Object To study the structural characteristics of protein bond polysaccharide Le 3 from the fruitbody of Lentinus edodos (Berk ) Pegler Methods Relative molecular weights were determined by gel permeation chromatography Structural characteristics were expounded by infra red scan, gas chromatography, ? elimination reaction and Sepharose gel electrophoresis The content of total polysaccharide and protein were determined by colorimetry Results Mean molecular weight and molecular number of Le 3 were 13 700 and 12 500 Typical absorption peak of the polysaccharide as shown in infra red spectrum was of ? type glycosidic linkage Le 3 was composed of arabinose, xylose, mannose, galactose, glucose and glucuronic acid The molar ratio of the neutral saccharides was Ara∶Xyl∶Man∶Gal∶Glu=0 31∶0 47∶1 00∶1 15∶8 92 The content of total saccharide and protein were 70 62% and 25 31% The saccharides were not linked to peptides chain through O glycosidic linkage Le 3 contained RNA Conclusion Le 3 was a kind of new type polysaccharide isolated from the fruitbody of L edodes

Objective To evaluate the effects of immobilization of flexor tendon with elastic rubber band after the repair in 58 cases with flexor tendon completely severed in Zone Ⅱ . Methods The severed flexor tendon was repaired with the modified Kessler method. The rubber band was linked to the end of the nail with silk thread. The forearm and hand joint were immobilized by dorsal plaster cast.The rubber band was fixed with short tube plaster at the lower part of forearm so as to keep the interphalangeal joints in flexion. 48 hours post operation, exercises were done to extend fingers initiatively and flex fingers passively with the help of the elastic rubber band. The practice lasted for 3 weeks. Results The patients were followed up for three to six months.The digital functions of flexion and extension were normal. Conclusion Using elastic rubber band to immobilize flexor tendon post operation proves to have a remarkable effect on the functional recovery.

AIM: To find whether lipoxin A_4 (LXA_4) inhibits cell proliferation induced by TNF-? in rat mesangial cells, and to explore the molecular mechanisms of signal pathways of LXA_4 actions. METHODS: Cultured rat mesangial cells were growth-arrested and exposed to TNF-? with or without preincubation with LXA_4. Proliferation of mesangial cells was measured by MTT methods. Activities of STAT_3 were analyzed by electrophoretic mobility shift assay. Expression of cyclin E mRNA was assessed by RT-PCR. Cyclin E proteins were determined by Western blotting analysis. RESULTS: TNF-?-induced proliferation and increased mRNA and protein expression of cyclin E in mesangial cells were inhibited by LXA_4 in a dose-dependant manner. TNF-?-stimulation of the STAT_3-binding activities in mesangial cells was down-regulated by lipoxin A_4. CONCLUSION: Inhibitory effect of LXA_4 on TNF-?-induced mesangial cell proliferation is mediated by Jak_1/STAT_3 signal pathway.

AIM: To examine whether lipoxin A_4 (LXA_4) induces apoptosis of rat renal interstitial fibroblasts and explore the mechanisms. METHODS: Rat renal interstitial fibroblasts (NRK-49F cells) were incubated in RPMI-1640 medium supplemented with 5% fetal calf serum and exposed to LXA_4 at the concentration of 10 nmol/L, 100 nmol/L or 1 ?mol/L for 24 hours. Prior to experiment, some cells were transfected with calpain 10 antisense oligodeoxynucleotide. Apoptosis of cells was recognized by double staining using fluorescent dye acridine orange and ethidium bromide, observed under laser scanning confocal microscopy and counted by flow cytometry following propidium iodide and annexin staining. Activity of caspase-3 was measured by colorimetric assay. The expression of calpain 10 mRNA was determined by RT-PCR. RESULTS: LXA_4 at the concentration of 100 nmol/L or 1 ?mol/L induced 9.83% or 33.82% apoptosis of cells, respectively. Treatment of cells with LXA_4 up-regulated the expression of calpain 10 mRNA and increased the activity of caspase-3. The transfection of the cells with calpain 10 antisense oligodeoxynucleotide inhibited the LXA_4-induced apoptosis, activity of caspase-3 and expression of calpain 10. CONCLUSION: LXA_4 at high concentration induceds apoptosis in rat renal interstitial fibroblasts via up-regulating of calpain 10 mRNA expression.

Objective To investigate the role of vascular endothelial growth factor-C (VEGF-C) and its receptors in the formation of lymphatic vessels and lymphatic metastasis in gastric cancer. Methods By the domestic and overseas literatures review,the expressions of VEGF-C and its receptors in gastric cancer,their role in tumor lymphatic metastasis and prospect in treatment of gastric cancer were summarized. Results There was a significant correlation between VEGF-C and its receptors and the formation of lymphatic vessels and lymphatic metastasis in gastric cancer. VEGF-C high expression might be an early event in lymphatic metastasis and could be considered as an independent predictive factor of lymphaticmicrometastasis. By inhibition of gastric cancer cell from secrete VEGF-C or blockage of the interaction of VEGF-C with VEGFR-3,it was possible to inhibit tumor angiogenesis and the invasion and distant spread of cancer cells,thereby decreased mortality and improve survival. Conclusion VEGF-C and its receptors may promote the formation of lymphatic vessels and lymphatic metastasis in gastric cancer. It may be an effective way to gastric cancer for the treatments against VEGF-C and its receptors.

Rabbits were intoxicated with hypodermic injection of 0.8% CdCl2 in normal saline every other day for 3 months,and the blood level of urea nitrogen (BUN) and creatinine (Cr) and the activities of Na/K-ATpase,Ca-ATPase,and ?-GT in the renal cortex were determined.It was found that there were no remarkable changes of BUN and Cr level but significant reduction of the activities of the 3 enzymes.It is believed that the reduction of the enzyme activities is one of the factors to initiate the functional disturbances and morphological damages of the kidneys and plays an important role in the mechanism of renal failure if it is not promptly corrected.

Objective To explore the inhibitory effect of mutant influenza A viruses to the activation of interferon regulatory factor 3 (IRF-3). Methods HEK293 cells were infected with A/FM/1/47,A/HK/1/68, A/HK/1/68-MA20, A/HK/1/68-MA20C and positive control Sendai virus (SV). Whether the slowly moved phosphorylation form Ⅲ and Ⅳ of IRF-3 appeared or not was compared by Western blot in cells infected with these viruses. Wild type of NS1 from A/HK/1/68 and mutant NS1 from A/HK/1/68-MA20 were subcloned into pcDNA3.1-flag respectively. They were transfected in HEK 293 cells respectively. At 16 hours posttransfection, cells were infected with Sendai virus for 8 hours. Whole cell extracts were analyzed by Western blot and then probed with monoclonal flag antibody to check the expression of NS1, or with anti-IRF-3 to observe the inhibitory effects of the wild and mutant NS1 to the activated IRF-3. Luciferase assay was carried out by co-transfection with reporter plasmid, pGL2B with interferon ? promoter, and wild or mutant NS1 cDNA expression plasmid. SV was used to infect these cells after the co-transfection. Results Only less virulent A/HK/1/68-MA20 and positive control SV can activate IRF-3. Activated form Ⅲ and Ⅳ of IRF-3 began to appear 9 hours post infection (h.p.i), and most significant activated IRF-3 appeared 23 and 26 h.p.i. Sequence analysis of NS1 of MA20 revealed that nucleotide position number 94 is mutated from T to C, and amino acid at position number 23 is changed from valine to alanine. Co-transfected with wild type NS1 made form Ⅲ and Ⅳ of IRF-3 almost disappear, but not mutant NS1. In the luciferase functional analysis, wild type NS1 can inhibit the luciferase activity of IFN-? promoter, which was induced by SV, to around 1/10. Again no inhibitory effects was observed of mutant NS1 in the luciferase assay. Conclusion The mechanism that A/HK/1/68-MA20 can activate IRF-3 is that point mutant NS1 abolished the inhibitory function of NS1.

Objective Interferon regulatory factors 3 (IRF-3) is a key transcription factor to regulate gene expression of interferon after virus infection. This study aims to look for new spliced isoforms of IRF-3 and to investigate their structures and functions. Methods RNA extracts from human embryonic kidney 293 cells were amplified by RACE and RT-PCR. New sequences were compared with published sequences of IRF-3 and murine EST database using bioinformatics method. A new sequence, IRF-3c, was subcloned into pcDNA3.1-flag. The IRF-3c/pcDNA3.1-flag plasmid was transfected in HEK 293 cells. Whole cell extract was analysed by Western blot and then probed with monoclonal Flag antibody. Luciferase assay was carried out by co-transfection with reporter plasmid, pGL2B with interferon ? promoter, and IRF-3c cDNA expression plasmid. At 16 hours posttransfection, cells were infected with Sendai virus for 8 hours. Cells were collected and assayed for luciferase activity. Results A novel spliced isoform of IRF-3, named IRF-3c was discovered. The new isoform is almost the same as IRF-3, except for the utilization of the 180 bp bases in intron 6 adjacent to exon 6. The first 2,3 and 4 bases are a stop codon, which may produce a protein with a truncated C-terminal stoped at amino acids 327. Western blot analysis confirmed an expected 44 kDa strong band. The new inserted bases can be found in murine EST database, suggesting a conservative function in evolution. The functional luciferase assay showed that IRF-3c inhibited the IFN? promoter activity to (around) 40%～50% as that of control after Sendai virus infection. Conclusions The discovery of a new isoform of IRF-3 provides a new insight into the functional regulation of IRF-3 family. It is a dominant-negative inhibitor for interferon ? promoter activity in the virus infection pathway, provides a mechanism for the fine-tuning of the virus-induced activation of the interferon response, and prevents interferon ? from its overexpression and its toxic effects. It is worthwhile to explore the role of IRF-3c in the pathogenesis of human diseases using IRF-3c’s specific sequence.

Objective To explore the related factors of early hematoma enlargement in patients with spontaneous intracerebral hemorrhage.Methods The clinical data of 142 patients with spontaneous intracerebral hemorrhage were analyzed.The first CT was performed within 6 h of onset and the second within 24 h of onset.Single factor analysis and multiple Logistic regression analysis were performed to determine the related factors of early hematoma enlargement.Results The incidence of early hematoma enlargement was 24.6％ (35/142).Multiple Logistic regression analysis revealed that the following four factors were independently associated with early hematoma enlargement:age (OR =1.069,P =0.003),systolic pressure (OR =1.865,P =0.026),sharp of hematoma (OR =2.712,P =0.028),using mannitol (OR =2.939,P =0.020).Location of hemorrhage and volume of hemorrhage were not associated with early hematoma enlargement (P ＞ 0.05).Conclusions Age,systolic pressure,sharp of hematoma and using mannitol are the important predictors of early hematoma enlargement with spontaneous intracerebral hemorrhage.In patients with older age,higher systolic pressure,irregularly shaped hematoma,close observation of hematoma enlargement should be made CT-scanning check.And the caution in the early use of dehydrating agent should be careful.

Objective To examine whether lipoxin A4(LXA4) induces apoptosis of rat renal interstitial fibroblasts and explore the mechanism concerned.Methods Rat renal interstitial fibroblasts (NRK 49F cells)were incubated in RPMI 1640 medium supplemented with 5%fetal calf serum and exposed to LXA4 at the concentration of 10 nmol/L, 100 nmol/L or 1 ?mol/L for 24 hours. Prior to experiment,some NRK 49F cells were transfected with Smac antisense oligodeoxynucleotide. Apoptosis of NRK 49F cells was recognized by double staining using fluorescent dye acridine orange and ethidium bromide,and observed under laser scanning confocal microscopy and counted by flow cytometry following propidium iodide and annexin staining. Activity of caspase 3 was measured by colorimetric assay. The expression of Smac was determined by Western blotting analysis.Results LXA4 at the concentration of 100 nmol/L or 1 ?mol/L induced apoptosis of 9 83%or 33 82%of NRK 49F cells respectively, and reduced the cells of S and G2～M phase and increased the cells of G0～G1 phase in a dose dependent manner. Treatment of NRK 49F cells with LXA4 up regulated the expression of Smac protein and increased the activity of caspase 3. The transfection with Smac antisense oligodeoxynucleotide inhibited the LXA4 induced apoptosis and expression of Smac in NRK 49F cells. Conclusion LXA4 at high concentration can induce apoptosis of rat renal interstitial fibroblasts via the up regulation of Smac expression.