Bone marrow-derived mesenchymal stem cells (MSCs) have potentials of proliferation, multilineage differentiation and homing to injured tissue and organs. Therefore, many researchers consider them as seed cells and attempt to use them to repair injured tissues and organs. In the allo- even xero-transplantation of these seed cells, graft rejection is an important problem. A large amount of data show that MSCs have specific immune characteristics; they block dendritic cell maturation, decrease immune response of T cells, do not express class Ⅱ major histocompatibility complex molecules, and mediate immunosuppression by secreting some dissoluble factors. The specific low immunogenicity of MSCs could avoid rejection in xero-trensplantation for human and animals in vivo, showing advantages in treatment. Therefore, they are promising to be used as seed cells for kidney construction.

Objective To establish the experimental animal model of acute tubular necrosis (ATN) in rats induced by caulis aristolochiae manshuriensis (CAM) containting aristolochic acid (AA) and compare the interventional effects among ligustrazine, prednisone and benazepril. Methods Male SD rats were divided randomly into six groups, 12 rats in each group. Control group, model group, prednisone group, benazepril group, ligustrazineⅠgroup and ligustrazineⅡgroup were given respectively by gavage with 3 ml/d distilled water,5 g?kg-1?d-1 CAM decoction (CAM 2 g/ml, AA 0.54 mg/ml, AA-Ⅰ0.46 mg/ml) for 60 days, then 3 ml/d distilled water, 10 g?g-1?d-1 CAM decoction for 30 days. Two hours after CAM gavage, control group and model group were given with normal saline. Prednisone group, benazepril group, ligustrazine group and ligustrazineⅡgroup were given with prednisone 5 mg?kg-1?d-1, benazepril 1.7 mg?kg-1?d-1, ligustrazine 50 mg?kg-1?d-1, ligustrazine 150 mg?kg-1?d-1 respectively by gavage for 90 days. Histopathology of kidney tissue was examined after 90 days. Results The renal tissue of control group was normal. Light microscopy of model group revealed patchy vacuolar changes of cells from proximal convoluted tubular epithelium, disorder and loss of brush border, exfoliated epithelial cells in the lumina, exposure of areas of denuded and rupture and thickness and atrophy of tubular basement membrane (TBM), edema and infiltration of inflammatory cells in the interstitium, focal segmental proliferation of glomerular mesangial cells and increase of mesangial matrix, part thickness of interlobular arterial walls. The above abnormalities of other four groups were significantly attenuated compared to model group. Electron microscopy of model group revealed patchy vacuolar changes and fatty degeneration of cells from proximal convoluted tubular epithelium, swelling of mitonchondria, reduce of organelle, karyorrhexis, apoptosis, infiltration of inflammatory cells (phagocytes and lymphocytes) in the interstitium and infiltration of lymphocytes in the epithelium, thickness of interlobular arterial walls, stenosis of lumina. The above abnormalities of electron microscopy in other four groups were remarkably improved compared to model group as well, especially in ligustrazine II group and prednisone group. Conclusions Pathological change of ATN is confirmed in kidney tissue and the rat ATN model induced of AA is successfully established. Benazepril, prednisone and ligustrazine can attenuate the toxic effects by AA. Prednisone and ligustrazine have a better efficacy.

Objective To investigate the pathological characters of capillaries in tubulointerstitial nephropathy,and to study the expression of vascular endothelial growth factor(VEGF)and proliferation cell nuclear antigen(PCNA)in capillary endothelial cells in aristolochic acid-induced nephropathy in rat.Methods 46 male Wistar rats were randomly divided into 2 groups,the model group was composed of 26 rats which were gavaged with the extract of Caulis Aristolochiae Manshuriensis(aristolochic acid AA 20mg/kg?d),and the control group consisted of 20 rats which were treated with equal volume of potable water.The rats were sacrificed in batches at the end of 4th,8th and 12th weeks,and the blood samples were collected from abdominal aorta for the tests of renal functions.The kidney of each rat was harvested.The renal tissues were stained with HE,PAS,and Masson's technique for the analysis of degree of tubulointerstitial injury,interstitial fibrosis,and peritubular capillary(PTC),and the expressions of both VEGF and PCNA were morphologically observed and immunohistochemically analyzed.Results In the model group,the level of serum creatinine/body weight increased markedly,the renal pathological changes consisted of severe acute renal tubulointerstitial damage with cloudy swelling of tubule,degeneration,and exfoliation.With prolongation of feeding time,the damage progressively aggravated,and showed a remarkable tubulointerstitial injury.The interstitial fibrosis area was 31.36% at the end of 12th week.The renal capillaries showed thickening of vessel wall,narrowing of the vessel cavity,obstruction or hyalinization of some capillaries.There was focal infiltration of inflammatory cells around the injured vessels.But there was no apparent change in the globules compared with that in control group.The PTC dwindled in caliber or distorted,and the PTC density decreased significantly in models,especially in the region of tubular damage or interstitial fibrosis.The expression of VEGF showed compensatory up-regulation at the 4th week,but it was down-regulated gradually.The expression of PCNA was up-regulated at the 4th week,but down-regulated after 8th week,and only a few basement membrane naked tubular cells showed positive expression at 12th week.Conclusion AA could induce injury and loss of capillaries of the kidney.The decrease in capillary density might contribute to the impairment of renal function and progressive interstitial fibrosis,and the relative deficiency of VEGF expression may be related to PTC injury,which is one of the causes of chronic progression of aristolochic acid nephropathy.

Mesenchymal stem cells express major histocompability complex (MHC)Ⅰin low levels, but do not express MHC Ⅱ, so it has a low immunogenicity. It can block maturity of dendritic cells, depress immune response of T lymphocytes, and also mediate immune suppression by secreting several soluble cytokines. In light of these characters, mesenchymal stem cell can avoid allograft rejection in human and animal transplantation. It has been reported previously that human mesenchymal stem cells cultivated in growing rodent embryos can grew out of new kidney and ureter. Furthermore, urea nitrogen and creatinine levels were significantly higher in liquid collected from the new ureter compared with the animal serum levels, which suggested that the new kidney had generated urine. This native kidney has low immunogenicity, little side-effect after transplantation, extensive sources, so it may have wide space in self-kidney transplantation.

Objective To investigate the expression and significance of VEGF and NF-κB/p65 in the two subtypes of pleomorphic adenoma. Methods Immunohistochemistry was applied to detect the expression of VEGF and NF-κB/p65 in pleomorphic adenoma of 60 patients including 31 cases of cell-rich and 29 cases of stroma-rich as well as normal salivary gland tissues of 30 cases from adjacent tumor. Results VEGF positive staining was mainly found in tumor epithelia,while NF-κB/p65 positive was detected in gland alveolus cell and ductal epithelia. The Mean optical density( MA ) values of VEGF were 955.67 ± 305.79,149. 13 ± 60. 85 and 53.46 ± 9. 66, respectively, in cell-rich adenoma, stroma-rich adenoma and normal control. The difference in VEGF expression between the groups was significant (Ps ＜ 0. 05 ) . The MA values of NF-κB/p65 were 529. 80 ± 164. 81,43.40 ±5.46 and 6. 84 ± 1.91 ,respectively,in three groups mentioned above. The difference in NF-κB/p65 expression between the groups was significant ( Ps ＜ 0. 05 ). In pleomorphic adenoma, the expression level of NF-κB/p65 was positively correlated with VEGF. Conclusion ( 1 ) The angiogenesis and proliferation potential of carcinomas increased with the cell component in pleomorphic adenoma. Stroma-poor adenoma is more frequently subjected to malignant transformation than stroma-rich adenoma. (2) NF-κB/p65 may have effects on angiogenesis by activating VEGF. Detecting the expression of VEGF and NF-κB/p65 may be helpful to predict the biological behavior of pleomorphic adenoma and prognosis of the patients, which couldprovide useful information for future targeted therapy of pleomorphic adenoma.

Objective To explore the effect of VEGF-C,VEGFR-3 gene in primary culture in vitro of laryngeal squamous carcinoma tissue by dexamethasone (DEX). Methods Applying the technique of Drimary tissue cuiture in vitro,twelve specimens,which were identified pathologically as laryngeal squamous cell carcinoma,were cultured.After succeeded culture of twelve specimens,the cells were divided in to two groups.One continued to cuhum,another gave 10-7 mol/L of DEX.VEGF-C and VEGFR-3 gene express in 12 cases of cell were examined by reverse transcription polymerase chain reactions(RT-PCR).Results The frequency of VEGF-C(83%) and VEGFR-3(58%) in primary culture in wtro of laryngeal squamous carcinoma tissue were higher than that of the primary culture in vitro of laryngeal squamous carcinoma tissue by dexamethasone (DEX) (25%,8%P＜0.05).Conclusion The expressions of VEGF-C and VEGFR-3 in Drimary culture in vitro of laryngeal squamous carcinoma tissue by DEX are decreased.It may provide guide to apply DEX in clinic laryngeal squamous carcinoma chemotherapy and laryngeal squamous carcinoma laryngeal obstruction.

Objective To evaluate the efficacy and safety of immunosuppressive strategy of leflunomide (LEF) combined with calcineurin inhibitor and prednisone in Chinese renal transplant recipients.Methods Prospective clinical study was performed in 16 stable renal transplant recipients treated with calcineurin inhibitor, prednisone and either mycophenolate mofetil (MMF) or azathioprine (Aza), and MMF or Aza was replaced by LEF afterward. The loading dose of LEF was 50 mg/day for 3-5 days, and the maintenance dose was 20 mg/day. Prednisone was maintained 10 mg/day. The doses of CsA were adjusted according to its blood concentration.Results One patient dropped due to economic reason 1 mouth later, and continued to take Aza. Another one patient switched back to MMF due to a slight increase of serum creatinine (from 130 ?mol/L to 143 ?mol/L) 6 months after conversion. The remaining 14 patients were still on LEF. The mean observation time under LEF was (8.7?7.2) months (1～18 month). The mean serum creatinine was (103?24) ?mol/L before conversion, (95?26) ?mol/L at 6th month, and (108?27) ?mol/L at one year post-conversion. There was one episode of biopsy proved reversible acute rejection. The observed side effect was tolerable alopecia in 2 cases and rash in 1 case, but there were no significant increases in liver function, decrease of blood WBC count, deterioration of anemia, or any serious infection in any of the patients.Conclusion Immunosuppressive strategy of LEF+calcineurin inhibitor and prednisone is effective and safe in stable Chinese renal transplant recipients.

Objective To observe the effect of different calcium concentration peritoneal dialysate on calcium-phosphorus metabohsm in uremia patients under peritoneal dialysis. Methods Forty uremia patients who the first entered peritoneal dialysis were divided into two groups: standard calcium concentration group (20 cases), low calcium concentration group (20 cases). Treated with continuous ambulatory peritoneal dialysis (CAPD), combining with oral calcium carbonate, observing 12 months. Results When the first entering CAPD, serum calcium, phosphorus, calcium-phosphorus product, immunoreactive parathyroid hormone (iPTH) of the two groups had no differences. Hyperphosphatemia, lower iPTH were universal. After 6 months' treatment, hypercalcemia and hyperphosphatemia in standard calcium concentration group got increasingly serious.While in low calcium concentration group, serum calcium, phosphorus, calcium-phosphorus product all decreased significantly, moreover, they were also lower than post-treatment standard calcium concentration group(P<0.01). In the following 6 months, in low calcium concentration group,serum calcium, calcium-phosphorus product maintained at the normal level serum phosphorus also decreased to the normal level iPTH maintained about 150 ng/L. During the treatment, all patients had no hypotension and hyperspasmia. Conclusion Low calcium concentration peritoneal dialysate can ameliorate disorder of calcium-phosphorus metabohsm, delay the appearance of its comphcations.

ObjectiveTo study the effect of berberine on diabetes or insulin resistance accompanied with reduction of skeletal moscle and wasting in db/db mice.Methods db/db mouse-a model of diabetes/insulin-resistance was studied,with the wild type mouse as control.After being treated with berberine ( 5 mg · kg-1 · d-1 ) for 3 weeks,the muscle size of tibia anterior (TA) of the animals was measured after staining with laminine/Hyosin Heavy chain-Slow using immunochemistry,then observed under fluorescent microscope and calculated with software.The rate of [ 14 C ] -Phenalanine incorporation into the uuscle was measured to analyze the protein synthesis,and the [ 3 H ] -Tyrosine released into the medium was determined in order to analyze protein degradation.The mRNA expressions of muscle atrophy Fbox-1 ( Atrogin-1 ) and muscle ring finger-1 ( MuRF-1 ) were measured by Northern blot.Results With berberine treatment,blood glucose and fat levels were lowered [ ( 18.55 ± 3.79 vs 26.32 ± 4.02 ) mmol/L,P＜0.01 ; ( 2.75 ± 0.30 vs 3.77 ± 0.52 ) g,P＜0.05 ],but tibia anterior muscle weight/length ratio and cross-section area were decreased,rates of protein synthesis in isolated muscles of db/db mice were decreased by 18％ -22％,and the rates of degradation were significantly raised by 24％ -26％ after berberine treatment.There also was increased the transcription and translation of Atrogin-1 and MuRF-1,accompanied with decreased eukaryotic initiation factor 3 subunit (eIF3-f)protein level simultaneously. ConclusionBerberine improves hyperglycemia and insulin resistance by down-regulating blood sugar and body fat,but it causes reduced protein synthesis and minimally enhanced protein degradation.The mechanism might be related to berberine-induced up-regulating Atrogin-1 、MurF-1 and down-regulating eIF3-f.

BACKGROUND:The rapidly developed transplantation of peripheral blood stem cells have been successfully used to substitute bone marrow transplantation and become the first choice method for transplantation of hemopoietic stem cells.It is relatively difficult to collect peripheral blood stem cells from young age and low body mass infants.OBJECTIVE:To investigate the safety and adverse effects of peripheral blood stem cell collection from young age and low body mass through the use of blood cell separator.METHODS:Two type 1 diabetes mellitus infants,aged younger than 2 years old and with body mass less than 15 kg,were treated using autologous hemopoietic stem cell transplantation.The two infants were adequately comforted to lesion the fear of collection.At 1 week prior to collection,calcium agent was orally taken to reduce the incidences of low calcium.Within 24 hours prior to collection,oily food was forbidden,and on the day of collection,milk was forbidden,to avoid chylemia,which influences blood collection.Prior to collection,200 mL 25 Gy y-ray radiated red blood cells suspension was injected into the tube,which was routinely placed in the subclavian vein,to avoid hypovolaemic syndrome and the effects on hematocrit.Individualized collection parameters were set,During collection,blood circulation volume was 3,4 times of systemic blood circulation to ensure sufficient total circulation volume.During isolation,the ratio of ACD to whole blood was kept between 1:11 and 1:13 to prevent sodium citrate poisoning.RESULTS AND CONCLUSION:Peripheral blood stem cells were successfully collected during first intention in each infant.During collection,stable vital signs but no adverse effects were observed.After collection,mononuclear cells weighted 14.71×108/kg and 18.82×108/kg respectively,and CD34+ cells were about 34.13×108/kg and 32.38×106/kg,respectively in each infant.Therefore,it is feasible to collect peripheral blood stem cells from infants of young age and low body mass under sufficient psychological preparation.