The study was performed to detect the binding proteins for advanced glycation end products (AGEs) on human joint synovial cells (HSCs). Normal human synovial cells (type A and type B cells) were isolated and cultured in vitro. Binding assay was performed with radiolabeled human serum albumin modified by AGE (AGE HSA). Specific binding was defined as total binding minus binding in the presence of excess unlabeled AGE HSA. The result showed that: specific dose dependent binding of 125 I AGE HSA to immobilized HSCs was observed with R=4.90 0.75 10 4 /cell , Kd = 1.27 0.19 10 -6 M in type A HSCs , and R= 3.48 0.32 10 5 /cell, Kd= 1.38?0.16 10 -7 M in type B HSCs. TNF ?,IL 1? and AGE HSA upregulated the expression of AGE binding proteins on HSCs. Normal HSCs express specific AGE binding proteins. TNF ?, IL 1? and AGE HSA upregulate the expression of these proteins, suggesting that joint resident cells may be involved in the pathogenesis of dialysis related amyloidosis.

Objective To observe the clinical efficacy of staphylococcal protein A immunoadsorption plus nonmyeloablative chemotherapy with CD34+ autologous peripheral blood stem cell transplantation in the treatment of refractory systemic lupus erythematosus (SLE). Methods Three patients with active SLE were enrolled into this study. All patients were diagnosed with lupus nephritis by renal biopsy and poorly responded to routine therapy. Before transplantation, patients were given 6 sessions of immunoadsorption apheresis using columns of staphylococcal protein A-silica with an interval of 3 days; each session processed 3 L plasma and a total of 18 L plasma was processed over the 6 treatments. Three days following the immunoadsorption apheresis, the mobilization of stem cells was realized by intravenous cyclophosphamide at a dose of 2 g per square meter of body surface area and subcutaneous recombinant human granulocyte colony-stimulating factor (G-CSF) at a dose of 5 g per kilogram of body weight per day for 5 days. Then, peripheral blood raonoclonal cells were obtained by CS-3000 Cell Separator, and passed through the Clini Macs CD34+ cell selection device, with the final concentration of CD34+ cells being 2.6×106, 2.1×106 and 2.4×106 per kilogram of body weight respectively, and that of CD3+ cells being 3×105, 2.1×105, and 2.0×105 per kilogram of body weight, respectively, in these three patients. The conditioning regimen consisted of oral fludarabine of 50 mg/d for 5 days plus intravenous pig anti-human thymocyte immunoglobulin (ATG) at a daily dose of 90 mg/kg for 5 days. After 72-hour treatment with ATG, the frozen stem cells were infused back to the patients. Clinical manifestations and lupus-correlated immune parameters were compared in patients at baseline and after transplantation. Results Following immunoadsorption apheresis, an obvious decrease was observed in the level of serum anti-dsDNA, antinuclear antibody and IgG antibodies, while an increase in the level of serum complement 3. All patients achieved the reconstruction of hemopoiesis 2-3 days after the transplantation. Also, an apparent clinical remission was achieved with the SLEDAI score being less than 3. Six months after the transplantation, serum anti-dsDNA and antinuclear antibodies as well as urine protein were undetectable, the level of complement 3 reached the normal range, and renal function was restored. Conclusions Staphylococcal protein A immunoadsorption plus nonmyeloablative CD34+ autologous peripheral blood stem cell transplantation are effective and safe for refractory SLE, but the long-term effect remains to be connfirmed by further studies.

ObjectiveTo explore the effect of carotid artery sympathetic net exfoliation combined with hyperbaric oxygen therapy on athetosic type cerebral palsy (CP).Methods46 CP children (athetosic type) were divided into the treatment group (n=25) and control group (n=21). All cases of two groups were treated with carotid artery sympathetic net exfoliation as well as children of the treatment group were added with drug and hyperbaric oxygen therapy. Therapeutic effect of two groups was compared.ResultsOne week after operation, the effect rate was 66.67% in control group and 64% in treatment group with no significant difference between two groups. 3 months after operation, the effect rate of the treatment group increased to 96.00%, but that of the control group was not changed, there was a significant difference between two groups (P<0.05).ConclusionCarotid artery sympathetic nerve net exfoliation can improve the symptoms of athetosic type CP children, if combined with hyperbaric oxygen therapy, the curative effect will be further improved.

Objective To investigate how mobility training affects the synapses and their functioning in the region around a cerebral infarct.Methods One hundred and fifty rats were randomly divided into 5 groups:a mo- tor training group,a normal saline group,an Ara-c inhibition group,a mevastatin group and a control group.Cere- bral infarcts were surgically induced in all 150 rats,and the level of either glial fibrillary acidic protein(GFAP)or synaptophysin,and the cholesterol content around the infarct were observed at 7,21 and 42 days using immunohisto- chemical staining and high performance liquid chromatography.Results At 7,21 and 42 days after the infarct model was induced,significant differences in the optical density of either GFAP or synaptophysin and in cholesterol levels were noted between the motor training group and the control group.Ara-c inhibition was also significantly high- er in the controls.The optical density of synaptophysin and the cholesterol levels were significantly lower in the me- vastatin group than in the motor training group.Conclusion Motor skill training can improve synapse redefinition in rats with a model of acute cerebral infarct.Astrocytes may play a crucial role by means of the secretion of choles- terol in the region around the infarct.

Effects of 7 fungus strains on tobacco quality by applying fungal leavens on upper leaves were studied. Results showed that BF03, BF06 and BF63 can remarkably change the chemical components of tobacco, such as soluble sugar, protein, nicotine etc., and make them more harmonious. And the smoking quality of the flue-cured tobacco leaves treated was much better than the control.

Objective: To establish a multiplex PCR-microarray method for detecting important enteropahogens.Methods: Uniplex and multiplex PCR were performed to obtain the best primer sets for identifying the target bacteria at species and multi-species level.Fluorescent dyes were mixed into PCR reaction to determine whether it can affect the efficiency of amplification.To improve the efficiency of microarray,a 35 pairs primer-labeling system was optimized based on the hybridization results to find the best combination to avoid false negative results.Results: Specific PCR products were all obtained using species-specific primer sets.More preferential amplification may happen when more primer pairs were added to the reaction.The hybridization results showed a positive association between the efficiency of multiplex-PCR and signal intensity.Conventional PCR yielded more products than fluorescent dyes labeled PCR.Thirty-five primers were divided into three different combinations to label target respectively,hybridization results showed a high specificity.Conclusion: Mixing fluorescent dyes into PCR may reduce the efficiency of amplification and hybridization,but may have no effect on the analysis of hybridization results.The hybridization efficiency of microarray depends on the amplification efficiency of multiplex PCR.For microarray target labeling,three primer sets could be used to avoid negative hybridization led by preferential amplification of multiplex-PCR.It indicates that the multiplex PCR-microarray method is an attractive diagnosis tool for the high-throughput identification of enteropathogenic organisms especially for multiple causative agents and epidemiological investigations.

Objective To understand the genetic characteristics of Enterovirus 71 ( EVT1 ) circu-lating strains of Guangdong province in 2008. Methods We isolated an EV71 strain from the fatal case of the hand, foot and mouth disease during an epidemic of Guangdong in 2008. Its complete genome was se-quenced and analyzed comparatively. Results The results showed that the full length of EV71 GDFS-3 ge-nome( not including poly A tail ) is 7405 bp. No insertion or deletion is detected in the coding region. There are several insertions and deletions in 5'and 3'UTR. Phylogenetic analysis of GDFS-3 and reference strains showed GDFS-3 strain shares the highest nueleotide homology with TW984 strain(96.0% ) but low homology with SIN5865, MS and BrCr( about 81.0% ). GDFS-3 strain also shares the highest amino acid homology with TW984 strain(99.0% ). It clustered with reference strains of CA subgenotype in the phylogenetie tree. The nucleotide identity with CA reference strains is 91.0% -95.0%. Conclusion The phylogenetic analysis based on the entire genome demonstrates that GDFS-3 strain has the nearest genetic relationship with TW984 strains ( isolated in 2004). GDFS-3 may belong to the same subgenogroup ( CA ) with Taiwan predominant strains. Otherwise,Some mutations in 5'UTR of EV71 may play the very important role in heightened viru-lence.

OBJECTIVETo analyze effects of fracture of processus styloideus ulnae on prognosis in the treatment of distal radial fracture of type C according to AO classification.

METHODSThis was a retrospective case-control study, and the information was got ten through case evaluation and follow-up, including sex, age, patient satisfaction, Gartland & Werley score and radiographic score. There were 76 patient treated with open reduction and plate fixation in People's Hospital Affiliated to Peking University from July 2006 to July 2011. All the patients were divided into two groups: no combination with fracture of processus styloideus ulnae (group A, 56 cases), combination with fracture of processus styloideus ulnae (group B, 20 cases). The patients in group A and B were treated with open reduction and internal fixation; however the fracture of processus styloideus ulnae was not fixed. The indexes such as clinical data, bone grafting, joint movement, Gartland & Werley score and radiographic score were compared between two groups.

RESULTSThe ulnaris pain of patients in group B was more obvious than that in group A. The local VAS, palmar and dorsal flexion degree of wrist joint, motion VAS, patients satisfaction score, radial and ulnar deviation degree, pronation and supination of forearm degree, Gartland & Werley score and radiographic score were separately 0.1 ± 0.1, (51.1 ± 1.9)°, (60.2 ± 1.9)°, 0.6 ± 0.1 (23.1 ± 0.9)°, (28.7 ± 1.3)° (81.5 ± 2.6)°, (68.2 ± 2.7)° 1.9 ± 0.3, 89.6 ± 12.3 in group A; and separately 0.3 ± 0.3, (51.4 ± 2.3)°, (66.6 ± 1.7)°, 0.5 ± 0.2, (24.5 ± 2.0)°, (26.9 ± 1.8)°, (80.3 ± 2.5)°, (70.3 ± 3.7)°, 1.2 ± 0.4, 92.5 ± 7.5 in group B; there were no statistical differences in above indexes between two groups.

CONCLUSIONWhether the distal radial fracture with a concomitant unrepaired ulnar styloid fracture or not exerts no influence on mainly outcomes including function, radiography and motion of the wrist.

Bone Plates ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Prognosis ; Radius Fractures ; surgery ; Retrospective Studies ; Ulna Fractures ; surgery

Objective To construct a recombinant baculovirus dual expression vector containing NIS gene under the control of human telomerase reverse transcriptase (hTERT) promoter and plasminogen kringle 5 (K5) gene driven by early growth response 1 (Egr1) promoter,and to explore the feasibility of targeting both tumor and tumor vessel with combination of radioiodide and antiangiogenic therapy.Methods The hTERT-NIS gene and Egr1-K5 gene fragments were subcloned into baculovirus vector,then packaged and amplified in the sf9 cells to obtain recombinant baculovirus Bac-hTERT-NIS-Egr1-K5.Bac-CMV-NISEgr1-K5,Bac-hTERT-0-Egr1-K5 and Bac-hTERT-NIS-Egr1-0 were constructed as controls.The expression of NIS and K5 genes in human cervix cancers cells (HeLa) was examined by Western blot and quantitative real-time PCR.Functional NIS activity was confirmed by the uptake of 125I,the inhibition of NaClO4 and the cytotoxicity of 131I.The apoptotic effect of 131I-inducedK5 on human umbilical veins endothelial cells (HUVEC)was analyzed by an apoptosis assay using flow cytometry.Statistical analysis was performed using the analysis of variance.Results The recombinant baculovirus Bac-hTERT-NIS-Egr1-K5 was successfully constructed.The NIS gene under the control of hTERT promoter was specifically expressed in HeLa cells.The baculovirusinfected HeLa cells showed a significant increase of 125I uptake,which was significantly inhibited by NaClO4(F199.296,P＜0.05).Furthermore,a notable decreased cell survival rate (38.3％) was found after 131I treatment.The expression of K5 gene induced by 131I was elevated in a dose or time dependent manner and resulted in obvious inhibition with cell survival rate of 30.8％ in baculovirus-infected HUVEC cells,which was significantly higher than that in the control groups (11.2％ and 10.9％ respectively,F=19.926,45.409;both P＜0.05).Conclusions A recombinant baculovirus dual expression vector containing the NIS and K5 genes has been successfully constructed.This study suggests the feasibility of a synergistic strategy of NISbased raidoiodide therapy and K5-based antiangiogenic therapy in vitro,and make it possible to perform in vivo study in the near future.