Objective: To explore the relationship between the activity of Wnt/β-catenin signaling pathway and prostate cancer stem cells by utilizing a Wnt/β-catenin signaling reporter system. Methods: Immunofluorescence staining was used to explore the activation of Wnt/β-catenin signaling pathway in prostate cancer. Wnt/β-catenin signaling reporter plasmid 7TGP carrying T cell specific transcription factor (TCF) binding site and enhanced green fluorescent protein (GFP) was transfected into prostate cancer PC3 and DU145 cells by liposomes, respectively. Furthermore, the top 5% (GFP+) and bottom 5% (GFP-) groups of PC3 and DU145 cells according to the fluorescent intensity of GFP were collected via flow cytometry. Then Western blotting assay was used to compare the expression level of activated β-catenin protein in the nucleus of GFP+ and GFP- cell groups. The real-time fluorescent quantitative PCR was used to detect the expressions of Wnt/β-catenin signaling downstream target genes in GFP+ and GFPcell groups. Sphere formation assay and real-time fluorescent quantitative PCR were used to compare the stemness of GFP+ and GFP- cells. Results: The activity of canonical Wnt signaling pathway was low in most of prostate cancer cells, in which β-catenin was not activated and translocated to the nucleus. While β-catenin was increasingly translocated to the nucleus in a small percentage of prostate cancer cells resulting in high activation of Wnt signaling pathway. PC3-GFP+ and DU145-GFP+ cells had more expression of nuclear β-catenin comparing with the corresponding GFP- cells (both P < 0.05), and the expression level of Wnt signaling pathway down-stream target gene Axis inhibition protein 2 (AXIN2) was up-regulated (both P < 0.05). In addition, the expression levels of cancer stem cell markers including B-cell-specific moloney leukemia virusinsert site 1 (BMI1) and aldehyde dehydrogenase family 1 member A1 (ALDH1A1) were up-regulated in PC3-GFP+ and DU145-GFP+ cells comparing with the corresponding GFP- cells (all P < 0.05). The sphere formation capacity was remarkedly up-regulated in PC3-GFP+ and DU145-GFP+ cells comparing with the corresponding GFP- cells (both P < 0.05). Conclusion: Prostate cancer cells with higher activity of Wnt/β-catenin signaling show the enhanced stemness.

Objective To evaluate the diagnosis value of urodynamics in patients with benign prostate hypertrophy(BPH).Methods With urodynamic device,the full set of urodynamic exam was administrated in 427 patients with BPH,and the externalsphincter urethral myogram was monitored simultaneously in pressure-flow studies(PFS).The umdynamic finding such as Qmax 、Pdet-Qmax、Popen、DS(descending slope) and post-voiding residual(PVR)were recorded,as well as the situation of bladder detrusor constraction and bladder compliance and urethral sphincter coordination. The bladder outflow obstruction was diagnosed by A-G nomogram，P-Q plot and DS.The IPSS score and prostate volume were also acquired. Results The diagnostic rate of BOO is 81.5％,among them concomitantly detrusor muscle impair in 117 cases(27.4％), decreased bladder compliance in 162 case (37.9％)，urethral sphincter dyssynergia in 148 cases(34.7％)，and unstable bladder in l64 cases (38.4％). The increase degree of BOO show an increasing tendency with urodynamic findings such as Qmax ,Pdet-Qmax,Popen,DS and IPSS score and prostatic volume respectively，however a decreasing tendency with Qmax and bladder compliance. Conclusions The urodynamic exam plays an important role in diagnosis of BOO.There is a positive relation among degree of BOO with urodynamic finding such as Pdet-Qmax,Popen,DS and IPSS score and prostatic volume,however，negative relation with Qmax and bladder compliance respectively.

As a male sexual organ, the penis can not only meet the requirements for sexual life between sexual partners, but also increase and improve sexual satisfaction and enhance male self-confidence.The treatment of micropenis is a high-frequency trending topic.This paper reviews the different causes of micropenis, the treatment methods and progress of "penile elongation, " aiming to improve the understanding and provide solutions.

Objective To investigate the influence of thyroid hormone T3 preconditioning on interleukin-10 (IL-10) and interleukin-1 receptor antagonist (IL-IRa) modulation and neutrophil infiltration after renal ishchemia/reperfusion (IR) in mice, so as to study the protective effect of T3 on IR kidney. Methods Totally 120 male C57BL/6 mice were randomly divided into four groups (n = 30), namely, control group (sham operation), IR group (only received renal IR), T3 + IR group (T3 preconditioning for 48 h before renal IR), and NaOH + IR group (received equivalent 0. 1 mol/L NaOH soltuion 48 h before renal IR). The serum creatinine and blood urea nitrogen (BUN) were determined 24 h after reperfusion in each group; renal histological damages were scored using PAS staining; the levels of neutrophil infiltration were evaluated by MPO staining, and IL-10, IL-IRa mRNA expression was examined by real-time PCR at 1, 3, 6, 12, 24, and 48 h after reperfusion. Results The serum creatinine and BUN levels of T3+IR group were significantly lower than those of IR group 24 h after reperfusion (P< 0. 05), which was accompanied by lower histological score and significantly less neutrophil infiltration (P<0. 05). Real-time PCR results showed that IL-10 and IL-IRa mRNA expression in T3 +IR group was significantly higher than that in the IR group (P<0. 05) 12 h after reperfusion, which lasted for 48 h aft er reperfusion. The above parameters were similar between IR group and NaOH+IR group. Conclusion Thyroid hormone T3 preconditioning can alleviate renal IR injury, partly by increasing expression of IL-10 and IL-IRa and subsequently reducing neutron phil infiltration at the late phase of renal IR.

Objective: To investigate the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) combined with arsenic trioxide (ATO) (molecular formula: As2O3) on the proliferation and apoptosis of acute myeloid leukemia (AML) cells, and to explore the possible mechanisms. Methods: AML cells HL-60 and THP-1 were pre-treated with rhG-CSF (100 ng/mL), and then treated with different concentrations of As2O3. The relative proliferation rate was detected by CCK-8 method, while the apoptosis and cell cycle distribution were measured by FCM method. The expression levels of aquaporin 9 (AQP9) mRNA and protein in HL-60, THP-1 and acute promyelocytic leukemia NB4 cells as well as HL-60 and THP-1 cells treated with rhG-CSF (100 ng/ mL) were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. Results: RhG-CSF promoted the proliferation of HL-60 and THP-1 cells (both P < 0.01). Compared with the As2O3 group, rhG-CSF pre-treatment combined with 2 μmol/L As2O3 inhibited the proliferation of HL-60 and THP-1 cells (both P < 0.05), rhG-CSF combined with different concentrations of As2O3 increased the apoptotic rates of HL-60 and THP-1 cells (both P < 0.05). As2O3 caused G0/G1 arrest in HL-60 and THP-1 cells (both P < 0.05). rhG-CSF caused S-phase arrest in HL-60 and THP-1 cells (both P < 0.01), the effect was more obvious in rhG-CSF combined with As2O3 group (both P < 0.05). The expressions of AQP9 mRNA and protein in HL-60 and THP-1 cells were lower than those in NB4 cells (all P < 0.01). Compared with the untreated control group, 100 ng/mL rhG-CSF up-regulated the expression levels of AQP9 mRNA and protein in HL-60 and THP-1 cells (all P < 0.05). Conclusion: RhG-CSF can increase the sensitivity of non-M3 AML cells to As2O3, which may be associated with the up-regulation of AQP9 expression.

Objective: To explore the etiology and risk factors related to prognosis of primary natural killer (NK)/T cell lymphoma with secondary hemophagocytic syndrome (NK/T-LAHS) and review the advances in treatment of NK/T-LAHS. Methods: One case of primary NK/T cell lymphoma with secondary hemophagocytic syndrome was discussed in combination with a review of related literatures. Results: The patient was a 21-year old female who presented with high fever, hepato-splenomegaly, pancytopenia and jaundice. She was diagnosed with primary extranodal NK/T cell lymphoma (nasal type, ?B) with secondary hemophagocytic syndrome. A combined chemotherapy of etoposide, dexamethasone, L -asparaginase and high-dose methotrexate with vigorous supportive therapy was given. The patient's clinical condition was improved transiently after chemotherapy, but relapsed in a very short period and markedly deteriorated. Ultimately the patient refused treatment and took her own discharge againstadvice. Conclusion: Primary NK/T-LAHS is rare and has a rapid progression with poor prognosis. There are not effective treatments for NK/T-LAHS and novel therapeutic regimens should be investigated.

Objective • To investigate changes of immune thrombocytopenia (ITP) patients-derived bone marrow mesenchymal cells (BMCs) in cells survival, cytokines expression as well as the effects of BMCs on the biological behaviors of megakaryocytes. Methods • BMCs were collected from 7 ITP patients and 5 normal controls (NC), and cultivated by the whole marrow adherent method. Surface markers and basal apoptosis rate of BMCs were analyzed by flow cytometry (FCM). Proliferation of BMCs was assessed by CCK-8 method. Phorbol 12-myristate 13-acetate (PMA) was used to stimulate differentiation of HEL cells. The induced HEL cells (inHEL) were divided into 3 groups: inHEL cultured alone (group a), inHEL co-cultured with BMCs derived from ITP patients (group b), inHEL co-cultured with BMCs derived from NC (group c). After 72 h incubation, the expression of cell surface proteins (CD41a, CD42b) and cell apoptosis rate were analyzed by FCM. The mRNA and proteins expression levels of cytokines IL6, IL11, TPO, SCF were detected by real-time fluorescent quantitative PCR (RT-qPCR) and enzyme linked immunosorbent assay (ELISA), respectively. Results • Compared with NC, BMCs from ITP patients grew progressively slowly (Day 4, P=0.039; Day 6, 10, P=0.009; Day 8, P=0.007), cell basal apoptosis rates were increased [AV+PI- (early apoptosis rate), P=0.036; AV+PI+ (late apoptosis rate), P=0.003; AV+PI-/+ (total apoptosis rate), P=0.004]. Compared with group a, the expression of CD41a in group c was much higher (P=0.000). The expression of CD41a in group b was higher than that in group a (P=0.015), but still much less than that in group c (P=0.000). Compared with group a, the early and total apoptosis rate in group b, c and the late apoptosis rate in group c were decreased obviously (all P=0.000), whereas there was no obvious change of the late apoptosis rate in group b. However, compared with group c, the late and total apoptosis rate in group b were significantly increased (both P=0.000). The expression levels of IL6, SCF mRNA and IL6 protein were significantly decreased in ITP BMCs (all P=0.000), but there was no obvious difference in the expression levels of IL11 and TPO between ITP BMCs and NC BMCs. Conclusion • BMCs from ITP patients show some defects in supporting megakaryocytic differentiation and survival under co-culture conditions, which mechanisms are related to the reduction of IL6 and SCF expression.

Objective • To evaluate the value of texture analysis in the discrimination of colorectal cancer (CRC) and ulcerative colitis (UC). Methods • The CT images of 61 CRC patients, 62 UC patients and 42 control objects were retrospectively analyzed. All the patients were pathologically proved and performed triphasic contrast-enhanced CT scan: non-enhanced phase (NP), the arterial phase (AP) and the enteric phase (EP). The region of interest was drawn along the abnormal bowel wall's edge in each scan phase and texture features were generated by MaZda software. Based on 3 texture feature selection methods, the optimal subsets were generated and analyzed by 6 texture feature classification methods. The results were shown by misclassification rate (MCR). To compare the performance of texture-based classification and human visual classification, two radiologists with more than 10 years of gastrointestinal disease diagnostic experience analyzed the data. Results • The texture analysis based average MCR of differentiation between CRC and UC was (28.42±6.89)%, (28.19±4.07)%, (19.10±3.58)% in NP, AP, EP respectively. Compared with other texture feature classification methods, nonlinear discriminant analysis (NDA) was more accurate. In EP, NDA achieved an excellent classification result (MCR=12.61%). The average MCR between CRC and normally dilated bowel wall (NOR) was (13.33±7.21)%, (15.49±5.47)%, (6.74±3.02)%, while the average MCR between UC and NOR was (19.26±4.68)%, (20.04±6.63)%, (16.74±6.36)% in NP, AP and EP respectively. For visual classification between CRC and UC, the average MCR was (40.48±3.21)%, (35.71±1.60)%, (26.43±1.15)% in NP, AP, EP respectively. But the MCR of texture classification was lower than that of human vision classification, and computer texture classification had higher differential diagnosis rate. Conclusion • The CT-based texture analysis could be a feasible supplementary method to differentiate CRC from UC. NDA is more accurate than other classification methods, especially in EP.

Objective: To investigate the mechanism of fructose-induced monocyte chemoattratant protein-1(MCP-1) production in HK-2 cells. Methods: The HK-2 cells were divided into fructose incubated (1, 5 and 10 mmol/L) group, fructose and ketohexo-kinase inhibitor (KHK-IN) coincubation (fructose 5 mmol/L, KHK-IN was 12, 100 and 1000 nmol/L, respectively) group, uric acid incubation (5, 15 and 50 mg/dL) group, fructose and allopurinol co-incubation (fructose 5 mmol/L, allopurinol were 0.01, 0.1 and 0.5 mmol/L) group, uric acid and allopurinol co-incubation (uric acid 50 mg/dL, allopurinol respectively 0.01, 0.1and 0.5 mmol/L) group, H2O2 incubation (0.1 and 0.3 mmol/L) group, fructose and N-acetylcysteine (NAC) coincubation (fructose 5 mmol/L, NAC respectively 5, 10 and 50 mmol/L) group, and uric acid and NAC co-incubation (uric acid 50 mg/dL, NAC was 5, 10 and 50 mmol/L, respectively) group. The quantitative PCR method and Western blotting method were used to observe the expression of MCP-1 mRNA and protein. The effects of fructose and uric acid on the production of ROS in HK-2 cells were observed by using a fluorescent probe. Results: Fructose doseand time-dependently induced MCP-1 gene transcription and protein production in HK-2 cells, which could be blocked by the ketohexo-kinase blockers. Exogenous uric acid induced MCP-1 production in HK-2 cells. Allopurinol inhibited fructose, but not exogenous uric acid-induced MCP-1 expression. Both fructose and uric acid induced ROS generation. Incubation with H2O2 promoted MCP-1 production in HK-2 cells. NAC completely inhibited MCP-1 production induced by fructose and H2O2. Conclusion: Catalyzed by the ketohexo-kinase, fructose resultes the production of MCP-1 through uric acid and reactive oxygen species.

OBJECTIVE: Using radioactive Nano tracer at different sizes and doses in the Larynx of rabbits, to study the roles of them in the sentinel lymph node (SLN) biopsy in rabbits and to provide experimental evidences for the choices of ideal size and dose of radioactive Nano tracer of the sentinel lymph node biopsy in Laryngeal cancer patients. METHOD: thirty rabbits were randomly divided into six groups with five rabbits in each group. After 50 nm--0.01 ml, 50 nm--0.02 ml, 80 nm--0.01 ml, 80 nm--0.02 ml,100 nm--0.01 ml, 100 nm--0.02 ml of 99mTc-sulfur Colloid were separately injected into the Larynx, the number of SLNs, the initial and strongest radioactive time of SLNs, and the lasting time of radioactivity was obtained. RESULT: One to three SLNs were identified in one rabbit, so there were totally forty-five SLNs, which in the areas of II, III and IV. The group of 50 nm--0.02 ml had the largest number of SLNs and there were significant differences between the group of 50 nm-0.02 ml and 100 nm--0.01 ml. In the six groups, the group of 50 nm--0.02 ml was the earliest group of detecting the initial and strongest radioactivity of SLNs,which the time were 49.20 s and 178.60 s; the group of 100 nm--0.01 ml was the latest group of detecting the initial and strongest radioactivity of SLNs, which the time were 235.80 s and 311.20 s. Each group had radioactivity more than 30 minutes. CONCLUSION The group of 50 nm--0.02 ml was the best group, because it moved fast and had a higher rate of uptake in lymphangio. Moreover, the radioactivity time was more than 30 minutes. It was the ideal size and dose of SLN biopsy in larynx.
Animals ; Larynx ; diagnostic imaging ; Lymph Nodes ; diagnostic imaging ; pathology ; Male ; Nanostructures ; Rabbits ; Radioactive Tracers ; Radionuclide Imaging ; Sentinel Lymph Node Biopsy ; methods