Mechanism of exogenous nitric oxide in attenuating endotoxin-induced increase of rat pulmonary microvascular permeability

AIM and METHODS:The animal model of acute lung injury (ALI) caused by intratracheal instillation of lipopolysaccharides(LPS) in vivo and human peripheral blood polymorphonuclear neutrophil (PMN) in vitro were used to study the effects of sodium nitroprusside (SNP), nitric oxide (NO) donor, on LPS-induced PMN accumulation, microvascular permeability and PMN apoptosis. RESULTS:①In vivo, PMN accumulation in lung, the protein content in bronchoalveolar lavage fluid (BALF) and the Evans blue dye and monastral blue dye extravasation in lung tissue of LPS group were markedly higher than those of both sham operation group and LPS+SNP group. ②In vitro, the apoptotic percentage of SNP group was much higher than that of control group, while compared with LPS group, SNP+LPS group has significantly higher apoptotic percentage. CONCLUSIONS:SNP intratracheal instillation attenuated LPS-induced microvascular permeability and alleviated ALI. PMN apoptosis induced by SNP may be one of the potential mechanisms underlying the decrease of PMN accumulation in lung tissue.     

Preparation of gfp-bcl-XL-contained recombinant adenovirus vector by the homologous recombination in bacteria

AIM: To prepare gfp-bcl-X_L-contained recombinant adenovirus(rAd-gfp-bcl-X_L).METHODS: Bcl-X_L gene was amplified from pEGFP-C₃-bcl-X_L, subcloned into shuttle plasmid and formed transfer plasmid of pAdTrack-CMV-bcl-X_L. Then pAdTrack-CMV-bcl-X_L was linealinzed with PmeI and co-transformed into BJ5183 bacteria with adenovirus genomic plasmid of pAdEasy-1. The identified recombinant adenovirus plasmid was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles. The target gene was detected by PCR.RESULTS: There were about 35% positive recombinant bacterial clones after the co-transformation of pAdTrack-CMV-bcl-X_L and pAdEasy-1 into BJ5183. Recombinant adenovirus particle were produced and further amplified after the transfection of pAdEasy-1-gfp-bcl-X_L into 293 cells. PCR test indicated that the recombinant Ad contained bcl-X_L gene. The titer of the purified rAd-gfp-bcl-X_L was 6.5×10¹² PFU/L. CONCLUSIONS: The homologous recombination in bacteria is a convenient and high efficient method to prepare rAd-gfp-bcl-X_L. This affords a good gene transfer vector for the gene therapy in human’s diseases.     

Effects of riboflavin and ascorbic acid on apoptosis and proliferative inhibition of mouse thymocytes induced by deoxynivalenol in vivo

AIM: To explore the effects of riboflavin and ascorbic acid on the apoptosis induced by deoxynivalenol(DON) in mouse thymocytes. METHODS: The effects of riboflavin and ascorbic acid on the apoptosis and proliferation inhibition of thymocytes induced by DON in KM mice were studied with animal experiment, DNA agarose gel electrophoresis and flow cytometric DNA content analysis. RESULTS: Apoptosis rate of thymocytes in DON (4 mg/kg) treated group was13.73%±15.3% The percentages of apoptosis in riboflavin (1.25 mg/kg-10mg/kg) and ascorbic acid (25 mg/kg-100mg/kg) pretreated thymocytes groups were significantly lower than that in DON group (P <0.05). The result of DNA agarose gel electrophoresis showed that the characteristic ladder pattern of apoptosis was found in DON-treated thymocytes, but not in control and riboflavin pretreatment and ascorbic acid pretreatment groups The significant differences in proliferation index were not found among DON-treated thymocytes and riboflavin and ascorbic acid-pretreated thymocytes CONCLUSION: Pretreatment with riboflavin and ascorbic acid inhibit apoptosis of mouse thymocytes induced by DON in certain extent and have no effect on proliferation inhibition by DON.     

Effects of nitric oxide on hepatic encephalopathy in cirrhotic rats induced by LPS

AIM: To investigate the effects of nitric oxide (NO) on hepatic encephalopathy in cirrhotic rats induced by LPS. METHODS: The cirrhotic model of rats was established by complex pathogeny. Since the end of the 8 th week, the rats were intragastrically-infused with 0.9% salt, L-arginine(L-arg) and LNNA respectively for 2 weeks.The hepatic encephalopathy in cirrhotic rats were induced by 3 mg/kg LPS (ip) 4 hours before the rats were sacrificed. RESULTS: The normal behaviors and electroencephalograph were appeared in L-arg group. LNNA group showed hepatic encephalopathy. The content of NO₂^-/NO₃^- of brain tissue was markedly higher in L-arg group than LNNA group(P<0.05), but the content of histamine in brain tissue was lower in L-arg group than LNNA group(P<0.05). There was a negative correlation between the content of histamine in brain tissue and the content of NO₂^-/NO₃^- of brain tissue. CONCLUSION: NO can prevent hepatic encephalopathy in cirrhotic rats induced by LPS.     

Study on integrin-mediated albumin microbubbles adherence to activated leukocytes

AIM:To investigate the mechanism responsible for albumin microbubbles adherence to activated leukocytes. METHODS: In vitro studies were performed in which activated or nonactivated leukocytes were incubated with albumin microbubbles and observed under microscopy. The suspensions of leukocytes and microbubbles which contained or absented of integrins were analyzed with flow cytometry.RESULTS: A minimum of 50cells were identified under transillumination. 5 min after microbubbles were incubated with leukocytes, the number of cells interacting with microbubbles was greater for activated cells than for nonactivated cells(20.30±2.67 vs 4.50±1.43, P <0.01).Microbubbles attached to the surface of activated leukocytes were phagocytosed and remained intact for up to 30min. Microbubble attachment was inhibited notably by blocking the leukocyte β₂-integrin Mac-1(P <0.01) and by VLA-4mAb slightly(P <0.05) CONCLUSION: The mechanism of albumin microbubbles attaching to and phagocytosed by leukocytes was due to β₂-integrin and VLA-4 mediation. Phagocytosed microbubbles can remain at the regions of inflammation for15 min, also responsible to ultrasound.     

Protein expression of cyclin D2 and p16 in proliferation and differentiation of cultured cardiac myocytes

AIM:To investigate the protein expression of cyclin D₂ and p16 in proliferation and differentiation of cultured cardiac myocytes.METHODS:One-day-old Sparague-Dawley rats were used. Cardiac myocytes(CM) were collected by a trypsin-dispersal method and cultured. Cell growth line and fluorescence activated cell sorting (FACS) were used to investigate the proliferation of CM. Ultra-thin sections were made to observe the ultrastructure of CM under transmission electron microscope. The expression of cyclin D₂ and p16 in CM were measured using immunocytochemistry and image analysis.RESULTS:①Results of cell growth line and FACS analysis showed that cultured CM could proliferate in the first 3 cultured days, but the ability decreased quickly, concomitant with differentiation. CM was obseved quiescent in cell cycle three days later. The ultrastructure of CM showed the large amount of myofilaments and mitochondrion. ②The protein expression of cyclin D₂ in 3,4,5 day CM group was 0.89 times(P＜0.05),0.80 times (P＜0.05) and 0.56 times (P＜0.01) of that in 1 day group, respectively. The expression of p16 in CM was increased during the culture process, 2,3,4,5 day group were 1.63 times, 1.72 times, 1.99 times and 2.84 times (P＜0.01) of that in 1 day group, respectively.CONCLUSION:Cultured neonatal rat cardiac myocytes could proliferate during the first 3 days after incubation, but the ability of proliferation decreased, from the fourth day, concomitant with differentiation. Cyclin D₂ and p16 play the key roles in CM postnatal development. Downregulation of cyclin D₂ and upregulation of p16 may induce CM differentiation.     

Changes of intracellular potassium activity and membrane potential in guinea pig ventricular myocardium during superfusion with low sodium solutions and effects of low pH

AIM:To investigate the changes in intracellular potassium activity(aⁱK) and membrane potential(V_m) induced by low external sodium infusion (Low [Na]_o) and to detect the mechanisms involved and the relationship between aⁱK and V_m. METHODS:aⁱK and V_m were measured in infusion with different sodium concentrations using methods of convenient and ion selective microelectrodes in guinea pig ventricular myocardium. RESULTS:Low [Na]_o resulted in a decrease in aⁱK and an increase in V_m in a Na⁺ concentration-dependent manner.At the same time,contraction and resting potential increased, and action potential duration decreased markedly,but action potential amplitude was not affected. A change of the pH from 7.4 to 7.0 in low [Na]_o solution reduced the decrease in aⁱK, but did not affect the increase in V_m.CONCLUSION:A better linear relationship appeared between the changes in aⁱk and [Na]_o or in V_m and [Na]_o,while during each low [Na]_o the change in both aⁱK or V_m may reach a new balance level.     

中国的一个新记录属——冠生刺菌属

基于采自滇中哀牢山区的标本 ,报道我国的一个新记录属———冠生刺菌属CreolophusP .Karst.,并根据其主要特征把齿耳属SteccherinumGray的肉齿耳S .cir rhatum (Pers.exFr .)Teng归并到Creolophus属中。同时对有争议的相似种C .cirrhatus (Pers .exFr .)Karst.和C .corrugatum (Fr.)P .Karst.(=HydnumcorrugatumFr.)进行了比较研究 ,根据盖缘是否具有生殖能力的菌刺 ,提出两者应作为不同的种来处理。     

蔗渣滤泥培养料栽培平菇试验

以不同比例滤泥代替栽培料中的麦麸进行平菇栽培试验。结果表明 ,滤泥能替代栽培料中的麦麸。     

两系稻制种穗期病害发生较重的原因及防治措施

调查分析稻粒黑粉病、穗颈稻瘟等两系杂交稻制种穗期主要病害的发生原因,提出了清除菌源、轮换制种、健身栽培、对口药剂防治等防治措施。