Object To separate and characterize the chemical constituents of a plateau plant Halenia elliptica D Don Methods Elemental analysis (EA), 1HNMR and 13 CNMR, MS, FTIR and UV spectrometry, as well as DSC were employed Results Two needle shaped crystal chemical constituents obtained from H elliptica were confirmed to be 1 hydroxy 3, 7, 8 trimethoxyxanthone and 1, 7 dihydroxy 3, 8 dimethoxyxanthone, respectively Conclusion This is the first time for these two chemical constituents to be separated from this Tibetan medicinal plant

Objective To construct the multiple antigenic peptide (MAP) gene and E. coli ex-pression system, based on the out membrane protein OmpL1, LipL21 and LipL32 from Leptospira interro-gans, and better understanding of the immunological activity of the recombinant protein. Methods Using MI3KE display and Western blot, the advantage epitopes of OmpL1, LipL21 and LipL32 were identified and used to synthesize a new gene, then its prokaryotic expression system was constructed. The expression of re-combinant protein was determined by SDS-PAGE. The immunity activity of the recombinant protein was iden-tified by Western blot and ELISA. Results The synthetic gene was effectively expressed in E. coli and mainly presented in soluble form. The expression protein could react with the antileptospirosis antibodies in rabbit and human sera, which contained different serogroups. Conclusion The recombinant MAP gene of leptospires was successfully constructedand and expressed in E. coli. The recombinant protein had a good immune activity, and could cross-reacted with antileptospirosis antibodies from different serogroups.

Objective To summarize the imaging features of intra-pancreatic accessory spleen (IPAS)with multidetector computed tomography (MDCT) and improve the awareness and correct diagnosis of IPAS.Methods MDCT images of seven consecutive patients with surgically and pathologically confirmed IPAS were reviewed retrospectively.The investigated features included the location,size,shape,margin,density,and enhancement of the lesions.Results Four patients were male and three were female with a mean age of 49 years old.All the lesions were located at the dorsal side of parenchyma under the capsule of pancreatic tail.Three lesions were in round-like shape,and 4 in oval shape and all were well-defined.All the lesions were mass-like without necrosis and calcification.The maximum diameter of lesion ranged from 0.9 ～ 1.8 cm with a mean value of 1.4 cm.Compared with pancreatic parenchyma,the density of lesions were homogeneous on unenhanced CT,in arterial phase,slightly increased heterogeneous density was observed in 3 patients,slightly increased homogeneous density was observed in 4 patients.All the lesions appeared as slightly increased homogeneous density in portal phase.The CT value in unenhanced phase ranged from 50 ～ 61 Hu with a mean number of 55 Hu; and it ranged from 80 ～ 110 Hu with a mean number of 97 Hu in arterial phase; and the corresponding value was from 99 ～ 120 Hu with a mean number of 102 Hu in portal phase.Among the three patients underwent MDCT angiography,neither artery nor vein was compressed or invaded,and there was no vessel connected with lesions.Conclusions IPAS has some MDCT characteristics.For small solid mass in pancreatic tail,if the density and enhancement pattern is similar to that of spleen,the diagnosis of IPAS should be considered.

Objective To study the expression of hypoxia inducible factor-1 (HIF-1 ) and Notch signaling pathway downstream gene HES 1 in the hippocampus of pubertal rats with status epilepsy (SE), and to explore the regulation site of HIF-1αin Notch signaling pathway. Methods One hundred and seventy-six 21-day-old SD rats were randomly divided into control group (NS group), pentetrazole (PTZ)-induced SE group (PTZ group), and Notch signaling pathway speciifc inhibitor (DAPT) intervention group (DAPT group). In PTZ group PTZ was intraperitoneally injected to build SE model and in NS group normal saline was injected as control. The intraperitoneal injection of diazepam was used to terminate SE seizures. After successful modeling, the bilateral hippocampuses were isolated after the rats were sacriifced at 0.5, 1, 2, 4 and 8 h, respectively, and RT-PCR was performed to detect the mRNA expression of HES 1 and HIF-1α. The Western Blot was performed to detect protein expression in hippocampuses which were collected at 2 , 4 , 8 , 12 , and 24 h after successful modeling. DAPT group received intraperitoneal injection of DAPT 30 min before the start of molding, then the hippocampuses were isolated at 2 and 8 h after successful modeling. RT-PCR was performed to detect the mRNA expression of HES 1 and HIF-1αat 2 h, and Western blot was performed to detect protein expression at 8 h. Results At each time point after SE, the expression of mRNA of HES 1 and HIF-1αand the expression of protein were higher than the same time point of NS group (P

Objective To analyze and summarize the efficacy and the experience in the application of type Ⅰ and type Ⅱ bundled pancreaticojejunostomy in pancreaticoduodenectomy. Methods Between Jan.2005 and Dec. 2009, a total of 38 patients who underwent bundled pancreaticojejunostomy was enrolled, and their clinical data were retrospectively analyzed. 20 patients received type Ⅰ bundled pancreaticojejunostomy and 18 patients received type Ⅱ bundled pancreaticojejunostomy. The operative time, postoperative hospital stay, mortality and complications were compared. Results The operative time of type Ⅰ bundled pancreaticojejunostomy was (91 ± 20) min, and it was (63 ± 21) min in type Ⅱ procedure, and the difference was statistically significant (P ＜ 0. 05). The mortality and complications, postoperative hospital stay were 10.0%(2/20), 45.0% (9/20) and (20 ±2)d in type Ⅰ procedure, while they were 5.6% (1/18),38.9% (7/18) and(23 ±2)d in type Ⅱ procedure, and the difference was not statistically significant.Conclusions There was no significant difference in the effects between type Ⅰ and type Ⅱ bundled pancreaticojejunostomy. Carefully selective application of type Ⅰ and type Ⅱ bundled pancreaticojejunostomy helps complete these procedures.

Objective To screen the efficient antigenic epitopes in genus-specific envelope proteins OmpL1 and LipL21 of Leptospira interrogans for further development of multiple antigenic peptide (MAP)vaccine.Methods Based on bioinformatic technique,the combined epitopes of T and B lymphcytes in OmpL1 and LipL21 molecules were screened.Nucleotide fragments of each epitopes were amplified by PCR and then constructed their phage display systems.Using antisera against rOmpL1,rLipL21,L.interrogans serogroup Icterohaemorrhagiae strain Lai and leptospirosis patients' sera as the first antibodies.respectively,Western blot assays were performed to determine the immunoreaetivity and reactive ability of the epitopes with different antisera.Resuits Four combined epitopes of OmpL1 and two combined epitopes of LipL21 were selected out by the predicting procedure.All the amplified epitope fragments were accurately inserted into the region at N end of phage PⅢ protein and successfully expressed.All of the antisera could recognize each of the epitopes.Based on the results of Western blot,the two LipL21 epitopes at 97-112 and 176-184 showed similar strong hybridization signals with any of the antisera,and the hybridization signals of four OmpL1 epitopes with the three antisera were 173-191,87-98,297-320 and 59-78,from strong to weak.Conclusion The six combined epitopes in this study are efficiently antigenic.And the epitopes at positions 97-112 and 176-184 in LipL21 as well as the epitopes at position 87-98 and 173-191 in OmpL1 have a potential for developing leptospiral MAP vaccine.

Objective To clone mcp1, mcp2 and mop3 genes that encoding methyl-accepting chemotaxis proteins(MCP) of Campy/obacter jejuni for construction of their prokaryotic expression systems, and to establish chemotactic model in vitro of the microbe for determining chemotaxis-inducing substances and to understand relationship among MCPs and chemotactic inducers. Methods The segments of mep1, mcp2 and mcp3 genes were amplified by PCR and then sequenced after T-A cloning. Prokaryofic expression systems of the genes were subsequently constructed. SDS-PAGE pins Bin-Rod Gel Image Analyzer were used to examine the expression of target recombinant proteins rMCP1, rMCP2 and rMCP3, and Ni-NTA affinity chromatography was performed to purify the rMCPs. Rabbits were immunized with each of the three rMCPs to obtain antisera. Immunodiffusion assay was performed to measure the titers of antisera. IgG in each of the antisera were extracted by ammonium sulfate precipitation and DEAE-32 ion exchange chromatography, and IgG F(ab')2 were then prepared by pepsin enzymolysis and Sephadex G-100 chromatography. Chemotactic model in vitro of C. jejuni based on HAP( hard-agar plus) method was established to determine the chemo-taxis-inducing effect of eight candidate substances. Chemotaxis inhibition test based on IgG F(ab')2 bloc-king was applied to determine the function and diversity of MCPs. Results The segments with expected si-zes amplified from mcp1, racp2 and mcp3 genes were obtained by PCRs, and their nucleotide and putative amino acid sequences were completely same as the reported. The constructed prokaryotic systems could effi-ciently express rMCPs with the yields about 10% of the total bacterial proteins. Immunization with rMCP1, MCP2 and rMCP3 enables the rabbits to produce specific antibody. All the antisera had 1: 4 immunodiffusion titers. Both bovine bile and sodium deoxycholate(DOC) were able to induce ehemotactie movement of C. je-juni in a dosage-dependent manner ( P < 0.05 ). When MCP1 and MCP2 were blocked with their IgG F(ab')2, the ehemotaetic ability of C. jejurd were remarkably decreased (P <0. 05). However, MCP3 blocking did not affect the chemotaxis ( P > 0.05 ). Conclusion The C. jejuni MCPs are successfully ex-pressed in this study. Bovine bile and DOC are the inducers for chemotaxis of C. jejuni, and MCP1 and MCP2 are involved in the process of ehemotaxis iadueed by DOC.

Objective To explore clinical effect of repairing soft tissue defect in forearm, hand and foot with free super-thin anterolateral thigh perforator flaps. Methods At first the site of perforator vessels were determined by Doppler, then the flaps were designed and harvested with the site as center; the fascia lata and subcutaneous fat were removed by sandhill-likely only the 4.0 cm × 3.0 cm - 3.0 cm×2.5 cm disc-like fascia lata and dermis layer were reserved. 15 traumatic soft tissue defects including forearm, hand and foot were repaired with the ree super-thin antemlateral thigh perforator flaps. Results No vascular crisis happened and all skin grafts survived in donor sites. 2.0 cm×1.2 cm of the distal of flap was necrosis in 1 case and it was healed by dress changing. 15 cases were followed up 3 months-2 years and the average is 6 months. The contour and texture of all flaps were good and two point discrimination (2-PD) was about 8-10 mm of. Conclusions The contour and texture of free super-thin anterolateral thigh perforator flap are good, the feeling of recipient site recovered well, it's less injury for donor site and there is no reshaping for flap. It is a fineness donor site for repairing soft tissue defects in hand and foot.

AIM: Cystostomy is the traditionary method for detecting urodynamic indexes in mice, which de-stroys the continuity of the bladder, and there are significant differences between this method and the clinically used trans-urethral method.This study aims to develop an appropriate urethral catheter to investigate the advantages and application val-ue of transurethral method for urodynamic test.METHODS:A pediatric intravenous catheter was used for urethral catheter-ization on 8 female mice, and linked to connect the catheter to baroreceptor and micropump.The epidural catheter was also used as manometry tube.RESULTS:Using this method, the following urodynamic indicators has been successfully cap-tured:basal bladder pressure (BBP), bladder leak point pressure (BLPP), maximum voiding pressure (MVP), maxi-mum bladder capacity ( MBC ) , post-void residual urine volume ( PVR ) , voiding volume ( VV ) , efficiency of voiding ( EV) and bladder compliance ( BC) .CONCLUSION:This is the first successful simulation used in human body to a-chieve mouse urodynamic testing through the urethra catheter, which avoids the impact of cystostomy on urodynamics in mice, and the mice are able to keep long-term survival after tests for the follow-up molecular and genetic experiments.

Objective To investigate the expression of Musashi-2 and CD133 in colonic adenocarcinoma,and their correlation with the occurrence and development of colonic adenocarcinoma thereof. Methods The expressions of Musashi-2 and CD133 protein were detected by immunohistochemical method in 40 colonic adenocarcinoma samples and 15 normal colonic structure samples. The different histological types, TNM staging, lymph node metastasis, serous infiltration, distant metastasis and expressions of Musashi-2 and CD133 proteins in colonic adenocarcinoma tissues were analyzed. Results The positive expression rates of Musashi-2 and CD133 protein were 60%and 27.5%in 40 colonic adenocarcinoma samples and 26.7%and 0 in 15 normal colonic samples, which showed the significantly higher expression rates in colonic adenocarci-noma group than those of control group (χ2=4.850 and 5.156,P<0.05). There were significant differences in expressions of Musashi-2 proteins between different histological stages and TNM staging (P<0.05). The positive expression rate of Musa-hi-2 protein increased as the degree of differentiation decreased and TNM stage increased. There were significant differenc-es in expressions of CD133 proteins between different histological stages and lymph node metastasis (P<0.05). The positive expression rate of CD133 protein increased as the degree of differentiation decreased and lymph node metastasis occurred. Conclusion The expression of Musashi-2 and CD133 may be related with the initiation and development of colonic can-cer, which can be used as the stem cell markers of colonic adenocarcinoma for the further study.