Objective To compare the anti-inflammatory effects between methotrexate(MTX) and Yangqixue Qufengshi Recipe(YQXQFSR) on mice arthritis induced by collagen II.Methods The model of rheumatoid arthritis was induced by using collagen II in C_(57)BL/6?DBA/1 mice.The rats with arthritis were treated with MTX or YQXQFSR respectively.The indexes such as the onset day of collagen induced arthritis(CIA),the level of CII-reactive antibodies and the pathological scores of CIA were assessed.Results The onset day of CIA and the level of CII-reactive antibodies were not influenced in the treatment of mice arthritis with MTX and YQXQFSR respectively.But MTX could alleviate the damage of cartilage [(1.60?1.51) vs(3.56?1.33) scores,P

Objective To investigate the effect of CYP3A5 and C5 gene polymorphisms on the concentration/dose ratio of tacrolimus in Chinese liver transplant patients during the early posttransplantation period.Methods A total of 100 adult patients who underwent primary liver transplantation (LT) were enrolled.Tacrolimus dosage and trough blood concentration were detemined at first week after liver transplantation.Two single-nucleotide polymorphisms were genotyped and analyzed in both donor and recipient groups.The relationship between gene polymorphisms and tacrolimus concentration/dose ratio (C/ D ratio) was analyzed.Results The distribution of allele A in C5 rs17611 was 63.5 ％ among donors and 58.5％ among recipients.For CYP3A5,the rs776746 allele G represented the major alleles in both donors and recipients (71％ and 72％,respectively).The tacrolimus C/D ratio of recipients carrying allele AA in C5 rs17611 was significantly higher than that of recipients carrying the C5 rs17611 allele G.Both donor and recipient CYP3A5 rs776746 polymorphisms were highly correlated with the tacrolimus C/D ratio at first week after liver transplantation.No linkage disequilibrium between CYP3A5 rs776746 and C5 rs17611 polymorphisms was found (D'max =0.392,r2max =0.034).Recipient CYP3A5 rs776746 allele A,C5 rs17611 genotype AA,and donor CYP3A5 rs776746 allele A were associated with rapid tacrolimus metabolism.With increasing number of these alldes,tacrolimus C/D ratio was reduced during the one week after transplantation.Conclusion Recipient C5 rs17611 polymorphism is a new genetic locus that influences tacrolimus metabolism in patients after OLT during the early post-transplantation periocd.

A 68?year?old female patient was admitted to the hospital for multiple masses in the mouth and lungs as well as on dorsal hands for more than 20 days without obvious subjective symptoms. No abnormalities were found by physical examination. Dermatological examination showed two bean?sized dark?red nodules on the upper jaw as well as one pigeon egg?sized dark?red nodule on the left dorsal hand, and all the nodules were hard with smooth surfaces and limited mobility. Positron emission tomography?computed tomography (PetCT) revealed multiple metastases to the brain, lymph nodes, lungs, gastrointestinal tract, both kidneys, multiple bones and intermuscular tissues. Pathology of nodules from the upper jaw showed lowly differentiated tumor cells with osteoid matrix, chondroid structures and tumor bone in local areas, and immunohistochemical examination of tumor cells found positive staining for S100(focally), vimentin, CD99, P63 and Ki?67(60%), but negative staining for keratin. A diagnosis of osteosarcoma of the right side of the upper jaw was considered. Pathology of nodules from the dorsal hand revealed no obvious abnormalities in the epidermis, while there was a diffuse infiltration of medium?to large?sized histiocyte?like cells in the whole dermis with cell atypia and irregularly red?stained bone matrix and tumor bone in some regions. Immunopathology showed positive staining for Ki67(60%), and negative staining for CD3, CD10, CD20, Bcl?2, and Bcl?6. A diagnosis of cutaneous metastasis of osteosarcoma was made. The patient refused further treatment and died 6 months after the onset of lesions.

OBJECTIVE:To investigate the association of Cytochrome P4503A family polypeptide 5 [(CYP)3A5] and IL-10 poly-morphisms with the concentration/dose(C/D) ratio of tacrolimus in lung transplantation recipients. METHODS:CYP3A5 polymor-phisms at rs774746 and IL-10 polymorphisms at rs1800896 from peripheral blood cell of 51 lung transplant recipients was detected by Se-quenom MassArray iPLEX GOLD system. The relationship between gene polymorphism and tacrolimus C/D ratio was analyzed. RE-SULTS:The tacrolimus C/D ratio of the recipients who carried CYP3A5 rs776746 GG genotype were higher than those with AA+GG genotype,with statistical significance(P<0.01). The tacrolimus C/D ratio of the recipients who carried IL-10 rs1800896 AA genotype were higher than those with AG genotype during 1 week and 2 weeks after transplantation,with statistical significance(P<0.05). Com-bined analysis of these two genes,the tacrolimus C/D ratio of the recipients who carried more fast metabolic alleles were higher than those who didn't carry fast metabolic alleles,with statistical significance(P<0.01). CONCLUSIONS:Determination of recipients CYP3A5 at rs 776746and IL-10 at rs800896 genotype may be performed prospectively to help individualizing tacrolimus dose regimen.

Objective:To observe the effect of icariin on the osteoporosis of the ovariectomized rats,and to explore the mechanism.Methods:Fifty female rats aged 6 months were randomly divided into sham operation group, model group,positive drug group,low dose of icariin group and high dose of icariin group (n= 10).The rats in positive drug group were given with 1 mg· kg-1 nilestriol every week;the rats in low and high doses of icariin groups were given with 50 and 100 mg · kg-1 icariin every day.The bilateral ovaries of rats were excised by operation to establish the osteoporosis models.2 weeks after operation,the rats were treated with icariin for 12 weeks,and then they were sacrificed by drawing blood from abdominal aorta under anesthesia condion.The bone mineral density (BMD),serum biochemical indicators,bone histomorphology and the expressions of apoptosis-related proteins were detected.Results:Compared with sham operation group,the BMD of the rats in model group was decreased (P <0.01),the serum calcium level was decreased (P <0.05),and the serum phosphorus level was increased (P <0.05),and the serum BGP,ALP,NO and NOS levels were significantly decreased (P <0.05 or P <0.01).Compared with model group,the BMD of the rats in positive drug group and high dose of icariin group were obviously increased (P < 0.01 ), the serum calcium levels were increased, the phosphorus levels were decreased (P <0.05),and the BGP and ALP levels were increased significantly (P < 0.05 ). Compared with control group,the cortical in model group was thinned,the width of bone trabecula was reduced,the Bax and Caspase-3 expression levels in bone tissue were increased,and the Bcl-2 expression level was decreased.Compared with model group,the cortical in icariin group was thicked,the width of bone trabecula was increased,the Bax and Caspase-3 expression levels were decreased,and the Bcl-2 expression level was increased (P <0.05 or P <0.01). Conclusion:Icariin can protect the osteoporosis of ovariectomized rats,the function may be related to the inhibition of apoptosis.

Objective:To evaluate the correlation of inter-individual variation of cyclosporine dosage and blood concentration and CYP3A4 and CYP3A5 polymorphism in renal transplant recipients. Methods:Two hundred and twenty-one renal transplant recipients treated with cyclosporine were genotyped for CYP3A4 rs4646437C>T and CYP3A5 6986G>A using ligase detection reactions. The effects of genetic polymorphisms of CYP3A4 and CYP3A5 on cyclosporine trough concentration (C0/D) and 2 h post-dose concentra-tion (C2/D) during the period of 6 months, 6-24 months and above 24 months after renal transplant were studied. Results:CYP3A5 6986G>A genotype affected C0/D during the period of 6 months, 6-24 months and beyond 24 months (P<0. 05) with the order of CYP3A5 6986GG>GA>AA. CYP3A4 rs4646437C>T and CYP3A5 6986G>A genotype affected C2/D during the period above 24 months (P<0. 05) with the order of CYP3A4 rs4646437 CC>CT>TT and CYP3A5 6986GG>GA>AA. Conclusion: CYP3A5 6986G>A genotype affects C0/D and C2/D of cyclosporine, and CYP3A4 rs4646437 C>T genotype affects C2/D of cyclosporine. The effects of genotypes are varied in different stages.

Objective To screen for and validate unknown tumor suppressor genes (TSGs) in sporadic colorectal cancer (CRC) patients.Methods Through loss of heterozygosity (LOH) analysis on chromosome 10 in sporadic CRC,we have found D10S185 (10q23.31-24.33 ) exhibit a higher LOH frequency in our previous study.In present study,we screen for unknown TSGs in this region through the microarray.The expression of the new gene was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR).RT-PCR,immunohistochemistry and Western blot were done in colorectal cancer tissues with their pair-matched normal tissues in 50 cases to validate the results of microarray.Results Through the microarray-based high throughput screening,we found 4 significant down-regulated genes:PLCE1,CPEB3,NKX2-3 and SEMA4G,among them the down-regulation of PLCE1 was most significant.The results of qRTPCR were in relative agreement with the DNA microarray data.RT-PCR,immunohistochemistry and Western blot also showed that the expression of PLCE1 was at low levels in 46％ cancer tissues compared with normal tissues,more frequent in the poor differentiation tumor in patients under age 60 years (P ＜ 0.05 ).Conclusions This study demonstrated that down-regulation of PLCE1 was related to the tumorigenesis of sporadic colorectal cancer.PLCE1 might play a suppressive role in the development of colorectal cancer.

Objective To investigate the relationship between high-glucose-induced fibronectin(FN) expression and up-regulation of integrin-linked kinase(ILK) in human kidney tubular epithelial cells (HKC) and kidney of CD-1 mice. Methods Cultured human kidney tubular epithelial cells and streptozotocin (STZ)-indueed diabetic model of CD-1 mice were enrolled in this study.Western blot was used to detect the expression of FN and ILK.The kinase dead ILK plasmid (pCMV-kdlLK) were transferred to HKC. Results Four weeks after injection of STZ,CD-1 mice had higher blood glucose level as compared to the control [(20.3±2.7) mmol/L vs (6.1±1.4) mmol/L,P＜0.01].Meanwhile,expression of FN and ILK was significantly increased in diabetic mice as compared to the control (P＜0.01).There was positive correlation between the expression of FN and ILK (r=0.899,P＜0.01).High-glucose could up-regulate FN and ILK expression in cultured HKC in a time- and dose-dependent manner.Blockage of ILK activation by pCMV-kdILK abrogated high-glucose-incuced FN expression in HKC. Conclusions Highglucose can induce FN expression through up-regulating ILK expression.Blockage of ILK activation abrogates this effect.

Objective To investigate the effect of high glucose on renal tubular epithelial-mesenchymal transition,and to analyze the relationship between high glucose and transforming growth factor β1(TGF-β1)and the mechanism of renal interstitial fibrosis. Methods HKC and Smad7-overexpression HKC cells were grown in DMEM/F12 medium containing 5%～10%newborn calf serum.They were cultured for 16 h in free serum medium after 80%cells were adhered onto the surface of the flask.Afterwards,they were stimulated by high glucose(glucose concentration:25 mmol/L and 50 mmol/L).The expression of α-SMA,E-cadherin and fibronectin was detected by Western blot while the supernatant level of TGF-β1 was detected by ELISA.Cell motility and migration was evaluated using Boyden chamber motogenicity assay. Results In HKC induced by high glucose,the expression of α-SMA and fibronectin protein was highly upregulated while the expression of E-cadhefin protein was down-regulated.The expression of TGF-β1was up-regulated in a dose-dependent manner.These above-mentioned effects could be obviously inhibited by anti-TGF-β1 antibody.The protein expression of α-SMA,fibronectin and E-cadherin had no obvious change in Smad7-overexpression HKC induced by high glucose.HKC exhibited enhanced motility and invasive capacity in high glucose groups,compared to that in control group.Migrated cell counting was(12.4±3.7)and(18.6±4.4)cell/HP in 25 and 50 mmol/L glucose groups respectively. Conclusion High glucose may induce renal tubular epithelialmesenchymal transition through TGF-β1 pathway,which can be inhibited by blocking the Smad signal pathway.

Nonalcoholic fatty liver disease (NAFLD) is the result of the interaction between many factors, and “second hit” focusing on oxidative stress is a key factor for the progression of NAFLD. Autophagy has been a research hotspot in recent years, and more and more studies have shown that autophagy plays an important role in lipid metabolism in hepatocytes, inflammatory response, and liver fibrosis and is closely associated with oxidative stress. With reference to the latest research findings in China and foreign countries, this article analyzes the interrelation between autophagy and oxidative stress in the pathogenesis of NAFLD from the aspects of the association between autophagy and NAFLD and the association between oxidative stress and NAFLD. It is pointed out that the molecular mechanism of the regulation of oxidative stress by autophagy in the pathogenesis of NAFLD may be a research hotspot in future, and blockade or activation of a key pathway involved in the regulation of oxidative stress by autophagy may provide a new method for the treatment of NAFLD.