Objective:Vascular cell adhesion molecule 1(VCAM1)is involved in a series of physiological and pathological processes,such as immune and inflammatory response,tumor cell metastasis and invasion.But under trau-matic brain injury(TBI),specific types of cells with VC AM1 expression and the related functions are not clear.In order to further explore the specific functions of VCAM1 involved in TBI,this study constructed reporter mice of VCAM1 to explore the response of VCAM1 to TBI in detail.Methods:VCAM1-Cre/ERT2::Ai14 reporter mice were constructed by gene targeting technology and Cre/loxP system,the labeled cell types and labeling efficiency were validated by im-munofluorescence staining and reporter mice hybridization.The stab wound model was used to simulate TBI to induce the changes of VCAM1 expression in cells,and the characteristics of VCAM1 positive astrocytes were detected by immu-nofluorescence staining and fluorescent probe labeling.Results:The labeling efficiency of VCAM1-Cre/ERT2::Ai14 re-porter mice was higher than that of VCAM1 antibody as was seen by labeling of more endothelial cells of blood vessels and unique astrocytes.The distribution of these astrocytes was specific,for example in the nucleus accumbens,amygda-la,hypothalamus,and paraventricular fiber systems.TBI could significantly induce the expression of VCAM1 in astro-cytes(P＜0.0001).These induced astrocytes developed reactive qualities,including somal hypertrophy,GFAP ex-pression and proliferative ability.Conclusion:VCAM1-Cre/ERT2::Ai14 reporter mice could label cells with VCAM1 expression more sensitively,so they were more effective tools for observing expression and function of VCAM1.The up-regulation of VCAM1 expression in astrocytes after TBI surgery suggested that VCAM1 was an inflammatory response molecule in astrocytes,we recommended it as a new molecular indicator of reactive astrocytes.

Objective:To study the whole-brain distribution of vasoactive intestinal peptide(VIP)-positive neurons in the mouse brain and provide assistance for anatomical and functional studies of VIP neurons.Methods:VIP-Cre::Ai47 mice were used to label VIP neurons in the whole brain.Then,fluorescent imaging of the whole brain slices from VIP-Cre::Ai47 mice was calibrated using the standard brain map.Finally,the distribution density of VIP-positive neurons in different regions of the whole brain was statistically analyzed.Results:The overall distribution densities of VIP neurons in the cortex,olfactory bulb,and hippocampus were all greater than 5 neurons/mm2,and the distribution densities varied greatly in different subregions.The overall distribution density of VIP in the amygdala in the subcortical region was 4 neurons/mm2,and the distribution densities of VIP in the thalamus,hypothalamus,midbrain,and hind-brain regions were less than 2 neurons/mm2 on average,except for that in the supraoptic nucleus of the hypothalamus,where the density of VIP neuron distribution reached 20 neurons/mm2.Conclusion:VIP-positive neurons are mainly distributed in the cortex,hippocampus,and amygdala,which are highly associated with cognition and affection,and are rarely distributed in the thalamus and midbrain.These results suggest that VIP neurons play an essential role in emo-tional and cognitive functions.

Objective:Vascular cell adhesion molecule 1(VCAM1)is involved in a series of physiological and pathological processes,such as immune and inflammatory response,tumor cell metastasis and invasion.But under trau-matic brain injury(TBI),specific types of cells with VC AM1 expression and the related functions are not clear.In order to further explore the specific functions of VCAM1 involved in TBI,this study constructed reporter mice of VCAM1 to explore the response of VCAM1 to TBI in detail.Methods:VCAM1-Cre/ERT2::Ai14 reporter mice were constructed by gene targeting technology and Cre/loxP system,the labeled cell types and labeling efficiency were validated by im-munofluorescence staining and reporter mice hybridization.The stab wound model was used to simulate TBI to induce the changes of VCAM1 expression in cells,and the characteristics of VCAM1 positive astrocytes were detected by immu-nofluorescence staining and fluorescent probe labeling.Results:The labeling efficiency of VCAM1-Cre/ERT2::Ai14 re-porter mice was higher than that of VCAM1 antibody as was seen by labeling of more endothelial cells of blood vessels and unique astrocytes.The distribution of these astrocytes was specific,for example in the nucleus accumbens,amygda-la,hypothalamus,and paraventricular fiber systems.TBI could significantly induce the expression of VCAM1 in astro-cytes(P＜0.0001).These induced astrocytes developed reactive qualities,including somal hypertrophy,GFAP ex-pression and proliferative ability.Conclusion:VCAM1-Cre/ERT2::Ai14 reporter mice could label cells with VCAM1 expression more sensitively,so they were more effective tools for observing expression and function of VCAM1.The up-regulation of VCAM1 expression in astrocytes after TBI surgery suggested that VCAM1 was an inflammatory response molecule in astrocytes,we recommended it as a new molecular indicator of reactive astrocytes.

Objective:To study the whole-brain distribution of vasoactive intestinal peptide(VIP)-positive neurons in the mouse brain and provide assistance for anatomical and functional studies of VIP neurons.Methods:VIP-Cre::Ai47 mice were used to label VIP neurons in the whole brain.Then,fluorescent imaging of the whole brain slices from VIP-Cre::Ai47 mice was calibrated using the standard brain map.Finally,the distribution density of VIP-positive neurons in different regions of the whole brain was statistically analyzed.Results:The overall distribution densities of VIP neurons in the cortex,olfactory bulb,and hippocampus were all greater than 5 neurons/mm2,and the distribution densities varied greatly in different subregions.The overall distribution density of VIP in the amygdala in the subcortical region was 4 neurons/mm2,and the distribution densities of VIP in the thalamus,hypothalamus,midbrain,and hind-brain regions were less than 2 neurons/mm2 on average,except for that in the supraoptic nucleus of the hypothalamus,where the density of VIP neuron distribution reached 20 neurons/mm2.Conclusion:VIP-positive neurons are mainly distributed in the cortex,hippocampus,and amygdala,which are highly associated with cognition and affection,and are rarely distributed in the thalamus and midbrain.These results suggest that VIP neurons play an essential role in emo-tional and cognitive functions.

AIM: To extract and isolate polysaccharide from Poacynum Hendersonii leaves,determine its content and analyze the monosaccharide composition.METHODS: Poacynum Hendersonii leaves was extracted with hot water,crude polysaccharide was precipitated with ethanol,deproteinated according to Sevage method,coloured with acticarbon.Then of polysaccharide contents were measured by anthrone-H_2SO_4 colorimetry at the wavelength of 620 nm.The monosaccharide composition was determined by HPCE.RESULTS: The polysaccharide content was 0.97% of leaf weight,and Gal,Ara,and Man contents were three higher monosaccharides.CONCLUSION : The method is easy to carry out the baseline resolution in HPCE and has highly sensitivity.

Temporins are a kind of small,hydrophobic and C-terminus amidated antimicrobial peptides from Rana species.They are effective against bacteria,fungi,yeast,protozoa and viruses.Two novel temporins named as temporin-La(LLRHWKILEKYLanifc) and temporin-Lb(LFRHVVKIFEK.Ylamid.) were cloned from Lithobates catesbeianus.Synthetic peptides of temporin-La and temporin-Lb showed strong antimicrobial activities against bacteria tested,especially Gram-positive bacteria.Besides,temporin-La showed no haemolytic activity to rabbit erythrocytes at the concentration of 250 mg/L while temporin-Lb showed weak haemolytic activity(LC50≈ 230 μmol/L).Transmission electron microscopy showed that temporin-La and temporin-Lb induced different effects on bacterial structure of Staphylococcus aureus.

Aim To establish a rapid and sensitive diagnostic markerfor adjusting gut absorptive capacity after trauma. Methods A mi-cromethod for D-xylose test with phloroglucinal was established according toEberts with modification. The standard curves were repeated for fivetimes. Then the blood contents of fasting and orally taking 0. 5 g/kgD-xylose at 2nd and 4th hour were measured and the blood D-xylose con-tents of seven refilling rats with ischemia of intestine were mea-sured. Results The maximal absorption spectra was read at 554 nm byscanming of DU-7 Beckman. Good linearity of the D-xylose standard curvewas found with the range of 0 to 4 mmol/L(r=0. 9979 ±0. 0017) . Theblood content of rats at 2nd hour after orally taking D-xylose was ( 154 ± 6)mg/L amd that at 4th hour was decreased to (87 ± 11) mg/L. And theblood content at 2nd and 4th day after ischemic refilling was (162 ± 5)mg/L and (80 ± 8) mg/L respectively. The variation coefficients within andamong batches were 1.98% (n=6) and 3.10% (n=6), respective-ly. And D-xylose recovery rates were from 97.2% to 104. 3%. ConclusionMicromethod for D-xylose test appears to be simple, rapid and sensitive,and is an available index for estimating intestinal absorption after severetrauma.

OBJECTIVETo describe the methods which were used to develop collagen-based materials for wound dressing.

METHODSFresh frozen bovine tendon was treated with 0.05 mol/L acetic acid at pH 3.2 for 48-72 hours, homogenized, filtered, mixed with 8% chondroitin sulphate, for creating a deaerated 1.5%-2.5% collagen solution. The solution was lyophilized in either a pre-frozen or non-pre-frozen mould. The collagen sponge was then cross-linked with 0.25% glutaraldehyde for 24 hours. Three other types of wound dressings were developed using a similar method: collagen membrane with a polyurethane membrane onlay, polyurethane-coated collagen membrane and collagen membrane on gauze.

RESULTSIt was demonstrated that the use of frozen bovine tendon was stable, and that the prepared collagen sponge contained pores of 50-400 microm in diameter.

CONCLUSIONSCollagen could be used as wound dressing.

Amino Acids ; analysis ; Animals ; Biological Dressings ; Cattle ; Collagen ; analysis ; chemistry ; isolation & purification ; Freeze Drying ; Polyurethanes

Objective To study the injury factors, pathogenic process and clinical features of delay two-phase multiple organ dysfunction syndrome (MODS) in severe burned patients and to replicate a standardized animal model that would accurately imitate the clinical features of MODS.Methods Forty-five human patients with burn size larger than 30% total body surface area (TBSA) were analyzed. All of them underwent severe burn shock in early stage and sepsis in late stage. Thirty-two goats were randomly divided into three groups: 1) hemorrhagic shock (group H, n=6); 2) endotoxemia (group E, n=6); and 3) hemorrhagic shock plus endotoxemia (group M, n=20). Hemorrhagic shock was produced according to the method of Wigger (6.7 kPa for an hour, 1 kPa=7.5 mmHg). Endotoxin (E. coli O111 B4) was given via the portal vein 24 hours after the resuscitation of hemorrhagic shock, in a dose of 30 ng/kg/min for 5 consecutive days. During the observation period of 10 days, all animals were hemodynamically monitored, given standard metabolic support and due cardiac and pulmonary support according to human intensive care.Results All the patients showed burn shock at 1-3 days and hyperdynamic circulation, hypermetabolism and systemic inflammatory responses over two weeks post-injury. Thirteen cases were found to develop MODS according to the prevailing diagnostic criteria, and 10 of them died with a mortality of 77%. Eighteen animals died in group M with a mortality of 90%, 12 of the 18 developed MODS, with overall incidence of 60%. Most animals in group M showed changes similar to that observed in human cases. The experimentation proved that in the pathogenic process of MODS, there was a two-hit phenomenon in the dvelopment of the syndrome. To prevent the development of MODS, it therefore was imperative to blunt the first hit or the second hit, so that an excessive inflammatory response was alleviated. This postulation has been verified in the treatment of extensive burns. Two patients with burn extent reaching 100% TBSA survived with only mild acute respiratory distress syndrome (ARDS) and renal dysfunction after comprehensive treatment of burn shock, including adequate fluid resuscitation, drugs to remove oxygen free radicals, rapid restoration of pHi, and early extensive excision of burn eschars.Conclusion Both in human patients or animal experimentation, the typical delay two-phase MODS is shown to be produced by two successive insults in the forms of hypovolemic shock and sepsis. This postulation is helpful in formulating the prevention and treatment modality of MODS.