Objective:To study the effect of portal vein occlusion on intestinal mucosal barrier in rats and the protection of ulinastatin to the injury,to present the experimental data for the clinical surgery.Methods:70 Sprague-Dawley rats were randomly divided into controlled group (n=10),operation group (n=30) and operation+medication group (n=30).The portal vein were occlused 40 min in the operation groups and operation+medication groups.2ml blood from portal vein,lymph nodes around appendix,1cm small intestine wall were taken for endotoxin levels,bacterial translocation and pathiology examinations in the all rats 280 mins after operation.The mocusal barrier and microscopic structure of intestine were observed.Results:Compared between the control group and the operation group,endotoxin levels,bacterial translocation rates rise greatly and gut structure change obviously in the latter.Compared between the operation group and operation+medication group,the former changes is also obvious.Conclusion:The occlusion of portal vein can leads the decrease of intestine mocusal barrier and the increase of its permeability.Ulinastatin has a good protective effect on the damages above.

In order to clarify the metabolism pathways of scandoside methyl ester, the analysis of metabolites profiling in four kinds of liver microsomes was performed by using an ultra-performance liquid chromatography/ electrospray-tandem mass spectrometry (UPLC-ESI-MS). The data obtained from the 0 h-incubation and the 2 h-incubation were compared and analyzed. After incubation, 5 metabolites of scandoside methyl ester were found in rat, Beagles, rhesus monkey and human liver microsome. The results showed that scandoside methyl ester's major metabolic pathway in the liver microsomes is hydrolysis, oxidation and reduction reactions, and there are certain kinds differences between species. The study provides a research base for further research about iridoid compounds in vivo metabolic pathways.

To identify the active components in Bufo melanostictus Schneider and clarify the difference between fresh and dried Venenum Bufonis, a UPLC-Orbitrap MS method has been established. The separation was performed with gradient elution of acetonitrile and water (with 0.1% formic acid) as mobile phase. By comparing their retention time and high resolution mass data of Venenum Bufonis extracts, 39 effective components were primarily identified by MS/MS analysis in positive ion mode. Twenty-six of them were bufadienolides. There were significant differences in the main composition between fresh and dried Venenum Bufonis. There are fewer bufadienolides in fresh toad venom.

Objective: To observe the expressions of NMDAR2 and GLAST in the hippocampal gyrus and supraoptic nucleus(SON) under sleep deprivation in young rats.Methods: The changes in expressions of NMDAR2 and GLAST in SON and hippocampal gyrus under sleep deprivation in younger rats were observed by immunohistochemistry method. Results:The expressions of NMDAR2 and GLAST in SON and the hippocampal gyrus were significantly increased on the 3rd day of sleep deprivation,even more sigificantly on the 5th day,decreased gradually on 7th day,and became similar to those of control group on the 14th day.Conclusion: Sleep deprivation can affect the expressions of NMDAR2 and GLAST in the hippocampal gyrus and supraoptic nucleus,but the effect diminisshes gradually with prolonged time,which may be associated with self-regulation and self-protection of the central nervous system.

Objective To study the clinical value of orbital magnetic resonance imaging(MRI) in diagnosis and making a judgment about the curative effect of glucocorticoid in patients with Graves' ophthalmopathy (GO).Methods A total of 106 patients with GO were divided into active GO group or inactive GO group according to whether clinical activity score(CAS) ≥3 or not,respectively.T2 relaxation times(T2RTs) (ms) and areas(mm2) of four extra-ocular muscles (EOMs) (superior,inferior,medial,and lateral rectus) were directly measured by MRI T2-Mapping.Correlation analysis of T2 RTs and areas with eyeball motility,diplopia,exophthalmus,visual acuity and intraocular pressure (IOP) were also performed.The T2RTs and areas of EOMs before and after intravenous methylprednisolone(iv MP) pulse therapy were recorded,respectively.Results The mean T2RTs (88.9 ± 13.8)and mean areas(58.1 ± 23.8) of EOMs in active GO were significantly greater than those in inactive GO (80.6 ± 10.0and 46.2 ± 18.6,respectively),bing both significantly greater than those in volunteers with normal eyes(76.2 ±4.7and 30.3 ± 6.1,respectively) (P＜0.01).Maximal T2RTs and maximal areas of EOMs showed significant positive correlation(P＜0.01),and both were positively correlated weakly with CAS,reduced movement of eyeball,diplopia,exophthalmus and loss of visual acuity (P＜0.01).On the contrary,maximal T2RTs and maximal areas of EOMs showed negative correlation or no correlation with spontaneous retrobulbar pain,painful eye movement,redness of eyelids and abnormal IOP.Even if 1 ≤ CAS＜3,ophthalmic symptoms and signs also showed a positive responses to iv MP in patients with moderate to severe GO as long as prolonged T2RTs occurred at least in one of EOMs.The maximal T2RTs,maximal areas,mean T2RTs and mean areas of EOMs were significantly decreased at the end of iv MP treatment,meanwhile ophthalmic symptoms and signs were obviously improved.Conclusions Prolonged T2RTs,especially accompanied by enlarged area on the identical EOMs is an remarkable symbols of acute inflamed muscles.The findings suggested that the use of prolonged T2RTs can improve diagnostic sensitivity and predict the curative effect of iv MP in patients with moderate to severe GO and CAS ≥ 1.

Objective To establish a gata4 gene knockout zebrafish model of congenital heart disease, and construct transcription activator-like effector nuclease ( TALEN) vectors targeting gata4 gene.Method We construct TALEN vectors targeting zabrafish gata4 gene using unit assembly method and the in vitro-transcribed TALEN mRNAs were microinjected into one-cell stage zebrafish embryos.The efficiency of TALEN was identified by injected embryos, and mutations of zebrafish were screened and confirmed the different types through PCR and enzyme digestion.Results We successfully constructed correct targeting vectors by enzyme digestion and sequencing, and the gene knockout efficiency was 35.18%.We screened the mutant zebrafish and confirmed different types of gata4 gene mutations.Conclusions A gata4 knockout zebrafish model is successfully established, it can provide a good animal model for further research of congenital heart diseases.

Objective:To investigate the frequency of myeloid dendritic cells (mDC) and plasmacytoid dendritic cells (pDC) in peripheral blood of patients with systemic lupus erythematosus (SLE) and their relationship with renal injury.Methods:The frequency of peripheral mDC and pDC in 102 SLE patients and 10 healthy controls were detected by flow cytometry. The quantitative data were expressed by [ M( P25, P75)]. The measurement data of the two groups with non-normal distribution was analyzed by Mann Whitney U test. The correlation between the two groups was analyzed by Spearman rank correlation analysis and multiple linear regression. Results:The frequency of pDC [14.00%(7.92%, 19.65%) vs 24.55%(19.68%, 32.90%), Z=-3.163, P<0.01] and mDC [21.25%(13.28%, 32.83%) vs 34.85%(24.58%, 41.93%), Z=-2.607, P<0.01] in the peripheral blood of 102 patients with SLE were significantly lower than those of healthy controls. The frequency of pDC [9.09%(7.31%, 17.38%) vs 24.55%(19.68%, 32.90%), Z=-3.033, P=<0.01] and mDC [9.40%(7.88%, 21.60%) vs 34.85%(24.58%, 41.93%), Z=-3.231, P<0.01] in 12 patients with newly diagnosed SLE were also significantly lower than those in healthy controls. After adjustedfor confounding factors, multivariate analysis showed that SLEDAI level was the main factor influencing the frequency of pDC ( P=0.019) and mDC ( P<0.01). In addition, pDC[8.02%(2.25%, 9.97%) vs 16.70%(11.80%, 24.60%), Z=-2.490, P=0.015] and mDC[8.80%(5.99%, 12.80%) vs 20.20%(11.20%, 42.80%), Z=-2.226, P=0.029] in patients with active LN were also significantly lower than that of patients with stable LN. The mDC frequency was positively correlated with the levels of complement C3 ( r=0.455, P<0.01) and C4 ( r= 0.289, P, P<0.01). Conclusion:The frequency of mDC and pDC in SLE patients is significantly abnormal, which is closely related to disease activity. In addition, pDC and mDC may be involved in the occurrence and development of LN.

In order to clarify the metabolism pathways of scandoside methyl ester, the analysis of metabolites profiling in four kinds of liver microsomes was performed by using an ultra-performance liquid chromatography/ electrospray-tandem mass spectrometry (UPLC-ESI-MS). The data obtained from the 0 h-incubation and the 2 h-incubation were compared and analyzed. After incubation, 5 metabolites of scandoside methyl ester were found in rat, Beagles, rhesus monkey and human liver microsome. The results showed that scandoside methyl ester's major metabolic pathway in the liver microsomes is hydrolysis, oxidation and reduction reactions, and there are certain kinds differences between species. The study provides a research base for further research about iridoid compounds in vivo metabolic pathways.
Animals ; Chromatography, High Pressure Liquid ; Dogs ; Esters ; metabolism ; Humans ; Iridoids ; metabolism ; Macaca mulatta ; Microsomes, Liver ; metabolism ; Rats ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo predict part of medicinal properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces by using the traditional Chinese medicinal property data prediction platform, in order to establish the relationship between properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces.

METHODThe properties of traditional Chinese medicine components were predicted by using the medicinal property data prediction platform based on the pharmacological effects of the components.

RESULTThe total sum of identical or similar results of the prediction for the properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces accounted for over 75%.

CONCLUSIONThe self-similarity exists between properties of traditional Chinese medicine components and traditional Chinese medicine decoction pieces, which reflects the inheritance, additivity and emergence among different properties of traditional Chinese medicines.

Objective To investigate the effects of different culture conditions on the isolation and expansion of stem cells from second-trimester amniotic fluids.Methods Amniotic fluids were obtained from 15 pregnant women undergone amniocenteses for medical indications between 16-24 gestation weeks by transabdominal amniocenteses from September 2007 to June 2008.Amniotic fluids(10-20 ml)samples were collected and each WaS cultured under different conditions or groups.(1)Low-glucose DMEM(LD) medium supplemented with 10%of fetal bovine serum(group of 10% FBS);(2)LD medium with 20%of FBS(group of 20%FBS);(3)LD medium with 15%of FBS and 4 ng/ml of basic fibroblast growth factor (group of bFGF);(4)LD medium with 10%of FBS as well ag the culture plate coated with gelatin(group of gelatin).The effects of different conditions were evaluated by comparing the number of primary colonies,the cell morphology and the ability of expansion.The isolated stem cells were identified by flow cytometry,RT-PCR and differentiation ability to edipocyte.Resuits (1)The success rates of primary culture of the group of 10%FBS,20%FBS,bFGF and gelatin were 60%,73%,73%and 60% respectively(P>0.05).The numbers of colonies were 0.9±0.5,2.6±1.5,2.9±1.5,1.1±0.8(P<0.01 when group of 10%FBS and gelatin compared with group of 20%FBS and bFGF);among the primary colonies,fibroblast-like colonies accounted for 46%,49%,64%.44%respectively(P>0.05).(2)The second passage cells obtained from all of these four groups could difierentiate into adipocyte after induction.(3)In the group of bFGF,stem cells were isolated from 5 samples and expanded to nearly 107 cells after 5 passages(P<0.01 compared with other groups).(4)Karyotype were normal in all samples.(5)Stem cells from bFGF group showed positive expression of SSEA-4.Oct-4 and Nanog gene detected by flow cytometry and RT-PCR.Conclusion Stem cells can be isolated from second-trimester amniotic fluids;moderate serum concentration and supplementation of bFGF can improve the efficiency of isolation and expansion of amniotic fluid of stem cells.