OBJECTIVETo explore protective effects of acupuncture at "Zusanli" (ST 36) on cerebral tissue in rats with sepsis.

METHODSCecal ligation and puncture (CLP) was applied to duplicate the rat model of sepsis. According to random number table, thirty SD rats were divided into a sepsis model group (group A), a sepsis model plus electroacupuncture (EA) group (group B), and a sepsis model plus non-acupoint EA group (group C), ten rats in each one. EA with the same frequency and intensity at "Zusanli" (ST 36) and non-acupoint (0.5 cm laterally to "Zusanli") for 30 min was applied in the group B and group C, respectively. No treatment was given in the goup A. 6 hours after CLP, blood was acquired from abdominal aorta to measure the levels of neuron specific enolase (NSE). Then the rats were sacrificed by abdominal aorta exsanguination to take their cerebral tissue for measuring the levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6).

RESULTSSix hours after CLP, the level of NSE was (3.51 +/- 0.39) ng/mL in group B, which was significantly lower than (7.72 +/- 0.64) ng/mL in group A (P<0.05). The level of NSE was (8.02 +/- 0.72) ng/mL in the group C, which had no statistical significance with group A (P>0.05). The levels of TNF-alpha, IL-6 in cerebral tissue in group B were significantly lower than that of group A and C (all P>0.05).

CONCLUSIONEA at "Zusanli" (ST 36) has certain protective effect on septic rat's brain, which has some relationship with decreasing levels of cerebral tissue proinflammatory cytokine and plasma NSE. EA at non-acupoint has no the same action.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Humans ; Interleukin-6 ; immunology ; Male ; Phosphopyruvate Hydratase ; immunology ; Rats ; Rats, Sprague-Dawley ; Sepsis ; enzymology ; immunology ; therapy ; Tumor Necrosis Factor-alpha ; immunology

Bilirubin ; blood ; Child Development ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Neuropsychological Tests

Aged ; Angioplasty, Balloon, Coronary ; adverse effects ; Drug-Eluting Stents ; adverse effects ; Humans ; Male ; Plaque, Atherosclerotic ; complications ; Rupture ; Thrombosis ; complications

Objective:To construct a DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the anti-HBV siRNA genes, and to express this DNA vaccine in HepG2 cells. Methods:The HBV multi-epitope antigen gene was designed and synthesized before it was fused with enhanced green fluorescent protein(EGFP) gene, and cloned into the multi-clone site(MCS) of the eukaryotic expression vector pVAX1. The expressinig units of hIL-12 and siRNA were cloned into the BspH I and Mlu I site of pVAX1 respectively. Then the recombinant plasmid pVAX1-siHBV-HB-EGFP-hIL12 was transiently transfected HepG2 cells. The expression of HBV compound multi-epitope gene was observed through EGFP report gene. The expression of hIL-12 was analyzed by ELISA and the effects of anti-HBV siRNA was confirmed with rtPCR . Results: The analysis of enzyme digestion and sequencing both demonstrated that the trible-expressing HBV DNA vaccine has been constructed successfully. The green fluorescent image was detected in the transfected cells which could confirm the expression of the multi-epitope antigen gene. The amount of hIL-12 secretion was 1289pg/ml in supernatant at 48h after transfection and 1712pg/ml at 72h after transfection. The mRNA amount of HBV S gene, which was the siRNA target, had been obviously knockdown. Conclusion: The DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the siRNA genes was constructed and transiently expressed in HepG2 cells, and siRNA had shown us a good anti-HBV effect. It laid a foundation of further study on anti-HBV effect of the new DNA vaccine.

OBJECTIVETo investigate the absorption mechanism of genistein self-microemulsifying system in rat intestines.

METHODThe concentrations of phenol red and genistein by in situ perfusion in rats were determined by UV and HPLC, respectively. The effects of drug concentrations, pH, various intestinal segments and P-glycoprotein (P-gp) inhibitor verapamil on the absorption had been studied.

RESULTThe absorption rate constant (Ka) of genistein had no significant difference at concentrations of 0.05-0.5 mg x mL(-1) and pH of 5.4-7.8 in perfusion. It was Ka of jejunum > ileum > duodenum > colon. The absorption of genistein in jejunum had significant difference (P < 0.05) compared with other parts of intestines. Ka was increased obviously when verapamil was coper-fused with genistein (P < 0. 05).

CONCLUSIONThe absorption of genistein self-microemulsifying system is a first order process with passive diffusion mechanism related to P-gp efflux. It can be absorbed at all segments of rat intestine, and the jejunum is the best absorption segment, pH had no special effect on the absorption of genistein self-microemulsifying system in rat intestine.

ATP Binding Cassette Transporter, Sub-Family B ; antagonists & inhibitors ; Animals ; Chromatography, High Pressure Liquid ; Emulsions ; Genistein ; analysis ; metabolism ; pharmacokinetics ; Hydrogen-Ion Concentration ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Organ Specificity ; Rats ; Rats, Wistar ; Temperature ; Verapamil ; pharmacology

OBJECTIVETo investigate the effects of electroacupuncture at "Zusanli" (ST 36) on the volume of hepatic blood flow, water ratio and plasma alanine aminotransferase (ALT) in rats with delayed fluid replacement after hemorrhagic shock and to provide the references for electroacupuncture at Zusanli (ST 36) in treating hemorrhagic shock.

METHODSForty SD rats with hemorrhagic shock induced by bloodletting 40% of whole blood volume were randomly divided into a hemorrhage with no treatment (NT) group, an immediate fluid replacement (IFR) group, an electroacupuncture at Zusanli (ST 36) and delayed fluid resuscitation (EA/DFR) group and a sham electroacupuncture and delayed fluid replacement (SEA/DFR) group, 10 rats in each group. No treatment was performed in NT group. IFR group was treated with fluid replacement at 10 minutes after blood loss, and EA/DFR group was treated with electroacupuncture at "Zusanli" (ST 36) at 10 minutes after blood loss, while non-acupoint was punctured in SEA/DFR group. Two EA groups were received delayed fluid replacement at 3 hours after blood loss. The volume of hepatic blood flow and ALT before blood loss and 3 h and 12 h after blood loss, and water ratio 12 h after blood loss were measured.

RESULTSAfter blood loss, all parameters in IFR group and EA/DFR group were improved significantly in contrast with those in NT group (all P < 0.05). There was no significant difference between SEA/DFR group and NT group. Three hours after blood loss, the hepatic blood flow of IFR group was significant higher than those of NT group, EA/DFR group and SEA/DFR group (all P < 0.05), while the plasma ALT of IFR group was significant lower than those of NT group, EA/DFR group and SEA/DFR group (all P < 0.05), and the plasma ALT of EA/DFR group was lower than those of NT group and SEA/DFR group (both P < 0.05), the hepatic blood flow of EA/DFR group showed no significant difference compared with that of SEA/DFR group (P > 0.05). Twelve hours after blood loss, the plasma ALT and the water ratio of EA/DFR group and IFR group were significant lower than those of NT group and SEA/DFR group (all P < 0.05), and the hepatic blood flow of EA/DFR group and IFR group was significant higher than those of NT group and SEA/DFR group (all P < 0.05), while the plasma ALT of IFR group was significant lower than that of EA/DFR group (P < 0.05), and the hepatic blood flow of IFR group was significant higher than that of EA/DFR group (P < 0.05).

CONCLUSIONElectroacupuncture at "Zusanli" (ST 36) has a protective effects for hepatic ischemic injury in rats with delayed fluid replacement after hemorrhagic shock.

Acupuncture Points ; Animals ; Electroacupuncture ; Humans ; Ischemia ; therapy ; Liver ; blood supply ; Male ; Rats ; Rats, Sprague-Dawley ; Shock, Hemorrhagic ; therapy

Animals ; Cytochrome P-450 CYP1A2 ; Cytochrome P-450 CYP2C19 ; metabolism ; Cytochrome P-450 Enzyme System ; metabolism ; Cytochromes ; metabolism ; Liver Cirrhosis ; enzymology ; Rats

OBJECTIVETo study the anti-endotoxin activity and mechanism of F022 from Radix Isatidis.

METHODThe production of TNF-alpha and IL-6 of murine peritoneal macrophages stimulated by LPS was measured by ELISA. The temperature in rabbits was tested after i.v. administration of LPS. The lethality of BCG-primed mice was induced by LPS.

RESULTIf F022 was added to macrophages culture simultaneously with LPS or 1 h before addition of LPS, production of TNF-alpha and IL-6 by macrophages was remarkably inhibited in vitro. F022 inhibited the fever induced by LPS in rabbits and protected BCG-primed mice from LPS induced lethality if given before administration of LPS.

CONCLUSIONThe anti-endotoxin effect of F022 may inhibit LPS binding to its receptor, and it may be a LPS receptor antagonist.

Animals ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Fever ; chemically induced ; drug therapy ; Interleukin-6 ; secretion ; Isatis ; chemistry ; Lipopolysaccharides ; antagonists & inhibitors ; Macrophages, Peritoneal ; secretion ; Male ; Mice ; Mice, Inbred C57BL ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rabbits ; Tumor Necrosis Factor-alpha ; secretion

AIMTo investigate glutamate-induced [Ca2+]i changes in cultured rat neonatal cortical astrocytes after hypoxia/reoxygenation, H2O2 or high concentration of L-glutamate injury. In the meantime, the effects of Gingko biloba extract (GbE) were examined.

METHODS[Ca2+]i changes in astrocytes were monitored by laser scanning confocal microscopy with the Ca2+ sensitive fluorescent probe fluo-3.

RESULTSAfter astrocytes were impaired by hypoxia/reoxygenation, H2O2 (50 micromol x L(-1)) or L-glutamate (0.25 mmol x L(-)), the exogenous glutamate (27 micromol x L(-1)) could not induce increase of [Ca2+]i, but decrease by (3.3 +/- 1.6)%, (81 +/- 11)% and (81 +/- 7)%, respectively. Pretreatment with GbE (10 mg x L(-1)) could not improve injured astrocytic glutamate response. But after pretreatment with GbE (100 mg x L(-1)), glutamate-induced [Ca2+]i elevation of astrocytes after hypoxia/reoxygenation, H2O2 or high concentration of L-glutamate injury were (135 +/- 98)%, (117 +/- 93)% and (89 +/- 36)%, respectively. Nimodipine (1.6 mg x L(-1)) could also reverse the abnormal response of astrocytes after different injury.

CONCLUSIONHypoxia/reoxygenation, H2O2 and high concentration of L-glutamate impaired astrocytes' response to exogenous L-glutamate, and then bidirectional communication between astrocytes and neurons could not take place. GbE could improve the abnormal responses and maintain the normal function of astroglical network. These effects support that GbE has potential beneficial actions against brain injury.

Animals ; Astrocytes ; cytology ; metabolism ; Calcium ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Cerebral Cortex ; cytology ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ginkgo biloba ; chemistry ; Glutamic Acid ; toxicity ; Hydrogen Peroxide ; toxicity ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Reperfusion Injury

BACKGROUND AND OBJECTIVEEndothelial progenitor cells (EPCs) play an important role in hypoxia-triggered tumor vasculogenesis. However, the homing of exogenous EPCs in tumors is still unclear. In this study, we investigated the recruitment of exogenous EPCs in human lung adenocarcinoma model of nude mice.

METHODSEPCs labeled with green fluorescence protein (GFP) were transplanted into nude mice bearing human lung adenocarcinoma. The growth of tumor was observed. After the mice were killed, GFP-EPCs in different tissues were examined by fluorescence. The tumor tissues were stained for CD133, hypoxia-inducible factor-1alpha (HIF-1α), stromal cell-derived factor-1α (SDF-1α), and vascular endothelial growth factor receptor (KDR). Real-time polymerase chain reaction of CD133, HIF-1α, SDF-1α, and VEGF-1 were also performed.

RESULTSThe growth of tumor in EPC group was significantly faster than that in saline solution group (P <0.05). Under fluorescence microscope, GFP-EPCs were strongly expressed in both tumor and bone marrow. EPCs were recruited to the tumor periphery to participate in tumor vasculogenesis. The expression of CD133, HIF-1α, and SDF-1 mRNA in tumor and bone marrow were significantly higher than that in the liver, spleen, and skin (P<0.05).

CONCLUSIONSExogenous EPCs can be recruited to tumor and accelerate tumor growth. Except tumor, bone marrow can also recruit EPCs.

AC133 Antigen ; Adenocarcinoma ; metabolism ; pathology ; Animals ; Antigens, CD ; genetics ; metabolism ; Bone Marrow ; metabolism ; pathology ; Cell Line, Tumor ; Chemokine CXCL12 ; genetics ; metabolism ; Endothelial Cells ; pathology ; transplantation ; Female ; Glycoproteins ; genetics ; metabolism ; Green Fluorescent Proteins ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; Peptides ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Stem Cell Transplantation ; Stem Cells ; pathology ; Transfection ; Tumor Burden ; Vascular Endothelial Growth Factor A ; genetics ; metabolism