Diagnostic Value of MRI Features in Dual-phenotype Hepatocellular Carcinoma: A Preliminary Study.

This study aimed to explore the magnetic resonance imaging (MRI) features of dual-phenotype hepatocellular carcinoma (DPHCC) and their diagnostic value.The data of 208 patients with primary liver cancer were retrospectively analysed between January 2016 and June 2021. Based on the pathological diagnostic criteria, 27 patients were classified into the DPHCC group, 113 patients into the noncholangiocyte-phenotype hepatocellular carcinoma (NCPHCC) group, and 68 patients with intrahepatic cholangiocarcinoma (ICC) were classified into the ICC group. Two abdominal radiologists reviewed the preoperative MRI features by a double-blind method. The MRI features and key laboratory and clinical indicators were compared between the groups. The potentially valuable MRI features and key laboratory and clinical characteristics for predicting DPHCC were identified by univariate and multivariate analyses, and the odds ratios (ORs) were recorded. In multivariate analysis, tumour without capsule (P = 0.046, OR = 9.777), dynamic persistent enhancement (P = 0.006, OR = 46.941), and targetoid appearance on diffusion-weighted imaging (DWI) (P = 0.021, OR = 30.566) were independently significant factors in the detection of DPHCC compared to NCPHCC. Serum alpha-fetoprotein (AFP) > 20 µg/L (P = 0.036, OR = 67.097) and prevalence of hepatitis B virus (HBV) infection (P = 0.020, OR = 153.633) were independent significant factors in predicting DPHCC compared to ICC. The differences in other tumour marker levels and imaging features between the groups were not significant. In MR enhanced and diffusion imaging, tumour without capsule, persistent enhancement and DWI targetoid findings, combined with AFP > 20 µg/L and HBV infection-positive laboratory results, can help to diagnose DPHCC and differentiate it from NCPHCC and ICC. These results suggest that clinical, laboratory and MRI features should be integrated to construct an AI diagnostic model for DPHCC.

[Risk factors for massive hemorrhage after radical nephrectomy and removal of venous tumor thrombus].

OBJECTIVE: To investigate and analyze the risk factors of massive hemorrhage in patients with renal cell carcinoma and venous tumor thrombus undergoing radical nephrectomy and removal of venous tumor thrombus. METHODS: From January 2014 to June 2020, 241 patients with renal cancer and tumor thrombus in a single center of urology at Peking University Third Hospital were retrospectively analyzed. All patients underwent radical nephrectomy and removal of venous tumor thrombus. The relevant preoperative indicators, intraoperative conditions, and postoperative data were statistically analyzed by using statistical software of SPSS 18.0. The main end point of the study was intraoperative bleeding volume greater than 2 000 mL. Logistic regression analysis was used to determine the relevant influencing factors. First, single factor Logistic regression was used for preliminary screening of influencing factors, and variables with single factor Logistic regression analysis P < 0.05 were included in multivariate Logistic regression. In all statistical analyses, P < 0.05 is considered statistically significant. RESULTS: Among the 241 patients included, there were 60 cases of massive hemorrhage, 48 males and 12 females, with a median age of 62 years. The number of non-massive hemorrhage was 181. There were 136 males and 45 females, with a median age of 59 years. Univariate analysis showed that the clinical symptoms (both systemic and local symptoms, OR 2.794, 95%CI 1.087-7.181, P=0.033), surgical approach (open surgery, OR 9.365, 95%CI 4.447-19.72, P < 0.001), Mayo grade (Mayo 3-4, OR 5.257, 95%CI 2.806-10.886, P < 0.001), American Society of Anesthesiologists (ASA) score (ASA level 3, OR 2.842, 95%CI 1.338-6.036, P=0.007), preoperative hemoglobin (OR 0.978, 95%CI 0.965-0.991, P=0.001), preoperative platelet count (OR 0.996, 95%CI 0.992-1.000, P=0.037), maximum tumor thrombus width (OR 1.061, 95%CI 1.033-1.091, P < 0.001), Complicated with bland thrombus (OR 4.493, 95%CI 2.264-8.915, P < 0.001), adrenalectomy (OR 3.101, 95%CI 1.614-5.958, P=0.001), segmental resection of the inferior vena cava (OR 2.857, 95%CI 1.395-5.852, P=0.004). There was a statistically significant difference in these aspects(P < 0.05). Multivariate Logistic regression analysis showed that there was a statistically significant difference in surgical approach (open surgery, OR 6.730, 95%CI 2.947-15.368;P < 0.001), Mayo grade (Mayo 3-4, OR 2.294, 95%CI 1.064-4.948, P=0.034), Complicated with bland thrombus (OR 3.236, 95%CI 1.492-7.020, P=0.003). CONCLUSION: Combining the results of univariate and multivariate Logistic regression analysis, the surgical approach, Mayo grade, and tumor thrombus combined with conventional thrombus were associated risk factors for massive hemorrhage during surgery for renal cell carcinoma with tumor thrombus. Patients who undergo open surgery, high Mayo grade, and tumor thrombus combined with conventional thrombus are at a relatively higher risk of massive hemorrhage.

WASHC1 interacts with MCM2-7 complex to promote cell survival under replication stress.

BACKGROUND: WASHC1 is a member of the Wiskott-Aldrich syndrome protein (WASP) family and is involved in endosomal protein sorting and trafficking through the generation of filamentous actin (F-actin) via activation of the Arp2/3 complex. There is increasing evidence that WASHC1 is present in the nucleus and nuclear WASHC1 plays important roles in regulating gene transcription, DNA repair as well as maintaining nuclear organization. However, the multi-faceted functions of nuclear WASHC1 still need to be clarified. METHODS AND RESULTS: We show here that WASHC1 interacts with several components of the minichromosome maintenance (MCM) 2-7 complex by using co-immunoprecipitation and in situ proximity ligation assay. WASHC1-depleted cells display normal DNA replication and S-phase progression. However, loss of WASHC1 sensitizes HeLa cells to DNA replication inhibitor hydroxyurea (HU) and increases chromosome instability of HeLa and 3T3 cells under condition of HU-induced replication stress. Re-expression of nuclear WASHC1 in WASHC1KO 3T3 cells rescues the deficiency of WASHC1KO cells in the chromosomal stability after HU treatment. Moreover, chromatin immunoprecipitation assay indicates that WASHC1 associates with DNA replication origins, and knockdown of WASHC1 inhibits MCM protein loading at origins. CONCLUSIONS: Since efficient loading of excess MCM2-7 complexes is required for cells to survive replicative stress, these results demonstrate that WASHC1 promotes cell survival and maintain chromosomal stability under replication stress through recruitment of excess MCM complex to origins.

Distribution, co-existing metals, and potential health risk of fluorine in farmland soil in different anthropogenic activity dominated districts in a county-level city in Sichuan province, Southwest China, in 2015.

Continuous fluorine (F) accumulation in soil by anthropogenic activities leads to variously global environmental and health issues. Herein, 300 farmland soil samples were collected from different anthropogenic activity dominated districts for studying the distribution and related health risk of F in soils. Co-existing metal concentrations in soil samples were also analysed to evaluate the relationship between the distribution of F and metals in soil. The median value of the total F concentration of 488 mg kg-1 in the present samples was higher than the median background F concentration in topsoil in Sichuan province of China (261 mg kg-1). Concentration of water-soluble F (1.33-26.2 mg kg-1) was two or three orders of magnitude less than that of total F in soil. Levels of total and water-soluble F in soils collected from the district with longer contamination history were higher than that from other districts with shorter contamination period, indicating a historical contribution of anthropogenic activities to F accumulation in soil. Notable positive correlation between the total F and vanadium (V) concentration in soil can be partly linked to the usually negative charged form or a common source of F and V in soil (e.g. coal combustion). Compared with inhalation and dermal contact, present human exposure of F in soil was mainly caused by oral ingestion, and the health risks posed by F in soil for both children and adults were acceptable. However, considering the higher potential risk for children than adults, the accumulation of F in soil induced by anthropogenic activities should not be neglect.

Effects of ER-resident and secreted AGR2 on cell proliferation, migration, invasion, and survival in PANC-1 pancreatic cancer cells.

BACKGROUND: Anterior gradient-2 (AGR2) is a proto-oncogene involved in tumorigenesis and cancer progression. AGR2, predominantly localized in the endoplasmic reticulum (ER), is also a secreted protein detected in the extracellular compartment in multiple cancers. However, the biological functions of intracellular and extracellular AGR2 remain to be elucidated. METHODS: Based on the biochemical structure of AGR2 protein, PANC-1 pancreatic cancer cells stably expressing ER-resident or secreted AGR2 were generated by a lentivirus-mediated stable overexpression system. The capacities of cell proliferation, migration, invasion and survival were assessed in PANC-1 stable cells. Moreover, EGFR expression and activation were determined to explore the possible mechanism of AGR2 roles in pancreatic cancer tumorigenesis. RESULTS: It was discovered that secreted AGR2, but not ER-resident AGR2, promotes cell proliferation, migration and invasion of PANC-1 cells. Moreover, the data indicated that both the ER-resident and the secreted AGR2 enhance the survival capacity of PANC-1 cells after tunicamycin-induced ER stress and gemcitabine treatment. However, EGFR expression and activation were not found to be involved in AGR2-dependent oncogenic phenotypes in PANC-1 cells. CONCLUSIONS: Secreted AGR2 is predominantly involved in cell proliferation, migration and invasion in PANC-1 pancreatic cancer cells. Both secreted and ER-resident AGR2 contribute to the survival of PANC-1 cells under the challenging conditions. These findings provide insight into how different localizations of AGR2 have contributed to pancreatic cancer growth, metastasis, and drug sensitivity.

Clinical experience with the treatment of retroperitoneal vascular leiomyosarcoma originating from large veins.

BACKGROUND: Retroperitoneal vascular leiomyosarcoma (RVLMS) is an extremely rare disease in clinical practice, and it has poor prognosis. This article is to explore the diagnosis and treatment of RVLMS and present our experience. METHODS: Data of RVLMS patients were continuously collected in our hospital from August 2018 to February 2020: two males and two females with a median age of 56 (min-max = 33-61) years were included. Patients in whom paraganglioma could not be excluded were asked to take phenoxybenzamine before surgery. A multi-disciplinary team (MDT) meeting had been held and surgery was recommended. The operation procedures varied based on the tumor location, shape, and stage, and the core steps were "exposure of the retroperitoneum and tumor, identification of vital blood vessels, blocking the bloodstream, complete removal of the tumor and tumor thrombus, and release of blood flow". A Satinsky clamp was used to partially block the blood vessels. Follow-up was conveyed by revisits and phone calls. RESULTS: One patient underwent open surgery, and three patients underwent laparoscopic surgery, one of whom underwent conversion to open surgery. The procedures were finished successfully, with a median operative time of 314.5 (min-max = 224-467) mins. The median amount of intraoperative bleeding was 550 (min-max = 200-1500) ml, and three patients had transfusion during the operation. The mass was irregular in shape, with a median maximum size of 7.45 (min-max = 4.2-10.7) cm, and the pathological examination confirmed RVLMS, which has spindle-shape, high mitotic activity and atypia. One week after the operation, the median serum creatinine level was 85 (min-max = 70-99) µmol/L. The median follow-up time was 16 (min-max = 13-21) months, and 1 case reported asymptomatic recurrence. CONCLUSION: Uncharacteristic manifestations and imaging features contribute to the problematic diagnosis of RVLMS. Comprehensive preoperative evaluation and careful surgical planning are essential. Multicenter research is needed in the future to reach a dominant consensus.

Correlation of lower 2 h C-peptide and elevated evening cortisol with high levels of depression in type 2 diabetes mellitus.

BACKGROUND: A number of studies have explored the association between depression and ghrelin, leptin, and cortisol; further, postprandial C-peptide levels have a therapeutic effect on type 2 diabetes mellitus (T2DM). However, the relationship between C-peptide and depression in patients with diabetes, remains unclear. The aim of this study was to explore the association between depression and ghrelin, leptin, cortisol, and C-peptide in patients with diabetes. METHODS: We enrolled 50 adults without T2DM, 77 non-depressed adults with T2DM (free of Axis-I psychiatric disorders as assessed using the Mental Illness Needs Index (MINI), Patient Health Questionnaire (PHQ-9 score ≤ 4)) and 59 patients with T2DM and depression (PHQ-9 ≥ 7 and positive by the Structured Clinical Interview for DSM-5). The age range of the participants was 45-59 years of age. We compared the above three groups and explored the association between ghrelin, leptin, cortisol, C-peptide, and depression in patients with diabetes. A post-hoc power-analysis was finished. RESULTS: Compared with the non-depression T2DM group, the depression T2DM group had significantly higher blood glucose fluctuations. Further, compared with the non-depression T2DM and non-diabetic groups, the depression T2DM group had significantly lower levels of post-meal 2-h C-peptide and elevated evening cortisol (p < 0.01). Regression analysis revealed a significant negative correlation between depression severity and 2-h postprandial C-peptide in patients with diabetes (p < 0.01) and a significant positive correlation with midnight cortisol levels (p < 0.01). A post hoc power analysis showed that we had an adequate sample size and met the minimum requirement to attain 80% power. A post hoc power calculation also demonstrated that this study basically achieved power of 80% at 5% alpha level. CONCLUSIONS: Our findings indicate a correlation of low fasting levels of 2-h C-peptide as well as higher midnight cortisol levels with higher depression severity in middle-aged patients with T2DM.

Skeletal Extracellular Matrix Supports Cardiac Differentiation of Embryonic Stem Cells: a Potential Scaffold for Engineered Cardiac Tissue.

BACKGROUND/AIMS: Decellularized cardiac extracellular matrix (cECM) has been widely considered as an attractive scaffold for engineered cardiac tissue (ECT), however, its application is limited by immunogenicity and shortage of organ donation. Skeletal ECM (sECM) is readily available and shows similarities with cECM. Here we hypothesized that sECM might be an alternative scaffold for ECT strategies. METHODS: Murine ventricular tissue and anterior tibial muscles were sectioned into 300 mm-thick, and then cECM and sECM were acquired by pretreatment/SDS/TritonX-100 three-step-method. Acellularity and morphological properties of ECM was assessed. SECM was recellularized with murine embryonic stem cells (mESCs) or mESC-derived cardiomyocytes (mESC-CMs), and was further studied by biocompatibility assessment, immunofluorescent staining, quantitative real-time PCR and electrophysiological experiment. RESULTS: The relative residual contents of DNA, protein and RNA of sECM were comparable with cECM. The morphological properties and microstructure of sECM were similar to cECM. SECM supported mESCs to adhere, survive, proliferate and differentiate into functional cardiac microtissue with both electrical stimulated response and normal adrenergic response. Purified mESC-CMs also could adhere, survive, proliferate and form a sECM-based ECT with synchronized contraction within 6 days of recellularization. CONCLUSION: ECMs from murine skeletal muscle support survival and cardiac differentiation of mESCs, and are suitable to produce functional ECT patch. This study highlights the potential of patient specific of sECM to replace cECM for bioengineering ECT.

The bHLH142 Transcription Factor Coordinates with TDR1 to Modulate the Expression of EAT1 and Regulate Pollen Development in Rice.

Male sterility plays an important role in F1 hybrid seed production. We identified a male-sterile rice (Oryza sativa) mutant with impaired pollen development and a single T-DNA insertion in the transcription factor gene bHLH142. Knockout mutants of bHLH142 exhibited retarded meiosis and defects in tapetal programmed cell death. RT-PCR and in situ hybridization analyses showed that bHLH142 is specifically expressed in the anther, in the tapetum, and in meiocytes during early meiosis. Three basic helix-loop-helix transcription factors, UDT1 (bHLH164), TDR1 (bHLH5), and EAT1/DTD1 (bHLH141) are known to function in rice pollen development. bHLH142 acts downstream of UDT1 and GAMYB but upstream of TDR1 and EAT1 in pollen development. In vivo and in vitro assays demonstrated that bHLH142 and TDR1 proteins interact. Transient promoter assays demonstrated that regulation of the EAT1 promoter requires bHLH142 and TDR1. Consistent with these results, 3D protein structure modeling predicted that bHLH142 and TDR1 form a heterodimer to bind to the EAT1 promoter. EAT1 positively regulates the expression of AP37 and AP25, which induce tapetal programmed cell death. Thus, in this study, we identified bHLH142 as having a pivotal role in tapetal programmed cell death and pollen development.

Development of a novel individualized warfarin dose algorithm based on a population pharmacokinetic model with improved prediction accuracy for Chinese patients after heart valve replacement.

The gene-guided dosing strategy of warfarin generally leads to over-dose in patients at doses lower than 2 mg/kg, and only 50% of individual variability in daily stable doses can be explained. In this study, we developed a novel population pharmacokinetic (PK) model based on a warfarin dose algorithm for Han Chinese patients with valve replacement for improving the dose prediction accuracy, especially in patients with low doses. The individual pharmacokinetic (PK) parameter - apparent clearance of S- and R-warfarin (CLs) was obtained after establishing and validating the population PK model from 296 recruited patients with valve replacement. Then, the individual estimation of CLs, VKORC1 genotypes, the steady-state international normalized ratio (INR) values and age were used to describe the maintenance doses by multiple linear regression for 144 steady-state patients. The newly established dosing algorithm was then validated in an independent group of 42 patients and was compared with other dosing algorithms for the accuracy and precision of prediction. The final regression model developed was as follows: Dose=-0.023×AGE+1.834×VKORC1+0.952×INR+2.156×CLs (the target INR value ranges from 1.8 to 2.5). The validation of the algorithm in another group of 42 patients showed that the individual variation rate (71.6%) was higher than in the gene-guided dosing models. The over-estimation rate in patients with low doses (<2 mg/kg) was lower than the other dosing methods. This novel dosing algorithm based on a population PK model improves the predictive performance of the maintenance dose of warfarin, especially for low dose (<2 mg/d) patients.

Puerarin Exerts a Delayed Inhibitory Effect on the Proliferation of Cardiomyocytes Derived from Murine ES Cells via Slowing Progression through G2/M Phase.

OBJECTIVE: Puerarin, which shows beneficial and protective effects on cardiovascular diseases, is the main isoflavone extracted from Pueraria lobata (kudzu) root. The aim of this study was to investigate the effects of puerarin on in vitro myocardial proliferation and its underlying mechanism. METHODS: Myocardial differentiation of transgenic embryonic stem (ES) cells was performed by embryoid body-based differentiation method. The proliferation assay of cardiomyocytes (CMs) derived from ES cells (ES-CMs) was performed by EdU (5-Ethynyl-2'-deoxyuridine) staining. Flow cytometry was employed to determine the cell cycle distribution and apoptosis of purified ES-CMs. Quantitative real-time PCR was utilized to study the transcription of genes related to cell cycle progression. Signaling pathways relating to proliferation were studied by western blot analysis and application of specific inhibitors. RESULTS: Puerarin exerted a delayed inhibitory effect on the proliferation of ES-CMs at the early-stage differentiation. Meanwhile, puerarin slowed progression through G2/M phase without inducing apoptosis of ES-CMs. Further assays showed that puerarin up-regulated the transcription of Cyclin A2, Cyclin B1 and Cdk1 in ES-CMs. The ERK1/2 specific inhibitor PD0325901 and the PI3K specific inhibitor Wortmannin successfully reversed puerarin-induced up-regulation of Cdk1 but not Cyclin A2 and B1. CONCLUSION: These findings suggest that puerarin inhibits CM proliferation via slowing progression through G2/M phase during early-stage differentiation.

Camptothecin-Polysaccharide Co-assembly and Its Controlled Release.

ß-Cyclodextrin modified camptothecin (CPT-CD) was synthesized through esterification reaction and "click chemistry" to greatly improve the solubility of CPT in aqueous solution, and then, a supramolecular nanoparticle was constructed by strong noncovalent interaction between ß-cyclodextrin and adamantane and amphiphilic interaction by simply mixing CPT-CD and adamantane modified hyaluronic acid (HA-ADA) together. The obtained nanoparticle had a hydrophilic HA shell, which could target and recognize HA receptors overexpressed on the surface of cancer cells, and a hydrophobic CPT core, which could protect CPT from hydrolyzation. The results of cytotoxicity experiments showed that the nanoparticle we have designed in this work exhibited similar anticancer activities to, but with much lower side effects than, the commercial chemotherapeutic drug CPT in vitro. We believe that this work might provide a strategy for improving the treatment performance of CPT in laboratory and clinical settings.

Puerarin Suppresses the Self-Renewal of Murine Embryonic Stem Cells by Inhibition of REST-MiR-21 Regulatory Pathway.

BACKGROUND/AIMS: Puerarin shows a wide range of biological activities, including affecting the cardiac differentiation from murine embryonic stem (mES) cells. However, little is known about its effect and mechanism of action on the self-renewal of mES cells. This study aimed to determine the effect of puerarin on the self-renewal and pluripotency of mES cells and its underlying mechanisms. METHODS: RT-PCR and real-time PCR were used to detect the transcripts of core transcription factors, specific markers for multiple lineages, REST and microRNA-21 (miR-21). Colony-forming assay was performed to estimate the self-renewal capacity of mES cells. Western blotting and wortmannin were employed to explore the role of PI3K/Akt signaling pathway in the inhibitory action of puerarin on REST transcript. Transfected mES cells with antagomir21 were used to confirm the role of miR-21 in the action of puerarin on cell self-renewal. RESULTS: Puerarin significantly decreased the percentage of the self-renewal colonies, and suppressed the transcripts of Oct4, Nanog, Sox2, c-Myc and REST. Besides, PECAM, NCAM and miR-21 were up-regulated both under the self-renewal conditions and at day 4 of differentiation. The PI3K inhibitor wortmannin successfully reversed the mRNA expression changes of REST, Nanog and Sox2. Transfection of antagomir21 efficiently reversed the effects of puerarin on mES cells self-renewal. CONCLUSION: Inhibition of REST-miR-21 regulatory pathway may be the key mechanism of puerarin-induced suppression of mES cells self-renewal.

Endoplasmic reticulum stress may not be involved in intestinal epithelial cell apoptosis in experimental acute pancreatitis.

OBJECTIVES: To investigate whether endoplasmic reticulum (ER) stress is activated in the intestinal epithelium of acute pancreatitis (AP), and whether it is one of the inducing factors of the intestinal epithelial cell apoptosis in AP. METHODS: Twenty-four rats were randomly divided into two groups. AP was induced via retrograde injection of 3 % sodium taurocholate into the pancreatic duct. As a control group, rats received a sham operation. Forty-eight hours after the operation, the ultrastructural changes of ileal epithelial cells were investigated by transmission electron microscope. The protein expressions of GRP78, CHOP, caspase-12, and JNK in the ileal epithelium were determined by immunohistochemistry, and apoptosis was determined by TdT-mediated dUTP nick end labeling. The mRNA expressions of GRP78, CHOP, caspase-12, and JNK in the ileal epithelium were determined using quantitative RT-PCR. RESULTS: The ileal epithelium in rats with AP had significantly higher apoptotic cells compared with that of the control rats (P < 0.05). ER stress was activated in the ileal epithelium, which was characterized by dilated, irregular ER and upregulated expressions of GRP78 mRNA and protein. The mRNA and protein expressions of CHOP, caspase-12, and JNK in rats with AP were similar to that in the control rats (P > 0.05). CONCLUSIONS: ER stress is induced in intestinal epithelium during AP; however, ER stress is not likely to be involved in the apoptosis of the intestinal epithelium during AP.

[Efficacy and outcome of palliative TURP in patients with bladder outlet obstruction induced by advanced prostate cancer].

OBJECTIVE: To evaluate the efficacy and outcome of palliative transurethral resection of the prostate (pTURP) in patients with server bladder outlet obstruction (BOO) due to prostatic obstruction induced by advanced prostate cancer. METHODS: All the 16 patients who had a pTURP between November 2007 and January 2015 due to BOO (high residual urine volume combined with hydronephrosis or urinary retention refractory to medical treatment) at our institution were retrospectively assessed. All the patients were diagnosed with advanced prostate cancer (III stage or IV stage). The clinical data, functional and oncological follow-up results were evaluated. The cancer specific survivals were estimated by Kaplan-Meier analysis. RESULTS: The mean age of the patients was 73.8 years (63-81 years). Five cases were graded in stage III of prostate cancer and 11 in stage IV. The indications for pTURP were refractory urinary retention in 12 cases, and high residual urine volume with hydronephrosis in 4 cases. The mean prostate volume at pTURP was 43.2 mL (28-78 mL) and the mean PSA (prostate specific antigen) level before pTURP was 48.2 µg/L (2-107 µg/L). The patients had mean residual urine volume 166.4 mL (50-450 mL) and mean urinary flow rate 3.6 mL/s (0-6 mL/s, n=7) before pTURP. It took mean 62.9 min (35-94 min) in pTURP with mean estimated blood loss 126.9 mL (30-263 mL) and mean resected tissue 14.1 g (10-22 g). There were no transfusion cases. Postoperative mean serum PSA 20.5 µg/L (1-41 µg/L), residual urine volume 43.4 mL (0-400 mL) and urinary flow rate 10.1 mL/s (7-16 mL/s, n=7) were shown in these cases. A patient encountered persistent hematuria needing irrigation. Compared with preoperation, the patients had significantly lower serum PSA level (P<0.001), less residual urine volume (P<0.001) and more urinary flow rate (P=0.001) after pTURP. The mean follow-up after pTURP was 36 months (1-86 months). In addition, 2 patients received repeated pTURP. At the time of the latest analysis, 3 patients died from prostate cancer progression. As estimated by Kaplan-Meier analysis, the 2-, 3- and 5-year cancer specific survival rates after pTURP were 91%, 78% and 58%, respectively. CONCLUSION: Despite less resected tissue, greater delay in urination and reoperation rates, pTURP is a fairly effective procedure in patients with server BOO. Although a potential negative impact of pTURP on survival cannot be excluded, the estimated 3- and 5-year cancer specific survival rates in this series seem to justify this intervention.

[Protective effect of phloroglucinol on renal ischemia and reperfusion injury].

OBJECTIVE: To investigate the effect and mechanisms of Phloroglucinol (PG) on renal ischemia and reperfusion injury (IRI). METHODS: Forty-eight male Wistar rats were divided into 3 groups (16 rats per group): sham operated, saline-treated I/R (I/R), and PG-treated I/R (PG). I/R model: After removing the right kidney, renal I/R injury was induced by clamping the left renal artery for 45 min followed by reperfusion. The rats were administered with PG (30 mg/kg, intraperitoneally) or saline 15 min before renal ischemia. The blood and kidneys were harvested 6 and 24 h after reperfusion. Renal function and histologic changes of serum creatinine (SCr) and blood urea nitrogen(BUN)were assessed. Malondialdehyde (MDA),catalase (CAT),superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px)were measured. Nuclear factor-kapa B (NF-κB) and caspase-3 in the kidneys were also measured. RESULTS: SCr and BUN were (103.9±10.4) µmol/L and (15.2±1.0) mmol/L in I/R group, and (81.8±13.4) µmol/L and (11.5±1.2) mmol/L in PG group 6 h after reperfusion. SCr and BUN were (154.9±12.1) µmol/L and (28.1±1.4) mmol/L in I/R group, (103.8±5.9) µmol/L and (16.0±1.0) mmol/L in PG group 24 h after reperfusion.PG treatment significantly attenuated renal dysfunction and histologic damage caused by I/R injury (P<0.05).The I/R-induced elevation in kidney MDA level decreased, where as reduced kidney SOD,CAT and GSH-Px were increased. What is more, the apoptotic tubular cells, the levels of active caspase-3,and active nuclear factor kappa B dramatically decreased after PG treatment. CONCLUSION: PG protects murine kidney I/R injury by suppressing oxidative stress, inflammation, and cell apoptosis.

Penehyclidine hydrochloride ameliorates renal ischemia-reperfusion injury in rats.

BACKGROUND: Renal ischemia-reperfusion (I/R) injury is a major cause of acute kidney failure by mechanisms that involve oxidative stress, inflammation, and apoptosis. Penehyclidine hydrochloride (PHC), a selective anticholinergic agent, possesses anti-inflammatory, antioxidative stress, and antiapoptotic effects. Therefore, we investigated the ability of PHC to ameliorate renal I/R injury in Sprague-Dawley rats. MATERIALS AND METHODS: Rats were randomly assigned to three groups (35 rats per group): sham operated, saline-treated I/R, and PHC-treated I/R. After removing the right kidney, renal I/R injury was induced by clamping the left renal artery for 45 min followed by reperfusion. The rats were administered PHC (0.45 mg/kg, intravenously) or saline 30 min before renal ischemia. The blood and kidneys were harvested at 1, 3, 6, 12, or 24 h after reperfusion. Renal function and histologic changes were assessed. Markers of oxidative stress, inflammation, and apoptosis in the kidneys were also measured. RESULTS: PHC treatment significantly attenuated renal dysfunction and histologic damage caused by I/R injury. The treatment also decreased malondialdehyde level and attenuated the reduction in superoxide dismutase activity in the kidney. Moreover, the levels of activated p38 mitogen-activated protein kinase, nuclear factor kappa B, and caspase 3 were lower in the PHC-treated animals. CONCLUSIONS: PHC protected rat kidneys from I/R injury by attenuating oxidative stress, inflammatory response, and apoptosis. Thus, PHC may represent a novel practical strategy for the treatment of renal I/R injury.

[Ureteral stricture following renal transplantation: risk factors and surgical management].

OBJECTIVE: To evaluate the prevalence and the risk factors of the ureteral stricture following renal transplantation and outcomes of surgical managements. METHODS: By a retrospective analysis, we assessed 1 293 patients who underwent renal transplantation between January 2001 and December 2013 at our center. The patients with ureteral stricture had undergone surgical management which included neoureterocystostomy, ureteral anastomosis with the native ureter and endoscopic therapy. RESULTS: Ureteral stricture following renal transplantation was diagnosed in 17 cases (1.3%) including 6 males and 11 females. The mean age of the patients at the time of surgery was 44 (29 to 64) years. The median interval between ureteral stricture obstruction and kidney transplantation was 4 (1 to 120) months. The ureteral stricture was significantly correlated with multiple renal arteries, donor age and delayed graft function. The effective rate of surgical management was 65.75%. CONCLUSION: The incidence of ureteral stricture as a urologic complication after renal transplantation is low. The advanced techniques that preserve the ureteric blood supply should be applied. The surgical managements have respective advantages and disadvantages. The neoureterocystostomy shows best outcome, while ureteral anastomosis with the native ureter has poor outcome.

[Transabdominal laparoscopic radical nephrectomy of modified Pfannenstiel incision].

OBJECTIVE: To evaluate the efficacy and safety of transperitoneal laparoscopic radical nephrectomy with the modified Pfannenstiel incision. METHODS: Between Aug. 2012 and Jul. 2013, the same surgeon performed transperitoneal laparoscopic radical nephrectomy for 12 patients with renal masses. The approach was usually performed through 3 ports or 4 ports placed in a traditional manner. After laparoscopic radical nephrectomy was completed, and the specimen was entrapped in a specimen retrieval bag, a 7 cm modified Pfannenstiel skin incision was made over the symphysis pubis, lateralized slightly toward the side of surgery. RESULTS: All the procedures were completed without conversion to open radical nephrectomy. The operative time was 106 to 234 minutes and the blood loss was minimal. There was no intra- or post-operative complications. The pathological result was renal cell carcinoma. One patient died of multiple organ metastasis after 5 months. No recurrence was seen after 9 to 20 months' follow-up. CONCLUSION: Laparoscopic radical nephrectomy with the modified Pfannenstiel incision where the kidney is removed offers the benefits of improved cosmesis over the traditional muscle-cutting extension of an upper abdominal, lateral port site. The modified Pfannenstiel incision combines the advantages of a low abdominal incision and improved cosmesis, and can be considered a potential alternative for traditional laparoscopic nephrectomy.

Analysis of Nonhomologous End Joining and Homologous Recombination Efficiency in HEK-293T Cells using GFP Based Reporter Systems.

DNA double-strand breaks (DSBs) represent the most perilous DNA lesions, capable of inducing substantial genetic information loss and cellular demise. In response, cells employ two primary mechanisms for DSB repair: nonhomologous end joining (NHEJ) and homologous recombination (HR). Quantifying the efficiency of NHEJ and HR separately is crucial for exploring the relevant mechanisms and factors associated with each. The NHEJ assay and HR assay are established methods used to measure the efficiency of their respective repair pathways. These methods rely on meticulously designed plasmids containing a disrupted green fluorescent protein (GFP) gene with recognition sites for endonuclease I-SceI, which induces DSBs. Here, we describe the extrachromosomal NHEJ assay and HR assay, which involve co-transfecting HEK-293T cells with EJ5-GFP/DR-GFP plasmids, an I-SceI expressing plasmid, and an mCherry expressing plasmid. Quantitative results of NHEJ and HR efficiency are obtained by calculating the ratio of GFP-positive cells to mCherry-positive cells, as counted by flow cytometry. In contrast to chromosomally integrated assays, these extrachromosomal assays are more suitable for conducting comparative investigations involving multiple established stable cell lines.