Objective To study the effects of deep hypothermia and L-arginine(NO precursor) pretreatment on cerebral protection after circulatory arrest and resuscitation in a canine model. Methods Ten healthy adult dogs of both sexes, weighing 14.45?2.3kg, were randomly divided into two groups(n=5): sham treatment group and L-arginine pretreatment group. Extracorporeal circulation was established routinely,circulatory arrest was induced when the nasopharyngeal temperature was reduced to 18℃, then the animals were resuscitated 90min later with extracorporeal circulation. SjvO2 was measured at 30min before circulatory arrest, 0min, 45min, 90min after circulatory arrest and at 60minutes after resuscitation. The ultrastructure changes in cerabral cortex were observed with transmission electron microscope (HITACHI 7500). The brain water content was also determined after the dogs were sacrificed. Results SjvO2 had decreased obviously after circulatory arrest, attaining the lowest level in CA 90min.After resuscitation it increased again in both groups. The levels of SjvO2 in dogs of L-arginine pretreatment group were markedly higher than that of control group at 45min and 90min after circulatory arrest (P

Objective To investigate mechanism of netrin-1 regulating invasion of extra villous trophoblasts. Methods RT-PCR was used to detect six receptors expression including UNC5A, UNC5B, UNC5C, UNC5D, DCC and neogenin in extra villous trophoblast cell line TEV-1. The TEV-1 cells were cultured and devided into seven groups according to the concentration of netrin-1 adding into the medium, which include 10 μg/L, 50 μg/L, 100 μg/L, 500 μg/L, 1000 μg/L, 5000 μg/L and the control(the concentration of netrin-1 was 0 μg/L) groups. The proliferation and invasion of TEV-1 induced by netrin-1 were determined by CCK-8 assay and transwell invasion assay respectively. Results (1) Only neogenin and UNC5B were found to be expressed on TEV-1 by RT-PCR method. (2) In CCK-8 proliferation assay, after 72 hours culture, the proliferation of TEV-1 were 1.55 ±0.29 in 10 μg/L, 1.72±0. 31 in 50 μg/L, 2.15 ±0.35 in 100 μg/L, 1.42 ±0. 25 in 500 μg/L, 1.50±0. 27 in 1000 μg/L, and 1.38±0.23 in 5000 μg/L group, which were all higher than 1.00 ± 0.16 in control group significantly ( P<0.05 ). (3) In matfigel invasion assay, after 6 hours culture, the number of the trans-membrane cells in various netrin-1 group, including 41 ±4 in 10 μg/L, 47 ±5 in 50 μg/L, 55±6 in 100 μg/L, 44 3=5 in 500 μg/L, 43±5 in 1000 μg/L and 42 ±5 in 5000 μg/L group, were all higher than 30 ±4 in control group with statistical significance( P<0.05 ). (4) The fold changes of neogenin were 1.50 ± 0.16 in 10 μg/L, 1.83 ± 0.19 in 50 μg/L, 2.24 ± 0.25 in 100 μg/L, 2.12 ±0.24 in 500 μg/L, 2.12±0.23 in 1000 μg/L and 2.13 ± O. 23 in 5000 μg/L group, which were all higher than 1.00 ±0.11 in control group significantly( P <0.05 ). There were significant difference between group 10 μg/L and 50 μg/L, group 50 μg/L and 100 μg/L (P < 0.05). There were no significant difference between group 100 μg/L and 500 μg/L, group 1000 μg/L and 5000 μg/L (P>0.05). (5) The fold changes of UNC5 B 1.09 ± 0.11 in 10 μg/L, 1.47±0.14 in 50 μg/ L, 1.61 ±0.16 in 100 μg/L, 1.85±0.19 in 500 μg/L, 2.21±0.21 in 1000 μg/L and 2.42±0.23 in 5000 μg/L group, were all higher significantly when compared with 1.00 ± 0.07 in control group (P<0.05 ). There were significant difference between all groups ( P<0.05). Conclusion Netfin-1 can promote the potential of proliferation and invasion of extravillous trophohlasts in vitro through its receptors including neogenin and UNC5 B.

OBJECTIVE: To investigate the effects of compound preparation of Cordyceps sinensis and Tripterygium hypoglaucum (CSTHC) on survival time of grafted pigskin after allogeneic transplantation and its mechanism. METHODS: The pigskin was treated with CSTHC solution before allogeneic transplantation, and CSTHC ointment was applied for external use on the grafted pigskin after skin transplantation. Cyclosporine A (CsA) and normal saline were served as control. The survival time, the appearance and the histomorphological changes of the grafted pigskin were observed. The histomorphological changes of testicles in pigs were also examined. The CD4 and CD8 expressions in the grafted pigskins were measured by immunohistochemical method. The white blood cell count in peripheral blood and the liver and renal functions were also examined. RESULTS: The survival time of the grafted pigskin in the CSTHC-treated group was (28.50+/-3.26)d, which was much longer as compared with (10.60+/-1.52)d in the untreated group (P<0.01). The survival time of the grafted pigskin in the CsA-treated group was (28.33+/-3.50)d, and there was no remarkable difference in the survival time of the grafted pigskin between the CsA-treated group and the CSTHC-treated group. The expressions of CD4 and CD8 were lower in the CSTHC-treated group than those in the untreated group on the 7th and 14th day after skin graft (P<0.05), while there was no significant difference in the indices between the CSTHC-treated group and the CsA-treated group. The WBC count was higher in the untreated group than that in the CSTHC-treated group or CsA-treated group on the 7th day after skin graft (P<0.05). CONCLUSION: CSTHC can prolong the survival time of allogeneic grafted pigskin. Its mechanism of inhibiting the immunological rejection may relate to decreasing the expressions of CD4(+) and CD8(+) in the grafted pigskin and reducing the local inflammatory reaction.

Objectives To compare prospective memory (PM) deficits with retrospective memory (RM) deficits and to explore the correlation between PM and RM in chronic schizophrenia. Methods Fifty chronic schizophrenia pa?tients and fifty healthy controls were recruited. The PM performance [event-based PM (EBPM) and time-based PM (TB?PM)] were evaluated by the Chinese version of the Cambridge Prospective Memory Test (C-CAMPROMPT); working memory (WM) was evaluated by the digital span subtest (DS);immediate auditory logical memory (IALM), delayed audito?ry logical memory (DALM), immediate visual reproduction memory (IVRM) and delayed visual reproduction memory (DVRM) were evaluated by the logical memory and visual reproduction subtest. The score of each test was transformed to comparable standard score. Results Patients performed significantly worse on EBPM [(7.9 ± 3.4) vs. (13.7 ± 2.9)], TBPM [(6.9±3.6) vs. (13.0±3.2)], DS [sequence:(5.8±2.0) vs. (7.5±2.2);backward:(6.5±1.9) vs. (8.2±2.8)], IALM [(8.3±3.1) vs. (11.9 ± 2.5)], DALM [(7.4 ± 3.7) vs. (11.8 ± 2.6)], IVRM [(8.0 ± 2.7) vs. (11.2 ± 3.8)], and DVRM [(7.7 ± 3.5) vs. (10.8 ± 2.7)] scores than controls (P<0.05). The extent of deficits of EBPM and TBPM were greater than those of DS (sequence and backward), IALM, DALM, IVRM and DVRM (P<0.05), but not DALM (P>0.05). The performance of PM in chronic schizophrenia was significantly related to DS (sequence and backward), IALM, DALM and DVRM (P<0.05), but not IVRM (P=0.155). Conclusion:There are greater prospective memory deficits than retrospective memory deficits in chron?ic schizophrenia and the prospective memory deficits are correlated with the retrospective memory deficits in chronic schizophrenia.

Objective To explore the reliability and validity of the computerized Chinese version of Cambridge Prospective Memory Test (C-CAMPROMPT) for assessment of prospective memory (PM) in chronic schizophrenia patients. Methods 50 patients and 50 healthy controls formed the study sample. PM performance was measured with computerized C-CAMPROMPT, while the Wechsler Adult Memory Scale-Forth Edition (WMS-IV), Wisconsin Card Sorting Test (WCST) and Category Fluency Test (CFT) were administered to assess logical memory (LM), visual representation (VR), executive function and processing speed. Results The test- retest reliability (0.981, P<0.001), split half reliability (0.627, P<0.001) and internal consistency reliability (0.742) of C-CAMPROMPT were satisfied. The scores of C-CAMPROMPT and its subtest in schizophrenia were lower than that in healthy control (P<0.001). The performance of PM in patients with schizophrenia closely related to the scores of LM, VR, WCST-CC and CFT (r=0.34~0.89, P<0.05). The sensitivity (86%) and specificity (92%) of the scale were satisfied. Factor analysis extracted 2 factors. Conclusion The computerized C-CAMPROMPT shows a good reliability and validity for assessment of PM function in chronic schizophrenia, and is a sensitive, adaptable, stable instrument.

Objective To develop Prospective Memory Test and explore its reliability and validity in normal and schizophrenic population. Methods According to the structure of prospective memory, Prospective Memory Test was developed based on the previous researches. 40 inpatients with schizophrenia and 30 healthy subjects were assessed with Prospective Memory Test and Wechsler Memory Scale-Fourth Edition, Chinese (WMS-Ⅳ). Results Factor analysis extracted 2 main factors, named time-based prospective memory (TBPM) and event-based prospective memory (EBPM), including 10 items. The average score of Prospective Memory Test was lower in the patients than in the healthy controls by 1~2 standard deviations (P<0.001). Discriminant analysis showed that the specificity, sensitivity, and diagnostic consistency were 93.3%, 72.5% and 81.4%, respectively. Prospective Memory Test scores and two subtest scores positively correlated with total score of WMS-Ⅳ (r=0.44~0.53, P<0.001). The Cronbach's α among all the items was 0.76, and was 0.68 in the TBPM and 0.59 in the EBPM. Split-half reliability of the scale was 0.65 (P<0.001). Conclusion The reliability and validity of Prospective Memory Test are acceptable for schizophrenics.

OBJECTIVETo evaluate the effect of L-arginine pretreatment on cerebral metabolism for cerebral protection during deep hypothermic circulatory arrest (DHCA).

METHODSFifteen healthy adult canines of either sex weighing 14.7-/+2.4 kg were randomly divided into 3 groups (n=5), namely the normal saline group, L-arginine pretreatment group (pretreated with 100 mg/kg L-arginine 60 min before DHCA), and L-arginine combined with 7- nitroindazole treatment group (pretreated with 100 mg/kg L-arginine and 25 mg/kg7-Ni 60 min before DHCA). For all the canines, extracorporeal circulation was established routinely to allow nasopharyngeal temperature reduction to 18 degrees celsius;, at which point DHCA commenced followed 90 min later by reperfusion. At 30 min before DHCA and 0, 45 and 90 min after DHCA as well as at 60 min after reperfusion initiation, blood samples were collected from the jugular vein and arterial to measure the plasma lactic acid, and the cerebral cortex of the parietal lobe was sampled determine the activity of Na(+)-K(+)ATPase. The cerebral water content was also determined after execution of the canines.

RESULTSIn the two pretreatment groups, the level of lactic acid production (shown by the difference in lactic acid levels between the jugular venous and arterial blood) and the cerebral ATP consumption were similar (P>0.05), but both were significantly lower than those of the control group (P<0.05). The cerebral water content was the lowest in the combined treatment group, followed by exclusive L-arginine group, and the highest in the control group (P<0.05), with significant difference between the 3 groups (P<0.05).

CONCLUSIONL-arginine pretreatment can lower cerebral metabolism during DHCA to offer protective effect on the brain.

Animals ; Arginine ; pharmacology ; therapeutic use ; Brain ; drug effects ; metabolism ; Brain Ischemia ; etiology ; metabolism ; prevention & control ; Circulatory Arrest, Deep Hypothermia Induced ; adverse effects ; methods ; Dogs ; Female ; Indazoles ; pharmacology ; therapeutic use ; Male

The opening of laboratories in universities is one of important parts in teaching reform and it is necessary for bringing up high-qualified students.Combined with the practical teaching,we have a primary discussion with the problems of the laborato- ry opening and put forward some suggestions and measures.

OBJECTIVETo study the changes in the levels of autophagy-related proteins, Atg1, Atg5, and Beclin1, in organophosphate-induced delayed neuropathy (OPIDN) caused by tri-ortho-cresyl phosphate (TOCP), and to investigate the molecular pathogenic mechanism of OPIDN.

METHODSThirty adult Roman hens were randomly and equally divided into control group and 1, 5, 10, and 21 d intoxication groups. Each hen in the intoxication group was administered TOCP by gavage at a single dose of 750 mg/kg, while each hen in the control group was administered the same volume of corn oil. The hens were killed at the corresponding time points, and their tibial nerves and spinal cords were collected. The levels of Atg1, Atg5, and Beclin1 in the tibial nerves and spinal cords were measured by immunoblotting.

RESULTSCompared with those in the control group, the levels of Atg1 in tibial nerves decreased by 29.8%, 64.4%, 43.5%, and 19.8% at 1, 5, 10, and 21 d, respectively, after intoxication ((P < 0.05); the levels of Atg5 in tibial nerves decreased by 36.8%, 49.6%, 51.2%, and 31.5% at 1, 5, 10, and 21 d, respectively, after intoxication (P < 0.05); the levels of Beclin1 in tibial nerves decreased by 68.5%, 66.3%, and 32.2% at 1, 5, and 10 d, respectively, after intoxication (P < 0.05). Compared with those in the control group, the levels of Atg1 in spinal cords decreased by 23.5%, 48.7%, and 20% at 1, 5, and 10 d, respectively, after intoxication (P < 0.05); the levels of Atg5 in spinal cords decreased by 32.7%, 51.5%, 47.3%, and 39.6% at 1, 5, 10, and 21 d, respectively, after intoxication (P < 0.05); the levels of Beclin1 in spinal cords decreased by 28.9%, 50.2%, 43.2%, and 28.3% at 1, 5, 10, and 21 d, respectively, after intoxication (P < 0.05).

CONCLUSIONThe intoxication of TOCP is associated with the significant changes in the levels of autophagy-related proteins in the nervous tissues of hens, which might be involved in the pathogenesis of OPIDN.

Animals ; Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; drug effects ; Chickens ; Female ; Intracellular Signaling Peptides and Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism ; Nervous System Diseases ; chemically induced ; metabolism ; Neurofilament Proteins ; metabolism ; Spinal Cord ; metabolism ; Tibial Nerve ; metabolism ; Tritolyl Phosphates ; toxicity

OBJECTIVETo transfect antisense vector of human cyclooxygenase-2 (COX-2) gene into COX-2 highly expressing cholangiocarcinoma cell line QBC939 and explore its biological activities and role in carcinogenesis.

METHODSQBC939 cells were transfected with antisense vector of human COX-2 gene using LipoVec transfecting technique. Transfected cells were selected with G418; COX-2 mRNA was examined using reverse transcription polymerase chain reaction (RT-PCR) and COX-2 protein expression was detected by immunocytochemistry using isozyme selective antibodies. The proliferative status of transfected cells was measured by using methabenzthiazuron (MTT) assay; Cell cycle and apoptosis were analyzed by using flow cytometry.

RESULTSRT-PCR showed a lower COX-2 mRNA level in antisense vector transfected cells and immunocytochemistry showed a weaker COX-2 protein expression in antisense vector transfected cells. The antisense vector transfected cells proliferative index decreased significantly (P < 0.01), the percentage of S phase decreased remarkably (P < 0.05) in antisense vector transfected cells (9.27% +/- 1.91%) compared with that in QBC939 cells without transfection(16.35% +/- 2.87%), and the percentage of G0/G1 phase increased remarkably (P < 0.05) in antisense vector transfected cells (75.16% +/- 4.13%) compared with that in QBC939 cells without transfection (57.31% +/- 10.16%). Transfection with antisense vector of human COX-2 gene had no significant influence on the apoptosis in QBC939 cells (P > 0.05).

CONCLUSIONTransfection with antisense vector of human COX-2 gene could inhibit the proliferation of human cholangiocarcinoma QBC939 cells.

Apoptosis ; Bile Duct Neoplasms ; metabolism ; pathology ; Bile Ducts, Intrahepatic ; Cell Cycle ; Cell Division ; Cell Line, Tumor ; Cholangiocarcinoma ; metabolism ; pathology ; Cyclooxygenase 2 ; DNA, Antisense ; genetics ; Humans ; Isoenzymes ; biosynthesis ; genetics ; physiology ; Membrane Proteins ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; genetics ; physiology ; RNA, Messenger ; genetics ; Transfection