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.

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.

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 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.

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 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.

Objective To investigate the optimal dose of oxycodone for severe pain in cancer patients so as to try to alleviate the suffering rapidly.Methods Totally 135 cases with severe pain in terminal cancer patients were randomly divided into groups A,B,and C,45 cases in each group.Groups A,B,and C were treated with oxycodone hydrochloride sustained-release tablets 20 ～ 140 mg/d,150 ～ 290 mg/d,300 ～ 640 mg/d,respectively.Pain relief and incidence of adverse reactions were evaluated 2 weeks later.Results Pain intensity scores (numerical rating scale,NRS) in 3 groups dropped with statistical difference (P ＜ 0.05).Pain relief rates in groups B and C were statistically significant compared to group A (P ＜ 0.05).Quality of life score (appetite,mental status,sleep,daily activities,and interpersonal life) in group C had statistical significance compared to groups A and B (P ＜ 0.05).Group C had slightly higher incidence of adverse reactions than that of groups A and B without significant difference (P ＞ 0.05).Conclusions Large dose and extra large dose of oxycodone hydrochloride sustained-release tablets can effectively relieve severe pain in advanced cancer patients.Extra large dosage can improve patient's quality of life in spite of the higher incidence of adverse reactions,and patients can still be tolerated well.

Objective To investigate the effect of advanced oxidation protein products (AOPPs) on human renal tubular epithelial cells(HK-2) autophagy.Methods HK-2 cells were stimulated with AOPPs.RT-qPCR and Western blot were used to determine the expression of autophagy related protein LC3-Ⅱ/LC3-Ⅰ,Beclin1 and p62;Western blot was utilized to examine the activation of p38 MAPK pathway.Then p38 MAPK inhibitor (SB203580) was added and co-processed with AOPPs.The change of autophagy was observed Also,autophagy inducer rapamycin was added and co-processed with AOPPs.RT-qPCR and Western blot were used to detect the expression of cell cycle inhibitory protein p27.The cell total protein level was detected by the bicinchoninic acid (BCA) method.The hypertrophy change was observed.Results AOPPs down-regulated the expression of LC3-Ⅱ/LC3-Ⅰ and Beclin1,up-regulated expression of p62 and activated p38 MAPK pathway;in comparison with the AOPPs alone treatment group,the expression of LC3-Ⅱ/LC3-Ⅰ and Beclin in the SB203580 co-processing group was increased,while p62 was decreased;the p27 expression and cells total protein in the sirolimus co-processing group were down-regulated.Conclusion AOPPs inhibits the autophagy of HK-2 cells by activating p38 MAPK pathway and the autophagy inhibition participates in HK-2 cell hypertrophy.

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