OBJECTIVEImmunosuppressant tacrolimus (FK506) has shown neuroprotective effects on hypoxic-ischemic brain damage (HIBD) in the adult animal model. This study investigated whether FK506 has a protection against HIBD in neonatal rats by examining growthjassociated protein-43 (GAP-43) expression in the hippocampus.

METHODSNinety-six seven-day-old Sprague-Dawley rats were randomly divided into three groups: sham-operation, HIBD and FK506 intervention group. HIBD was induced in the later two groups. The FK506 intervention group was intraperitoneally injected with FK506 immediately after HIBD, at a dosage of 1 mg/kg daily, for three days. The HIBD group was injected with normal saline. Immunohistochemical technical was applied to examine GAP-43 expression in the hippocampus 24 and 72 hrs and 7 and 14 days after HIBD.

RESULTSCompared with the HIBD group, hematoxylin-eosin staining showed attenuated neuronal necrosis in the FK506 intervention group. In the HIBD group, the expression of GAP-43 increased significantly 72 hrs, and 7 and 14 days after HIBD compared with that in the sham-operation group. The GAP-43 expression in the FK506 intervention group was significantly higher than that in the HIBD group 72 hrs and 7 days after HIBD.

CONCLUSIONSFK506 might have neuroprotective effects against HIBD in neonatal rats.

Animals ; Animals, Newborn ; GAP-43 Protein ; analysis ; Hippocampus ; chemistry ; drug effects ; Hypoxia-Ischemia, Brain ; drug therapy ; metabolism ; pathology ; Immunosuppressive Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Tacrolimus ; pharmacology

OBJECTIVETo compare the efficacy and safety of midazolam combined with fentanyl and propofol combined with fentanyl as conscious sedation in oocyte retrieval of in vitro fertilization and embryo transplantation (IVF-ET).

METHODSEighty patients receiving IVE-ET were randomly divided into midazolam combined with fentanyl group (midazolam group) and propofol combined with fentanyl group (propofol group). Antalgic effects, circulation status (blood pressure, heart rate), respiration status (rate, oxygen saturation and respiration depression) during operation, nausea and vomiting, and amnestic effects after operation were compared.

RESULTNo differences of antalgic effects and circulation status between two groups were observed. Percentages of respiration depression,vomiting and amnesia of midazolam group were 5.0 %, 10.0 % and 25%, respectively, and those of propofol group were 25%, 27.5% and 7.5%, respectively, which had statistical significance.

CONCLUSIONAs conscious sedation, midazolam combined with fentanyl is better than propofol combined with fentanyl in oocyte retrieval of IVF-ET.

Adult ; Anesthetics, Combined ; administration & dosage ; Anesthetics, Intravenous ; administration & dosage ; Female ; Fentanyl ; administration & dosage ; Fertilization in Vitro ; Humans ; Midazolam ; administration & dosage ; Oocyte Retrieval ; methods ; Propofol ; administration & dosage

Objective To evaluate the efficacy and adverse effects of gefitinib in the treatment of fe- male patients with advanced adenocarcinoma of lung who had failed to previous chemotherapy.Methods These patients received 250mg of gefitinib orally,once daily until disease progression or development of intol- erable toxic reaction.They were evaluated one month after treatment and every other month thereafter.Results Among the 27 evaluable patients,there were 1 CR(3.7%),11 PR(40.8%),10 SD(37.0%)and 5 PD(18.5%). The overall response rate was 44.5%(95% CI 29%～68%);and 22 patients(81.5%)gained profit(CR+PR+ SD)from the clinical therapy(95% CI 62%～94%);the mean TTP was 7.2 months.Symptomatic improvement rate was 80.0%.The main adverse effects were mild rash and diarrhea.Conclusion gefitinib has significant efficacy in the treatment of female patients with advanced tung cancer who had failed to previous chemother- apy.Adverse effects are mild.gefitinib is a suitable therapy for these patients.

OBJECTIVE: In order to understand which kind of function genes play an important role for estimating wound age, the variation of difference genes' mRNA expression were compared after injury. METHODS: The mRNA expression levels of seven candidate genes (ICAM-1, NF-κB, MX2, MT1, MT2, sTnI, and Cox6c) were analyzed in contused rat skeletal muscle at different time points using real-time fluorescent quantitative PCR (RT-qPCR). The raw Ct values were normalized relative to that of RPL32 mRNA, and converted to standard Ct values. At each time point after injury, the standard deviations (SD) of the standard Ct values were calculated by SPSS. RESULTS: The expression trends of the seven genes were all found to be related to wound age, but there were lower variation coefficients and greater reliability of s TnI and Cox6c when compared with other genes. CONCLUSION The genes encoding structural proteins or proteins that perform basic functions can be suitable for wound age estimation.
Animals ; Contusions/genetics* ; Forensic Pathology ; Gene Expression Profiling ; Intercellular Adhesion Molecule-1 ; Muscle, Skeletal/metabolism* ; NF-kappa B ; Proteins ; RNA, Messenger/metabolism* ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Regression Analysis ; Reproducibility of Results ; Time Factors ; Wound Healing/genetics*

OBJECTIVETo investigate the mechanisms of memory impairment induced by pentylenetetrazole (PTZ)-kindled epilepsy in rats and the effects of endogenous histamine.

METHODSRats were injected i. p with a subconvulsive dose of PTZ every 48 h until fully kindled. Memory was tested by shuttle box with passive avoidance. Brain histamine was measured spectrofluorometrically. Neurons of hippocampus were investigated with HE stain.

RESULTPTZ-kindled epilepsy caused memory impairment in rats, i .e. latency of passive avoidance was shortened in shuttle box. Pretreatment of histidine, the precursor of histamine, showed an ameliorating effect on memory impairment induced by epilepsy. Decreased histamine contents in the hippocampus, thalamus and hypothalamus were observed after fully kindled in rat. In addition, intact neurons of the CA1 and CA3 regions in hippocampus decreased to 72.7 % and 78.9 % compared with those in control group.

CONCLUSIONPTZ-kindled epilepsy causes memory impairment, and it might be due to a decrease of brain histamine and loss of hippocampal neurons induced by epilepsy.

Animals ; Epilepsy ; chemically induced ; complications ; metabolism ; Hippocampus ; metabolism ; pathology ; Histamine ; metabolism ; Kindling, Neurologic ; metabolism ; Male ; Memory Disorders ; etiology ; metabolism ; Neurons ; metabolism ; pathology ; Pentylenetetrazole ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Fluorescence

OBJECTIVETo explore the role of superoxide dismutase (SOD) coenzyme in regulation of Fas/FasL signal transduction and apoptosis of alveolar macrophages in pneumoconiosis patients.

METHODS50 male and Han nationality cases, including the dust exposed workers, Phase I, II pneumoconiosis patients confirmed by local pneumoconiosis diagnosis group according to GBZ 70 - 2002 diagnosis standard, who underwent whole lung lavage treatment were chosen as subjects. Their alveolar macrophages (AMs) were collected and purified. The cells were divided into three groups: the untreated group, the Fas/FasL group and the SOD group. 5 x 10(6) purified AMs were added into incubating bottles containing DMEM for 2 hours for purifying, added with SOD coenzyme and other block reagents separately, and then incubated for 24 hours in CO(2) incubation. The cells were harvested and lysed. Western-blot were used to analyze the expressions of Fas, FasL, Caspases-8 and Caspases-3. Software of Quantity One 7.0 was used to analyze the relative quantity of Fas, FasL, Caspase-8 and Caspase-3. TUNEL and DNA fragment analysis were used to analyze AMs apoptosis.

RESULTSThe apoptosis index in SOD coenzyme group (9.50 +/- 2.76)% and Fas/FasL group (14.01 +/- 2.56)% was significantly lower than that of in untreated group (19.18 +/- 2.83)% (P < 0.05). The catachrestic DNA ladder appeared in untreated group, was looming in Fas/FasL group, and was not found in the SOD group. The expressions of Fas, FasL, Caspase-8 and Caspase-3 of phase I and II in SOD group were higher than in the other two groups (P < 0.05). There was no significant difference in the expression of Fas, FasL, Caspase-8 and Caspase-3 among different phases of pneumoconiosis (P > 0.05).

CONCLUSIONSOD coenzyme can effectively regulate Fas/FasL signal transduction and block AMs apoptosis.

Adult ; Apoptosis ; Cells, Cultured ; Fas Ligand Protein ; metabolism ; Humans ; Macrophages, Alveolar ; metabolism ; pathology ; Male ; Middle Aged ; Pneumoconiosis ; metabolism ; pathology ; Signal Transduction ; Superoxide Dismutase ; metabolism ; fas Receptor ; metabolism

OBJECTIVE: To investigate the relationship between the expression of secreted frizzled-related protein 5 (SFRP5) mRNA and the time interval after skeletal muscle injury in rats by real-time PCR. METHODS: A total of ninety SD rats were randomly divided into the contusion groups at different times including 4h, 8h, 12h, 16h, 20h, 24h, 28h, 32h, 36h, 40h, 44h, 48h after contusion, incision groups at different times including 4h and 8h after incision and the control group. The samples were taken from the contused zone at different time points. The total RNA was isolated from the samples and reversely transcribed to analyze the expression levels of SFRP5 mRNA. RESULTS: Compared to the control group, the expression of SFRP5 mRNA in contusion groups were down-regulated within 48 h after contusion and reached the lowest level at 20 h, and the expression of SFRP5 mRNA gradually increased from 20 h to 48 h after contusion. The expression of SFRP5 mRNA in the incised groups were significantly lower than that of the contusion groups at 4 h after injury. At the time of 8 h, the expression levels between the contusion and incision groups showed no statistically significant difference. CONCLUSION It is suggested that SFRP5 mRNA analysis may show regular expression and can be a marker for estimation of skeletal muscle injury age.
Animals ; Biomarkers/metabolism* ; Contusions/metabolism* ; Membrane Proteins/metabolism* ; Muscle, Skeletal/metabolism* ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction

BACKGROUNDValproic acid (VPA) improves early survival and organ function in a highly lethal poly-trauma and hemorrhagic shock model or other severe insults. We assessed whether VPA could improve organ function in a rat model of septic shock and illustrated the possible mechanisms.

METHODSForty Sprague-Dawley rats were randomly assigned to four groups (n = 10): control group, VPA group, LPS group, and LPS + VPA group. Lipopolysaccharide (LPS) (10 mg/kg) was injected intravenously to replicate the experimental model of septic shock. Rats were treated with VPA (300 mg/kg, i.v.) or saline. Six hours after LPS injection, blood was sampled for gas analysis, measurement of serum alanine aminotransferase, aspartate aminotransferase, urine nitrogen, creatinine and tumor necrosis factor-alpha. Lung, liver and kidney were collected for histopathological assessment. In addition, myeloperoxidase activity and tumor necrosis factor-a in pulmonary tissue were measured. Acetylation of histone H3 in lung was also evaluated by Western blotting.

RESULTSLPS resulted in a significant decrease in PaO2, which was increased by VPA administration followed LPS injection. In addition, LPS also induced an increase in the serum levels of alanine aminotransferase, aspartate aminotransferase, urine nitrogen, creatinine, and tumor necrosis factor-alpha. However, these increases were attenuated in the LPS + VPA group. The lungs, liver and kidneys from the LPS group were significantly damaged compared with the control group. However, the damage was attenuated in the LPS + VPA group. Myeloperoxidase activity and tumor necrosis factor-alpha levels in pulmonary tissue increased significantly in the LPS group compared with the control group. These increases were significantly inhibited in the LPS + VPA group. Acetylation of histone H3 in lung tissue in the LPS group was inhibited compared with the control. However, the level of acetylation of histone H3 in the LPS + VPA group was markedly elevated in contrast to the LPS group.

CONCLUSIONSTreatment with VPA can attenuate multiple organ damage caused by LPS induced septic shock. Our data also suggest that the beneficial effects are in part due to the decrease in inflammatory cytokines and restoration of normal acetylation homeostasis.

Acute Kidney Injury ; drug therapy ; metabolism ; Animals ; Blotting, Western ; Chemical and Drug Induced Liver Injury ; drug therapy ; metabolism ; Lung Injury ; drug therapy ; metabolism ; Male ; Multiple Organ Failure ; drug therapy ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Shock, Septic ; drug therapy ; metabolism ; Valproic Acid ; therapeutic use

BACKGROUNDErythropoietin elicits protective effects in lung tissue injury induced by ischaemic reperfusion and hyperoxia. We investigated the protective roles of erythropoietin in pulmonary inflammation and lung injury during acute endotoxaemia.

METHODSA total of 32 male Sprague-Dawley rats were randomly assigned to four groups: saline group, erythropoietin + saline group, saline + lipopolysaccharide group and erythropoietin + lipopolysaccharide group. Rats were treated with erythropoietin (3000 U/kg, i.p.) or saline, 30 minutes prior to lipopolysaccharide administration (6 mg/kg, i.v.). Four hours after lipopolysaccharide injection, samples of pulmonary tissue were collected. Optical microscopy was performed to examine pathological changes in lungs. Wet/dry (W/D) ratios, myeloperoxidase activity, malondialdehyde concentrations and tumour necrosis factor-alpha (TNF-alpha) as well as interleukin 1 beta (IL-1beta) levels in lungs were measured. The pulmonary expression of nuclear factor kappaB (NF-kappaB) p65 was evaluated by Western blotting. Differences between the different groups were analysed by one-way analysis of variance (ANOVA).

RESULTSThe lung tissues from the saline + lipopolysaccharide group were significantly damaged, which were less pronounced in the erythropoietin + lipopolysaccharide group. The W/D ratio increased significantly in the saline + lipopolysaccharide group (5.75 +/- 0.22) as compared with the saline group (3.85 +/- 0.20) (P < 0.01), which was significantly reduced in the erythropoietin + lipopolysaccharide group (4.50 +/- 0.35) (P < 0.01). Myeloperoxidase activity and malondialdehyde levels increased significantly in the saline + lipopolysaccharide group compared with the saline group, which was reduced in the erythropoietin + lipopolysaccharide group. The TNF-alpha level of pulmonary tissue increased significantly in the saline + lipopolysaccharide group ((9.80 +/- 0.82) pg/mg protein) compared with the saline group ((4.20 +/- 0.42) pg/mg protein, P < 0.01). However, the increase of TNF-alpha level of pulmonary tissue was significantly reduced in the erythropoietin + lipopolysaccharide group ((6.50 +/- 0.66) pg/mg protein, P < 0.01). Similarly, pulmonary IL-1beta levels were elevated markedly in the saline + lipopolysaccharide group in contrast to the saline group, whereas the elevation was much less in the erythropoietin + lipopolysaccharide group. The nuclear localization of p65 increased markedly in the saline + lipopolysaccharide group and this enhancement of nuclear p65 expression was much less in the erythropoietin + lipopolysaccharide group.

CONCLUSIONErythropoietin attenuates pulmonary inflammation and suppresses TNF-alpha and IL-1beta overproduction during acute endotoxaemia, which is partially mediated by inhibition of NF-kappaB.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Blotting, Western ; Endotoxemia ; immunology ; metabolism ; pathology ; Erythropoietin ; pharmacology ; Interleukin-1beta ; metabolism ; Lung ; drug effects ; immunology ; metabolism ; pathology ; Lung Injury ; chemically induced ; immunology ; Male ; Malondialdehyde ; metabolism ; NF-kappa B ; metabolism ; Organ Size ; Peroxidase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha

OBJECTIVEIsolation and expansion tumor spheres from colorectal cancer cell line Colo205 cultured in serum-free medium(SFM) supplemented with human recombinant EGF and bFGF.

METHODSColo205 cells were cultivated in SFM,while cells cultivated in serum-supplemented medium(SSM) served as the control. Cells morphology were observed by optical microscope, and expression of intestinal stem cells marker Musashi-1 was detected by immunocytochemical. To induce cell differentiation, tumour spheres were cultivated without EGF and bFGF in the presence of 10% serum. Then we analysed expressions of stem cell surface markers CD133 and CD44 among undifferentiated cell, post-differentiated cells and routine Colo205 cells under serum-supplemented culture condition by flow cytometry. At last we compared cell cycle and spectral karyotype between two groups.

RESULTSIn SFM consisting of EGF and bFGF, a minority of Colo205 cells could survive, proliferate and form the suspended tumor spheres. We detected high Musashi-1 expression in these cells. Compared with the SSM group and the post-differentiation SFM group, the expressions of CD133 and CD44 were significantly increased in the undifferentiated SFM group (P<0.05). There was no statistical difference in the expression of CD133 and CD44 between the post-differentiation SFM group and the SSM group (P>0.05). Cell cycle analysis indicated that tumor spheres were of a high proliferation state.We could not find any noticeable difference in the number of chromatosomes between the SFM group and the SSM group.

CONCLUSIONTumor spheres in which enriched cancer stem cells can be generated under serum-free culture condition with EGF and bFGF.

AC133 Antigen ; Antigens, CD ; metabolism ; Cell Culture Techniques ; methods ; Cell Line, Tumor ; Cell Proliferation ; Culture Media, Serum-Free ; Glycoproteins ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Neoplastic Stem Cells ; cytology ; metabolism ; Nerve Tissue Proteins ; metabolism ; Peptides ; metabolism ; RNA-Binding Proteins ; metabolism ; Spheroids, Cellular ; cytology