established method is simple and accurate ，and can be used to quickly screen and identify the main antioxidant components of G. uralensis decoction pieces ；the peak area of inverted peak can be used to evaluate the antioxidant active components of G. uralensis decoction pieces.