Animals ; Extremities ; blood supply ; Ischemia ; metabolism ; Male ; Myocardial Reperfusion Injury ; metabolism ; Nitric Oxide ; blood ; Oxidative Stress ; Rats ; Rats, Wistar

Adult ; Asthenopia ; epidemiology ; prevention & control ; Automobile Driving ; Color Perception ; Humans ; Middle Aged

OBJECTIVETo study the effect of Kanglaite injection on cyclooxygenase activity in lung carcinoma A549 cells.

METHODSThe expression of mRNA of COX-1 and COX-2 were measured by RT-PCR. The expression of COX-2 protein was measured by Western-blot.

RESULTKanglaite injection could selectively decreased COX-2 mRNA expression and protein expression while COX-1 mRNA expression was unchanged.

CONCLUSIONKanglaite injection is selectively inhibitory agent of COX-2, and possess inhibitory effects on cyclooxygenase 2.

Antineoplastic Agents, Phytogenic ; administration & dosage ; isolation & purification ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; enzymology ; pathology ; Cell Line, Tumor ; Coix ; chemistry ; Cyclooxygenase 1 ; biosynthesis ; genetics ; Cyclooxygenase 2 ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Gene Expression Regulation, Neoplastic ; Humans ; Injections ; Lung Neoplasms ; enzymology ; pathology ; Plant Oils ; administration & dosage ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Seeds ; chemistry

OBJECTIVETo investigate the relationship of lymphatic micovessel density (LMVD) detected by monoclonal antibody D2-40) with the VEGF-C expression in human breast cancer.

METHODSTissue samples of 102 breast cancers, 25 breast fibroadenomas and 10 normal breasts were collected. Immunohistochemical staining was used to detected the lymphatic micovessels with monoclonal antibody D2-40. The expression of VEGF-C was detect by SP immunohistochemistry, and VEGF-C mRNA by hybridization in situ.

RESULTSIn 102 breast cancers, the positive rate of D2-40 was 76.5% (78/102), higher than that in the breast fibroadenomas. LMVD in the periphery of breast cancer was 30.1 lymphatic microvessels per x 100 field of vision, which was significantly higher than that in the central area of the tumors (P = 0.000). The LMVD in the periphery of the breast cancers was correlated with the number of metastatic lymph nodes (r = 0.964, P < 0.01). The positive rates of VEGF-C protein and mRNA were 55.9% (57/102) and 59.8% (61/102), respectively, significantly higher than that in the breast fiberoadenomas and normal breast tissues (chi2 = 11.653, P = 0.003; chi2 = 10.345, P = 0.006), and were significantly correlated with the status of lymph node metastasis, clinical stage and the expressions of c-erbB-2 and p53 protein (P < 0.05). Both of VEGF-C protein and mRNA were significantly correlated with LMVD detected by D2-40 (P < 0.05), especially with the LMVD in the periphery of breast cancers (P < 0.05).

CONCLUSIONThe monoclonal antibody D240 can be used to detect the lymphatic endothelium in human breast cancer. The lymphatic microvessel density in the periphery of breast cancer is correlated with the lymph node metastasis and expression of VEGF-C. Therefore, VEGF-C may play a significant role in the lymphangiogenesis leading to metastasis of breast cancer.

Adult ; Aged ; Antibodies, Monoclonal ; Antibodies, Monoclonal, Murine-Derived ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Female ; Fibroadenoma ; metabolism ; pathology ; Humans ; Lymph Nodes ; pathology ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Microvessels ; pathology ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Vascular Endothelial Growth Factor C ; genetics ; metabolism ; Young Adult

Acute Disease ; Animals ; Ginkgo biloba ; Glutathione ; analysis ; Lung ; drug effects ; metabolism ; pathology ; Malondialdehyde ; analysis ; Paraquat ; toxicity ; Phytotherapy ; Plant Extracts ; therapeutic use ; Poisoning ; drug therapy ; Rats ; Superoxide Dismutase ; analysis ; Treatment Outcome

The effects of femoral nerve electrostimulation (FNES) on ischemia-reperfused myocardium were examined in the urethane- anesthetized rats to determine whether FNES may provide cardioprotection and to observe the possible mechanism. The area at risk (AR) and infarct area (IA) were determined using Evans blue and nitro-blue tetrazolium staining, respectively. Infarct size (IS) was defined as 100xIA/AR (%). The results are as follows: (1) During 30 min myocardial ischemia and subsequent 120 min reperfusion, the myocardial infarct size occupied (54.96+/-0.82)% of the area at risk. (2) FNES of high frequency (10 V, 100 Hz, 1 ms) significantly reduced myocardial infarct size to (36.94+/-1.34)% (P<0.01), indicating the cardioprotective effect FNES of high frequency on myocardial ischemia-reperfusion, while FNES of low frequency (10 V, 10 Hz, 1 ms) had no effect on myocardial infarct size. (3) Pretreatment with either naloxone (5 mg /kg, i.v), a nonselective opioid receptor antagonist, or glibenclamide (5 mg /kg, i.v), a K(ATP) channel antagonist, completely abolished the cardioprotection of FNES (100 Hz) from myocardial ischemia-reperfusion. It is suggested that FNES of high frequency can protect myocardium from ischemia-reperfusion injury. The possible mechanism is that FNES of high frequency may induce the release of opioids from the central nervous system, and the activation of opioid receptors in the heart results in an opening of myocardial K(ATP) channels which can protect myocardium.
Animals ; Electric Stimulation ; methods ; Femoral Nerve ; physiopathology ; Glyburide ; pharmacology ; Male ; Myocardial Infarction ; pathology ; Myocardial Reperfusion Injury ; pathology ; prevention & control ; Naloxone ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid ; metabolism

OBJECTIVETo investigate the effect of lymphadenectomy adjacent to inferior mesenteric artery root on the prognosis of rectal cancer.

METHODSClinicopathological data of 260 cases with rectal cancer undergone radical operation were analyzed retrospectively. The patients were divided into two groups. Group D(2): the lymph nodes adjacent to mesenteric artery root were not excised (n=188). Group D(3): the lymph nodes adjacent to mesenteric artery root were excised (n=72). Prognosis of two groups was compared during the follow-up period.

RESULTSIn group D(2), the 1-, 3-, 5-year total survival rates (TS) were 97.3%, 87.2% and 77.1%, and tumor-free survival rates (TFS) were 93.1%, 83.0% and 76.8% respectively. In group D(3 ), the 1-, 3-, 5-year total survival rates (TS) were 94.4%, 79.2% and 73.6%, and tumor-free survival rates (TFS) were 86.1%, 76.4% and 71.0% respectively. The differences of TS and TFS between two groups were not significant according to Kaplan-Meier analysis (P>0.05). Multivariate analysis revealed that the excision of lymph nodes adjacent to mesenteric artery root was not statistically correlated with the recurrence, metastasis and survival time after radical operation of rectal cancer.

CONCLUSIONExcision of lymph nodes adjacent to inferior mesenteric artery root has no significant impact on prognosis and it is unnecessary in the radical operation of rectal cancer.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymph Node Excision ; methods ; mortality ; Lymph Nodes ; surgery ; Lymphatic Metastasis ; Male ; Mesenteric Artery, Inferior ; surgery ; Middle Aged ; Prognosis ; Rectal Neoplasms ; mortality ; pathology ; surgery ; Survival Rate ; Treatment Outcome

OBJECTIVETo observe the expression and effect of human calcitonin gene-related peptide (hCGRP) gene mediated by recombinant adeno-associated virus (rAAV) in primary cultured corporal cavernosum smooth muscle cells of the rat and explore the possibility of using CGRP gene for gene therapy in erectile dysfunction.

METHODSThe primary cultured corporal cavernosum smooth muscle cells of the rat were randomly divided into 4 groups and infected with recombinant virus VssHGCMV-hCGRP, VssHGCMV, VssC-MV-GFP and the untreated, respectively. CGRP-like immunoreactivity was measured by protein dot blot assay in the 24 h-culture medium, and intracellular cAMP and cGMP levels in the cultured cells were also determined using radioimmunoassay to ascertain bioactivity of transduced CGRP.

RESULTSThe exogenous gene was transferred into primary corporal cavernosum smooth muscle cells by VssHGCMV-hCGRP infection and efficiently expressed. Compared with the control group, intracellular cAMP level in the cell infected by VssHGCMV-hCGRP was significantly increased (48.7 +/- 1.1 nmol/L vs 7.8 +/- 1.4 nmol/L, P < 0.01), whereas cGMP level remained unchanged in two groups, and CGRP-like immunoreactivity was also detected in the culture medium infected by VssHGCMV- hCGRP.

CONCLUSIONThe system of secretory expressing bioactive peptide rAAV mediated gene transfer may be used to express efficiently exogenous gene in corporal cavernosum smooth muscle cells and affect cAMP level in the corporal cavernosum smooth muscle cells of the rat.

Animals ; Calcitonin Gene-Related Peptide ; biosynthesis ; genetics ; Cells, Cultured ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Dependovirus ; genetics ; Male ; Muscle, Smooth ; cytology ; metabolism ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley ; Recombination, Genetic ; Transfection

OBJECTIVETo determine whether recombinant adeno-associated virus-mediated overexpression of hCGRP in the corpus cavernosum can affect the continuous production of hCGRP in the penile tissue and enhance erectile responses in STZ-induced diabetic rats.

METHODSDiabetes mellitus was induced by a single intraperitoneal injection of 60 mg/kg streptozotocin in male SD rats. VssHGCMV-hCGRP, VssCMV-GFP and rAAV solution were injected into the corporal cavernosum of STZ-induced diabetic rats, respectively. The corporal tissue was obtained from groups of 8 rats on day 5 post-injection, and the expressions of CGRP and GFP in cavernosal tissue were detected using immunohistochemistry and laser scanning confocal microscopy, respectively. Cavemosal tissue cAMP and cGMP levels were measured using radioimmunoassay. On day 5 post-injection, intracavernous pressure induced by electrostimulation of penile dorsal nerves was measured and recorded with a biological signal processing system in each group rat.

RESULTSrAAV transduction efficiency of GFP reporter gene was measured by laser scanning confocal microscopy and was observed in the penile tissue, especially in the corporal cavernosum and the vessel 5 days after transfection with VssCMV-GFP. Immunohistochemistry showed that the CGRP increased in the corporal cavernosum. In addition, both cAMP and cGMP levels in the corpora cavernosa transfected with VssHGCMV-hCGRP were significantly increased, compared with controls [(48.4 +/- 6.5) nmol/L and (21.2 +/- 13.6) nmol/L vs (16.7 +/- 2.5) nmol/L and (0.42 +/- 0.12) nmol/L, respectively]. More importantly, 5 days after administration of VssHGCMV-hCGRP,a significant increase was observed in the erectile response to penile dorsal nerve stimulation in the diabetic rat [(60.5 +/- 4.5) mm Hg vs (22. 3 +/- 1.3) mm Hg].

CONCLUSIONSThis results demonstrate that rAAV-mediated transfer of the CGRP gene can increase production of endogenous CGRP, cAMP and cGMP in corpora cavernosa of STZ-induced diabetic rats. Moreover, overexpression of CGRP enhances ICP and the erectile response to penile dorsal nerve stimulation in the diabetic rat.

Adenoviridae ; genetics ; Animals ; Calcitonin Gene-Related Peptide ; biosynthesis ; genetics ; Diabetes Mellitus, Experimental ; physiopathology ; Green Fluorescent Proteins ; biosynthesis ; Humans ; Male ; Penile Erection ; physiology ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transfection

The purpose of this study was to investigate the repair of the osteoarthritis(OA)-induced cartilage injury by transfecting the new TGF-β3 fusion protein (LAP-MMP-mTGF-β3) with targeted therapy function into the bone marrow-derived mesenchymal stem cells (MSCs) in rats. The recombinant of pIRES-EGFP-MMP was constructed by combination of DNA encoding MMP enzyme cutting site and eukaryotic expression vector pIRES-EGFP. LAP and mTGF-β3 fragments were obtained from rat embryos by RT-PCR and inserted into the upstream and downstream of MMP from pIRES-EGFP-MMP respectively, so as to construct the recombinant plasmid of pIRES-EGFP-LAP-MMP-mTGF-β3. pIRES-EGFP-LAP-MMP-mTGF-β3 was transfected into rat MSCs. The genetically modified MSCs were cultured in medium with MMP-1 or not. The transfected MSCs were transplanted in the rat OA models. The OA animal models were surgically induced by anterior cruciate ligament transaction (ACLT). The pathological changes were observed under a microscope by HE staining, Alcian blue, Safranin-fast Green and graded by Mankin's scale. pIRES-EGFP-LAP-MMP-mTGF-β3 was successfully constructed by means of enzyme cutting and sequencing, and the mTGF-β3 fusion protein (39 kD) was certified by Western blotting. Those genetically modified MSCs could differentiate into chondrocytes induced by MMP and secrete the relevant-matrix. The transfected MSCs could promote chondrogenesis and matrix production in rat OA models in vivo. It was concluded that a new fusion protein LAP-MMP-mTGF-β3 was constructed successfully by gene engineering, and could be used to repair the OA-induced cartilage injury.