Objective:To investigate the clinical effect of extraperitoneal robot-assisted laparoscopic modified Y-V plasty(LMYVP)in the treatment of refractory bladder neck contracture(BNC).Methods:We retrospectively analyzed the clinical data on 10 cases of refractory BNC after transurethral resection of the prostate between September 2020 and January 2023,all with a history of recurrent urethral dilatation and at least two failures in transurethral surgical treatment of scarring.After definite diagnosis and re-moval of some of the scar tissues to flatten the elevated bladder neck under the cystoscope,we performed robot-assisted LMYVP using the da Vinci Si robotic system and a four-port extraperitoneal approach.The surgical procedure involved an inverted T-shaped incision in the bladder neck and urethral stricture ring,an inverted V-shaped excision of the scar area at the 3-9 o'clock position on the ventral side of the prostatic urethra,continuous full-layer suturing of the bladder neck and inverted V-shaped urethra with 3-0 barbed thread,and indwelling of an F20 silicone catheter for 2 weeks.At 3 months after surgery,we performed cystoscopic examination,measured the maximum urinary flow rate(Qmax),and obtained the IPSS and quality of life(QOL)scores of the patients.Results:Operations were successfully completed in all the cases.At 3 months after surgery,the patients showed significantly increased Qmax([3.65±1.27]vs[20.3±1.77]ml/s,P＜0.05),IPSS(5.9±2.02 vs 30±1.15,P＜0.05)and QOL score(1.3±0.95 vs 5.2±0.79,P＜0.05)compared with the baseline.Cystoscopy revealed a wide and flat bladder neck with good survival and hemody-namics of the bladder flap.All the patients met the criteria for clinical cure at a median follow-up of 13.2 months.Conclusion:Extraper-itoneal robot-assisted LMYVP provides a new strategy for urinary tract reconstruction in the management of refractory BNC,with the ad-vantages of minimal invasiveness,high efficiency and high success rate.

AIM:To explore the effects of mammalian target of rapamycin (mTOR) double inhibitor AZD8055 on autophagy and apoptosis of human cholangiocarcinoma cell line HuCCT1. METHODS:The effect of AZD8055 on the viability of HuCCT1 cells was detected by MTT assay. Autophagosome was detected by acridine orange (AO) staining. Af-ter treated with AZD8055, the expression levels of apoptosis-related proteins Bcl-2, Bax and cleaved caspase-3 and auto-phagy marker proteins beclin 1, LC3 and p62 were determined by Western blot. Apoptotic rate was analyzed by flow cyto-metry with Annexin V-FITC/PI double staining. RESULTS:AZD8055 significantly inhibited the viability of HuCCT1 cells (P<0.05). AO staining showed that AZD8055 significantly increased orange granules in the cytoplasm. After treated with AZD8055, compared with the control group, the protein level of beclin 1 and the ratio of LC3-Ⅱ/LC3-Ⅰ were enhanced, while p62 was attenuated (P<0.05). The protein expression level of pro-apoptotic regulator Bax was down-regulated and anti-apoptotic regulator Bcl-2 was increased. The protein level of cleaved caspase-3 was reduced (P<0.05). The results of flow cytometry showed that AZD8055 inhibited cell apoptosis. CONCLUSION:AZD8055 inhibits the viability of cholangiocarcinoma cells, and the mechanism is closely related with autophagy induced by AZD8055.

OBJECTIVETo evaluate the correlation between programmed death-ligand 1 (PD-L1) expression in primary lung cancer cells, tumor associated macrophages (TAM) and patients' clinicopathological characteristics.

METHODSFrom 2008 to 2010, 208 non-small cell lung cancer patients who underwent surgery or CT-guided biopsy were recruited from Huadong Hospital, Fudan University. Immunohistochemistry staining was performed to evaluate the PD-L1 expression in both primary lung cancer cells and CD68 positive TAM. The relationship between PD-L1 expression and the clinical pathology was evaluated using χ(2) test. Spearman's rank correlations were used to determine the correlation between PD-L1 expression in tumor cells and macrophages.

RESULTSPositive PD-L1 expression in primary cancer cells was found in 136 (65.3%) patients, which were negatively correlated with lymph node metastasis (P=0.009) and smoking history (P=0.036). Besides, TAM with PD-L1 expression (found in 116 patients) was positively associated with smoking history (P=0.034), well-differentiation (P=0.029) and negative lymph node metastasis (P=0.0096). A correlation between PD-L1 expression in primary tumor cells and non-small cell lung cancer associated macrophages was found (r=0.228, P=0.021).

CONCLUSIONPD-L1, secreted from TAM, might induce cancer cells apoptosis, and decrease lymph node metastasis.

Adult ; Aged ; Aged, 80 and over ; Apoptosis ; B7-H1 Antigen ; secretion ; Carcinoma, Non-Small-Cell Lung ; pathology ; secretion ; Cell Line, Tumor ; Female ; Humans ; Lung Neoplasms ; pathology ; secretion ; Lymphatic Metastasis ; Macrophages ; pathology ; secretion ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo develop an alternative method for assessment of gene delivery systems in vivo.

METHODSMouse primary spleen lymphocytes were genetically modified in vitro by a retroviral vector harboring a Gaussia luciferase (Gluc) expression cassette. After implantation of these cells into recipient mice, the expression of Gluc was detected in whole blood or plasma collected.

RESULTSAs little as 10 muL whole blood drawn from the recipient mice could guarantee prompt reading of Gluc activity with a luminometer. And the reading was found in good correlation with the number of genetically modified spleen lymphocytes implanted to the mice.

CONCLUSIONSGluc may be useful as an in vivo reporter for gene therapy researches, and Gluc blood assay could provide an alternative method for assessment of gene delivery systems in vivo.

Animals ; Arecaceae ; enzymology ; Cell Line ; Gene Transfer Techniques ; Genes, Reporter ; Humans ; Luciferases ; genetics ; Mice

OBJECTIVETo screen and identify the peptides that specifically bind to CD13 on monocytes.

METHODSThe phages capable of specific binding to CD13 were screened in the phage-displayed 12-peptide library. The affinity of the selected phages with CD13 was verified with enzyme-linked immunosorbent assay (ELISA). The sequences of the peptides bound to the phages were deduced according to the phage DNA sequences, and the functional peptides aligned using the BLASTP on the Website NCBI were synthesized. To analyze the biological function of the screened peptides, the location of the peptides bound to THP-1 cells was detected using immunofluorescence assay. The blocking effect of WM15 on the peptide binding to THP-1 cells was assessed by immunofluorescence assay.

RESULTSThe phages that specifically bound to CD13 were effectively enriched to approach saturation after 4 rounds of panning. The recovery rate in the fourth round was 30 times that in the first round. Twenty selected phages were verified by ELISA, and the signals of 10 phages were higher than the control. The sequences of the peptides P9 and P7 showed 83% and 100% identity with those of human cytomegalovirus (HCMV) UL38 and UL105, respectively. The peptides bound to the cell membrane of THP-1 cells as shown by immunofluorescence assay. The binding of the peptides P9 and P7 to THP-1 cells was blocked by CD13-specific monoclonal antibody WM15 at different levels.

CONCLUSIONTwo peptides (P7 and P9) that can specifically bind to CD13 have been screened successfully, and these two peptides show specific binding to CD13 on the membrane of THP-1 cells.

Amino Acid Sequence ; Binding, Competitive ; CD13 Antigens ; analysis ; metabolism ; Cell Line ; Humans ; Molecular Sequence Data ; Peptide Library ; Peptides ; metabolism ; Protein Binding

OBJECTIVETo investigate the effectiveness of injectable tissue engineering to repair full-thickness meniscal defects.

METHODSFrom June 2008 to February 2009 full-thickness of meniscal defects were created in the anterior corner of goats, which with no blood supply, in a diameter of 2 mm. Then bone marrow stem cells (BMSCs) was mixed with injectable calcium alginate gel to fill the defects. Other groups include the calcium alginate gel and empty group were served as control groups. At different time points, the animals were sacrificed and macroscopy, microscopy determination, electroscopy and MRI detection were performed to assess the outcomes of repairing.

RESULTSThe meniscal defects had been filled thoroughly in 16 weeks after operation with white, tough and elastic repair tissue similar to normal meniscal fibrocartilage in the tissue engineering groups. The repair tissue was mainly fibrochondrocytes in line with the calcium alginate fiber. Thick matrix secreted by the cells crammed the space between fibers. The view under electroscopy demonstrated that the microstructure of the repair tissue was normal and cells were in a fibrocartilage phenotype.

CONCLUSIONThe full-thickness meniscal defects in regions without blood supply can be reconstructed effectively with injectable tissue engineering.

Alginates ; Animals ; Bone Marrow Cells ; Cells, Cultured ; Disease Models, Animal ; Gels ; Glucuronic Acid ; Goats ; Hexuronic Acids ; Injections ; Male ; Stem Cells ; Tibial Meniscus Injuries ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo introduce the biplanar opening high tibial osteotomy with rigid fixation for adult varus knee.

METHODSTwelve patients with varus knee and degeneration of medial compartment received the biplanar opening high tibial osteotomy between June 2001 and July 2008. The pre-operative deformity was about 11.5 degrees without osteoarthritis changes in other compartments. The average range of motion was more than 90 degrees . One of the patients had ruptures of LCL and ACL, who received osteotomy after reconstruction of the ligaments. The main symptom before the operation was pain in medial compartment. The range of motion, Lysholm score and subjective satisfactory examinations were assessed before and after the osteotomy.

RESULTSAll of the osteotomy sites were healed at twelve to sixteen weeks after operation. No complications such as plate broken or injury of nerve or blood vessel had occurred. The mean correct angle was 9.5 degrees . No degenerative changes had developed in other compartments of the knee. The mechanical axis of the lower extremities was maintained during the follow-up. The overall satisfactory rate was 83.3%. Statistically significant changes exist in the Lysholm score and varus degree.

CONCLUSIONThe open-wedge high tibial osteotomy is suitable for the symptomatic genu varum in younger patients with good short-term and mid-term results.

Adult ; Female ; Humans ; Joint Deformities, Acquired ; surgery ; Knee Joint ; Male ; Middle Aged ; Osteotomy ; methods ; Retrospective Studies ; Tibia ; surgery ; Treatment Outcome

OBJECTIVETo investigate the effect of pH value and fluoride ions on the corrosion resistance of pure Ti and Ni-Cr-Ti alloy in the artificial saliva.

METHODSElectrochemical technique was used to measure the electric potential of corrosion (Ecorr), current density of corrosion (Icorr) and polarization resistance (Rp) of pure titanium and Ti-Ni-Cr alloy in the artificial saliva with different pH value and fluoride concentrations. After electrochemical analysis, microstructure and phase diffraction were examined by FSEM.

RESULTSWith the lower pH value, the Ecorr and Icorr of pure titanium and Ti-Ni-Cr alloy increased, the Rp decreased, there was a significant difference (P<0.05). The Ecorr and Icorr increased markedly, the Rp significantly reduced in the artificial saliva containing 0.2% NaF (P<0.01). FSEM showed that pure titanium and Ti-Ni-Cr alloy surface corrosion, pure titanium in the artificial saliva containing 0.2% NaF was most serious.

CONCLUSIONLower pH value decreases the corrosion resistance of pure titanium and Ti-Ni-Cr alloy and the artificial saliva containing fluoride ions decreases the corrosion resistance of pure titanium.

Chromium Alloys ; chemistry ; Corrosion ; Dental Alloys ; chemistry ; Electrochemistry ; Fluorides ; chemistry ; Hydrogen-Ion Concentration ; Materials Testing ; Metal Ceramic Alloys ; chemistry ; Nickel ; chemistry ; Saliva, Artificial ; chemistry ; Surface Properties ; Titanium ; chemistry

Objective To study the mechanism of the protective effect of α_(1A)-AR antagonist Tamsulosin (Tam) on renal ischemic injury in hepatorenal syndrome (HRS) rats. Methods HRS was induced in male Sprague-Dawley (SD) rats by intraperitoneal injection of D-(+)-galactosamine hydrochloride (GalN). Thirty-two HRS rats were divided into 3 groups randomly. Then surgical occlusions of the infrahepatic inferior vena cave (OIVC) were performed; Tam, an α_(1A)-AR antagonist, was administered daily before injection of GalN. During the operation, hemodynamic changes of the kidney and liver were measured by laser Doppler flowmetry (LDF). Serum samples were collected to measure serum levels of liver and renal function. At the same time, renal and liver samples were harvested and stained by HE and immunohistochemistry. Other parts of the sample were fixed with 2.5% glutaral to observe the cellular ultrastructure of renal and vascular endothelium by electron microscope. Results OIVC developed much severe ischemic injury in the kidney and liver of HRS rats. The renal cortex blood perfusion (RCBP) of HRS rats decreased rapidly after OIVC, and did not return to baseline after reflow. Pathological study showed severe injury of the liver and kidney. However, expressions of alpha-1 AR on renal artery and kidney were reduced in those rats that had received Tam before OIVC. Histological examination of the kidney also showed few abnormalities. Conclusion Marked increases of contractive response of renal artery of HRS rats induced by up-regulation of α_1-AR may be associated with a high risk of progression to acute renal failure in HRS rats after ischemia. Tamsulosin has a protective effect on renal ischemic injury through reducing α_(1A)-AR overexpression in HRS rats.

Adolescent ; Coronary Angiography ; Coronary Vessel Anomalies ; pathology ; surgery ; Female ; Fistula ; congenital ; pathology ; surgery ; Humans ; Male ; Middle Aged