BACKGROUND: At present, there is little related report about producing a whole-kidney acellular matrix (ACM) scaffold in rats using perfusion. The cellular biocompatibility of the ACM is poorly understood. OBJECTIVE: To produce a whole-kidney ACM scaffold in rats by perfusion, to evaluate the cytocompatibility of ACM with the L929 cells in vitro, and to assess the possibility of ACM as the cytoskeleton and tissue-engineered urinary organ construction. DESIGN, TIME AND SETTING: An in vitro observation was performed at the Central Laboratory of Zhujiang Hospital, Southern Medical University from February to May 2009. MATERIALS: Kidneys were obtained from 12-week-old Whista rats, while ureter, renal veins and renal artery were reserved. Intravenous catheters were inserted through renal arteries to establish channels for perfusion. Whole-kidney retrograde perfusion was performed with successively heparinized PBS, 1% SDS, deionized water, 1% TritonX-100 and antibiotic-containing PBS under a pressure of 9.81 kPa to prepare whole-kindney acellular matrix scaffolds. METHODS: ① Samples were randomly divided into blank group (without any cells), negative control group (culture media), experimental group (rat kidney ACM leaching liquor), and positive control group (culture media containing 0.64% phenol). L929 cells in the logarithmic phase were seeded in 96-well plates at the density of 4×10~3/well, with 5 wells in each group. At 24, 72, and 120 hours after incubation, cells were stained with MTT method to detect absorbance at 490 nm and calculate relative growth rate. ② Control group (culture medium), experimental group (rat kidney ACM leaching liquor), and positive control group (culture media containing 0.64% phenol) were set up to detect cell apoptosis at 48 hours after culture using flow cytometry. MAIN OUTCOME MEASURES: Microstructure of the scaffold, cytotoxicity and cell apoptotic rate. RESULTS: After SDS and TritonX-100 union processing, reticulate structures made of basilar membrane and collagen were shown under scanning electron microscope rather than normal structures of cells. At every time intervals (24, 72, and 120 hours), there was no significant difference in the absorbance between experimental group and negative control group (P > 0.05). The grade of the cytotoxicity of the ACM was .0-1. There was no significant difference in cell apoptotic rate between experimental group and negative control group (P > 0.05). CONCLUSION: The whole-kidney acellular matrix scaffolds in rat by perfusion have good biocompatibility.

Objective To present our initial experience of pure laparoendoscopic single-site surgery (LESS) for radical cystectomy and bilateral pelvic lymph node dissections. Methods 10 patients with pathology confined bladder urothelial carcinoma underwent laparoendoscopic single-site radical cystectomy, including 9 males and 1 female. After a 3-4 cm lower median abdominal incision was made, quadport or homemade single multichannel port was inserted, and conventional and prebent laparoscopic instruments were utilized. The surgical procedure included bilateral pelvic lymphadenectomies, radical cystectomy and building with a sigmoid orthotopic neobladder by open surgery.Results No extra port needed, neither conversion to open or conventional laparoscopic surgery. The time of LESS procedure ranged from 130 to 330 min (mean 243 nin). Estimated blood loss ranged from 50 to 600 ml (mean 270 ml). 5 patients needed blood transfusion of 2 to 4 units. The pathologic evaluation revealed bladder urothelial carcinoma, negative margins and negative pelvic lymph node involvement. No mortality or severe complications were observed perioperatively. After followup of more than 6 months, all revealed controllable urination at daytime, while 4 revealed nocturnal incontinence and needed one or two pads during nighttime. No evidence of recurrent or metastatic disease was detected. Conclusions LESS radical cystectomy and bilateral lymphadenectomies was safe and feasible, and short-term follow-up showed good tumor control outcomes. Homemade single multichannel port made of two elastic ring and glove was simple and effective.

Objective To discuss the feasibility of percutaneous nephrolithotomy (PCNL) for treating upper urinary calculi under local anesthesia.Methods One thousand three hundred and sixty-three patients who suffered with upper urinary calculi were treated with PCNL, the puncture and tracts were created using local anesthesia and guided through ultrasound.Of the 1363 patients, 475 patients had complicated renal caluli, 520 patients had kidney pelvic calculi and 368 had upper uretere calculi.Results All of the patients successfully received PCNL under the local anesthesia.Of the 1363 patients five tracts were used in two patients, four tracts were used in four patients, three tracts were used in nine patients, double tracts were used in 25 patents and one tract was used in the remaining patients.The stone-free rate was 96.0% in the kidney pelvic calculi patients, 100.0% in the upper uretere calculi patients, and 90.1% in the complicated renal caluli patients.90.0% patients were find well throught the operation, 10.0% patients find a little pain and solved by another more 5 - 10 ml lidocaine local injection or 50 - 75 mg pethidine hydrochloride intramuscular injection.No case stop operation because of pain or position changed.All without any severe complications such as damaged of liver, spleen, thorax and intestine.Conclusion The PCNL handled under local anesthesia was simple safe and effective, deserved clinical popularizing use.

Objective To explore the inhibitory effect of tetramethylpyrazine (TMP) on ultraviolet A-induced senescence as well as matrix metalloproteinase-1 (MMP-1) and-3 (MMP-3) mRNA expressions in human dermal fibroblasts (HDFs).Methods HDFs were isolated from the prepuce by enzymatic digestion, and subjected to primary culture.Cultured HDFs were randomly divided into several groups: control group cultured in high-glucose DMEM medium and receiving no treatment, three TMP groups treated with 20, 50 and 100 mg/L TMP respectively, UVA group receiving UVA radiation alone, UVA + TMP groups pretreated with 20, 50 and 100 mg/L TMP respectively for different durations followed by UVA radiation.UVA radiation was given once daily for 5 consecutive days.The 55th passage HDFs served as the P55 group (senescence control group).Subsequently, CCK-8 assay was performed to evaluate the proliferative activity of HDFs in vitro, optical microscopy to observe the morphologic changes of HDFs after UVA radiation, β-galactosidase staining to estimate the senescence in HDFs, and real-time fluorescence-based quantitative PCR to quantify the mRNA expressions of MMP-1 and MMP-3 in HDFs.Statistical analysis was carried out by one-way analysis of variance (ANOVA) followed by least significant difference (LSD)-t test or Dunnett's T3 test.Results Compared with the control group, the proliferation of HDFs was significantly but transiently inhibited in vitro after the treatment with 100 mg/L TMP for 48 hours (P ＜ 0.05), but showed no significant changes after the treatment with 20 or 50 mg/L TMP for 24, 48 or 72 hours or after the treatment with 100 mg/L TMP for 24 or 72 hours (all P ＜ 0.05).The pretreatments with TMP of 20, 50 and 100 mg/L for 24, 48 and 72 hours all promoted the proliferation of HDFs to a certain degree in the UVA + TMP groups compared with the UVA group, with significant differences in cellular proliferative activity among the UVA group, UVA + TMP groups and control group at 24, 48 and 72 hours (F =17.451,15.231, 23.535, all P ＜ 0.01).Compared with the UVA group, the proliferative activity of HDFs was significantly increased in UVA + 100-mg/L TMP group at 24, 48, 72 hours, UVA + 50-mg/L TMP group at 24 and 72 hours and UVA + 20-mg/L TMP group at 72 hours.After repetitive UVA radiation, HDFs in the UVA group experienced an increase in cell volume, granule acount, and β-galactosidase expression, which was similar to the changes in the P55 group, while the pretreatments with 20, 50 and 100 mg/L TMP for 24 hours suppressed these UVA-induced changes in HDFs.The percentage of β-galactosidase-positive HDFs was 68.417％ ± 1.181％ in the UVA group, 58.167％ ± 5.620％ in the UVA + 20-mg/L TMP group, 45.167％ ± 5.502％ in the UVA + 50-mg/L TMP group, 43.000％ ± 2.000％ in the UVA + 100-mg/L TMP group, 33.667％ ± 5.865％ in the control group, and 76.000％ ± 6.557％ in the P55 group, with significant differences among these groups (F =45.918, P ＜ 0.01).Furthermore, the UVA group significantly differed from the UVA + TMP groups and control group in the percentage of β-galactosidase-positive HDFs and mRNA expressions of MMP-1 and MMP-3 (all P ＜ 0.05).Conclusion TMP can protect HDFs against senescence induced by repetitive UVA radiation, and down-regulate the mRNA expressions of MMP-1 and MMP-3 during senescence.

Aim To study toxic effect of cyclophosphamide on the embryonic cerebral and skeleton development of rats.Methods The pregnant rats were randomly divided into control group and experimental group(5,10,15,20 mg?kg-1 body wt).The experimental group was given cyclophosphamide and control group was given normal saline(sc)d 8~10 after becoming pregnant,and all rats were cut open the belly under anesthesia with ether at d 20 of gestation,the effects of cyclophosphamide on the embryonic body,four limbs form,cerebral and skeleton development of rat were observed.Results The embryos of 5 mg?kg-1 group were normal;the aqueduct of midbrain and left and right lateral ventricle expanded,cerebral hemisphere,lumbar vertebrae,rib,metacarpal bone were hypoplasia of embryos of 10 mg?kg-1 group and the symptoms of 15 mg?kg-1 group were graver than those of 10 mg?kg-1 group;20 mg?kg-1 group did not form embryos.Conclusion Cyclophosphamide has significant toxic effect on the embryonic cerebral and skeleton development of rats.

OBJECTIVETo investigate the effect of docetaxel on the interaction between C-jun and androgen receptor (AR) in prostate cancer LNCaP cells and its androgen-independence subtype LNCaP-bic cells.

METHODSLNCaP and LNCaP-bic cells were treated with docetaxel and the changes in AR and AP-1 expression were evaluated using luciferase assay. Western blotting and immunoprecipitation assay were employed to analyze the effects of docetaxel on the expressions of C-jun and AR and their interaction.

RESULTSLuciferase assay showed that LNCaP and LNCaP-bic cells expressed higher levels of AR and C-jun after docetaxel treatment. Docetaxel induced a higher level of p-c-jun expression in LNCaP-bic cells than in the parental LNCaP cells. Western blotting showed a strong PSA protein expression in LNCaP-bic cell after docetaxel treatment. Docetaxel caused a stronger interaction between AR and C-jun in LNCaP-bic cells.

CONCLUSIONDocetaxel activates ligand-independent AR transcription, and the interaction between AR and C-jun may affect the outcome of docetaxel chemotherapy.

Cell Line, Tumor ; Humans ; Male ; Prostate ; metabolism ; Prostatic Neoplasms ; metabolism ; Proto-Oncogene Proteins c-jun ; metabolism ; Receptors, Androgen ; metabolism ; Taxoids ; pharmacology

OBJECTIVETo construct three-dimensional (3D) models of renal stones and perform percutaneous nephrolithotomy (PCNL) virtual surgery simulation. Methods CT images were obtained from 8 patients with renal stones. Images segmentation and reconstruction were performed using MIMICS 10.0 software to construct the 3D model of the renal stones, which provided the anatomical relationships between the kidney and the adjacent organs. The optimal PCNL virtual surgery simulation for each individual case was performed using FreeForm Modeling System on the basis of the 3D model.

RESULTSEight 3D models of renal stone were constructed. The 3D model of the renal stones represented the interrelationships of the stones, intrarenal vessel, and the collecting system with the adjacent anatomical structures. Individualized PCNL virtual surgery simulations including percutaneous puncture, dilatation and pneumatic lithotripsy were performed successfully in all the 8 3D models.

CONCLUSIONDigital 3D model of renal stone provides the reliable and comprehensive imaging information for surgical design, and PCNL virtual surgery simulation has important clinical significance to improve the stone clearance rate and reduce the surgical complications.

Adult ; Female ; Humans ; Imaging, Three-Dimensional ; Kidney Calculi ; diagnostic imaging ; surgery ; Male ; Middle Aged ; Nephrostomy, Percutaneous ; methods ; Software ; Tomography, Spiral Computed ; User-Computer Interface

Background: Aberrant Bcl-2 transcription is closely related with nodal diffuse large B-cell lymphoma (DLBCL), however, the relationship between Bcl-2 and primary gastrointestinal DLBCL (PGI-DLBCL) was not fully studied.Aims: To investigate the relationship between Bcl-2 gene amplification and protein expression and clinicopathological characteristics, immunophenotype and prognosis of PGI-DLBCL.Methods: Clinical data was collected from 136 PGI-DLBCL patients receiving surgical treatment, and a telephone interview was conducted for survival information.Bcl-2 gene amplification and protein expression in tumor tissue were determined by fluorescence in situ hybridization and immuno-histochemistry, respectively, and relationships between Bcl-2 and clinicopathological characteristics, immunophenotype and prognosis of PGI-DLBCL were analyzed.Results: Among 136 PGI-DLBCL patients, 33 (24.3%) showing gene amplification and 90 (66.2%) showing protein expression of Bcl-2;gene amplification was correlated with primary tumor location, Ann Arbor stage, serum lactate dehydrogenase level, B symptom and International Prognostic Index (IPI) score (P<0.05), while protein expression was correlated with primary tumor location and immunophenotype (P<0.05).5-year overall survival (OS) in patients positive for Bcl-2 gene amplification and patients with non-GCB immunophenotype and positive for Bcl-2 protein expression were inferior to those negative ones (41.5%vs.71.5%, P<0.05;54.6% vs.84.6%, P<0.05).In Bcl-2 gene amplification or protein expression positive patients, 5-year OS of CHOP chemotherapy was inferior to that of rituximab combined with CHOP chemotherapy (48.6%vs.80.3%, P<0.05;66.4%vs.83.4%, P<0.05).Conclusions: Detection of Bcl-2 gene amplification is useful for prediction of prognosis in PGI-DLBCL.Both patients with Bcl-2 gene amplification and non-GCB patients with Bcl-2 protein expression have a poorer prognosis.Rituximab may improve the prognosis in patients with Bcl-2 gene amplification or protein expression.

OBJECTIVE[corrected] To evaluate the method and technique of single-port laparoscopic radical nephrectomy.

METHODSForm January 2009 to September 2011, 22 patients with renal carcinoma were treated with single-port laparoscopic radical nephrectomy. An incision about 5 cm in length was made through the umbilicus or in the postaxillary line under the 12th rib to establish the peritoneal or retroperitoneal working space. A single-port cannulation was deployed and the operation was carried out using standard and crooked laparoscopic equipment.

RESULTSThe operations were completed successfully in all the 22 cases without conversion to open surgery, but additional trocar was needed in 5 cases. The mean operative time of radical nephrectomy was 150 min (90-240 min). The mean postoperative hospital stay was 7.6 days (3-15 days). The operation left a roughly 5-cm-long scar in all the cases.

CONCLUSIONSingle-port laparoscopic radical nephrectomy is safe and feasible with good cosmetic effect and shows much potential in radical resection of renal carcinoma.

Adult ; Aged ; Female ; Humans ; Kidney Neoplasms ; surgery ; Laparoscopy ; methods ; Male ; Middle Aged ; Nephrectomy ; methods ; Young Adult

OBJECTIVETo investigate the effect of stable knockdown of DNA methyltransferase 3b (DNMT3b) on the proliferation and apoptosis of bladder cancer cells.

METHODSLentivirus expressing DNMT3b siRNA or the negative control siRNA was infected in human bladder cancer BIU-87 cells. MTT assay and flow cytometry were used to detect cell proliferation and apoptosis, respectively. The inhibitory effect of DNMT3b knockdown on xenograft tumors in nude mice was observed. Real-time PCR and Western blotting were carried out to investigate the expression level of cell apoptosis related genes. Methylation specific PCR was used to examine the methylation in the promoter region of the cell apoptosis related genes.

RESULTSThe results of real-time PCR and Western blotting showed that DNMT3b mRNA and protein level were stably knocked down in BIU-87 cells. Stable DNMT3b knockdown suppressed BIU-87 cell growth and the tumor formation ability of the cells in nude mice. DNMT3b knockdown promoted the apoptosis of BIU-87 cells, increased the mRNA and protein expression of the cell growth and apoptosis related genes including DAPK, Bax and RASSF1A, and significantly decreased the methylation of these genes.

CONCLUSIONStable DNMT3b knockdown can affect the methylation of the cell growth and apoptosis related genes to regulate their expression, which might be a possible mechanism for suppressed cell growth and enhanced apoptosis of BIU-87 cells.

Animals ; Apoptosis ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; Gene Knockdown Techniques ; Humans ; Mice ; Mice, Nude ; Neoplasm Transplantation ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Urinary Bladder Neoplasms ; genetics ; pathology