The root of Caragana sinica (Buc' hoz)Rehd' is a Chinese folk medicine which is said to be useful for thetreatment of asthenia syndrome,vascular hypertension, leukorrhagia, arthrodynia, and as antiinflammatory agent. In the course of our search for inhibitors of protein kinase C from Chinese herbs,chemical constituentsof the root of C. sinica was examined. Frorn ethanol extract of this crude drug,five isof1avones were isolatedand identified as flemichapparnin B, formononetin, pseudobaptigenin, 5-hydroxy-7, 4'-dimethoxy isoflavone,and 5-hydroxy-7-methoxy-3', 4' -methylenedioxy isoflavone.

BACKGROUND:Bone marrow mesenchymal stem cells (BMSCs) of mice are different from human and rats.The difficulties of culture,sustaining impermanency activity of BMSCs after passage and short-term effect of stem cells tracking limited the study of mice.OBJECTIVE:To obverse modified isolated culture method of BMSCs of mice and the feasibility of long-term fluorescent labeling stem cells.DESIGN,TIME AND SETTING:The experiments were conducted at the Affiliated Hospital of Academy of Military Medicine Science from June to December in 2008.MATERIALS:C57BL/6 mice,males and females,4-6 weeks of age,mean weighing 20 g,were used.METHODS:Stem cell culture fluid serum and liquid change manner were optimized using adherent screening and Percoll separation method.Rat BMSCs were incubated.In accordance with previous experiences of MSCs between human and macaque,Hyclonehigh-grade fetal bovine serum was selected for mouse MSC incubation.Serum was 10% of the medium.Bone marrow cells were washed out using LG-DMEM to filter bone dregs and small muscle blocks.Subsequently,samples were added on the Percoll separating medium at relative density of 1.082,and then incubated in 75 cm~2 culture flask at the concentration of 1.5×10~6/cm~2.BMSCs at the second passage were labeled with CM-Dil.MAIN OUTCOME MEASURES:Morphological changes in primary cultured and subcultured mouse BMSCs were measured.BMSC surface antigen of the second passage of mice (CD105,CD44,CD25,CD34) were determined using flow cytometry.The modified method was assessed to harvest the purity of stem cells.Activity of mouse BMSCs was identified using adipogenic and osteoplastic differentiation.The strength of fluorescent cells following multiple passage was observed.RESULTS:①The attached cells were observed 48 hours after primary cells culture and changed shuttles,triangles and flats at 7 days after culture.The figure become bunches and radial pattam at 3 passages.②MSCs highly expressed CD105,CD44 phenotypes and seldom expressed CD34 and CD45.③Spindle shape of cells gradually disappeared,with increased cell body.Some cells grew in cluster.MSCs changed figures to multilayer knots 10 days after inducing osteoplastic differentiation.MSCs became roundness and appeared fat drops in cells 9 days after inducing adipogenic differentiation.Red fat particles were shown following oil O staining.④The labeling cells gave out oranges light,and marked rates were over 80% in MSCs under the fluorescent microscope.The labeling cells were over 47% in 4 passages MSCs using flow cytometry.CONCLUSION:The modified method gained high-dosage cells in shorten culture time at passage 2 and made CM-Dil long lime labeling cells,which made more convenient for MSCs experiment on mice and stem cells tracer experiment in vivo.

Objective To investigate the diagnostic significance of synchronous observation on bone marrow biopsy,bone marrow smear and peripheral blood smear in stubborn hematonosis.Methods Thirty-seven patients with difficult and complicated hematologic diseases were detected by peripheral blood smear,bone marrow smear and bone marrow biopsy.Those slides and smears were observed.Results The 37 cases with blood disease was diagnosed of aplastic anemia in 3 cases,5 cases of myelodysplastic syndrome,6 cases of primary myelofibrosis,13 cases of secondary myelofibrosis,7 cases of lymphoma with bone marrow infiltration and 3 cases of metastatic carcinoma of bone marrow.In the diagnosis of aplastic anemia,myelodysplastic syndrome,primary and secondary myelofibrosis,lymphoma and metastatic carcinoma of bone marrow diagnosis,bone marrow biopsy is superior to smear.Conclusion Bone marrow biopsy,bone marrow smear and peripheral blood smear synchronous observation can effectively increase the diagnostic rate,decrease misdiagnosis rate,and correctly identify the stage of the disease.

Objective To develop a measurement method for determination of optical parameters of a red blood cell ( RBC) suspension based on the measurement of spatial scattered light signals without using an integrating sphere.Methods Multiple independent photoelectric sensors and light intensity modulation were used to obtain the measured values of diffuse reflectance,diffuse transmittance and collimated transmittance.The measured data results were imported into a Monte Carlo simulation based RTE to inversely determine the absorption coefficient, scattering coefficient and anisotropy factor of the measured sample using a new perturbation method.Results The described measurement method was applied to determine the optical parameters of a polystyrene microsphere suspension with a mean diameter of 2.6μm,and the results were essen-tially consistent with the calculated optical parameters by Mie code.Then, the RBC suspension was used for testing optical parameters,and the results were basically consistent with the parameters in the literature.Conclusion The system based on the measurement of spatial scattered light signals without using an integrating sphere will provide a quick and accurate approach for quantitative analysis of free hemoglobins and RBC suspensions.

Objective To investigate the role and possible mechanism of P120 catenin involving in the hemodynamic changes by inducing vascular endothelial cells injury through an in vitro experiment. Methods The hemodynamic environment under the different hemodynamic conditions at the vascular bifurcations was simulated through a T-shaped flow chamber system designed by ourselves. The human umbilical vein endothelial cells (HUVEC)cultured in vitro were stimulated and used the HUVEC cells of the small interfering RNA (SiRNA)after P120ctn gene fragments being knocked out. After loading flow rate of 250 and 500 ml/ min respectively and acting on for 12 h,the HUVEC morphology,growth pattern,and expression of P120ctn and other proteins were observed. Results (1)Normal HUVEC:500 ml/ min was loaded for 12 h,the cells were fused excessively at the impinging point,the cell gaps became narrowed,the cell density decreased and the morphology was elongated in the high wall shear stress (WSS)and wall
shear stress gradient (WSSG)regions. A part of cells migrated downstreamly,and their arrangement direction was consistent with the direction of impinging flow. Compared with the unloaded impinging flow field,after the 2 kinds of impinging flows being loaded for 12 h,the expression levels of P120ctn,vascular endothelial calcium (VEC),Kaiso,α-catenin,and other proteins were decreased. The expression level of matrix metalloproteinase 2 (MMP-2)was increased. There were significant differences (all P < 0. 05). (2)HUVEC after P120ctn being knocked out:Under the impact of the impinging flow,the cell growth time was reduced to 60 min. 250 ml/ min being loaded for 60 min,the impinging point and its surrounding cells still maintained the polygon,but some cells in the high WSS and high WSSG regions began to move downstreamly and aggregated,the cell arrangement mode partly arranged along with the direction of the flow;500 ml / min being loaded for 60 min,the cell density in the high WSS and high WSSG regions were decreased significantly and the morphology was elongated. A large number of cells migrated downstreamly and aggregated. The arrangement mode was parallel and consistent with the direction of the impinging flow. Compared with the unloaded impinging flow field,after the 2 kinds of velocities being loaded for 60 min,the expression levels of VEC,Kaiso,α-catenin proteins were decreased. The expression level of MMP-2 was increased,There were significant differences (all P < 0. 05) Conclusions The hemodynamic change may induce the changes in vascular endothelial cell morphology,growth pattern,and expression of P120ctn and other related proteins, leading to the decrease of vascular endothelial cell adhesion connection stability and the expression changes of related proinflammatory factors. The knockout of P120ctn may result in a further decrease of the vascular endothelial cell adhesion connection stability.

Objective To compare the efficacy of pantoprazoh based short-term quadruple regimens in the treatment of Helicobacter pylori (H. Pylori) infection for 4 days,5 days or 7days. Methods 166 patients with H. Pylori associated severe gastritis were randomly divided into pantoprozole quadruple regimens of 4-day group (n =61) ,5-day group (n = 54) or 7-day group (n = 51). The regimen was pantoprazole 40 mg,bismuth potassium citrate 220 mg,elarithromycin 250 mg and amoxicillin 1 g twice daily. The patients received pantoprazole for 1 week, bismuth potassium citrate for 2 weeks,clarithromycin and amoxicillin for 4 days,5 days or 7 days respectively. The H. Pylori eradication and symptomatic relief was determinded by 13C-UBT at least 4 weeks after the therapy. Results The H. Pylori eradication rates of 4-day,5-day or 7-day panteprazole quadruple regimens were 73.8% (45/61) ,75.9% (41/54) and 80.4% (41/51) respectively. The pain relief rates were 82.4% (42/51) ,85.1% (40/47) and 88.9% (40/45) in 4-day,5-day and 7 day group. Conclusion The 4-day and 5- day pantoprazoh based quadrual therapy is a short- term, effective, safe and lower therapeutic- cost regimen for H. Pylori eradication.

Objective To investigate the effects of Chinese herbal medication combined with nutrition intervention on perioperative nutrition in patients with colon carcinoma.Methods Ninety-six patients with colon carcinoma were divided according to their odd or even number into the control group and the intervention group.The former were given routine nutrition and the latter Chinese herbal medication combined with nutrition intervention.Then the mini-nutritional assessment(MNA)was used to assess the changes before and after operation and the complications.Results The intervention group was significantly better than the control one in terms of all nutrition factors but hematoglobin(P<0.05).The incidences of complication and dystrophy in the intervention group were lower than in control group(P<0.05).Conclusion The application of Chinese herbal medication combined with nursing intervention is important for the improvement of their nutrition and the reduction of postoperative complications.

Objective:To construct NMB0315 eukaryotic expression recombinant vector ,detect specific humoral and cellular immune response induced by the recombint DNA vaccine intramuscularly in female BALB /c mice,evaluate the immunocompetence and immunoprotection of the vaccine , so as to provide experimental basis for the development of a novel nucleic acid vaccine against N.meningitidis serogroup B .Methods: The whole NMB0315 gene was amplified by PCR from the standard strains MC 58 genomic DNA,cloned into a plasmid pcDNA3.1(+),identified by double digestion of the recombinant plasmid with restriction enzymes and se -quencing.The recombinant vector pcDNA 3.1 (+)/NMB0315 was transfected into eukaryotic COS-7 cells and RAW264.7 cells, the NMB0315 protein was detected by immunocytochemical method and Western blot respectively .The levels of specific humoral and cellular immune response were detected after inoculating in female BALB /c mice intramuscularly with the recombinant plasmid .The immune protective effect was investigated with the DNA vaccine and the bactericidal titer of the immune serum was deter mined by serum bactericidal assay ( SBA ) in vitro.Results: The recombinant pcDNA3.1 (+)/NMB0315 was effectively transcripted and expressed in eukaryotic cells and the specific humoral and cellular immune responses were induced in the inoculated mice .In the re-combinant pcDNA3.1(+)/NMB0315 group ,the levels of serum IgG,IgG1,IgG2a,IgG2b and IgG3 and genital tract sIgA were significantly higher than in controls ( P<0.001 ) .The stimulation index in the culture supernatant of the spleen lymphocytes of the vaccine group was higher than that of the control group (P<0.05).The ratios of serum IgG2a/IgG1 in the DNA vaccine group were less than 1.The bactericidal titer of the NMB 0315+CpG group reached 1:128 following three immunizations , the protection rate of the vaccine group was 70%against the N.meningitidis strain MC58.Conclusion:The NMB0315 nucleic acid vaccine could induce higher levels of humoral immunity and cellular immunity and showed effective protection against N .meningitidis serogroup B , the immune serum had strong bactericidal activity in vitro .

Objective To evaluate the effect of sevoflurane anesthesia on activation of gliocytes in aged rat hippocampi.Methods One hundred and four male Wistar rats,weighing 600-650 g,were randomly divided into 2 groups (n =52 each) using a random number table:control group (group C) and sevoflurane group (group S).Group S inhaled 3.6％ sevoflurane for 2 h,and open reduction and internal fixation was performed after tibial fracture was induced.Y-maze test and fear conditioning test were carried out on days 1,3 and 7 after surgery.Six rats of each group were chosen on days 1,3 and 7 after surgery and hippocampi were removed to detect the activation of microglias and astrocytes (by immunofluorescence) and expression of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) (by Western blot).Results Compared with group C,the number of entries into each arm was significantly reduced,the duration of N arm stay was shortened,the percentage of time spent freezing induced by condition and the percentage of time spent freezing induced by context were decreased,and the activation of microglias and astrocytes was increased,and the expression of IL-6 and TNF-α was up-regulated in S groups (P ＜ 0.05).Conclusion Sevoflurane-induced postoperative cognitive dysfunction may be related to enhanced activation of gliocytes in aged rat hippocampi.

Autoimmune Diseases ; drug therapy ; Hepacivirus ; Hepatitis C ; drug therapy ; Humans ; Interferons ; therapeutic use