Aim To establish an allogenetic mouse skin trans-plant model,in order to provide a research model for immunosup-pressive drugs. Methods Skins from the ears of C57BL/6 mice were transplanted to the back of BALB/c mice and skin isografts ( BALB/c mice to BALB/c mice) were used as control. Cyclos-porin A( CsA) was used as a model compound to test the imm-nosuppresive effect on allogenetic graft rejection. Following the transplation and CsA treatment, the graft rejection score and graft skin survival rate were quantified. Four and nine days after transplantation,serum IL-4,IL-12 and IFN-γ levels were meas-ured using ELISA kits. Twelve days after transplantation, mice were sacrificed. The weight of spleen and thymus was obtained, and CD4 + and CD8 + population of spleenic T cells were ana-lyzed using flow cytometer. Histological features were assessed by hematoxylin-eosin( HE) staining of formalin-fixed, paraffin-em-bedded graft skins. Results After transplantion, the graft rejec-tion score increased and graft skin survival rate decreased gradu-allly. Serum IL-12 and IFN-γ levels of allograft mice increased markedly. Compared with those of isograft mice, mice with skin allograft displayed a significant increase in the percentage of the CD8 + T cell subpopulation. Remarkable inflammation, such as edema, inflammatory cell infiltration were observed in allograft mice. Compared with saline treated mice, CsA significantly re-duced the graft rejection score and improved survival rate of skin grafts. And also, CsA treated mice had smaller spleen and thy-mus. Mice that received high doses of CsA had significantly less CD8 + T cells than those treated with saline. Moreover, allograft skins in mice that received CsA had less inflammation. Conclu-sions Allogenetic mouse skin transplantation exhibits acute graft rejection. CsA can inhibit the rejection in a dose dependent manner.

Aim To investigate the impact of CYP3 A5 genetic polymorphism on modified releasing tacrolimus pharmacokinetics in Chinese stable renal transplant re-cipients. Methods Pharmacokinetics of once daily-ta-crolimus( tac-q. d. ) and twice daily-tacrolimus( tac-b. i. d. ) were determined by CLIA, CYP3A5 genotype was measured by PCR-RFLP. Each 10 patients receiv-ing tac-q. d. and tac-b. i. d. respectively were en-rolled, and each 5 patients receiving tac-q. d. were matched to poor metabolizer ( PM ) and extensive me-tabolizer ( EM ) group respectively according to CYP3A5 genotypes. Results AUC0~24 h for tac-q. d. was 1. 78 folds higher than AUC0~12 h for tac-b. i. d. , and dose-adjusted C0 was 40% lower for tac-q. d. than for tac-b. i. d. There were no significant differences for other parameters between the two groups; Cmax, AUC0~24 h and C0 were 1. 75, 1. 96 and 2. 49 folds higher for PM than for EM, and dose-adjusted Cmax, AUC0~24 h and C0 were 1. 80, 2. 34 and 2. 64 folds higher for PM than for EM. There were good correla-tions between AUC0~24 h and C0 for tac-q. d. Conclu-sion Conversion from tac-b. i. d. to tac-q. d. results in requirement of increased tacrolimus dose and detec-tion of CYP3A5 genotype, which is necessary for ensu-ring C0 in the range of therapeutic window.

Aim To investigate the effects of berberine combined with bone marrow mesenchymal stem cells ( BM-MSCs) on the energy metabolism of human um-bilical vein endothelial cells ( HUVECs ) under the condition of high glucose. Methods ①The state of cell reproduction and cell proliferating activity were de-termined by MTT assay. ②The cell cycle was detected by flow cytometry ( FCM ) . ③DNA damage of cells was measured by comet tail assay. ④The contents of ATP, ADP and AMP were determined by high perform-ance liquid chromatography ( HPLC ) and the level of energy charge ( EC) was calculated. ⑤The expression of CCR and COX mRNA was detected by reverse tran-scription-polymerase chain reaction ( RT-PCR) . ⑥The expressions of COX and CCR were detected by Western blot. Results ①The proliferating activity of HUVECs declined apparently and the proliferation decreased af-ter high glucose intervention. Meanwhile, the quantity of cells during S +G2 dropped dramatically and there was certain degree of damage to DNA. The berberine and BM-MSCs respectively improved the proliferating activity and the proliferation in different degrees, in-creased the quantity of cells during S+G2 and promo-ted the repair of DNA ( P <0 . 01 ) , and so did the combination of the two, with a better effect than each of them alone. ②After high glucose intervention and the damage caused, the content of both ATP and ADP of HUVECs was reduced, and EC level also declined significantly, while the content of AMP increased. The berberine and BM-MSCs respectively up-regulated the content of ATP and ADP ( P<0. 01 ) , and so did the combination of the two, with a better effect than each of them alone. ③After high glucose intervention and the damage caused, the expression of COX, CCR mR-NA and protein decreased obviously. Yet, all of the three gained a dramatic increase when the berberine, BM-MSCs or the combination of the two were added ( P<0. 01 or P <0. 05 ) , among which the combination worked more effectively. Conclusions The berber-ine, BM-MSCs and the joint use of the two could im-prove the energy metabolism of HUVECs, which had been damaged by high glucose, probably because the berberine and BM-MSCs could up-regulate the expres-sion of COX, CCR mRNA and protein, which leads to the hydrolyzation of glucose oxide and thus the im-provement of blood environment and the enhancement of glucose's supply and intake of HUVECs. Then, here comes the final result: the improvement of the energy metabolism of damaged vascular endothelial cells by high glucose.

Aim To study the preventative effects of noninvasive delayed limb ischemic preconditioning ( NDLIP) on sudden cardiac death in rats with myocar-dial infarction. Methods Thirty healthy SD male rats weighting ( 250 ± 10 ) g were randomly divided into 3 groups:① myocardial infarction ( MI ) group: animal model of MI was established by making surgical ligation of animal LAD. ② MI plus NDLIP group: after the success of the animal model of MI, NDLIP was carried out every other day until 4th week. ③Sham group:as the negative control group, animals were taken heart LAD threading but no ligation. All rats were fed con-ventionally. At the end of 4 weeks, three groups of rats were administered with metaraminol ( 0. 2 mg · min-1 ) . ECG, drug cumulant of sudden death and death onset time were recorded. After sudden death, blood samples were withdrawn from abdominal aorta and serum was separated via centrifugation. ELISA method was used to measure serum caspase-3 , HSP70 and SOD concentration. Results While metaraminol led animal cardiac sudden death, the rats heart rate ( HR) kept declining with the increase of dosage of metaraminol during the administration period. Rat HR of MI+NDLIP group [ ( 479 ± 8 ) vs ( 416 ± 19 ) beat ·min-1 , ( 446 ± 32 ) vs ( 370 ± 20 ) beat · min-1 , (376 ± 53) vs (305 ± 29) beat·min-1, (307 ± 63) vs (244 ± 33) beat·min-1, (283 ± 45) vs (121 ± 35 ) beat · min-1 , P <0. 01 ] was markedly higher than that of MI group at 0 , 5 , 10 , 30 , 50 min before death. Compared with MI group, drugs cumulant to sudden death and death onset time of MI + NDLIP group [ ( 14. 58 ± 3. 03 ) vs ( 10. 76 ± 2. 73 ) mg, (72. 9 ± 15. 2 ) vs ( 53. 8 ± 13. 6 ) min, P <0. 01 ] were significantly increased. Compared with MI group, serum caspase-3 content of MI+NDLIP group was sig-nificantly reduced [ ( 2. 01 ± 0. 52 ) vs ( 2. 34 ± 0. 38 )μg·L-1 , P<0. 01 ]; HSP70 levels were remarkably increased [ ( 3. 01 ± 0. 58 ) vs ( 2. 70 ± 0. 43 ) μg · L-1 , P <0. 05 ]; SOD levels were significantly im-proved [(1. 99 ± 0. 65) vs (1. 70 ± 0. 58) mg·L-1, P<0. 01 ] . Conclusion NDLIP can prevent sudden cardiac death after myocardial infarction in rats, which may be mediated by reducing the myocardial cell apop-tosis, increasing protective protein expression and en-hancing antioxidant capacity.

Aim To investigate anti-hepatic fibrotic mechanism of Acanthus ilicifolius alkaloid A ( HBOA ) involved in high mobility group box 1 ( HMGB1 ) . Methods A hepatic fibrosis model of rat was estab-lished by the olive oil of CCl4 for 12 weeks. Then, at the 8th week,the successful model rats were randomly divided into model control group, colchicine group, HBOA high-dose group and HBOA low-dose group. From the 9th week,the rats in each group were treated with the drugs daily for 4 weeks respectively. The changes of liver histopathology and collagen were ob-served by HE staining and Masson staining, and the serum indicators including aspartate aminotransferase (AST),alanine aminotransferase(ALT) , total biliru-bin ( T-BIL ) , HMGB1 , interleukin-1β( IL-1β) and tumor necrosis factor-α( TNF-α) were determined. Moreover , the protein of HMGB1 in liver was examined by immunohistochemistry, and the expression of HMGB1 mRNA was measured by real-time fluores-cence quantitative PCR. Results Compared with the model control group,HBOA high-dose and HBOA low-dose groups significantly attenuated the fibrotic degree induced by CCl4 , markedly decreased the levels of ALT, AST, T-BIL, HMGB1, IL-1β, TNF-α. Moreo-ver, the expression of HMGB1 protein and mRNA in liver was decreased. And furthermore, serum HMGB1 level had significant positive correlation with IL-1β, TNF-α,ALT,AST and T-BIL. Conclusion HBOA has beneficial effects against liver fibrosis in rat which is induced by CCl4 , the mechanisms may be related to the inhibition of inflammatory response to HMGB1 .

Aim To observe the effects of glucose-stim-ulated insulin secretion ( GSIS ) on rat islets after S1 P treatments and the underlying molecular mechanisms. Methods Collagenase P and Histopaque 1077 were used to digest and isolate rat pancreatic islets, and Dispase II was used to digest pancreatic islet to obtain pancreatic cells. Insulin secretions were measured after S1P (0~20 μmol·L-1 ) treatment under low glucose ( LG, 2. 8 mmol·L-1 ) and high glucose ( HG, 16. 7 mmol·L-1 ) conditions. Patch-clamp recordings were applied to monitor voltage-dependent potassium chan-nel currents (Kv currents) after S1P treatment. Calci-um image technique was used to measure the changes of intracellular Ca2+ concentration after S1P ( 0 ~20μmol·L-1 ) treatments. Results HG group signifi-cantly increased insulin secretion compared to LG group ( P<0. 01 ) . S1 P had no effect on insulin se-cretion under LG condition ( P>0. 05 ) . S1 P increased insulin secretion significantly in a dose-dependent man-ner under HG condition ( P<0. 01 ) . Kv currents ofβcells were inhibited significantly after S1 P treatment ( P<0. 01 ) . S1 P increased the concentrations of in-tracellular Ca2+ in a dose-dependent manner under HG condition( P <0. 01) . Conclusion S1P may pro-mote GSIS by inhibiting Kv currents and increasing the level of intracellular Ca2+.

When senescence induction is based on DNA damage, senescent cells display a unique phenotype, which has been termed “senescence-associated secretory phenotype”( SASP ) . SASP, including proinflammatory cytokines, growth factors, chemokines, matrix remodeling enzymes and other cytokines, may be an important driver of chronic inflammation and therefore may be part of a vicious cycle of inflammation, DNA damage and senescence. Senescence-associated secretory products released by such cells can affect the neighboring cells and further exacer-bate their regenerative capacity. SASP is associated with many chronic age-related diseases.

Nicotinamide adenine dinucleotide phosphate oxidase ( NOXs) contributes to the production of reactive oxygen species ( ROS) in liver fibrosis, resulting in the activation of endoplas-mic reticulum stress ( ERS ) and IRE1α-XBP1 signaling path-way. ROS is a series of oxygen metabolites and its derivatives, produced by the single electron reduction of molecular oxygen ( O2 ) , including superoxide anion ( O2- ) , hydroxyl radical (-OH) , hydrogen peroxide ( H2 O2 ) , hypochlorite ion ( OCl-) and so on. They can interact with a large number of molecules, including small inorganic molecules, proteins, lipids, carbohy-drates and nucleic acids, resulting in lipid peroxidation of cell damaging molecules. And as a second messenger, ROS can also affect the proliferation and activation of HSC in liver fibrosis, and induce the hepatocyte apoptosis through a variety of cellular signal transduction. Here we review the current status of the study on the mechanism of NOXs in liver fibrosis.

Glypican-3 ( GPC3 ) plays very important role in the regulation of cell growth and differentiation in hepatocellular car-cinoma ( HCC ) . GPC3 is closely related to the occurrence and development of HCC. A dramatic elevation of GPC3 expression has been reported in a large proportion of HCC, which suggests that GPC3 is remarkably sensitive and specific to the diagnosis of HCC. GPC3 is a potential therapeutic target of HCC. This paper reviews the structure and function of GPC3, the progress of im-munotherapy based on GPC3 of HCC, and discusses the prospect of therapeutic target of liver cancer in the future.

Aim To establish ICR animal model with C6 glioma stem cells, to provide the ideal model for the further study of gli-oma stem cells in brain glioma model. Methods C6 glioma stem cell was cultured in vitro by suspension,and was identified with Nestin antibody. C6 stem cells of ICR mouse glioma model were used to investigate survival state and tumor volume in mice after the operation. HE staining and CD133 immunohistochemi-cal study were adopted to investigate the postoperative pathologi-cal changes in mice. Results The expression of Nestin was 96. 01% in C6 glioma stem cells, and Nestin was highly ex-pressed in the cultured C6 glioma stem cells. Mice were inocula-ted with tumor after loss of appetite, weight, behavior and slow, sluggish reaction. Tumor volume at day 21 after modeling was (9. 77 ± 6. 58) mm3 . After HE staining, the model showed the invasive growth, tumor cell shrinkage and derangement. Immu-nohistochemical CD133 staining revealed that tumor cytoplasm color was brown. Conclusion Glioma model can be established based on glioma stem cells into a high rate of tumor, the tumor cycle is short, which can be used as an ideal model for glioma.