Objective To study the effect of cyclophosphamide pulse therapy combined with leflunomide (LEF) on refractory lupus nephropathy (RLN).Methods Thirty-six patients diagnosed as RLN were selected.Several observation parameters were compared before and after cyclophosphamide pulse therapy combined with LEF.The observation parameters included serum albumin,serum complement C3 and C4,systemic lupus erythematosus (SLE) activity index and urinary protein change.The side effects of cyclophosphamide pulse therapy combined with LEF were observed.Results The serum albumin was (20.17 ± 4.09) g/L,complement C3 was (0.40 ± 0.19) g/L,complement C4 was (0.08 ± 0.03) g/L,SLE activity index was 16.06 ± 4.17,and urinary protein was ( 9.79 ± 3.42 ) g/24 h before treatment and (38.10 ± 5.16) g/L,(0.78 ± 0.11 ) g/L,(0.16 ± 0.13)g/L,4.01 ± 1.24,( 1.14 ± 0.59) g/24 h after treatment,and there were significant differences between before treatment and after treatment (P ＜ 0.01 ).During the therapy,side effects were reported in 22 patients.However,these side effects had no impact on the therapy.After further treatment,these side effects gradually decreased and eventually disappeared.Conclusions Cyclophosphamide pulse therapy combined with LEF is effective in treating RLN.Patients' tolerance to the therapy is generally very good.

Objective:To investigate the expressions and their clinical significance of vascular endothelial growth factor (VEGF) and extracellular matrix (ECM) components in non-small cell lung cancer (NSCLC). Methods:Expressions of VEGF and ECM components (fibronectin, FN and collagen Ⅳ, cⅣ) in 50 cases of NSCLC tissues and 20 cases of normal lung tissues were detected by immunohistological analysis. Their relationship with clinical features of NSCLC and the correlation of expression of VEGF and Fn and cⅣ were analyzed.Results:The positive expression rates of VEGF, Fn, and cⅣ were 96%, 78%, and 50% in NSCLC tissues. The expressions of VEGF and Fn were significantly higher than those in normal lung tissues (P<0.05). The expression of Fn and over-expression of VEGF were associated with lymph node metastasis (r=1.00, P<0.001). The survival rate of patients with over-expression of VEGF was greatly lower than that with weak expression of VEGF (P=0.022). The survival rate of Fn-negative patients was markedly higher than that of Fn-positive patients (P=0.046). Conclusion:VEGF and ECM component Fn were highly expressed in NSCLC, which correlated with lymph node metastasis and survival rate. Expression of ECM and VEGF had positive correlations, suggesting that ECM might be one of the anti-angiogenesis targets for tumor therapy.

Objective:To make a prospective study on the effectiveness and safety of toremifene (TOR) combined with novelbine/cisplatin (NP) in the treatment of patients with advanced non-small cell lung cancer (NSCLC) whose first line platinum-based chemotherapy was failure. Methods:Forty-four patients with stage ⅡB-Ⅳ NSCLC, who failed in the first line cisplatin-based chemotherapy from January 2004 to February 2006, were enrolled in this study. All the patients received TOR combined with NP second line chemotherapy for two cycles. The response rate and adverse reaction were evaluated. The survival rate was analyzed.Results:The 44 patients received average 1.8 cycles of chemotherapy (1-3 cycles). The response of 37 patients could be evaluated including 21 patients who received NP regimen before and 16 patients who received platinum-based chemotherapy. After second line therapy, 4 of the 37 patients had partial response (PR), 19 had stable disease (SD), 14 had progressive disease (PD), and no patient had complete response (CR). The total response rate (CR+PR) was 10.8% (4/37). The disease-controlling rate (CR+PR+SD) was 62.2% (23/37). The response rate and disease-controlling rate of squamous cell lung cancer (SCC) were 27.3% (3/12) and 72.7% (8/12), which were significantly higher than adenocarcinoma [0% (0/18) and 44.4% (8/18), P<0.05]. The median survival time was 8.2 months, the median time for SD was 4.0 months (1.0-10.2 months), and the 1-year survival rate was 24.4%. The median survival time and 1-year survival rate of SCC patients had no significant difference compared with adenocarcinoma patients (9.2 vs 7.1 months; 33.3% vs 27.7%, P＝0.72). There was no significant difference in survival rate between male and female patients. One patient stopped therapy for liver function injury (hyperbilirubinemia). The adverse reactions induced by chemotherapy mainly included gastrointestinal reaction, bone marrow suppression, and liver function injury. No serious adverse reaction occurred. Conclusion:The clinical efficacy of second line TOR combined with NP regimen is similar with the first line regimen for NSCLC patients, especially for SCC patients. The frequency of adverse reaction is not increased.

Objective To investigate the effect of fluoride on expression of Runx2 mRNA and protein in bone tissue of rats. Methods Fourteen SD rats were randomly divided into two groups: control group(tap water with fluoride ＜ 0.06 mg/L), and fluorosis group(fluoride 50 mg/L in water). After 4 moths, expressions of both mRNA and protein of Runx2 in rat bone tissue were determined by RT-PCR and Western blotting. Results The results showed that the expression of Runx2 mRNA and protein in fluoride-treated bone tissue were 2.287 ± 0.261 and 0.929 ± 0.229, respectively, both of which were significantly higher than those of control group(0.995 ± 0.123,0.317 ± 0.068, t = 11.85,6.78, P ＜ 0.05). Conclusions Fluoride can increase the expression of Runx2 mRNA and protein in bone tissue of rats, and Runx2 may be involved in the pathogenesis of bone injury caused by fluoride.

Objective To study the inhibitory effects of stachyose on the hyperacute rejection in pig-to-human heart xenotransplantation.Methods A pig-to-human xenogeneic heart transplantation model was established based on an in vitro free heart blood perfusion system.The pig hearts were di- vided into two groups:group A(pig hearts treated with human blood perfusion as control)and group B(pig hearts treated with human blood plus stachyose perfusion).After perfusion for 1h,the heart xenografts were examined for deposit of lgG and IgM by immunohistochemistry and pathological analy sis.Results The mean survival time of perfusion hearts in groups A and B was(9.5?2.5)min and (46.8?8.1)min respectively(P

Objective To explore a new simpler method for the preparation of recombinant alpha virus as a novel vaccine at the DNA level. Methods Plasmids expressing ? gal protein and helper plasmids were transfected into BHK cells. Virus in culture supernatant of the transfected BHK cells were collected and purified and used to infect BHK cells in vitro to identify the expression of target gene and the titre of the recombinant virus. Results Recombinant virus with high titre, prepared by this method, could be expressed well in mammalian cells in vitro . Conclusion High titre recombinant alpha virus can be produced at the DNA level and this method can be applied for vaccine preparation and gene therapy.

Objective To investigate the biofilm formation,the alginate biosynthetic gene ex- pression and analyze the mucA gene sequence of mucoid Pseudomonas aeruginosa PA17 and nonmu- cold Pseudomonas aeruginosa PA01.Methods The modified plate culture method was used to estab lish the biofilm model in vitro.Semi-quantitative RT-PCR was used to determine the expression level of algD in planktonic condition and during the formation of biofilm.The mucA gene of PA17 and PA01 was amplified and the products were sequenced.Results PA17 biofilm was mature at 6th day, and PA01 biofilm was mature at 3rd day.The structures of the biofilms were both like pellicle.In planktonic condition,the algD expression of PAl7 was higher than PA01;in biofilm formation,the algD expression was maximal when the biofilm was mature.There was a 166～333 deletion mutation and 342A→G in mueA gene of PA17,the mucA gene of PA01 was the same with the sequence of Genbank.Conclusions The mucA gene mutation of PA17 was a new type,which maybe the reason for the little expression difference of algD between PA17 and PA01 during the biofilm formation than it in planctonic condition and the same structure of PA17 and PA01 biofilm.

Background N-ethyl-N-nitrosourea (ENU)-induced mouse mutagenesis is a powerful approach for the study of gene function and the generation of human disease models.Objective This study was to create the corneal morphologic change and map the mutant gene of a kind of corneal opacity in ENU mutagenesis in mouse.Methods ENU was intraperitoneally injected in forty C57BL/ 6J (B6) male mice aged 8-10 weeks old.The male mice were mated with the same strain female mice.Their progenies were screened for visible eye mutation,and the mutant mice were mated with the same strain mice to confirm the heredity of mutation phenotypes.Hematoxylin & eosin staining was used to examine the histopathological change of cornea in one mouse with ENU-induced corneal opacity.To map the mutant gene,[(B6×D2)F1 ×B6] N2 mutant mice were bred,and the genome of the N2 mice was scanned by microsatellite markers distributed equally on the mouse chromosome.The microsatellite linked to the mutant gene was determined by the log odds score.This experimental procedure was approved by Ethic Committee about Experimental Animal Care and Use of Yangzhou University.Results The founder mouse,which was the progeny of an ENU-treated B6 male mouse and an untreated B6 female mouse,had a corneal opacity phenotype.After mating the mutant with B6 mice,19 of 59 descendants appeared corneal opacity phenotype.Thickening of corneal stroma,neoangiogenesis,infiltration of inflammatory cells and proliferation of fibroblasts were exhibited in cloudy cornea in ENU-induced mutated mice under the optical microscope.After linkage analysis between microsatellite markers and the mutant gene,the mutant gene was linked to D2Mi307,which was located at 63.42 cM.Three cases of 26 N2 mice underwent recombination with the LOD 3.79.The mutant gene associated with the cornea phenotype was located on chromosome 2.Conclusions This study map the mutant gene associated with the cornea phenotype on chromosome 2.The strain might be used as a mouse model for heritable human corneal opacity.

Objective: To investigate the pathological changes of human hepatocellular carcinoma (HCC) after continuous passaging in nude mice. Methods: The mice model of HCC SMMU-LTMN were continuously observed for 20 years (1987-2007) . The subcutaneously transplanted carcinoma had been passaged for 228 generations. The pathological data of abdominally transplanted HCC, orthotopically transplanted HCC in nude mice, and orthotopically transplanted HCC in NOD-SCID mice were recorded. The pathological studies were conducted by light microscope, electron microscope, image analysis, chromosomal analysis, and measurement of alpha-fetoprotein (AFP) in peripheral blood. Results: (1) The local invasion and metastasis of of tumors were present in all the above 4 models for a long time. The local invasion rate and the pulmonary metastasis rate of subcutaneously transplanted tumors were 59.70% (40/67) and 37.10% (23/62), respectively. The pulmonary metastasis rate of abdominally transplanted tumors was 59.02%(36/61). The intra-hepatic and pulmonary metastasis rate of the othotopically transplanted tumors were 18.18%(4/22) and 31.82% (7/22), respectively. The pulmonary metastasis rate of HCC in NOD-SCID mice was 53.85%. (2) The tissue structure and the differentiation of the 10th generation tumor cells was similar to those of primary HCC, with grade 2 differentiation and coarse trabecular pattern as the main characteristics. From the 11th generation to the 228th generation, the main characteristics of tumor cells were grade 3 differentiation and lump pattern. Electron microscope also showed worse differentiation. (3)The AFP level was 92 500 ?g/L in cells before the 32th generation; it decreased to 6 729?g/L from the 33th-130th generation cells; and the level of the 220th generation was 1 000-5 000 ?g/L.(4)The DNA contents had a wide distribution (from 2c to 6c) in abdominally transplanted tumors and the pulmonary metastatic tumors; the mean DNA index in the former tumors (2.60?0.20) was wider than the that in the latter (2.10?0.26) . (5)From the 55th generation to 206th generation, it was found that tumor cells had integrated into the chromosome of the nude mice. Conclusion: The subcutaneously transplanted HCC in nude mice can be stably expressed for 20 years, with no change in the local invasion and metastasis ability of HCC. The differentiation of the tumor cells worsenes and the AFP level is decreased in the blood; some chromosome of tumor cells integrate into the chromosome of nude mice, which may be related to the internal environment of nude mice and the multi-potential differentiation of the tumor cells.

Pseudomonas aeruginosa (PA) is a kind of opportunistic pathogen which can cause a wide range of serious infections clinically.These infections are often associated with the formation of biofilm, and are difficult to treat due to the complexity of mechanisms.Some studies have showed that genotype and phenotype of biofilm PA will change, and biofilm PA will aggregate together and secrete extracellular polymeric substance.Therefore, innate immune system could not recognize the camouflaged antigens.Besides, biofilm PA can secrete a variety of virulent factors to hamper the function of innate immune system.This article introduces the main chronic infection caused by PA biofilm and the relationship between biofilm PA and natural innate immune system.