Objective To present a modified technique of total cystectomy and ileal neobladder. Methods Radical cystectomy and ileal neobladder were performed for 25 male patients. The dome, the upper part or the lateral walls and the base of the bladder were dissected antegradly. After cutting the ureters, the mobilization was then made retrogradly.For the construction of the reservior, the isolated 40 cm ileal segment was opened along its antimesenteric border and then arranged into a W shape. The wall of ileal pouch was created by side-to-side blanket suture of the incised ileum. The ureters were implanted at the two corners of W shape with antireflux nipple. Results 24 of 25 cases of ileal neobladder have been followed up for 2 to 84 months with a mean of 24 months.The operation took 150 to 310 minutes with a mean of 240 minutes. The intraoperative blood transfusion was 0 to 1200 ml.1 patient died of cancer metastasis. 23 patients have achieved excellent daytime continence,whereas only 14 had nighttime continence. The mean bladder capacity for the entire group was 310 ml. Renal functions of 2 patients were mildly abnormal.Hypokalemia occurred in 2.There was no metabolic acidosis.There were 3 cases of hydroureter or hydronephrosis on one side. 1 calculus of neobladder was discovered on BUS. On voiding cystography, no reflux has been noted in all.And no patient suffered from urethral recurrence. Conclusions The modified total cystectomy and ileal neobladder takes less time and is more feasible.

AIM:To explore the impact of granulocyte colony-stimulating factor (G-CSF) on acute graft-ver-sus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in a murine model and its possible mechanisms.METHODS:Male C57BL/6 (H-2b) and BALB/c (H-2d) mice were used as the allogeneic and syngeneic donor mice , respectively .Moreover , female BALB/c mice were used as recipient mice .The recipient mice were conditioned by a single dose ( 8 Gy ) of total body irradiation ( TBI ) .The recipient mice were randomly divided into 7 groups:TBI group, Syn-BMST control group, post-Syn-BMST G-CSF administration (Syn-BMST+G-CSF) group, allo-BMT control group, post-allo-BMT G-CSF administration (allo-BMT+G-CSF)group, allo-BMST control group and post-al-lo-BMST G-CSF administration (allo-BMST+G-CSF) group.The mice in control groups and G-CSF administration groups were subcutaneous injected with 0.1 mL normal saline (NS) and 0.1 mL NS containing 2μg G-CSF per day from 1st day, respectively.The effect of G-CSF on aGVHD was evaluated by clinical manifestations and pathological changes , as well as survival time of the mice in different groups .The serum levels of IL-2, IL-4, IFN-γand TNF-αin allo-BMST and allo-BMST+G-CSF groups were detected by ELISA at 10th day.Flow cytometry was used to analyze the immunophenotypes of splenocytes at 10th day.RESULTS:The mice in TBI group were all died for hematologic failure on 9~15 d after TBI.No effect of G-CSF on the survival of the mice underwent Syn-BMST and transplantation of single allogeneic marrow cells was observed.The mean survival days in allo-BMST group and allo-BMST+G-CSF group were (34.8 ±4.5) d and (19.8 ± 6.1) d’respectively (P<0.01).Moreover, post-transplant administration of G-CSF increased the spleen total nucleated cells count (SpTNC), NK cells subset, and DC1/DC2 ratio in the spleen with over 99%of donor chimerism rate at 10th day.No difference in the levels of serum IL-2, IL-4, IFN-γand TNF-αbetween the 2 group at 10th day was found.CON-CLUSION:The administration of G-CSF after allo-BMST significantly aggravates mouse aGVHD .The expansion of NK cells stimulated by G-CSF may be involved in the mechanism of generating alloreactivity against host cells .These results imply there may be potential risk of evoking or aggravating acute GVHD if G-CSF is administered in the early stage of clini-cal allo-HSCT.

0.05).There was a positive correlation between the tempo of the engraftment and the progressive increase of the doses of MNC in the range of (3～5.99)?108/kg,but which was not found in the range of 6?108/kg.Conclusion These results indicate that MNC taken as an index for the measurement of the hematopoietic stem cell/progenitor content can reliably predict hematopoietic reconstitution after both HLA-matched and-mismatched sibling donor peripheral blood stem cell transplantation,with the rate and tempo of engraftment comparable with that in CD34+ cell group,and however,the target dose was reached with one apheresis in all 83 donors in MNC group(100%),as compared with 14 donors in CD34+ cell group(37.84%),suggesting that MNC may replace CD34+ cell as an independent index for the assessment of the stem cell/progenitor content in clinical practice.

AIM: To investigate the expression of SCL (stem cell leukemia) gene in bone marrow stromal cells (BMSCs) and bone marrow hematopoietic cells from patients with aplastic anemia (AA) and normal individuals. METHODS: Bone marrow stromal cells from AA (9 cases) and normal individuals (33 cases) were amplified by long-term in vitro culture. The adherent and nonadherent cells were collected respectively. RT-PCR-ELISA assay was then performed to detect the expression of SCL gene and the housekeeping gene ?_2 microglobulin (?_2M). The expression ratio of SCL gene were analyzed and its expression level was normalized by ?_2M gene acting as an internal calibration for the purpose of semi-quantitative analysis. RESULTS: The expression ratio of SCL gene was lower in BMSCs from AA (22.2%) than that in normal controls (69.7%, P

AIM: To investigate the expression of transcription factor GATA-3 gene in the bone marrow stromal cells (BMSCs) from patients with aplastic anemia (AA) and normal controls. METHODS: The expression of GATA-3 gene was analyzed by using RT-PCR-ELISA in BMSCs from 34 normal cases and 9 cases with AA. The standardized semi-quantitative expression level of GATA-3 gene in BMSCs from patients with AA was compared with normal controls. RESULTS: The expression of GATA-3 gene was detected in BMSCs from both normal controls and the cases with AA. The expression level of GATA-3 gene in BMSCs from AA was significant higher than that from the normal controls (P

BACKGROUND: Cancer stem-like cells (CSCs) are a small subset of cells in tumors that exhibit self-renewal and differentiation properties. CSCs play a vital role in tumor formation, progression, relapse, and therapeutic resistance. B7-H3, an immunoregulatory protein, has many protumor functions. However, little is known about the mechanism underlying the role of B7-H3 in regulating gastric cancer (GC) stemness. Our study aimed to explore the impacts of B7-H3 on GC stemness and its underlying mechanism. METHODS: GC stemness influenced by B7-H3 was detected both in vitro and in vivo . The expression of stemness-related markers was examined by reverse transcription quantitative polymerase chain reaction, Western blotting, and flow cytometry. Sphere formation assay was used to detect the sphere-forming ability. The underlying regulatory mechanism of B7-H3 on the stemness of GC was investigated by mass spectrometry and subsequent validation experiments. The signaling pathway (Protein kinase B [Akt]/Nuclear factor erythroid 2-related factor 2 [Nrf2] pathway) of B7-H3 on the regulation of glutathione (GSH) metabolism was examined by Western blotting assay. Multi-color immunohistochemistry (mIHC) was used to detect the expression of B7-H3, cluster of differentiation 44 (CD44), and Nrf2 on human GC tissues. Student's t -test was used to compare the difference between two groups. Pearson correlation analysis was used to analyze the relationship between two molecules. The Kaplan-Meier method was used for survival analysis. RESULTS: B7-H3 knockdown suppressed the stemness of GC cells both in vitro and in vivo . Mass spectrometric analysis showed the downregulation of GSH metabolism in short hairpin B7-H3 GC cells, which was further confirmed by the experimental results. Meanwhile, stemness characteristics in B7-H3 overexpressing cells were suppressed after the inhibition of GSH metabolism. Furthermore, Western blotting suggested that B7-H3-induced activation of GSH metabolism occurred through the AKT/Nrf2 pathway, and inhibition of AKT signaling pathway could suppress not only GSH metabolism but also GC stemness. mIHC showed that B7-H3 was highly expressed in GC tissues and was positively correlated with the expression of CD44 and Nrf2. Importantly, GC patients with high expression of B7-H3, CD44, and Nrf2 had worse prognosis ( P = 0.02). CONCLUSIONS B7-H3 has a regulatory effect on GC stemness and the regulatory effect is achieved through the AKT/Nrf2/GSH pathway. Inhibiting B7-H3 expression may be a new therapeutic strategy against GC.
Humans ; Cell Line, Tumor ; Neoplasm Recurrence, Local ; NF-E2-Related Factor 2/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Signal Transduction ; Stomach Neoplasms