Objective To investigate the dose-effect relationship of donor antigenic specificity CD4~+ CD25~+ Treg cells prolonging the survival of rat kidney allograft.Methods Sixty allograft kid- ney transplantation animal models were established with SD rats as donors and Wistar rats as recipi- ents.CD4~+ CD25~+ T cells were sorted from Wistar rats'spleens by way of MACS and the phenotypes of donor antigenic specificity were induced in vitro.According to the quantities of prepared CD4~+ CD25~+ T cells injected through tail vein in kidney transplantation,models were divided into four ex- perimental groups:2?10~5(group I),5?10~5(groupⅡ),1?10~6(groupⅢ),2?10~6(groupⅣ).The models which had no injection served as control group(n=12).The survival of the transplanted kid- ney was observed.The levels of blood serum creatinine were determined and the histopathological changes in the transplanted kidney were observed at day 4,9 and 15.The results of histopathology were evaluated according to the standard of Banff Schema and the semi-quantitative scores were gained by way of Watanabe.The reaction indexes of receptor spleen cells to the donor antigens were tested by way of MTT at day 15.Results The mean survival time of transplanted kidneys was the highest in groupⅢ[(31.4?4.6)days]and lowest in the control group[(11.7?6.2)days].There was signif- icant difference between groupⅢand control group in levels of serum cre'atinine(P

OBJECTIVETo observe the clinical effect of low-dose once-daily tadalafil combined with Shuganyiyang Capsules in the treatment of mild-to-moderate erectile dysfunction (ED).

METHODSNinety patients with mild-to-moderate ED were equally randomized to groups A, B and C to receive Shuganyiyang Capsules, tadalafil, and tadalafil + Shuganyiyang Capsules, respectively. The scores of the patients on IIEF-5 and SF-PAIRS (15-Item Short Form of Psychological Interpersonal Relationship Scales) were recorded before and at 1 and 3 months after treatment.

RESULTSThe IIEF-5 scores of groups A, B and C were 10.13 +/- 1.55, 11.00 + 1.60 and 10.73 +/- 1.91 before treatment, and 13.77 +/- 2.11, 17.77 +/- 2.13 and 17.17 +/- 3.84 at 1 month after treatment, significantly higher in B and C than in A (P <0. 001) , but with no remarkable difference between B and C (P =0. 411). At 3 months after treatment, the IIEF-5 scores were 15.77 +/- 2.05, 18.07 +/- 2.24 and 19.37 +/- 3.76 in the three groups, dramatically higher in B and C than in A (P <0.001) as well as in C than in B (P<0.05). The scores on sexual self-confidence, sexual spontaneity and time concerns in SF-PAIRS were 3.90 +/-0.80, 8.67 +/- 1.94 and 14.43 +/- 1.92 before medication, 5.83 +/- 1.02, 9.90 +/- 1.75 and 11.17 +/- 1.68 at 1 month and 6.73 +/- 0.98, 11.07 +/- 2.08 and 10.67 +/-1.60 at 3 months after medication in group A; 4.17 +/- 0.87, 9.37 +/-1.43 and 14.47 +/-1.57 before medication, 6.47 +/-0.78, 10.83 +/- 2.18 and 10.20 +/-1.56 at 1 month and 6.83 +/-0.91, 11.30 +/- 1.88 and 9.47 +/- 1.57 at 3 months in group B; and 4.23 +/-0. 94, 9.50 +/- 1.89 and 14.67 +/- 2.91 before medication, 8.03 +/- 1.67, 13.43 +/-1.10 and 9.70 +/-1.21 at 1 month and 8.93 +/- 1.78, 14.70 +/- 1.26 and 8. 87 +/- 0. 97 at 3 months in group C. Compared with the baseline, the SF-PAIRS scores of the three groups were all significantly improved after treatment (P <0. 05) , and markedly higher in C than in the other two groups (P <0.05).

CONCLUSIONLow-dose once-daily tadalafil combined with Shuganyiyang Capsules is obviously effective in the treatment of mild-to-moderate ED, which not only improves the patients'erectile function, sexual self-confidence and sexual spontaneity, but also reduces their time concerns.

Adult ; Carbolines ; administration & dosage ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Erectile Dysfunction ; drug therapy ; Humans ; Male ; Tadalafil ; Treatment Outcome

This study is aimed at investigating the potentials of ex vivo expansion and pluri-differentiation of cryopreservation of adult human bone marrow mesenchymal stem cells (hMSCs) into chondrocytes, adipocytes and neurocytes. Cryopreserved hMSCs were resuscitated and cultured for 15 passages, and then induced into chondrocytes, adipocytes and neurocytes with corresponding induction medium. The induced cells were observed for morphological properties and detected for expressions of type II collagen, triglyceride or neuron-specific enolase and nestin. The result showed that the resuscitated cells could differentiate into chondrocytes after exposure to transforming growth factor beta(1) (TGF-beta(1)), insulin-like growth factor I (IGF-I) and vitamin C (V(C)), and uniformly changed morphologically from a spindle-like fibroblastic appearance to a polygonal shape in three weeks. The induced cells were heterochromatic to safranin O and expressed cartilage matrix-procollagenal (II) mRNA. The resuscitated cells cultured in induction medium consisting of dexamethasone, 3-isobutyl-1-methylxanthine, indomethacin and IGF-I showed adipogenesis, and lipid vacuoles accumulation was detectable after 21 d. The resuscitated hMSCs were also induced into neurocytes and expressed nestin and neuron specific endolase (NSE) that were special surface markers associated with neural cells at different stage. This study suggested that the resuscitated hMSCs should be still a population of pluripotential cells and that it could be used for establishing an abundant hMSC reservoir for further experiment and treatment of various clinical diseases.
Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cells, Cultured ; Cryopreservation ; methods ; Humans ; Mesenchymal Stromal Cells ; cytology ; Pluripotent Stem Cells ; cytology ; Tissue Engineering ; methods

Objective To evaluate 1-week and 1-year outcomes of carotid artery stenting (CAS)using cerebral perfusion CT(PCT). Methods The clinical database of 20 patients with unilateral carotid artery stenosis( ≥60% ) who underwent CAS were retrospectively reviewed. Relative cerebral blood volume (rCBV), relative cerebral blood flow(rCBF) and relative mean transit time( rMTT) were measured by using cerebral PCT within one week before CAS and at one week and at one year after CAS. Cerebral MRI was performed within one week before CAS. The noncontrast CT was performed within one week before CAS and immediately after CAS. The arteriography was performed at one year after CAS. The variance analysis was performed to determine whether there were significant differences of rCBV, rCBF, rMTT in anterior cerebral artery area( ACA area), middle cerebral artery area( MCA area), posterior cerebral artery area( PCA area),basal ganglia area, front and back cortical watershed area( CWS area) and internal watershed area( IWS area) among the different time points. Results In the three measures, there was no significant difference of rCBV in all areas among the three time points( P ＞ 0. 05 ) , and there was no significant difference of rCBF and rMTT in PCA area( P ＞ 0. 05 ), but there were significant differences of rCBF and rMTT in all other areas among the three time ponits(P ＜0. 01). In one week before CAS, at one week and at one year after CAS, rCBF of 20 patients is 0. 86 ±0. 06, 0. 95 ±0. 04, 0. 98 ±0. 07 in ACA area, 0. 81 ±0. 04, 1.06 ±0. 04, 1.03 ±0.07 in MCA area, 0. 84 ±0. 06, 0. 97 ±0. 04, 0. 96 ±0. 04 in basal ganglia, 0. 78 ±0. 03,0. 97 ±0. 03, 0. 96 ±0. 02 in front CWS area, 0. 77 ±0. 03, 1.00 ±0. 02, 0. 98 ±0. 03 in back CWS area,and 0. 80 ± 0. 04, 0. 94 ± 0. 03, 0. 93 ± 0. 04 in IWS area ( F = 18. 95, 146. 41,63.03,540. 85,415.97,164.19, P＜0. 01). rMTT is 1.17 ±0.05, 1.04±0.04, 1.01 ±0.06 in ACA area, 1.41±0.06, 1.08±0.04, 1.07±0.04 in MCA area, 1.20±0.06, 1.06±0.04, 1.05±0.04 in basal ganglia, 1.41 ±0.05,1.10 ±0. 05, 1.09 ±0. 04 in front CWS area, 1.43 ±0. 10, 1.07 ±0. 06, 1.08 ±0. 06 in back CWS area,1.29±0.10, 1.09 ±0.05, 1.11 ±0.07 in IWS area (F=51.74, 248. 89, 70.08, 381.68, 288.94,41.53, P ＜0. 01 ). There were significant differences of rCBF and rMTT between those measured one week before CAS and one week or one year after CAS ( P ＜ 0. 01 ), but there were no significant differences of rCBF or rMTT in any area measured between those at 1 week after CAS and those measured at 1 year after CAS(P＞0.05). Conclusions Hemodynamic outcome at one year after CAS is good in the absence of contralateral carotid artery steno-occlusive disease. In addition, the coherence of results between 1-week and 1-year indicates that the outcome of one week after CAS could predict long-term hemodynamic outcome.

Objective To study the clinical feature,treatment,and prognosis of the cytomegalovirus (CMV)pneumonia patients treated with immunosuppressor against kidney disease.Mlethod The patients received immunosuppressor against kidney disease in The First Affiliated Hospital of Sun Yat-sen University from June 1999 to December 2006.CMV antigen of leucocyte in the peripheral blood and/or bronchoalveolar lavage fluid of these patients were detected with immunocytochemical methods,and 21 patients were found suffering from CMV pneumonia.The 21 patients were introvenously injected with ganciclovir 5～10 mg/(kg?d),and the immunosuppressive agent treatment suspended.Their clinical feature and prognosis were retrospectively analyzed. Results The 21 patients received corticosteroids before CMV pneumonia contracted,of them,13 patients had been intensively treated with Methyllprednisolone with mean total dose(3.2?0.6)g.Of them,15 had been treated with cyclophosphamide with mean total dose(3.8?1.3)g.The median time from the beginning of using immunosuppressor to the onset of CMV pneumonia was 25(13～92)days.All patients had fever,cough, shortness of breath and X-ray showed interstitial pneumonia,of them,19 patients developed hypoxemia,and 11 patients' CMV antigen was positive in the leucocyte from bronchial lavage fluid.The result showed 9 patients survived and 12 died.The average duration of treatment with ganciclovir was(26.2?6.3)days. CMV pneumonia is a serious complication in patients who were treated with immunosuppressor against kidney disease.The mortality is high.Ganciclovir is a medicine of choice to treat CMV pneumonia.

Porcine reproductive and respiratory syndrome virus (PRRSV) has been recognized as one of the most important pathogens of pigs throughout the world. In 2006, more than 10 provinces of China have experienced an epizootic outbreak of pig diseases characterized by high fever, reddened skin and high morbidity and mortality. From June 2006 to April 2007, we have investigated some clinical samples in Hubei province by RT-PCR and cloned several major genes, N, GP5 and NSP2 gene, shown in this study. Phylogenetic analysis of these genes revealed that the highly pathogenic PRRSV variant, ZB, was responsible for 2006 emergent outbreak of pig disease in Hubei province similar with those variants isolated from other provinces in China in 2006, and belongs to the NA-type PRRSV. In the PRRSV variants, the N and GP5 shear about 90% identity with prototypic ATCC VR-2332 and some typical NA-type Chinese isolates, except the 2850bp NSP2 gene (only shares 65% identity with ATCC VR-2332). But they all shear more than and 97% identity with other highly pathogenetic Chinese PRRSV strains. Additionally, there are extensive amino acid (aa) mutations in the GP5 protein and 2 deletions in the Nsp2 protein when compared with the previous isolates. Most of the variants found in 2006 epizootic outbreak of pig diseases in China were the farthest variants from the typical NA-type PRRSV in phylogenetic distance, and these diversities may be responsible for the differences in the pathogenicity observed between these variants and original Chinese PRRSV strains.

OBJECTIVETo determine the location of the vertebral artery foramens from C(3) to C(6) and their relationship to the point 1 mm medial to the center of the lateral mass and to identify the value of oblique radiograph for cervical lateral mass screw trajectory by a cadaveric study.

METHODS(1) Twenty-eight cervical specimens (C(3)-C(7)) of human cadavers aged from 28 to 79 years were analysed. The transverse radiographs of C(3)-C(6) vertebrae were taken and the angle between the parasagittal plane and the line connecting the point of the lateral mass with the lateral limit of the transverse process foramen of C(3)-C(6) were measured. (2) The K-wires were drilled into lateral mass of C(3)-C(6) starting 1 mm medial to the center of the lateral mass and exiting by the juncture between the transverse process and the facet in ten specimens. Four wire placements under direct visualization, including placement of the wire tip staying the ventral cortex and 2, 4, 6 mm over-penetration of the ventral cortex of lateral mass, were performed separately on each specimen. After each placement, radiographs were taken on 45 degrees oblique left and 45 degrees oblique right views. Each intervertebral foramen on the oblique radiographs was divided into two parts: superior and inferior parts. The former is the true intervertebral foramen, while the latter is the intertransverse foramen on the gross specimen. The number of wire tips in each part was quantified for each placement. All results on the radiographs were compared with those on the gross anatomy.

RESULTS(1) The angles between the parasagittal plane and the line connecting the posterior starting point of the lateral mass with the lateral limit of the transverse foramen (C(3)-C(6)) were lateral to the sagittal plane, ranging from 5 degrees to 12 degrees. Among the vertebrae, there were no statistically significant difference (P > 0.05). (2) 15% of the wires without over-penetration and 41.3% with 2 mm over-penetration were found in the inferior parts of the intervertebral foramen in oblique views, while the wires were not noted in the intervertebral foramen by gross anatomy. with 4 mm over-penetration of the ventral cortex, 35% and 65% of wires were noted in the superior and inferior parts of the intervertebral foramen respectively, while only 28.8% of wires were found in the inferior part approximating the nerve roots in gross specimens. With 6 mm over-penetration, the number in the intervertebral foramen were 63.8% superiorly and 36.2% inferiorly on the oblique radiographs while all the tips were at the inferior part (intertransverse foramens) in gross specimens. The tip of wire crossed the line connecting the posterior borders of the intervertebral foramens in oblique radiographs when it penetrated the ventral cortex of lateral mass 4 mm or more.

CONCLUSIONS(1) There is no risk of damaging the vertebral artery if a screw is directed more than 15 degrees lateral to the sagittal plane at C(3 approximately 6) starting 1 mm medial to the center of the lateral mass. (2) Ideal screw tip position on oblique radiograph may not cross the line connecting the posterior borders of the intervertebral foramen on radiograph. If the screw tip is noted in the superior part of intervertebral foramen on the oblique radiograph, the screw may be identified as dangerous.

Adult ; Aged ; Bone Screws ; Cadaver ; Cervical Vertebrae ; anatomy & histology ; surgery ; Humans ; Internal Fixators ; Middle Aged ; Models, Anatomic ; Spinal Fusion ; instrumentation ; methods ; Spinal Nerve Roots ; anatomy & histology ; Vertebral Artery ; anatomy & histology

Background Anterior chamber associated immune deviation (ACAID) is characterized by a Th2 cell response. GATA-3 has been shown to be necessary for the activation of Th2 cells. This study was designed to examine the expression of GATA-3 in the development of ACAID. Methods ACAID was induced by injection of 50 μg interphotoreceptor retinoid binding protein (IRBP) into the anterior chamber (AC) of Wistar rats. Delayed-type hypersensitivity (DTH) was evaluated on day 3, 7, 14, 21, 28 after IRBP inoculation. GATA-3 expression was detected using immunohistochemical staining. The expression of GATA-3 mRNA at different time points after AC injection of IRBP was assayed by reverse transcriptase polymerase chain reaction (RT-PCR). Results A significant DTH reaction was observed in Wistar rats on day 3 and 5 after IRBP inoculation. The DTH reaction was decreased 7 days after IRBP inoculation. GATA-3 expression was weak at both mRNA and protein levels in the normal spleen, but was significantly increased on day 5, 7, 14, and 21 after AC injection of IRBP. Conclusion The expression of GATA-3 is increased during ACAID, suggesting that GATA-3 may be involved in the development of ACAID.

The purpose of the present study was to investigate the effects of advanced glycation end products (AGEs) modified protein on the permeability of endothelium monolayers and morphological changes of actin cytoskeleton. The roles of receptor for AGEs (RAGE), oxidant stress and the activation of p38 MAPK pathway in this pathological procedure were elucidated. Human umbilical vein endothelial cells (HUVECs)-derived cell line (ECV304) were incubated with AGEs modified human serum albumin (AGE-HSA) in concentrations of 12.5, 25, 50, and 100 microg/ml respectively, for 2, 4, 8, 12 and 24 h. As control, HSA of the same concentration was administered to cells. Then TRITC-albumin was added to evaluate Pa value that reflects the permeability of endothelial monolayer. Furthermore, to visualize the morphological changes of actin cytoskeleton, the treated cells were incubated with rhodamine-phalloidin to stain F-actin. The results showed that the trans-endothelial membrane flux of albumin was significantly increased in a concentration- and time-dependent manner upon the stimulation of AGE-HSA, accompanying with actin reorganization. The blockage of AGE and RAGE binding with anti-RAGE IgG and the pharmacological inhibition of NADPH oxidase or p38 MAP kinase greatly attenuated the AGE-induced hyperpermeability response, respectively. These results indicate that RAGE, NADPH oxidase and p38 MAPK are possibly involved in the mediation of AGEs-induced barrier dysfunction and actin cytoskeleton reorganization in endothelial cells.
Actin Cytoskeleton ; physiology ; Capillary Permeability ; physiology ; Cell Line ; Cells, Cultured ; Endothelium, Vascular ; cytology ; Glycation End Products, Advanced ; physiology ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Oxidative Stress ; physiology ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; physiology ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo determine the association between urine transforming growth factor beta(1) (TGF-beta(1)) concentration and long-term renal allograft function.

METHODSPatients undergoing kidney transplantation between August 1, 1999 and June 30, 2001 and survived for one year with normal renal functions were investigated. The blood and urine TGF-beta(1) concentrations were tested at an interval of at least 6 months. Totally 134 patients completed the 3-year follow up investigation. Correlation between their renal functions (creatinine clearance rates) and their urine relative TGF-beta(1) concentrations 1 year after renal transplantation were determined. Of the 134 renal recipients, 16 were diagnosed to have chronic allograft nephropathy (CAN), and their blood and urine TGF-beta(1) concentrations 1 year after renal transplantation were compared with those of the recipients free of CAN.

RESULTSThere was a positive correlation between long-term renal functions (loss of creatinine clearance rates) and in relative concentration of TGF-beta(1) urine 1 year after renal transplantation. The urine TGF-beta(1) concentrations of CAN and CAN-free recipients 1 year after transplantation were 182.7-/+40.2 and 398-/+33.5 pg/mg.Cr, respectively, showing significant differences. The blood TGF-beta(1) concentrations of CAN and CAN-free recipients were comparable (32.1-/+4.7 and 31.9-/+4.8 ng/ml, respectively).

CONCLUSIONUrine TGF-beta(1) is significantly elevated even before the onset of renal dysfunction in patients with CAN, and urine TGF-beta(1) level in early stage after renal transplantation can help predict long-term renal function.

Adult ; Female ; Follow-Up Studies ; Humans ; Kidney Diseases ; etiology ; physiopathology ; Kidney Transplantation ; adverse effects ; methods ; Male ; Postoperative Complications ; blood ; physiopathology ; urine ; Time Factors ; Transforming Growth Factor beta1 ; blood ; urine