OBJECTIVETo investigate the effects of advanced glycosylation end products (AGEs) modified bovine serum albumin (AGE-BSA) on the expression of reactive oxygen species (ROS) and monocyte chemoattractant protein-1 (MCP-1) in human renal mesangial cells (HRMCs).

METHODSHRMCs were cultured in vitro with medium containing different doses of AGE-BSA or BSA (50,100, 200, 400 mg/L) for 48 hours, or with AGE-BSA (200 mg/L) for different times (12, 24, 48, 72 h). Immunocytochemistry assay was used to estimate the protein level of RAGE. The ROS in cells were measured by flow cytometry and the mRNA expression of MCP-1 were analyzed by semi-quantiative reverse transcription-polymerase chain reaction (RT-PCR) after treatment with AGE-BSA or BSA.

RESULTSThe protein level of RAGE was upregulated in the HRMCs with AGE-BSA. The expression of ROS and MCP-1 significantly enhanced by incubation of AGE-BSA in a time- and dose-dependent manner. The effects of AGE-BSA-induced up-regulation of ROS and MCP-1 level was significantly blocked by neutralizing antibodies to RAGE, while the expression of ROS and MCP-1 stood nearly unchanged after cultured with huamn IgG.

CONCLUSIONThe expression of ROS and MCP-1 in HRMCs is induced by AGE-BSA through RAGE, which may have potential effects in the pathgenic mechanism of diabetic nephropathy.

Cells, Cultured ; Chemokine CCL2 ; metabolism ; Glycation End Products, Advanced ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin, Bovine ; pharmacology

Objective To discuss the role of non-fusion without decompression in surgical treatment of unstable AO type A thoracolumbar fractures. Methods A retrospective study was performed on 42 patients with AO type A thoracolumbar fractures (T11-L2) treated with short segment pedicle screw fixation from February 2004 to February 2008. Patients were divided into two groups, ie, Croup A (treated with short segmental pedicle screw fixation without decompression or fusion) and Group B (treated with short segmental pedicle screw fixation without decompression but with fusion). The pre-operative, postoperative and follow-up local kyphotic angle, vertebrae compression rate were compared between two groups. Results In Croup A, average local kyphotic angle and average vertebrae compression rate were 19.1° (15. 4°-29. 8°) and 46% (30%-63%) respectively before operation, but 5. 0° (0. 3°-10.3°) and 10% (0-28%) respectively after operation. Twenty-one patients were followed up for average 21.2 months (12-46 months), which showed average local kyphotic angle of 7° (1.8°-10.7°) and average vertebrae compression rate of 10% (2% -22%) at final follow-up. In Croup B, average local kyphotic angle and average vertebrae compression rate were 25.8° (15.9°-34.5°) and 55% (30%-76%) respectively before operation, but 7.1° (1.5°-19. 1°) and 15% (0-28%) respectively after operation. Fifteen patients were followed up for mean 17.9 months (12-31 months) , which showed mean local kyphotic angle of 8.3° (0.7°-19.2°) and average vertebrae compression rate of 15% (l%-26%) at final follow-up. There was no pseudarthrosis, implant breakage, pedicle screw pull-out or severe back pain. There was statistical difference in local kyphotic angle and vertebrae compression rate between two groups.Conclusion Unstable AO type A thoracolumbar fractures with minor neurological deficit can be treated with pedicle screw fixation only without decompression or fusion.

?AlM: To evaluate the clinical effects and security of posterior chamber implantable Collamer lens ( lCL ) implantation in patients with extreme highly myopia.
?METHODS:ln this study, 18 patients ( 32 eyes ) with extreme highly myopic patients who had undergone posterior chamber lCLs implantation from July 2010 to July 2013 were evaluated. Diopter -10. 5 ~ 19. 0D, and astigmia -0. 5 ~4. 5DC. Changes in intraocular pressure ( lOP ) , refraction, visual acuity and corneal endothelium, anterior chamber depth, iris, high arch, lens were noted at 1d, 1wk, 1, 3mo and 1a after surgery respectively, and follow-up was of 1a.
? RESULTS: Before surgery, the uncorrected visual acuity (UCVA) were 0. 01~0. 05, and the best spectacle-corrected visual acuity ( BSCVA) were 0. 4 ~ 1. 0. One month after surgery, the UCVA were 0. 5~1. 2. The mean vault were 547±222 μm (95%CI 442~672μm) and 528±268μm (95%CI 354 ~635μm) for 1mo and 1a, respectively (P = 0. 81), and there was no significant difference. Anterior subcapsular opacities in 1 eye, mild and transient increase in lOP in 3 eyes, and chronic pigment dispersion in 2 eyes were observed. There was no serious complication.
?CONCLUSlON: Posterior chamber phakic intraocular lens implantation is an effective and safe method for correcting patients with extreme highly myopia.

Objectives:To study working memory deficit after stroke.Methods:118 patients with stroke di- agnosed by CT scans(study group)and 118 healthy people(control group)were collected from April to December in 2003.All objects were evaluated with Wechsler Memory Scale(WMS),Trail Making Test A and B,and Wisconsin Card Test(WSCT).Results:There were significant difference between the study group and control group in execu- tive function,attention function,visual recognition and visual recall;The study group had more error in Trail making Test B,A and executive error((?)?SE was 200.0?15.2,76.3?10.4,23.9?0.9)than control group(133.3?7.2, 24.2?1.6,9.9?0.5).Compared with control,patients with stroke of different locations had different deficits in work- ing memory:①for frontal stroke,the deficit manifested in executive function,digital span(P0.05).④for thalamic stroke,the deficit manifested in executive function,digital span,visual recognition and visual recall,(P

Objective To investigate the clinical effect of uterine artery embolization chemotherapy in the treatment of special site pregnancy .Methods 36 patients with special site pregnacy were randomly divided into the observation group and control group .The control group received uterine curettage after uterine artery embolization treatment,the observation group received bilateral uterine artery perfusion of methotrexate combined with uterine cu -rettage after embolization therapy .Time of-HCG returned to normal ,intraoperative blood loss ,hospitalization time and incidence rate of complications were compared between the two groups .Results After treatment,the time of -HCG returned to normal,intraoperative blood loss and hospitalization time in the observation group were (27.3 ±3.2)h, (120.5 ±18.3) mL,(85.6 ±8.4) h,which were significantly less than (76.4 ±8.2) h,(375.1 ±68.4) ml and (147.1 ±10.5)h in the control group (t=6.75,4.54,4.43,all P<0.05).The incidence rate of complication in the observation group was 16.8%,which was significantly lower than 50.1% in the control group (χ2 =4.49,P <0.05).Conclusion The uterine artery chemotherapy and embolization in the treatment of special parts of pregnancy can significantly shorten the hospitalization time and time of -HCG returned to normal ,reduce the intraoperative blood loss,the clinical effect is good ,it can be used as an effective method for the treatment of special site pregnancy .

In order to find the cardiotonic constituents of lateral roots of Aconitum carmichaelii Debx., the investigation was carried out. Silica gel column chromatography, Sephadex LH-20, medium-pressure MCI and reverse phase ODS column chromatography were used to separate the 90% EtOH extract of the lateral roots of Aconitum carmichaelii Debx. The structures of the isolated compounds have been identified by chemical properties and spectroscopic analyses. Ten compounds were isolated and their structures were elucidated as benzoic acid-5-hydroxy-2-benzoyl-amino methyl ester (1), honokiol (2), pinoresinol (3), salicylic acid (4), p-hydroxy-cinnamic acid (5), songorine (6), karakoline (7), mesaconitine (8), hypaconitine (9) and 14-benzoylhypaconitine (10), separetely. Compound 1 is a new compound and its structure has been established by NMR, HR-ESI-MS, UV, IR and X-Ray. Compound 2-5 are isolated from the lateral roots of Aconitum carmichaelii Debx. for the first time.
Aconitum ; chemistry ; Cardiotonic Agents ; chemistry ; isolation & purification ; Plant Roots ; chemistry

Thermostable alpha-amylase from Pyrococcus furious is an important industrial enzyme in brewing and alcohol production. Eexpression of the thermostable a-amylase in plants can reduce greatly costs in the production of alcohol using crop plants. A chloroplast expression vector, p64A, containing the thermostable alpha-amylase gene from Pyrococcus furious, was constructed with clpP-trnL-petB-chlL-rp123-rpl2 as Chlamydomonas reinhardtii plastid homologous recombinant fragments and spetinomycin-resistant aadA gene as select marker. The plasmid p64A was transferred into the chloroplast genome of C. reinhardtii by the biolistic method. Nine independently transformed lines were obtained by 100 mg/L spectinomycin selection. PCR amplification, Southern blot analysis of the transgene and cultivation in the dark all showed that the a-amylase gene had been integrated into chloroplast genome of C. reinhardtii. The activity of amylase expressed in the chloroplast of C. reinhardtii was detected by amylase activity assay and found to be as high as 77.5 u/g fresh weight of cells. These experimental results demonstrated the possibility of using transgenic chloroplasts of plant as bioreactors for production of industrial enzymes.
Chlamydomonas reinhardtii ; genetics ; Chloroplasts ; genetics ; Enzyme Stability ; Plasmids ; Polymerase Chain Reaction ; Pyrococcus furiosus ; enzymology ; alpha-Amylases ; chemistry ; genetics ; metabolism

This study was purposed to investigate the effect of bortezomib (Bor) and arsenic trioxide (As(2)O(3)) combination on multiple myeloma cell line KM3 and its mechanisms. KM3 cells were cultured with different concentration of Bor or As(2)O(3) as well as both for a certain time. The cell proliferation was analysed by MTT assay and the concentration of 50% proliferation inhibition (IC(50)) was calculated. Early apoptosis and late apoptosis of KM3 cells were detected by Annexin-V-FITC Kit, and the change of transmembrane potential was measured by flow cytometry. mRNA of Caspase-3, Bim and Bcl-xL were detected by RT-PCR. The results showed that the proliferation inhibitory rate of KM3 cells treated by Bor plus As(2)O(3) was much higher than that of KM3 cells treated by Bor only for 72 h [ (27.64 ± 0.81)% vs (21.67 ± 2.20)%, P < 0.05]. There were more KM3 cells treated by Bor plus As(2)O(3) in early apoptosis at 48 h and late apoptosis at 72 h than that of KM3 cells treated only by Bor [ (53.20 ± 3.70)% vs (35.40 ± 2.58)%, P < 0.01; (63.96 ± 2.97)% vs (54.08 ± 3.76)%, P < 0.01]. Transmembrane potential (Δψm) of KM3 cells treated by Bor plus As(2)O(3) decreased more at 48 h, as compared with Bor alone. The expression levels of caspase-3 mRNA and Bim mRNA in KM3 cells treated with Bor plus As(2)O(3) were higher than that in KM3 cells treated with Bor alone. But the expression level of Bcl-xL mRNA was lower than that in KM3 cells treated with Bor alone. It is concluded that As(2)O(3) can enhance the apoptosis-inducing effect of Bor on multiple myeloma cell line KM3, which is associated with decreasing the expression of Bcl-xl mRNA and increasing the expression of Caspase-3 and Bim mRNA.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Arsenicals ; administration & dosage ; pharmacology ; Bcl-2-Like Protein 11 ; Boronic Acids ; administration & dosage ; pharmacology ; Bortezomib ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Humans ; Membrane Proteins ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Oxides ; administration & dosage ; pharmacology ; Proto-Oncogene Proteins ; metabolism ; Pyrazines ; administration & dosage ; pharmacology ; bcl-X Protein ; metabolism

OBJECTIVETo establish a murine model of sensitization, and investigate the effect and mechanism of sensitization on allogeneic donor bone marrow cells (BMCs).

METHODSSensitized BALB/c mice were established by transfusions of allogeneic splenocytes. The donor reactive antibodies were detected by binding and complement-dependent cytotoxicity assays. After irradiation, 1 × 10(7) BMCs of C57BL/6 donor mice were injected into non-sensitized or sensitized BALB/c recipient mice. The distribution pattern of donor BMCs in peripheral blood, spleen and bone marrow of recipient mice were analyzed at different time points (2 h, 12 h and 48 h) post transplantation. Hematopoietic recovery post transplantation was assessed, and survival was monitored. Moreover, sera and splenocytes derived from non-sensitized or sensitized recipients were incubated with allogeneic BMCs in vitro, and the cytotoxic indexes were calculated in the immune experiments.

RESULTSThe binding and complement-dependent cytotoxicity assays showed that a high level of donor reactive antibodies was presented in sensitized sera. Compared with the non-sensitized recipients, the homing assay showed significantly decreased distributions of allogeneic donor BMCs in peripheral blood, spleen and femur of sensitized recipients. Non-sensitized recipients survived long term after irradiation, while all the sensitized recipients died within 12-15 days. Fourteen days post transplantation, the white blood cells and BMCs of non-sensitized recipients were (3240 ± 300) × 10(6)/L and (396 ± 27) × 10(6)/femur, respectively; while the white blood cells and BMCs of sensitized recipients were (320 ± 80) × 10(6)/L and (6 ± 2) × 10(6)/femur, respectively; the differences were statistically significant between this two groups (P < 0.05). Seven days post transplantation, the percentage of donor cells in bone marrow of non-sensitized and sensitized recipients was (48.07 ± 4.70)% and (0.77 ± 0.11)%, respectively, and the differences were statistically significant (P < 0.05). Furthermore, the white blood cells and BMCs following transplantation decreased along with time in sensitized recipients. The immune experiments of complement-dependent cytotoxicity reaction, cytotoxic T lymphocytes reaction and antibody-dependent cellular cytotoxicity showed the cytotoxic indexes were higher in sensitized group than the non-sensitized group.

CONCLUSIONA sensitized model was established by transfusions of allogeneic spleen cells. Allogeneic donor BMCs were rejected in sensitized recipients, and its mechanism might be through immune impairment pathways.

Animals ; Bone Marrow Cells ; Bone Marrow Transplantation ; Disease Models, Animal ; Graft Rejection ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Tissue Donors ; Transplantation, Homologous

OBJECTIVEStudying the gene expression profiling of different stage hairy root of Salvia miltiorrhiza, in order to find functional genes.

METHODThe contents of second metabolites were determined by HPLC and gene expression profiling was detected by cDNA microarray. cDNA labeled with a fluorescent dye (Cy5 and Cy3-dCTP) was produced by Eberwine's linear RNA amplification method and subsequent enzymatic reaction. The microarrays were scanned with a ScanArray Express scanner using ScanArray 2.0 software and quantified by signal intensities of individual spots from the 16-bit TIFF images using GenePix Pro 4.0. The linear normalization method was used for data analyze. Northern blot was used to test the gene expression results obtained by microarray. Different expressed genes were sequenced and analyzed by gap4 software, and then they were analyzed with BLASTX, BLASTN, GO and KEGG.

RESULTGrowth rate and second metabolites analysis indicated that the stage from 30 d to 45 d was the growth stage, while the stage from 45 d to 60 d was the second metabolites accumulation stage. Accordingly 30 d hairy root was chosen as a reference, which was hybridized with 45 d and 60 d hairy root separately. Total 203 different expressed genes were obtained. Northern blot showed that the result was identical with the microarray result. After sequenced, there were 172 genes clustered into 114 clusters (Unigenes). Among them, 62 unigenes had known functions, 34 unigenes were hypothetical protein, 9 unigenes were homologues with no similarity and 9 unigenes were unidentified protein with low similarity. Total 67 genes were classified into cellular component ontology, molecular function ontology and biological process ontology based on GO analysis. Total 26 genes, which represented 29 metabolic-related enzymes, were located in metabolic maps based on KEGG pathway classification.

CONCLUSIONSeveral important functional genes related to second metabolite synthesis were cloned such as P450 and copalyl diphosphate synthase genes. cDNA microarray was a useful tool for functional genomics of traditional Chinese medicine.

Alkyl and Aryl Transferases ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Plant ; Genomics ; methods ; Oligonucleotide Array Sequence Analysis ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; genetics ; growth & development ; metabolism ; Plants, Medicinal ; genetics ; growth & development ; metabolism ; Salvia miltiorrhiza ; genetics ; growth & development ; metabolism