Structural shifts in the intestinal microbiota of rats treated with cyclosporine A after orthotropic liver transplantation / 医学前沿

Understanding the effect of immunosuppressive agents on intestinal microbiota is important to reduce the mortality and morbidity from orthotopic liver transplantation (OLT). We investigated the relationship between the commonly used immunosuppressive agent cyclosporine A (CSA) and the intestinal microbial variation in an OLT model. The rat samples were divided as follows: (1) N group (normal control); (2) I group (isograft LT, Brown Norway [BN] rat to BN); (3) R group (allograft LT, Lewis to BN rat); and (4) CSA group (R group treated with CSA). The intestinal microbiota was assayed by denaturing gradient gel electrophoresis profiles and by using real-time polymerase chain reaction. The liver histopathology and the alanine/aspartate aminotransferase ratio after LT were both ameliorated by CSA. In the CSA group, the numbers of rDNA gene copies of Clostridium cluster I, Clostridium cluster XIV, and Enterobacteriaceae decreased, whereas those of Faecalibacterium prausnitzii increased compared with the R group. Cluster analysis indicated that the samples from the N, I, and CSA groups were clustered, whereas the other clusters contained the samples from the R group. Hence, CSA ameliorates hepatic graft injury and partially restores gut microbiota following LT, and these may benefit hepatic graft rejection.

Efficacy and Safety of a Steroid-Free Immunosuppressive Regimen after Liver Transplantation for Hepatocellular Carcinoma

BACKGROUND/AIMS: We aimed to evaluate the efficacy and safety of an immunosuppressive regimen without steroids after liver transplantation (LT) for hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). METHODS: Sixty-six HCC patients who underwent an immunosuppressive regimen without steroids after LT were enrolled in the steroid-free group. The preoperative characteristics and postoperative outcomes of these patients were compared with those of 132 HCC recipients who were placed on an immunosuppressive regimen using steroids (steroid group). The incidence of acute rejection, HBV recurrence, infection, and new-onset diabetes mellitus and the overall and tumor-free survival rates were compared between the two groups. RESULTS: Differences were not observed in the 1-year (83.3% vs 97.0%, p=0.067), 3-year (65.4% vs 75.8%, p=0.067) or 5-year (56.3% vs 70.7%, p=0.067) patient survival rates or in the 1-year (62.1% vs 72.7%, p=0.067), 3-year (49.8% vs 63.6%, p=0.067) or 5-year (48.6% vs 63.6%, p=0.067) tumor-free survival rates between the two groups, respectively. In the steroid-free group, the patients who fulfilled the Milan criteria had higher overall and tumor-free survival rates than those in the steroid group (p<0.001). The prevalence of HBV recurrence (3.0% vs 13.6%, p=0.02) was significantly lower in the steroid-free group compared with the steroid group. CONCLUSIONS: After LT, an immunosuppressive regimen without steroids could be a safe and feasible treatment for HBV-related HCC patients, thus resulting in the reduction of HBV recurrence. Based on the observed survival rates, patients who fulfill the Milan criteria may derive benefits from steroid-free immunosuppression.

Prediction of liver transplantation prognosis by transient elastography / 中华器官移植杂志

Objective To investigate the clinical value of transient elastography in adult after liver transplantation,by means of evaluating the correlation of liver stiffness measured by FibroScan with liver/renal functions.Method Forty-three patients received orthotopic liver transplant in our hospital during Dec.10,2013 and Mar.19,2014 were included in this study.Liver stiffness measurement (LSM) was performed after transplantation.Clinical data and laboratory tests including liver function and renal function were collected and analyzed.Result Bivariate correlation showed that body mass index (BMI),MELD score,graft-to-recipient weight ratio (GRWR) and warm ischemia time had no correlation with LSM.LSM at the 1st day after transplantation (LSM-1) showed no correlation with cold ischemia time,but LSM at the 7th day after transplantation (LSM-7) did,with R =0.335,P =0.028.LSM-1 showed positive correlation with the ICU time (R =0.488,P =0.001),but LSM-7 didn't.There was significantly positive correlation between LSM and aspartate aminotransferase,bile acid and creatinine,but no significant correlations were found between LSM and alanine arninotransferase,alkaline phosphatase,cholinesterase,gamma-glutamyl transferase,total bilirubin,direct bilirubin,indirect bilirubin,urea nitrogen and uric acid.The group with higher LSM-1 had longer ICU time than the lower group (9 d vs,7 d,P =0.013),and so was the hospital stay (34 d vs.23 d,P =0.023).For the LSM-7,there was no significant difference in ICU time and hospital stay between the two groups.The group with higher LSM-1 had higher serious complication incidence than the lower group (78.57％ vs.27.59％,P =0.002),but the two groups in LSM-7 showed no significant difference in serious complication incidence.Conclusion The LSM partially correlates with the liver function and renal function of liver transplantation recipient,and may have its clinical value for assessing the early prognosis after liver transplantation.

Preparation of Electrophoretic Microcolumn Packed with Quartz Microncrystal and Its Application to Electrophoretic Separation / 分析化学

The preparation of electrophoretic microcolumn and its application to the electrophoretic separation of amino acids with a 2-mm I.D. fused-silica microcolumn packed with uniform quartz microncrystal prepared by hydrothermal synthesis are reported. With 1.5 mmol/L disodium phosphate buffer solution (pH 11.5) containing 30% (V/V) methanol, the tryptophan, phenylalanine and tyrosine were separated by the microcolumn electrophoresis and detected by an UV spectrophotometer without derivatization. The limits of detection were 0.038, 0.21 and 0.20 mol/L, respectively. The separation efficiency of tryptophan was 4.4×104 plates/m. The sample capacity of the electrophoretic microcolumn achieved 35 μL. The precisions of the microcolumn electrophoresis were satisfactory. The thermal effects of the electrophoretic microcolumn that without packing, packed with 360 μm quartz sand and with 9 μm length quartz microncrystal were discussed, respectively. It was found that the electrophoretic microcolumn packed with quartz microncrystal was able to inhibit Joule heat, increase sample capacity and enhance detection sensitivity. The microcolumn electrophoresis is one of the high-performance separation techniques for an in-situ, real-time and portable electrokinetic flow analysis system.

The effect of fibronectin and arginine-glycine-aspartic-serine tetrepeptide on chemotherapy sensitivity of liver carcinoma cell lines / 中华普通外科杂志

Objecfive To study the effect of fibronectin(FN)and arginine-glycine-aspartateserine tetrapeptide (RGDS)on chemotherapy sensitivity of hepatocellular carcinoma(HCC)cell lines.Methods A HCC cell lines 7721 was divided into BSA group(BO),FN group(FO),RGDS group(RO)and FN+RGDS group(FR).Mitomycin-C(MMC)was added into each group with different concentrations for 2 h,12 h and 24 h.MTT assay was used to detect the survival rate of the tumor cell at different time,and fluorescent staining method and flow cytometry was used to detect the tumor cell's apoptosis.Results The survival rate and apoptosis rate was significantly different between each group with the treatment of MMC for 12 h or 24 h,and no statistical difference was found after incubation for 2 h.Conclusions With time FN down-regulates MMC induced apoptosis of the HCC cells.making HCC cells 1ess sensitive to chemotherapy,while RGDS enhances the sensitivity of HCC cells to chemotherapy.

Inhibitory effect of tea polyphenols on renal cell apoptosis in rat test subjects suffering from cyclosporine-induced chronic nephrotoxicity / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of tea polyphenols on renal cell apoptosis in rat test subjects suffering from cyclosporine A (CsA)-induced chronic nephrotoxicity.</p><p><b>METHODS</b>Four groups of rats with CsA-induced chronic nephrotoxicity were respectively treated with vehicle olive oil, tea polyphenols, CsA and tea polyphenols plus CsA. At the end of the 28th day of treatment, 24 hours urine and blood samples were obtained, and the animals were then sacrificed. The serum and urine samples were analysed for creatinine clearance, and kidney tissue was used for pathologic analysis of renal tubular injury and interstitial fibrosis. The TUNEL assay, apoptosis-related enzyme caspase-3 mRNA detected by RT-PCR, and its enzymatic activity were analysed for the possible detections of cell apoptosis.</p><p><b>RESULTS</b>CsA-treated rats displayed increased apoptosis of the tubular and interstitial cells, in comparison with vehicle-treated controls (18.3 +/- 4.6 vs 4.8 +/- 1.3 cells/mm(2), P < 0.05). In comparison with animals treated by CsA, animals treated with CsA plus tea polyphenols demonstrated significantly improved levels of creatinine clearance (0.12 +/- 0.03 vs 0.22 +/- 0.02 ml.min(-1).100 g(-1) body weight, P < 0.05), tubular injury (2.29 +/- 0.43 vs 1.42 +/- 0.26, P < 0.05), and interstitial fibrosis (2.83 +/- 0.20 vs 1.46 +/- 0.19, P < 0.05), and showed a statistically significant decrease in tubular and interstitial cell apoptosis (18.3 +/- 4.6 vs 7.7 +/- 2.1 cells/mm(2), P < 0.05). The expression of caspase-3 mRNA and caspase-3 activity was significantly higher in the CsA-treated group than that of the CsA plus tea polyphenols (TP)-treated group (P < 0.05).</p><p><b>CONCLUSION</b>These results suggested that tea polyphenols significantly inhibits apoptosis of the tubular and interstitial cells in rats with cyclosporine-induced chronic nephrotoxicity, and that tea polyphenols may be useful to prevent CsA-associated kidney toxicity.</p>

CsA downregulates IFN-gamma gene transcription after liver transplantation by inhibiting NF-kappa B activity / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the relationship between NF-kappa B activity and IFN-gamma gene expression, as well as the histopathological changes following liver transplants, both with and without cyclosporin A (CsA) treatment.</p><p><b>METHODS</b>Sixty male Wistar and Thirty male SD rats were subjected to orthotopic liver transplants. Fourty-five of the Wistar rats were used as recipients, and were divided into 3 groups: group I, syngeneic control (Wistar-to-Wistar); group II, acute rejection (SD-to-Wistar); and group III: acute rejection treated with cyclosporin A by intramuscular injection (SD-to-Wistar + CSA). After the liver transplants, electrophoretic gel mobility shift assay was used to analyze NF-kappa B activity in the splenocytes of recipient rats, and RT-PCR was used to measure IFN-gamma gene expression in grafted liver specimens. In addition, histopathological examinations were performed to assess the severity of acute liver rejection.</p><p><b>RESULTS</b>In group I, low levels of NF-kappa B activity were only detectable on day 5 and 7 post-transplant, and only weak IFN-gamma mRNA expression was observed at all time points. By contrast, both high NF-kappa B activity and high expression levels of IFN-gamma mRNA were detected at all time points in group II. In group III, NF-kappa B activity and IFN-gamma mRNA expression were significantly inhibited, as compared to group II (P < 0.05). A good correlation was found between NF-kappa B activity and IFN-gamma mRNA expression (r = 0.815). In addition, NF-kappa B activity and IFN-gamma mRNA expression mirrored histopathological changes in all three experimental groups.</p><p><b>CONCLUSIONS</b>Changes in IFN-gamma mRNA expression levels after liver transplantation are at least partially due to changes in levels of NF-kappa B activity. CsA appears to downregulate NF-kappa B activity, thus inhibiting IFN-gamma gene transcription. Blocking the NF-kappa B mediated transcription pathway may be of benefit in preventing liver transplant rejection.</p>

Effects of power-frequency magnetic fields exposure on phosphorylation and enzymatic activity of stress-activated protein kinase and its upstream kinase / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the effects of 50 Hz power-frequency magnetic fields on signal transduction pathway of stress-activated protein kinase(SAPK), and explore the cellular signal transduction mechanism of the biological effects induced by power-frequency magnetic fields.</p><p><b>METHODS</b>Chinese hamster lung(CHL) cell line was exposed to power-frequency magnetic fields with two intensities for different exposure durations. The cytoplasmic protein was extracted and the phosphorylated portion of SAPK and SEK1/MKK4 was measured with Western blotting analysis. The SAPK enzymatic activity was measured by the solid-phase kinase assay in cells exposed to power-frequency magnetic fields for 15 min.</p><p><b>RESULTS</b>Both 0.4 mT and 0.8 mT power-frequency magnetic fields could enhance the phosphorylation of SAPK in a time-relative course manner, and reached the maximum extent at 15 min, with an increase of 20% and 17% respectively. The solid-phase kinase assay showed that the enzymatic activities of SAPK were also increased, which were 2.9 +/- 0.4 and 2.1 +/- 0.9 times of control respectively. However, the duration of SAPK phosphorylation induced by 0.8 mT was longer than that of 0.4 mT, while the duration and extent of SAPK dephosphorylation was remarkably shorter than that of 0.4 mT. The power-frequency magnetic fields under equal conditions could not phosphorylate(activate) the SEK1/MKK4.</p><p><b>CONCLUSION</b>Power-frequency magnetic fields could activate the SAPK, but not SEK1/MKK4. It is suggested that power-frequency magnetic fields may activate SAPK signal transduction pathway through a kinase other than SEK1/MKK4. The activation mechanism of SAPK of power-frequency magnetic fields needs to be identified in more detail.</p>

The effect of an antioxidant tea polyphenols on cell apoptosis in rat model of cyclosporine-induced chronic nephrotoxicity / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effect of tea polyphenols on cell apoptosis in rat model of cyclosporine-induced chronic nephrotoxicity.</p><p><b>METHODS</b>Four groups of animals in rat model of cyclosporine-induced chronic nephrotoxicity were respectively treated by olive oil (n = 6), tea polyphenols (TP, n = 6), cyclosporine A (CsA, n = 8) and TP plus CsA (n = 8). At the end of 28th day of treatment, all animals were sacrificed and blood was analyzed for blood serum creatinine and creatinine clearance, kidney tissue for pathologic analysis. The TUNEL assay, caspase-3 mRNA expression detected by reverse transcription-polymerase chain reaction (RT-PCR) and caspase-3 activity were used for the analysis of cell apoptosis.</p><p><b>RESULTS</b>CsA plus TP ameliorated the CsA-induced decrease of renal function and interstitial fibrosis. There was a significant increase in the number of apoptosis-positive cells in the CsA-vs-CsA plus TP-treated group at four weeks (18.9 +/- 3.3 vs. 7.7 +/- 1.4, P < 0.05). The expression of caspase-3 mRNA and caspase-3 activity of CsA-treated group was significantly higher than that of CsA plus TP-treated group (P < 0.05).</p><p><b>CONCLUSION</b>These results indicate that antioxidant tea polyphenols significantly inhibit apoptosis of tubular and interstitial cells in rat model of chronic cyclosporine-induced nephrotoxicity, and suggest that the decrease of cell apoptosis exerted by tea polyphenols may be one of mechanisms to protect renal function and tissue structure.</p>

Pretreatment of hypertonic saline attenuates the hepatic ischemia reperfusion injury induced by neutrophils / 中国病理生理杂志

AIM:To explore the effect of the pretreatment of hypertonic saline(HTS) in hepatic ischemia reperfusion(I/R) injury.METHODS:The rats were divided into sham group(sham group),ischemia reperfusion group(IR group) and pretreatment of hypertonic saline group(HTS group).Partial hepatic ischemia reperfusion model was used.The rats were sacrificed at the time of 1 h,3 h,6 h,12 h and 24 h after reperfusion in each group,respectively.Blood samples were obtained to examine ALT.The expression of the CD11b/CD18(Mac-1) on the neutrophils was analyzed by flow cytometry.RT-PCR and Western blotting were used to examine the expression of intercellular adhesion molecule-1(ICAM-1) in livers and chromatometry was performed to detect the activity of myeloperoxidase(MPO) in livers.The morphology of hepatocytes and the structure of sinusoid were observed by histological examinations.RESULTS:① HTS pretreatment decreased the level of ALT at the time points of 3 h,6 h and 12 h after reperfusion(P

Mycophenolate acid inhibits hepatitis B virus replication in HepG2.2.15 cells / 中国病理生理杂志

AIM: To investigate the effect of mycophenolate acid(MPA) on hepatitis B virus(HBV) replication in vitro.METHODS: In the presence or absence of guanosine,the HepG2.2.15 cells were treated with different concentrations of MPA(1-20 mg/L) for 4 days.Hepatitis B surface antigen(HBsAg) and hepatitis Be antigen(HBeAg) in supernatant were detected by ELISA.Intracellular HBV core mRNA and HBV DNA were analyzed by reverse transcription-polymerase chain reaction(RT-PCR) and slot blot hybridization,respectively.RESULTS: MPA suppressed the expression of HBsAg and HBeAg,and inhibited the replication of HBV DNA.The effect of MPA on HBV replication was reversed by addition of exogenous guanosine.CONCLUSION: MPA suppresses the expression of HBsAg,HBeAg and replication of HBV DNA in HepG2.2.15 cells.Reducing the synthesis of guanosine nucleotides may be involved in the mechanism of the inhibitory activity of MPA on HBV replication.

Infusion of donor specific bone marrow alleviates acute rejection reaction in liver allotransplantation / 中国病理生理杂志

100 d),and the histological grade of rejection was significantly lower than that in group Ⅱ(P

Lymphotactin gene transcription is inhibited by cyclosporine A during acute cardiac allograft rejection in mice / 中国病理生理杂志

AIM: To investigate lymphotactin (Lptn) gene transcription during acute cardiac allograft rejection and the inhibitory effect of cyclosporine A (CsA). METHODS: Graft specimens were harvested at indicated time to determine morphological changes by pathological examination. The grade of acute cardiac allograft rejection was evaluated by using modified Banff scoring system. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine the Lptn mRNA expression in cardiac grafts. NFATc1 activity of splenocytes after transplantation was assessed by enzyme-linked immunoabsordent assay (ELISA). RESULTS: Prominent splenomegaly on day 3 posttransplantation was found in C57BL/6-Balb/c group. The extent of myocardial inflammatory infiltration was scored 2.667?0.577 at day 5 and 2.333?0.577 at day 7, respectively. Splenomegaly was ameliorated by CsA treatment, and the extent of myocardial infiltrate was scored 1.000?0.000 at day 5 and 1.333?0.577 at day 7, respectively. Lymphotactin mRNA was undetectable in cardiac isografts. Lymphotactin mRNA, which was inhibited partially by CsA, was upregulated strongly in acutely rejecting cardiac allografts at day 5 and day 7. Further studies demonstrated that NFATc1 activity in splenocytes, which markedly upregulated during acute rejection, was completely inhibited by CsA. CONCLUSION: Lptn appears to be a key chemokine of lymphocyte infiltration during acute allograft rejection. Inhibition of NFATc1 activity by CsA seems to decrease Lptn expression incompletely, suggesting that there was else mechanism to regulate Lptn expression other than NFAT pathway.

Cloning of a new human cytochrome P450 2A6 cDNA / 中国病理生理杂志

AIM:To clon human cytochrome P450 2A6 cDNA. METHODS:Using reverse transcription polymerase chain reaction(RT-PCR) and DNA recombinant technique, a full-length cDNA encoding cytochrome P450 2A6( CYP2A6 ) from human liver was cloned into pBluescript vector. The cDNA segment was identified by DNA sequencing.RESULTS: Comparing with the CYP2A6 sequence, the cloned CYP2A6 cDNA had two different bases, codon 8 CTG(Leu)→TTG(Leu), codon 479 GGC(Gly)→GTC(Val) and was quite different in their 3′end noncoding region. Comparing with CYP2A7 seqence reported by Fernandez-Salguero,the cloned CYP2A6 cDNA had some different in 5′ end coding sequence and several differencey in the 3′ end coding and noncoding sequence, but both codon 479 were GTC(Val).Comparing with the CYP2A7 seqence reported by Yamano,the cloned CYP2A6 cDNA had some difference in the coding sequence but the 3′ end no-coding area was the same. CONCLUSION: The cloned cDNA was a new cDNA of CYP2A6 which may be transcripted from a new allele of CYP2A6.

Phagocytosis of viable apoptotic cells inhibits the activation of T lymphocytes / 中国病理生理杂志

AIM: To investigate whether viable apoptotic cells and phagocytosis of them affect the activation of T lymphocytes. METHODS: Ultraviolet irradiation was used to induce apoptotic cells in vitro and the model of phagocytosis of these cells was established. Cytokine TGF ?1 was detected by ELISA. The rate of apoptotic cells and phagocytosis of them were assessed by flow cytometry. Furthermore, flow cytometry was also employed to examine the expression of activation signs, such as CD69, CD25, CD71, of T lymphocytes under the intervention of apoptotic cells and macrophage which ingested apoptotic cells, to reflect whether the apoptotic cells and the phagocytosis of these cells could influence the activation of lymphocytes stimulated by Con A. RESULTS: Ingestion of apoptotic cells increased TGF ?1 secretion. Only the macrophages that had ingested apoptotic cells could suppress the activation of lymphocytes. The expression of the markers of lymphocytes activation such as CD69, CD25, CD71 had been restrained. These inhibition effects were abolished by monoclonal anti-TGF ?1 antibody. CONCLUSION: The macrophages that have ingested apoptotic cells inhibit expression of CD69, CD25 and CD71 of T lymphocytes stimulated by ConA. This effect is dependent on the increase in TGF ?1 secretion in local site. [

Effects of NS-398 on the proliferation and apoptosis of HepG2 cells / 中国病理生理杂志

AIM: To evaluate the effects of NS-398, a cyclooxygenase-2(COX-2) inhibitor, on the proliferation and apoptosis in HepG2 cells. METHODS: The effects of NS-398 on the proliferation of HepG2 cells was evaluated by MTT. DNA fragmentation gel analysis was used to analyze the apoptotic cells; DNA ploidy and apoptotic cell percentage were examined by flow cytometry. Furthermore, the expression of COX-2 and Bcl-2 mRNA was identified by competitive RT-PCR. RESULTS: NS-398 inhibited cell proliferation and induced apoptosis in HepG2 in a concentration-dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased with NS-398 concentration increasing. The quiescent G_0/G_1 phase was accumulated with decreasing of Bcl-2 mRNA. Whereas NS-398 had no effect on the expression of COX-2 mRNA, no correlations were found between COX-2 mRNA and the HepG2 cell proliferation and apoptosis induced by NS-398 (r=0.056 and r=0.119, respectively). CONCLUSION: NS-398 significantly inhibits the proliferation and induces apoptosis in HepG2. Mechanisms may be involved in accumulation of quiescent G_0/G_1 phase and decrease in Bcl-2 mRNA expression, but independent to COX-2 mRNA expression. [

Molecular mechanism of etodolac-induced apoptosis in SMMC7721 cell line / 中国病理生理杂志

AIM: To investigate the possible role of nuclear transcription factor kappa B (NF-?B), Bcl-2, Bax and caspase-3 in etodolac-induced apoptosis of liver tumor SMMC7721 cell line. METHODS: Cell apoptosis was determined by flow cytometry analysis with PI staining and DNA laddering. Expression of Bcl-2 and Bax protein was measured by Western blotting. Caspase-3 activity was evaluated by active caspase-3 apoptosis kit with flow cytometry. NF-?B activation was detected by ELISA-based TransAM~(TM) NF-?B p65/p50 kit. RESULTS: Etodolac, a selective COX-2 inhibitor, stimulated apoptosis in liver tumor SMMC7721 cell line significantly. Flow cytometry showed that the apoptotic rate was 16.3%?3.1%, 19.9%?3.6%, 22.9%?3.2%, 31.2%?3.3% with different concentrations of etodolac (0.25, 0.50, 1.0 or 2.0 mmol/L), while the apoptotic peak did not appear in the control group (0 mmol/L) (P

Correlative analysis between renal TGF-?1 expression and cell apoptosis in a rat model of cyclosporine-induced chronic nephrotoxicity, and effect of tea polyphenols on TGF-?1 expression and apoptosis / 中国病理生理杂志

AIM: To investigate the correlation between renal transforming growth factor-?1 (TGF-?1) expression and cellular apoptosis, and effect of tea polyphenols (TP) on TGF-?1 expression and apoptosis. METHODS: Four group animals in a rat model of CsA-induced chronic nephrotoxicity were respectively treated by vehicle (olive oil, 0.1 mL?kg~(-1)?d~(-1), sc), TP (80 mg?kg~(-1)?d~(-1), ig), CsA (15 mg?kg~(-1)?d~(-1), sc) and TP (80 mg?kg~(-1)?d~(-1), ig) plus CsA (15 mg?kg~(-1)?d~(-1), sc). At the end of 28th day of treatment, renal creatinine clearance and tissue pathology were analyzed. The TGF-?1 expression was detected by immunohistochemistry and Western blotting. TUNEL assay, apoptosis-related enzymatic activity caspase-3 were also detected. RESULTS: Compared to CsA-treated rats, the animals treated with CsA plus TP showed a significant increase in the renal creatinine clearance (0.12?0.03 vs 0.22?0.02,P

Activation of JNK/SAPK pathway in vero cells induced by N-methyl-N'-nitro-N-nitrosoguanidine / 中国病理生理杂志

AIM and METHODES: To evaluate the possible signal transduction mechanism of nontargeted mutagenesis in vero cells induced by DNA damaging agent N-methyl-N-nitro-N-nitrosoguanidine(MNNG),the activation of c-Jun NH 2-terminal kinase/stress activated protein kinase(SAPK/JNK) pathway in vero cells induced by MNNG was studied. Western Blot analysis and Solid-phase kinase assay were used to measure the phosphorylation of JNK1 and kinase activity of JNKs, respectively. RESULTS: After 0.2 ?mol/L, 2.5 h MNNG or 1 mg/L, 1 h cycloheximide (CHM) treatment, the proportion of phosphorylated JNK1 in cell extract increased significantly, simultaneously the kinase activity of JNKs increased dramatically(6.7 and 3.0 folds respectively), as measured by the phosphorylation of c-Jun, a substrate of JNKs. CONCLUSION: Both 0.2 ?mol/L 2.5 h MNNG and 1 mg/L 1 h CHM treatment can induce the activation of JNK/SAPK pathway, one of the stress signal transduction pathways, in vero cells.

The in vitro immune effects of dendritoma formed by mouse hepatocellular carcinoma cells and lymphotactin gene modified dendritic cells / 中国病理生理杂志

AIM: To investigate the in vitro anti tumo r immune responses of dendritoma formed by mouse hepatocellular carcinoma cells and lymphotactin gene modified dendritic cells (DCs). METHODS: DCs prepared from mouse bone marrow were genetically mo dified by lymphotactin adenovirus, and fused with H22 cells by polyethylene glyc ol. RT-PCR and ELISA were employed to identify lymphotactin expression at mRNA a nd protein levels. The phenotypes and fusion efficiency were detected by FACS. T he stimulatory capacity of DC to T cells was detected by mixed leukocyte reactio n. The cytotoxicity activity against H22 cells was assayed by LDH method. RESULTS: Lymphotactin effectively expressed by DCLptn/H22 hybrid oma. DCLptn/H22 cells induced potent T cell proliferation effect and generated s trong CTL reaction against allogenic H22 cells. CONCLUSION: Lymphotactin genetic modification enhanced the in vitro immune activity of dendritoma.