Objective To investigate the mechanisms regarding protection of doxycycline from articular cartilage cataplasis in osteoarthritis,and provide experimental evidence for prevention and cure of degenerative osteoarthropathy and for treatment of osteoarthritis.Methods (Ⅰ) Models:Sixteen New Zealand rabbits underwent right anterior cruciate ligament riesction,and fed in each cage.The animals were injected with penicillin 400 thousands IU once a day for 3 days to prevent from infection.On the fourth day after operation,the animals were compelled to use their operated knee by driving 60 minutes per day for 4 weeks.(2) Animal groups:the animals were divided into two groups.The animals in experimental group were injected into the knees with 1.33％ doxycycline 0.3 ml (total 0.4 mg),once a day for 4 weeks.Rabbits in the control group were treated under the same condition using normal saline 0.3 ml.All rabbits were fed in their own cage and not drove.The total time was 4 weeks.(3)Example collection and data analysis:Rabbits were sacrificed by aeroembolism and their knee joints were examined by gross morphology.The mRNA expressions of MMP-3 and TIMP-1 in the cartilage and synovium were analyzed by a reverse transcription polymerase chain reaction ( RT - PCR).Results For the experimental group,condyles of femur cartilage inspection revealed that 6 cases were in Grade 0 and 2 cases were in Grade 1,and whirbone cartilage inspection showed that 5 cases were in Grade 0 and 3 cases were in Grade 1.For the control group,2 cases were in Grade 0 and 6 cases were in Grade 1.The extent of articular cartilage cataplasis was significantly different mRNA expression of MMP-3 ( 1.20 ± 0.32) (experimental) and ( 1.97 ± 0.54) (compared) ( t =3.47,P＜ 0.05).The expression of MMP-3 mRNA in cartilage and synovium was significantly decreased,respectively; while the expression of TIMP-1 mRNA was significantly increased in the experimental group(3.22 ±0.42) compared to the control group( 1.90 ±0.33) ( t =6.99,P ＜ 0.01 ).Conclusions Intra-articular injections of doxycycline inhibits the articular cartilage and synovial MMP-3 mRNA expression and increases TIMP-1 mRNA expression,thereby reducing the extent of degeneration of articular cartilage.

Mycotoxin covers a large number and variety of secondary metabolites, which can induce toxicological effects in mammals. T-2 toxin and citreoviridin (CIT) are common mycotoxins. Many agricultural products are polluted to varying degrees during harvest or storage. Their toxin levels depend on environmental factors such as temperature and humidity of the soil, storage conditions and so on. These two mycotoxins pose potential health hazards to human beings. Among them, T-2 toxin causes degeneration and necrosis of epiphyseal plate cartilage in the development stage, which produces pathological changes similar to Kashin-Beck disease; CIT causes myocardial damage, which is associated with the outbreak of endemic Keshan disease. Therefore, the pathogenicity of T-2 toxin and CIT has been widely concerned. In this paper, the production, physicochemical properties, toxicological effects, pathogenesis and control measures of T-2 toxin and CIT are reviewed.

AIM: Nur77 expression can lead to T cell apoptosis, and it is activated by the second messenger calcium. The calcium-responsive elements in the Nur77 promoter are two putative binding sites for the transcription factor, myocyte enhancer factor-2D (MEF2D). In this study, we explored whether MEF2D could be acetylated in Nur77-induced T cell apoptosis and whether MEF2D acetylation was affected by the regulation of Nur77 expression. METHODS: The experiment was performed at Department of Hematology-Oncology in the First Hospital of Jilin University from November 2006 to September 2007. ①Escherichia coli DH5?and plasmid pcDNA3 were held by the Research Center in the First Hospital of Jilin University. Plasmids pET32M, Flag-p300 and pSilencerTM-p300 RNAi were gifts from Professor Zhengguo Wu from Hong Kong University of Science and Technology. Plasmid NFATp was contributed by Yun Chen from Harvard University. Jurkat cells were provided by Department of Hematology- Oncology in the First Hospital of Jilin University. ②The full-length Nur77 gene, generated by PCR, and the Nur77-dependent luciferase reporter gene were subcloned into plasmid pcDNA. The variant sequences of MEF2D (1-514, 1-121, 1-300, 301-514) and the MEF2D (4KR) lysine-to-arginine mutants, at lysine sites K245/K250/K267/K279, had the Flag epitope at their amino termini. All of the above plasmids were constructed in bacterial expression vectors and the constructed clones were verified by sequencing and then transfected with liposome DMRIE-C. ③The impact of p300 on the trans-activation function of Nur77, mediated by MEF2D, was detected by luciferase reporter assays after Nur77-dependent reporter gene was transfected together with MEF2D, p300 or NFATp into Jurkat cells. In vivo acetylation of endogenous MEF2D and whether acetylation of MEF2D was affected by blocking the expression of p300 were detected by immunoprecipitation. The acetylated sites of MEF2D were detected by acetylation assays in vitro. The impacts of both MEF2D acetylation defects on trans-activation of Nur77 and on apoptosis of T cells induced by Nur77 were detected by luciferase reporter assays and flow cytometry, respectively. RESULTS: ①Although the dimeric complex of MEF2D and NFATp could not induce the transcription of Nur77, significant transcription was achieved with the ternary complex of p300, NFAT and MEF2D. The dimeric complex of p300 and MEF2D could also significantly induce Nur77transcription.②MEF2D was acetylated after the calcium signaling pathway was activated by PMA/Iono. The acetylation level of MEF2D was markedly reduced when p300 expression was blocked by p300 RNAi. ③The simultaneous mutation of K245/K250/K267/K279 lysine sites largely prevented the acetylation of MEF2D, which demonstrated that K245/K250/K267/K279 lysine sites in vitro were the major sites of MEF2D acetylation. ④The MEF2D acetylation defect increased by 2.48 fold the transcription f of Nur77, when compared with vacant vector. But compared with wild type MEF2D, transcription of Nur77 was decreased by 70.4%. ⑤When compared with vacant vector, Nur77 expression could increase total T cell apoptosis of early and late periods from 4.3% up to 16.3%; the co-expression of wild type MEF2D and Nur77 could further increase total T cells apoptosis of early and late periods from 16.3% to 31.0%. However, the co-expression of acetylation defective MEF2D mutant and Nur77 significantly decreased total T cells apoptosis of early and late periods from 16.3% to 9.2%. CONCLUSION : p300 has a major impact on the transactivation function of Nur77 mediated by MEF2D. MEF2D could be acetylated in Nur77-induced T cell apoptosis and acetylated MEF2D could promote T cell apoptosis by upregulating the transcription function of Nur77.

Objective To study the methods and efficacy of selective or superselective renal artery embolization for renal neoplasms,abcesses and bleeding. Methods Sixty cases of renal carcinomas and 4 cases of abcesses were all treated with selective renal artery embolization before surgery,and 3 cases of advanced renal carcinomas were treated with selective renal artery embolization alone.Embolization materials for carcinoma and abcesses were mainly gelfoam.Nine cases of renal angiomyolipomas,23 of traumatic renal hemorrhages,1 undergoing extracorporeal shock-wave lithotripsy (ESWL) and 2 nephrolithotomy,all of whom had severe hematuria,were treated with selective or superselective renal segment artery embolization.Embolization materials for hemorrhages were mainly self-blood clots and(or) gelfoam,and the materials for angiomyolipoma were pure alcohol and(or) gelfoam. Results The sizes of carcinomas and abcesses were decreased in all of the 60 cases of renal carcinomas and 4 cases of renal abcesses.The edema around the carcinomas and abcesses became obvious,and bleeding was reduced,which were convenient for operation and prolonged survivals.All the renal bleeding were cured with selective or superselective renal artery embolization except for one traumatic case treated by operation for continuous bleeding.All these cases were followed up for 1 month to 18 years.One case had mild atrophy of kidney and in one case IVU didn't develop.All the others were in good condition. Conclusions Selective or superselective renal artery embolization is a safe,effective approach with less complications and most probability to preserve the diseased kidney. Renal artery embolization as a valuable treatment should be studied and widely applied.

Objective To observe the efficacy of prostatic massage combined with α1 receptor blocker in the treatment of type Ⅲ prostatitis. Methods 86 cases were randomly divided into two groups. The treatment group( n =45 ) were given twice-weekly prostatic massage in combination with α1 receptor blocker( terazosin 2mg per night) for 6 weeks. Thc control group(n =41 )were only given terazosin 2mg per night for 6 weeks. AII the patients were asked to complete the National Institutes of Healthe Chronic Prostatitis Symptom Index(NIH-CPSI) before and after the treatment. Results 86 cases were evaluated after the treatment. The NIH-CPSI scores of the two groups decreased from ( 29.5 ± 4.2 ) to ( 9.6 ± 3.8), from ( 28.9 ± 3.4) to ( 14.6 ± 3.6) respectively, with statistically significant differences from pretreatment( t = 12.131、6.999, all P ＜ 0.05 )as well as between the combined therapy group and the control group(t = 4.649, P ＜ 0.05). There were no serious side effects during the therapy. Conclusion Prostatic massage combined with α1 receptor blocker can relieve the symptoms in type Ⅲ prostatitis patients and improve their life quality. The combined therapy can produce better efficacy than terazosin used alone.

Objective To investigate the effects of static magnetics on adhesion of neutrophil and endothelium and expression of intercellular adhesion molecule-1(ICAM-1) on human umbilical vein endothelial cell(HUVEC). Methods HUVEC were exposed to 0.05mT, 0.1mT, 1mT static magnetics adhesion of neutrophil was calculated after HUVEC was stimulated by LPS. Flow cytometry and ELSIA were used to study expression of ICAM-1 on endothelium with stimulating of LPS. Results The adhesion of neutrophil l on HUVEC was attenuated by 0.05mT, 0.1mT static magnetics(58% vs. 40%, 38%, P

The study was made on 3H-leucine incorporation into brain of offsprings born by their mothers who were fed in severe iodine deficiency area. Control group received an iodine-supplemented diet (1.7mg KIO3/kg food). Disintegration per minute (dpm) was used as an index to assess the 3H-leu-cine incorporation rate in comparision of the two groups.The results of 3H-leucine incorporation into brain showed that the dpm/mg of brain cortex and cerebellum was significantly lower in experimental group than that of control group at the time of 1-14 hours after injection of the preparation on 11-day offsprings (p

Objective To clarify the role of a cell cycle regulator, cyclin D1 and CDK6 in patients with gastric carcinoma. Method Tissue samples from 48 patients with gastric carcinoma were included in the current study. Expression levels of cyclin Dl and CDK6 in samples of normal mucosa and tumor tissue were analyzed by Western blot. Result Overexpression of cyclin Dl and CDK6 protein were demonstrated in 58% and 69% of gastric cancer tissues, respectively. Several clinicopathologic parameters, including depth of tumor invasion, pathologic lymph node status and tumor stage ( P

Objective To investigate the expression of matrix metalloproteinase-2 (MMP-2)and matrix metalloproteinase-9(MMP-9) in MIA PaCa-2 cells blocked by AS-ODN cultured in hypoxia.Methods Heparanase(Hpa) expression of MIA PaCa-2 cells was blocked by AS-ODN and cultured in hypoxia.The expression of MMP-2 and MMP-9 mRNA and proteins in cell lysate was evaluated by RT-PCR and Western blot respectively,and the enzymatic activities of MMP-2 and MMP-9 in supernatants were detected by gelatinase activity assay.Results Hypoxia stimulated mRNA and protein expression of MMP-9 in cultured MIA PaCa-2 cells and elevated at 6h,12 h(P ＜0.05)and 24 h(P ＜ 0.01).When Hpa expression was inhibited by AS-ODN,the expression of MMP-9 mRNA and protein as well as the gelatinase activity in supernatant decreased dramatically at 12 h and 24 h,especially at 24h(P ＜0.01),however,no significant difference of MMP-2 expression and gelatinase activity was observed after AS-ODN transfection.(P ＞ 0.05).Conclusion In hypoxia,MMP-9 expression,either mRNA or protein in cultured MIA PaCa-2 cells,increased gradually accompanied with elevated gelatinase activities.When the heparanase expression was inhibited,the MMP-9 mRNA and protein,as well as the gelatinase B activity in supernatant,were decreased dramatically at 12h and 24h,however,no significantly differences of MMP-2 expression and gelatinase A activity were observed after the AS-ODN transfection.

Objective To explore the expression of ininor histocompatibility antigen 13 (HM1 3) in gastric carcinoma and the relationship with clinicopathological parameters and prognosis.Methods The expression of HM 13 was detected by immunohistochemistry in a total of 90 pairs of paraffin-embedded gastric cancer tissue specimens and corresponding paraneoplastic tissues.The correlation between clinicopathological parameters and the expression of HM13 in gastric carcinoma were also analyzed.Downstream gene heme oxygenase-1 (HO-1) expression was detected by qRT-PCR after HM13 gene was interfered.Results High expression of HM13 protein was observed in 47％ gastric carcinoma compared with that in 61％ corresponding paraneoplastic tissues (x2 =3.78,P =0.052).HM13 expression has positive correlation with pathological TNM stage (x2 =5.022,P =0.025) and distant metastasis (P =0.033),but not with age (x2 =0.832,P =0.362),gender (x2 =0.779,P =0.378),tumor size (x2 =0.804,P =0.370),tumor differentiation (x2 =0.430,P =0.512),gross types (x2 =2.069,P =0.150),tumor stromal invasion (x2 =0.167,P =0.683) and lymph node status (x2 =0.396,P =0.529).The patients with low HM13 expression had worse overall survival [(OS):(30 ± 5) months vs.(47 ± 5) months,x2 =6.456,P =0.011] and progress free survival [(PFS):(29 ± 5) months vs.(46 ± 5) months,x2 =6.742,P =0.009].Expression of HO-1 was up-regulated after HM13 gene was interfered (0.532±0.013 vs.0.395±0.011,t=13.93,P＜0.05).Conclusion The low expression of HM13 is associated closely with advanced stage and distant metastases of gastric carcinoma.