AIM and METHOD: Human endothelial nitric oxide synthase (eNOS) gene was transfected into human phagocytic cell U937 and the effects of gene transfer on cytokines and cAMP production were observed. RESULTS: A functional eNOS was stably expressed in transfected U937 cells, but NO release was undetectable in intact transfectants. However, eNOS gene expression upregulated tumor necrosis factor - a release and downregulated interleukin - 10 and cAMP production in either presence or absence of NOS inhibitor N? - monomethyl - L - arginine. CONCLUSION: The function of tranfected eNOS gene product showed cellular speciality. The effector molecule that changed the produced pattern of cytokines and cAMP in phagocytic cells seems not likely the nitric oxide.

BACKGROUND: Mesenchymal stem cells are the stem cells that possess the capability for self-renewal and multi-directional differentiation. Umbilical cord is the tissue outside the embryos and would be fallen off after parturition. In addition, it has wide source and no ethical restriction, so it is promising to be the first choice for mesenchymal stem cells. OBJECTIVE: To detect the surface markers CD29, CD44, CD49e, CD73, CD90, CD34, CD45, and CD271 of human umbilical cord-derived mesenchymal stem cells (hUCMSCs) prior to and after cryopreservation and resuscitation. METHODS: After isolation and culture, morphology of the primary, P4 and P8 hUCMSCs was observed prior to cryopreservation and after resuscitation. Surface markers CD29, CD44, CD49e, CD73, CD90, CD34, CD45, and CD271 of primary, P4, and P8 hUCMSCs were detected through the use of flow cytometry prior to cryopreservation and after resuscitation RESULTS AND CONCLUSION: hUCMSCs prior to cryopreservation and hUCMSCs of different passages after resuscitation present the same phenotype, i.e., positive for CD29, CD44, CD49e, CD73, and CD90, and negative for CD34, CD45, CD271. These findings suggest that primary hUCMSCs do not present changes in surface markers after cryopreservation and resuscitation.

Objective: To investigate the changes of gene expression in CD34+ hematopoietic stem and progenitor cells (HSPCs) under different growth environments. Methods: Umbilical cord blood mononuclear cells (UCB MNCs) were cultured in static and stirred systems. After 7 days of culture, CD34+ cells were isolated and total RNA was extracted. Gene expression patterns of CD34+ cells from fresh, static and stirred cultures were compared using differential display (DD). Results: 30 gene fragments displayed differential expression levels based on the conditions of DD. One of differentially expressed genes was identified as RAN, which is a member of oncogene RAS family. This gene may be associated with proliferation of hematopoietic cells. Conclusion: Different growth environments induced differential gene expression patterns of CD34+ HSPCs. These differentially expressed genes would give new insights into optimizing in vitro environments for expanding hematopoietic cells.

Objective To investigate the value of ultrasound integrated backscatter (IBS) in quantitative analysis of in-traluminal thrombus in abdominal aortic aneurysm (AAA).Methods IBS of intraluminal thrombus in 29 patients were measured,including All (average image intensity) and SDI (standard deviation of image intensity).Meanwhile,different echoes and characteristics of IBS in thrombus were compared and pathologic analysis was performed.Results ①Individual All value of homogenous thrombus differed remarkably,SDI value was low.There was no significant difference about All in homogeneous group and adjacent cavity group (P>0.05).All and SDI value in adjacent wall group were higher than those in other groups (P<0.01).②According to pathologic analysis,cellulose contents were abundant in adjacent wall group,but fewer in adjacent cavity group and homogeneous group.Conclusion IBS might be regarded as a sensitive and specific method and a quantitative basis for estimating and predicting the rupture risk of AAA.

OBJECTIVETo study the pathological change of rats' benign hyperplastic prostate (BHP) after radical denervation.

METHODSA total of 65 male spontaneous hypertension rats (SHR) at 30 weeks age were randomly assigned into treatment group, sham surgery control group and normal control group. In surgery group, all the axonal branches of the major pelvic ganglion (MPG) supplying the bilateral prostate were truncated, followed performing of cystostomy; In sham surgery control group, only cystostomy was performed; In normal control group, no procedure was performed. The rats were sacrificed at 3, 7, 11, 15 and >or= 21 d post-operation respectively. The gross morphological changes of prostate in all animals were observed.

RESULTSIn treatment group, the prostate in 3 d post-operation showed granular solidification and shrunken volume and the changes occurred gradually over time. The glandular epithelial cells showed gradual degeneration, necrosis and detachment. The glandular epithelium became progressively thinner, the smooth muscles elongated and thinned progressively and the stromal components showed mild to moderate overgrowth. At the later stage, the glandular epithelium, glandular lumen and smooth muscles gradually disappeared and the prostate was largely replaced by connective tissues. Electron microscopic study showed that the glandular cells gradually underwent vacuolar degeneration and the structures of basement membrane became fuzzy. The smooth muscles cells degenerated overtime and the fibroblasts and collagenous fibers in the stroma overgrew slowly. At the late stage, most of the glandular cells became necrotic, the basal membrane and smooth muscle cells disappeared and collagenous fibers were highly hyperplasic. In surgery group in 3 d post-operation, the S-100 staining of nerve fiber was diffuse and disappeared after 11 d while it persisted normally in other groups. The two values in sham surgery control group showed no significant changes post-operatively.

CONCLUSIONSAfter radical denervation, the rat prostate with benign hyperplasia (gland and smooth muscles) undergoes dramatic atrophic changes and the volume decreases significantly. It suggests that this treatment may represent a novel therapy for BPH.

Animals ; Denervation ; methods ; Disease Models, Animal ; Male ; Microscopy, Electron, Transmission ; Prostate ; innervation ; pathology ; ultrastructure ; Prostatic Hyperplasia ; pathology ; surgery ; Random Allocation ; Rats ; Rats, Inbred SHR

Objective To investigate the effect of transplantation of bone marrow mesenchymal stem cells (MSCs) genetically modified with human hepatocyte growth factor gene (hHGF) on angiogenesis in the rat lung.Methods Twenty F344 rats,aged 2 months,weighing 200-250 g,were randomly divided into 2 groups ( n =10 each):HGF group and control group (group C).MSCs genetically modified with hHGF was injected through the external jugular vein in group HGF.While the equal volume of DMEM culture medium (1 ml) was given instead in group C.The mean pulmonary artery pressure was detected at 28 days after transplantation.Then the rats were sacrificed and the lungs were removed for determination of the content of hHGF,expression of proliferating cell nuclear antigen (to reflect the degree of endothelial cell proliferation showed by the small pulmonary vessels) and Ⅷ factor (to reflect the density of the small pulmonary vessels),and microscopic examination.Results Compared with group C,no significant change was found in mean pulmonary artery pressure ( P ＞ 0.05),while the content of hHGF,degree of endothelial cell proliferation,and density of the small pulmonary vessels were significantly increased in group HGF ( P ＜ 0.01).No change was found in the structure of the small pulmonary vessels in group HGF.Conclusion Transplantation of MSCs genetically modified with hHGF can promote angiogenesis in the rat lung.

Objective To investigate the effect of human hepatocyte growth factor (hHGF) genetic modification on the ameliorating effects of mesenchymal stem cells (MSCs) implantation on pulmonary microvascular rarefaction in a rat model of pulmonary hypertension (PH).Methods MSCs were obtained from F344 rats and transduced with lentiviral vector modified with human HGF (hHGF-MSCs) or empty vector (EGFP-MSCs).Sixty-six 7 week old male F344 rats weighing 180-250 g were used in this study.PH was induced by left pneumonectomy and subcutaneous monocrotaline (MCT) 60 mg/kg injected at 2 weeks after operation.The animals with PH were randomly divided into 3 groups:control group (group C),EGFP-MSCs group (group E) and HGF-MSCs group (group H).Groups H and E received hHGF-MSCs or EGFP-MSCs 5 × 105 in DMEM 1 ml iv at 3 weeks after subcutaneous MCT injection,while group C received plain DMEM 1 ml.Mean pulmonary arterial pressure (mPAP) was measured and right ventricular hypertrophy and angiogenesis in the lung were assessed and the content of rat HGF (rHGF) and hHGF protein in lung tissue and pulmonary capillary density (by immuno-histochemistry) was measured at 2 weeks after MSCs implantation.The survival rates within 45 days after MCT administration were compared among the 3 groups.Results No hHGF was detected in groups C and E.Both hHGF-MSCs and EGFP-MSCs significantly reduced MPAP and right ventricular hypertrophy and increased pulmonary capillary density and survival rates in groups H and E as compared with group C and the efficacy of hHGF-MSCs was significantly greater than that of EGFP-MSCs.Barium angiography revealed that distal pulmonary vasculature was significantly increased in group H as compared with groups E and C.The survival of the rats receiving hHGF-MSCs was significantly longer in group H than that in groups E and C.Conclusion hHGF genetic modification can improve the ameliorating effects of MSCs implantation on PH-related microvascular rarefaction.

OBJECTIVETo observe blood uric acid levels and Goldstein grading, as well as their correlation in Wilson's disease (WD) patients with different Chinese medical syndrome types.

METHODSTotally 906 WD patients in line with inclusive criteria were assigned to 6 groups, i.e., the heart spirit confused by phlegm group (HSCP, 26 cases), the phlegm-fire disturbing heart group (PFDH, 90 cases), the retention of damp-heat group (RDH, 113 cases), deficiency of qi and blood group (DQB, 168 cases), the deficiency of Gan-yin and Shen-yin group (DGYSY, 327 cases), the deficiency of Gan and Shen group (DGS, 182 cases) due to different Chinese medical syndrome types. Recruited were another 160 healthy subjects having similar ages and diet structures, who came for medical examinations, as the healthy control group. Venous blood was collected from the medial cubital vein of each-patient on an empty stomach in early mornings to detect blood uric acid levels. Results Blood uric acid levels were lower in each syndrome type group than in the healthy control group (146.08 +/- 67.24 micromol/L in the HSCP group; 157.08 +/- 69.77 micromol/L in the PFDH group; 162.58 +/- 97.72 micromol/L in the RDH group; 156.20 +/- 62.63 micromol/L in the DQB group; 161.83 +/- 111.23 micromol/L in the DGYSY group; 194.41 +/- 90.01 micromol/L in the DGS group; 242.39 +/- 87.55 micromol/L in the healthy control group, P < 0.01). Blood uric acid levels were higher in the DGYSY group than in the other 5 syndrome groups (P < 0.01). Correlation analyses between Goldstein grading and blood uric acid showed that, along with increased Goldstein grade (that was aggravating disease conditions), WD patients' blood uric acid levels decreased (P < 0.01).

CONCLUSIONSWD patient's blood uric acid levels decreased more. Blood uric acid levels and Goldstein grading were different in various Chinese medical syndrome types. Blood uric acid levels had certain value in assessing the severity of WD.

Asian Continental Ancestry Group ; Heart ; Hepatolenticular Degeneration ; blood ; classification ; diagnosis ; Humans ; Medicine, Chinese Traditional ; Syndrome ; Uric Acid ; blood

BACKGROUNDMinimally invasive flexible ureteroscopy techniques have widely adopted in the management of patients with renal stones. We performed this study to investigate the value of virtual reality simulator training in retrograde flexible ureteroscopy renal stone treatment for catechumen.

METHODSThirty catechumen, included 17 attending physicians and 13 associate chief physicians, were selected for study. The trainees first underwent 1-hour basic training to get familiar with the instrument and basic procedures, then followed by 4-hour practice on virtual reality simulators. Before and after the 4-hour training, all trainees undertake an assessment with task 7 program (right low pole calyces stone management). We documented for each trainee the total time of procedure, time of progressing from the orifice to stone, stone translocation and fragmentation time, laser operate proficiency scale, total laser energy, maximal size of residual stone fragments, number of trauma from the scopes and tools, damage to the scope and global rating scale (GRS). The proficiency of this training program was analyzed by the comparison of the first and second assessment outcomes.

RESULTSSignificant improvement was observed in retrograde flexible ureteroscopy management of renal stone on virtual reality simulators after finishing the 4 hour special-purpose training. This was demonstrated by improvement in total procedure time ((18.37±2.59) minutes vs. (38.67±1.94) minutes), progressing time from the orifice to stone ((4.00±1.08) minutes vs. (13.80±2.01) minutes), time of stone translocation ((1.80±0.71) minutes vs. (6.57±1.01) minutes), fragmentation time ((4.43±1.25) minutes vs. (13.53±1.46) minutes), laser operate proficiency scale (8.47±0.73 vs. 3.77±0.77), total laser energy ((3231.6±401.4) W vs. (5329.8±448.9) W), maximal size of residual stone fragments ((2.66±0.39) mm vs. (5.77±0.63) mm), number of trauma from the scopes and tools (3.27±1.01 vs. 10.37±3.02), damage to the scope (0 vs. 0.97±0.76) and GRS (29.27±2.95 vs. 9.87±2.21). The differences between the first and the second assessment were all statistically significant (all P < 0.01).

CONCLUSIONThe virtual reality simulator training program can help the trainees to rapidly improve their retrograde flexible ureteroscopy skill in renal stone treatment.

Adult ; Computer Simulation ; Humans ; Kidney Calculi ; Male ; Ureteroscopy ; education ; Urology ; education

OBJECTIVETo investigate the differential expression of the hyaluronic acid synthase (HAS) family in human bladder transitional cell carcinoma (BTCC) and its potential clinical significance.

METHODSThe relative quantitative detection of the expression of HAS isoforms (HASs) was performed in 78 human BTCC tissues (mRNA & protein) and 12 normal human bladder mucosa (protein) by real-time RT-PCR and Western blot, and the results were statistically analyzed according to the clinical data.

RESULTSAll the BTCC tissues expressed three HAS isoform mRNA and protein, but to a different extent, as for mRNA, HAS3 > HAS2 > HAS1 (P < 0.001), with a significant difference in HAS1/HAS2, HAS1/HAS3 and HAS2/HAS3 (P = 0.003, < 0.001, 0.006, respectively). Among the proteins, the HAS2 expression was the highest, with a significant difference in HAS1/HAS2, and HAS2/HAS3 (P = 0.004, 0.001, respectively), but not in HAS1/HAS3. The elevation of HAS1 mRNA and protein expression was significantly related with the tumor malignancy, tumor initial onset/recurrence, T1/T2 and T1/T3-4 stags, and tumor grading (P = 0.02, < 0.001, 0.038, < 0.001; 0.025, 0.031, 0.023, 0.002; respectively). The HAS2 mRNA expression was significantly related with tumor size (diameter ≤ 3.0 cm/> 3.0 cm), tumor number (single or multiple), tumor initial onset/recurrence, T-staging, and histopathological differentiation (low grade/high grade) (P = 0.012, 0.004, < 0.001, < 0.001, < 0.001, respectively), but its protein expression was not significantly different in all subgroups except with the tumor size (mass diameter > 3.0 cm/≤ 3.0 cm). However, HAS3 mRNA and protein expression had no significant difference among all the subgroups. In normal human bladder mucosa, no HAS expressions were detected.

CONCLUSIONSThe abnormally high expression of the HASs further indicate the reliability of hyaluronan as a urinary marker for human BTCC. Compared with HAS1 and HAS3, HAS2 as a marker may have more usefulness in studies on human BTCC carcinogenesis or development. The high expression of HAS1 protein seems to play a more important role in the BTCC tumorigenesis, and may indicate a poor prognosis of the BTCC patients.

Biomarkers, Tumor ; Blotting, Western ; Carcinoma, Transitional Cell ; genetics ; metabolism ; Glucuronosyltransferase ; metabolism ; Humans ; Hyaluronan Synthases ; Hyaluronic Acid ; metabolism ; Neoplasm Recurrence, Local ; RNA, Messenger ; biosynthesis ; Reproducibility of Results ; Urinary Bladder Neoplasms ; genetics ; metabolism