Objective:To observe the expression of microRNA (miRNA, miR) -7850 in renal cancer tissues, and to explore the effect of miR-7850 on the proliferation and migration of renal cancer cells and on the regulation of serine proteinase inhibitor B3 (SERPINB3) gene expression.Methods:Real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-7850 in renal cancer tissues and renal cancer cell lines. The renal cell carcinoma cell line with the lowest expression of miR-7850 was selected, and the negative control sequence (miR-NC) and miR-7850 mimics were transfected into renal cell carcinoma cells by Lipofectamine 2000 transfection reagent, respectively, which were defined as miR-NC group and miR-7850 group. qRT-PCR was used to detect the expression of miR-7850 in transfected renal cancer cells. The cell proliferation and migration ability after transfection were detected by cell counting kit-8 (CCK-8) method and transwell experiment. Bioinformatics prediction and dual luciferase reporter gene experiments were used to verify the target gene of miR-7850. qRT-PCR and Western blot were used to detect the expression of target genes in renal cancer cells after transfection.Results:Compared with adjacent tissues (5.95±0.44), the expression of miR-7850 in kidney cancer tissues (1.19±0.33) was lower ( P<0.01). Compared with immortalized proximal renal tubular epithelial cells (1.01±0.07), the expression of miR-7850 was lower in renal cancer cell lines ( P<0.05), and the lowest in A498 cells (0.13±0.01) ( P<0.01). The expression of miR-7850 in the miR-7850 group (7.46±0.93) was significantly higher than that in the miR-NC group (1.01±0.08) ( P<0.01), indicating successful transfection. Compared with the miR-NC group, the cell proliferation ability of the miR-7850 group was significantly reduced ( P<0.05). The number of migrating cells in miR-NC group and miR-7850 group were (139.50±12.31) and (75.09±16.05) cells, respectively, and the cell migration ability in miR-7850 group decreased significantly ( P<0.01). Bioinformatics technology shows that the target gene of miR-7850 was SERPINB3. The dual luciferase reporter gene experiment confirmed that miR-7850 can target the SERPINB3 gene ( P<0.05). Compared with the miR-NC group, the expression of SERPINB3 in cells of miR-7850 group was significantly reduced ( P<0.05), as well as the CDK4, CyclinD, Snail and Vimentin. Conclusions:miR-7850 is lowly expressed in renal cancer tissues and cell lines. miR-7850 can inhibit the proliferation and migration of renal cancer A498 cells, which may be related to its inhibition of SERPINB3 gene expression.

The optimized cultural medium of fermentation for Candida sp.mutant strain YQ5 to produce S-adenosylmethionine was studied.The results of single factor experiment showed that the most favorable pH value,carbon source,nitrogen source organic nutrient is 6.0,8% sucrose,1.5% tryptone and 2% yeast extract,respectively.As to inorganic salts,MgSO4?7H2O,CaCl2,FeSO4?7H2O,CoCl2,CuSO4?5H2O,H3BO3 could improve accumulation of the intercellular SAM.The ingredients of the culture medium are also opti-mized by the orthogonal experiment.Fermentation for 48 h under the optimal condition resulted in AdoMet production at 1740.0 mg/L.

Objective:To investigate the effects of musk-1, a glucoprotein component isolated from the water extract of musk, on some functions of rat polymorphonuclear leukocytes activated by IL-8 in vitro.Method：An in vitro incubation system was used. Superoxide anion production was determined by cytochrome C reduction. β-glucuronidase and lysozyme release was quantitated by enzyme reactions in which phenolphthaleinglucuronic acid and Micrococcus Lysodeikticus were as the substrates, respectively.Results:In comparison with control, musk-1 at concentration 1～100 μg*ml-1 can increase superoxide anion production by 91.7%～291%, and decrease β-glucuronidase and lysozyme release by 2.2%～58.1% and 3.9%～39.8%, respectively.Conclusion:Inhibition of lysosomel enzyme release might be considered as one of mechanisms of antiinflammatory action of musk.

AIMTo investigate the expression of matrix metalloproteinase-9 (MMP-9) in mouse ears induced with croton oil and the inhibitory effect of dexamethasone, indomethacin and resveratrol on MMP-9 expression, and further explore the relationship between anti-inflammation and MMP-9 inhibition of these three medicines.

METHODSImmuno-histochemistry was used to detect the expression of MMP-9 in mouse ears. Expression of MMP-9 in U937 cells was analyzed by gelatin zymography.

RESULTSMouse ear edema induced with croton oil was inhibited significantly by dexamethasone and indomethacin at the dose of 10 mg.kg-1 and resveratrol at 50 mg.kg-1 administered subcutaneously. The inhibitory rate was 76.2% (P < 0.001), 56.7% (P < 0.001) and 36.9% (P < 0.001) respectively. The MMP-9 expression increased in mouse ears induced with croton oil and inhibited by dexamethasone, indomethacin and resveratrol at above doses. Gelatin zymography results showed that MMP-9 expression in U937 cells increased significantly after exposed to PMA at 1 x 10(-8) mol.L-1 (P < 0.001); MMP-9 expression induced with phorbol myristate acetate(PMA) was inhibited by dexamethasone at 1 x 10(-9), 1 x 10(-7) and 1 x 10(-5) mol.L-1, indomethacin at 1 x 10(-6) and 1 x 10(-5) mol.L-1 and resveratrol at 1 x 10(-6) and 1 x 10(-5) mol.L-1.

CONCLUSIONThe inhibition of MMP-9 expression may be one of the anti-inflammatory mechanisms of dexamethasone, indomethacin and resveratrol.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Croton Oil ; Dexamethasone ; pharmacology ; Ear Diseases ; chemically induced ; metabolism ; Edema ; chemically induced ; metabolism ; Humans ; Indomethacin ; pharmacology ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Matrix Metalloproteinase Inhibitors ; Mice ; Mice, Inbred ICR ; Random Allocation ; Stilbenes ; pharmacology ; U937 Cells ; metabolism

AIMTo investigate the stepwise development and germ cell gene expression in allografted neonatal mouse testes and the differentiation of immature human testicular cells in xenografted human testes.

METHODSImmunodeficient nude mice were used as hosts for allografting of neonatal mouse testes and xenografting of human fetal testicular tissues. Stepwise development and stage-specific gene expression of germ cells in allografts were systematically evaluated and parallel compared with those in intact mice by periodically monitoring the graft status with measurement of graft weight, histological analysis and determination of five stage-specific genes. Human testicular tissues from 20 and 26 weeks fetuses were used for the xenografting study. Histological analysis of xenografts was performed 116 and 135 d after the grafting procedure.

RESULTSIn the allografting study, progressive increase in tissue volume and weight as well as in tubule diameter in grafts was observed; the appearance time of various germ cells in seminiferous tubules, including spermatogonia, spermatocytes, round and elongate spermatids and sperm, was comparable with that in intact donors; the initiation of gene transcription in grafts showed a similar trend as in normal mice. Graft weight ceased to increase after 7-8 weeks and degradation of grafts was observed after 5 weeks with progressive damage to seminiferous epithelium. In the xenografting study using immature human testicular tissues, graft survival and development was indicated by increasing graft weight, Sertoli cells differentiation into advanced stage, germ cells migration and location to the basal lamina and formation of a niche-like structure.

CONCLUSIONThe developmental course and gene expression pattern of germ cells in allografts were similar to those in intact mice. The best time point for retrieval of mouse sperm from grafts was 5-7 weeks after grafting procedure. An accelerated development of immature human testicular cells could be achieved by ectopic xenografting of human testes.

Animals ; Animals, Newborn ; Base Sequence ; DNA Primers ; Gene Expression Profiling ; Humans ; Immunologic Deficiency Syndromes ; physiopathology ; Male ; Mice ; Mice, Inbred BALB C ; Testis ; growth & development ; metabolism

AIMTo study the effects of indomethacin on interleukin-6 (IL-6) expression stimulated with lipopolysaccharide (LPS) in rheumatoid arthritic patients' synoviocyte.

METHODSFibroblast-like cells (FLS) from rheumatoid arthritic patients' joint tissue were cultured for 24 h and incubated 24 h with LPS (1 mg.L-1) or the supernatant of U937 cells stimulated by LPS (1 mg.L-1). After indomethacin or dexamethasone added into the supernatant of U937 cells, FLS was incubated with the super natant for 24 h. The expression of IL-6 protein was detected by radioimmunoassay. The mRNA expression of IL-6 was accessed by RT-PCR.

RESULTSLPS did not obviously affect the growth of FLS, and the protein secretion and mRNA expression of IL-6 were not changed in FLS treated with LPS. The IL-6 secretion and IL-6 mRNA expression were significantly increased in FLS cultured with the supernatant from U937 cell treated with LPS. Indomethacin at concentrations of 1 x 10(-7)-1 x 10(-5) mol.L-1 obviously inhibited the protein secretion and mRNA expression of IL-6 in FLS cultured with the supernatant from U937 cell stimulated with LPS, and the inhibitory effects increased as the concentrations of indomethacin increased.

CONCLUSIONIndomethacin can inhibit the increase of IL-6 expression caused by supernatant of U937 cells stimulated with LPS in FLS.

Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Arthritis, Rheumatoid ; metabolism ; pathology ; Cells, Cultured ; Fibroblasts ; metabolism ; pathology ; Humans ; Indomethacin ; pharmacology ; Interleukin-6 ; biosynthesis ; genetics ; Lipopolysaccharides ; pharmacology ; RNA, Messenger ; genetics ; Synovial Membrane ; metabolism ; pathology ; U937 Cells

The aim of this study was to investigate the expression of integrin linked kinase (ILK) and protein kinase B (PKB/Akt) in acute leukemia (AL), explore the possible effects of ILK/Akt pathway on AL pathogenesis. The expression level of ILK mRNA and Akt mRNA in different types and stages of AL bone marrow mononuclear cells was detected by reverse transcription polymerase chain reaction (RT-PCR). The results showed that the expression of ILK and Akt in do nove AL group was higher than that in AL-CR group and normal control group (P < 0.05), while there was no statistically difference between de nove AL and relapsed AL groups (P > 0.05). The expression of ILK positively correlates with Akt expression in de nove AL group (P < 0.05). It is concluded that the expression of ILK and Akt mRNA is abnormally higher in AL, ILK/Akt pathway may be involved in the pathogenesis and progression of AL and may be a potential therapeutic target for AL.
Adolescent ; Adult ; Aged ; Bone Marrow Cells ; metabolism ; Female ; Humans ; Leukemia ; metabolism ; pathology ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA, Messenger ; genetics ; Young Adult

Female ; Humans ; Liver Failure, Acute ; blood ; Liver Neoplasms ; blood ; Liver Transplantation ; Male ; Thrombelastography ; Whole Blood Coagulation Time

OBJECTIVETo establish a murine model of sensitization, and investigate the effect and mechanism of sensitization on allogeneic donor bone marrow cells (BMCs).

METHODSSensitized BALB/c mice were established by transfusions of allogeneic splenocytes. The donor reactive antibodies were detected by binding and complement-dependent cytotoxicity assays. After irradiation, 1 × 10(7) BMCs of C57BL/6 donor mice were injected into non-sensitized or sensitized BALB/c recipient mice. The distribution pattern of donor BMCs in peripheral blood, spleen and bone marrow of recipient mice were analyzed at different time points (2 h, 12 h and 48 h) post transplantation. Hematopoietic recovery post transplantation was assessed, and survival was monitored. Moreover, sera and splenocytes derived from non-sensitized or sensitized recipients were incubated with allogeneic BMCs in vitro, and the cytotoxic indexes were calculated in the immune experiments.

RESULTSThe binding and complement-dependent cytotoxicity assays showed that a high level of donor reactive antibodies was presented in sensitized sera. Compared with the non-sensitized recipients, the homing assay showed significantly decreased distributions of allogeneic donor BMCs in peripheral blood, spleen and femur of sensitized recipients. Non-sensitized recipients survived long term after irradiation, while all the sensitized recipients died within 12-15 days. Fourteen days post transplantation, the white blood cells and BMCs of non-sensitized recipients were (3240 ± 300) × 10(6)/L and (396 ± 27) × 10(6)/femur, respectively; while the white blood cells and BMCs of sensitized recipients were (320 ± 80) × 10(6)/L and (6 ± 2) × 10(6)/femur, respectively; the differences were statistically significant between this two groups (P < 0.05). Seven days post transplantation, the percentage of donor cells in bone marrow of non-sensitized and sensitized recipients was (48.07 ± 4.70)% and (0.77 ± 0.11)%, respectively, and the differences were statistically significant (P < 0.05). Furthermore, the white blood cells and BMCs following transplantation decreased along with time in sensitized recipients. The immune experiments of complement-dependent cytotoxicity reaction, cytotoxic T lymphocytes reaction and antibody-dependent cellular cytotoxicity showed the cytotoxic indexes were higher in sensitized group than the non-sensitized group.

CONCLUSIONA sensitized model was established by transfusions of allogeneic spleen cells. Allogeneic donor BMCs were rejected in sensitized recipients, and its mechanism might be through immune impairment pathways.

Animals ; Bone Marrow Cells ; Bone Marrow Transplantation ; Disease Models, Animal ; Graft Rejection ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Tissue Donors ; Transplantation, Homologous

This study was aimed to investigate the expression of SALL4 and BMI-1 mRNA in acute leukemia (AL) and its clinical significance. mRNA expression levels of SALL4 and BMI-1 in 62 AL patients and 10 controls were measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The results showed that (1) the expression of SALL4 mRNA were up-regulated in AL (except M3 and T-ALL), and the expression of of SALL4 mRNA in AML was significantly higher than that in B-ALL (p<0.05); expression was negative in 9 out of 10 controls, and expression level of SALL4 mRNA in one case was low; (2) the expression of SALL4 in de novo AL was higher than that in controls, which significantly decreased at complete remission (CR). The difference between CR group and de novo group was statistically significant (p<0.05). In relapsed patients, the expression of SALL4 increased, slightly higher than that in de novo AL group (p>0.05); (3) the expression pattern of BMI-1 was the same to that of SALL4 except the up-regulation in M3, and the expression of BMI-1 was positively correlated with that of SALL4 in acute leukemia (r=0.684, p<0.01). It is concluded that SALL4 and BMI-1 expressions are up-regulated significantly in acute leukemia, which may contribute to the development of leukemia, thus become an important index for evaluation of development, relapse and prognosis in acute leukemia.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Leukemia ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Nuclear Proteins ; genetics ; metabolism ; Polycomb Repressive Complex 1 ; Prognosis ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; Repressor Proteins ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism ; Young Adult