WWP2 overexpression inhibits the antitumor effects of doxorubicin in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a common liver cancer that accounts for 90% of cases. Doxorubicin exhibits a broad spectrum of antitumor activity and is one of the most active agents in HCC. WW domain-containing protein 2 (WWP2) is highly expressed in HCC tissues and activates protein kinase B (AKT) signaling pathway to enhance tumor metastasis. However, the role of WWP2 in the glycolysis and antitumor effects of doxorubicin and the epigenetic alterations of WWP2 in HCC remain to be elucidated. The levels of WWP2 and N6-methyladenosine methyltransferase-like 3 (METTL3) in clinical samples and cells were investigated. WWP2 were silenced or overexpressed to study the role of WWP2 in regulating cell proliferation, colony formation, and glycolysis. RNA immunoprecipitation was performed to test m6 A levels. Quantitative reverse-transcription polymerase chain reaction (RT-PCR) and Western blot were used to measure mRNA and protein, respectively. WWP2 silencing inhibits cell proliferation, colony formation, and glycolysis, while WWP2 overexpression has the inverse effects via the AKT signaling pathway. Silencing WWP2 enhances doxorubicin's antitumor effect, while WWP2 overexpression suppresses doxorubicin's antitumor effect. Data also support that METTL3 mediates WWP2 m6A modification, and m6A reader, IGF2BP2, binds to the methylated WWP2 to promote the stability of WWP2, leading to upregulation of WWP2. METTL3 mediates WWP2 m6A modification, which can be recognized and bound by IGF2BP2 to increase the stability of WWP2, leading to WWP2 overexpression which inhibits the antitumor effects of doxorubicin through METTL3/WWP2/AKT/glycolysis axis.

Silencing of WWP2 inhibits adhesion, invasion, and migration in liver cancer cells.

The role and clinical implication of the WWP2 E3 ubiquitin ligase in liver cancer are poorly understood. In the current study, we investigated the expression level of WWP2 and its functions in cell adhesion, invasion, and migration in liver cancer. We used real-time PCR to detect the expression of WWP2 in liver cancer and adjacent samples from the People's Hospital of Lishui and also analyzed The Cancer Genome Atlas (TCGA) RNA-seq data by bioinformatics. Migration and invasion were detected by transwell analysis. We detected a strong WWP2 expression in tumor tissues of the People's Hospital of Lishui, and the survival rate was significantly higher in patients with lower WWP2-expressing tumors. WWP2 small hairpin RNA (shRNA) lentivirus stably infected cells (shWWP2), Huh7, showed slower growth speed compared with scramble control-infected cells in a xenograft mouse model. Knockdown of WWP2 Huh7 and BEL-7404 cells demonstrated a reduction in adhesion, invasion, and migration. Gene set enrichment analysis (GSEA) showed that WWP2 is positively correlated to cancer-related pathways including the chemokine signaling pathway. WWP2 also regulated MMP-9, caspase-9, CXCR3, and CCR5 expression in liver cancer cells. In addition, knockdown of CXCR3 and CCR5 significantly inhibited cell proliferation, adhesion, invasion, and migration in Huh7 and BEL-7404 cells. Our data suggest that targeting of WWP2 may be a therapeutic strategy for liver cancer treatment.

Lysobacter erysipheiresistens sp. nov., an antagonist of powdery mildew, isolated from tobacco-cultivated soil.

A bacterial strain, RS-LYSO-3T, was isolated from tobacco-cultivated soil, collected near Chuxiong, Yunnan province, southwestern China. RS-LYSO-3T could effectively inhibit the invasion of powdery mildew on tobacco. The colonies of RS-LYSO-3T were pale yellow, and its cells were Gram-stain-negative and rod-shaped, with 68 mol% DNA G+C content. Gene sequence analysis for its 16S rRNA gene revealed the highest similarity (97.78 %) with that of Lysobacter spongiicolaKMM 329T. Chemotaxonomic data showed that RS-LYSO-3T possesses a quinone system with Q-8, and iso-C16 : 0, summed feature 9 and iso-C15 : 0 as the predominant fatty acids, all of which support the affiliation of RS-LYSO-3T to the genus Lysobacter. The results of DNA-DNA hybridization, physiological and biochemical tests clearly proved that RS-LYSO-3T is a representative of a novel species of the genus Lysobacter, for which the name Lysobacter erysipheresistens sp. nov. is proposed. The type strain is RS-LYSO-3T (=CCIC 23922T=JCM 31042T).

Within-island speciation with an exceptional case of distinct separation between two sibling lizard species divided by a narrow stream.

Delimitation of genetic and geographic boundaries between species is a focus of evolutionary biology. In this study, we demonstrated fine-scale differentiation of Takydromus formosanus species complex comprising four insular endemics on Taiwan Island. Phylogeny and ancestral range reconstruction based on mitochondrial DNA sequences of 430 Takydromus lizards (405 lizards of this complex throughout their distribution range, and 25 lizards from 11 other species) indicated that the major branching process occurred within Taiwan, which represented a solid evidence of within-island speciation on this small island. We further demonstrated an exceptional case of a pair of sister species, T. viridipunctatus and T. luyeanus, that were separated by a narrow stream with a width of only 15m. This pattern might be one of the narrowest contact zones ever documented in terrestrial vertebrates. To evaluate the level of genetic introgression between these sister species, a fine-scale collection of another 382 lizards was conducted along a transect line across the stream. A total of 13 microsatellite markers and mtDNA genotyping was used to detect a low proportion of hybrids (5.7-9.9% from STRUCTURE, and 2.3% from DAPC). Our results indicated that the two clades are highly differentiated across this extremely short distance.

[A survey of the quality of life among 740 men after surgical sterilization].

OBJECTIVE: To evaluate the life quality of men after surgical sterilization. METHODS: We investigated the incidence of psychosomatic reactions, their influential factors and the quality of life among 740 men after surgical sterilization. We also evaluated the effectiveness of mental intervention for those with psychosomatic reactions. RESULTS: Totally, 59 (8%) of the subjects were not satisfied with their life status, including 42 (5.67%) suffering from psychosomatic reactions, and their psychosomatic reactions were related to their education background, economic status and understanding of sterilization. After 2 - 24 months of mental intervention, 34 (80.95%) of the 42 cases were cured, 6 (14.29%) relieved, and 2 (4.76%) unimproved. CONCLUSION: Mental intervention is effective for psychosomatic reactions in men after surgical sterilization.

Mixed epithelial and stromal tumor of the kidney: report of eight cases and literature review.

Mixed epithelial and stromal tumor of the kidney (MESTK) is the term given to a class of uncommon biphasic tumors of the kidney, with few reported cases. We describe eight cases of MESTK with detailed clinicopathological data and follow-up information. With this report, we hope to increase clinical awareness that MESTK should be considered as one of the possible diagnoses for cystic renal mass, especially in peri-menopausal women or those who receive hormone therapy. In addition, regular follow-up is necessary for the any cases with malignant potential.

[Valuable experiences and analysis of causes of urine-induced sepsis].

OBJECTIVE: To explore the characteristics, early symptoms and valuable experiences in the diagnosis and treatment of urine-induced sepsis. METHODS: The clinical data of 30 cases of urine-induced sepsis between August 2008 and April 2012 were reviewed retrospectively. Among them, YAG laser lithotripsy (n = 3) and transurethral prostatic resection (TURP) (n = 2) were performed. One case was after transurethral resection of bladder (TURB). There were 3 cases after cystectomy and Bricker and another 18 during and after percutaneous nephrolithotomy (PCNL). RESULTS: A total of 12 PCNL cases have significant drop in blood pressure during operation with non-bleed causes. A total of 26 cases developed symptoms of chills and high fever after 2 hours of surgery. The earliest symptoms of urine-induced sepsis were decline in blood pressure and high fever. After resistance to infection and symptomatic treatment actively, 27 patients were rescued successful (90%), 3 patients died (10%). Among the 3 deceased patients, 1 case was after YAG laser lithotripsy under ureteroscope, 1 case was after PCNL, 1 case was after TURP. CONCLUSIONS: The onset of urine-induced sepsis onset may be insidious. But this condition deteriorates rapidly and it is rather difficult to reverse. Most cases could be managed when the onset characteristics were recognized early. Prevention and early treatment are keys of lowering the mortality.

[Management experience of acute renal failure induced by unilateral ureteral calculi obstruction].

OBJECTIVE: To explore the causes and treatment options of acute renal failure induced by unilateral ureteral calculi obstruction. METHODS: The clinical data of 12 cases of acute renal failure induced by unilateral ureteral calculi obstruction between August 2008 and July 2012 were reviewed retrospectively. There were 5 males and 7 females with an average age of 65.7 years. Their clinical data and treatment options were retrospectively analyzed and summarized. Seven cases showed right side ureteral calculus with hydronephrosis while another 5 presented left side ureteral calculus with hydronephrosis. Serum creatinine was higher than 310 µmol/L in 12 cases. Anuria appeared in 4 cases for 1-7 days while oliguria in 8 cases for 2-10 days. High fever was present in 11 cases, the highest of whom was 40 °C. White blood cell count increased in 10 cases (>10×10(9)/L) and decreased in 2 cases (<4 × 10(9)/L). RESULTS: The therapeutic options included insertion of double J stent for internal drainage (n = 1), percutaneous nephrostomy for external drainage (n = 10) and open operation (n = 1). Traditional treatments were performed to manage ureteral calculus in the above 11 cases with drainage. All cases had improved renal function after comprehensive treatment of anti-infection, antishock, rinsing stones and relieving obstruction. All 12 cases were treated successfully. CONCLUSIONS: Unilateral ureteral calculus may impair contralateral renal function and cause acute renal failure due to the absorption of toxin at obstructive side. The keys of management are eliminating toxin and relieving obstruction.

5-epi-ent-Kaurane diterpenoids from the aerial parts of Isodon eriocalyx and their anti-atherosclerotic potential.

The phytochemical investigation of the EtOAc extract from the aerial parts of Isodon eriocalyx afforded seventeen diterpenoids, including eight undescribed compounds. Eriocalyxins H-L have unique structural characteristics featuring a 5-epi-ent-kaurane diterpenoid scaffold with eriocalyxins H-K also possess an unusual 6,11-epoxyspiro-lactone ring while eriocalyxin L, a 1,7:3,20-diepoxy-ent kaurene, features an 1,7-oxygen linkage. The structures of these compounds were elucidated by spectroscopic data interpretation, and the absolute configurations of eriocalyxins H, I, L, and M were confirmed by single-crystal X-ray diffraction. The isolates were screened for their inhibitory activities against VCAM-1 and ICAM-1 at 5 µM. While eriocalyxin O, coetsoidin A and laxiflorin P were found to significantly inhibit both VCAM-1 and ICAM-1, 8 (17),13-ent-labdadien-15 â†’ 16-lactone-19-oic acid displayed evidently inhibitory effect against ICAM-1.

[Effects of huangkui capsule on renal inflammatory injury by intervening p38MAPK signaling pathway in rats with adriamycin-induced nephropathy].

OBJECTIVE: To explore the potential mechanisms of huangkui capsule (HKC), an extract from Abelmoschus manihot (AM), for ameliorating renal inflammatory injury by regulating p38 mitogen-activated protein kinase (MAPK) signaling pathway in rats with adriamycin-induced nephropathy (ADRN). METHOD: Nineteen Sprague-Dawley (SD) rats were randomly divided into three groups, the sham-operation group, the untreated model group,and the HKC-treated group. Rats in the untreated model group and the HKC-treated group were made into ADRN model by right nephrectomy and twice intravenous injections of adriamycin( ADR, 0.4 mL and 0.2 mL respectively within 4 weeks). After the model successfully established, rats in the HKC-treated group were orally given HKC (2 mg x kg(-1) per day), while rats in the untreated model group and the sham-operation group were intervened with distilled water respectively. The intervention for all rats was 4 weeks. Rats' body weight were weighted and 24 h urinary protein excretion (Upro) was detected at the end of the 1st, 2nd, 3rd, and 4th week after the intervention of HKC or distilled water. All rats were sacrificed at the end of the 8th week after nephrectomy, and then, to withdraw blood and kidney to examine the blood biochemical parameters, the glomerular morphological changes, alpha-smooth muscle actin (alpha-SMA) and collagen type I expressions,and the glomerular macrophages infiltration. Besides, the protein expression of transforming growth factor (TGF)-beta1, p38MAPK, as well as phosphorylated p38MAPK (p-p38MAPK) in renal tissues were detected by Western blotting. RESULT: As compared with rats in the untreated model group, in the HKC-treated group,the HKC treatment significantly improved Upro, serum albumin, mesangial cell proliferation, extracellular matrix (ECM) and collagen deposition,and decreased the expression of alpha-SMA and collagen type I and the infiltration of ED1+ and ED3+ cells in glomeruli. In addition, it significantly down-regulated the protein expression of TGF-beta1 and p-p38MAPK in renal tissues. CONCLUSION: HKC had the effects on ameliorating renal inflammatory injury in vivo. It could reduce the expression of TGF-beta1 and improve the infiltration and activation of inflammatory cells in glomeruli by way of intervening p38MAPK signaling pathway in kidney through down-regulating the protein expression of p-p38MAPK, as the key signal molecule.

Mathematical processing of trading strategy based on long short-term memory neural network model.

At present, gold and bitcoin have become mainstream assets in market transactions. Due to the volatility of gold and bitcoin prices, we can buy and sell assets like gold and bitcoin the same way we buy and sell stocks. The research goal of this article is to develop an optimal trading strategy that maximizes our post-trade returns. By studying the relationship between the two, on the one hand, it supplements and enriches the theoretical research on the rate of return of gold and Bitcoin, on the other hand, it provides a certain reference for investors to construct investment strategies. The research on the cointegration relationship between them has important practical significance. At the same time, it has important practical significance for the research on the cointegration relationship between bitcoin and gold.

The possible reduction pathways of 2,4,6-trinitrotoluene (TNT) by sulfide under simulated anaerobic conditions.

To predict the final fate of 2,4,6-trinitrotoluene (TNT) and its intermediates in an anaerobic fermentative solution containing reduced sulfur species and to provide a basis for the adoption of remediation methods, we investigated the pathways of TNT (TNT(0) = 50 mg/L) reduction by Na(2)S at 30 ± 1 °C in an acetic acid-sodium bicarbonate buffer. Liquid chromatography/mass spectrometry (LC/MS) was used to identify TNT metabolites at different reaction times. The law of growth and decline of TNT and its metabolites was determined with time. The LC/MS result, combined with the physicochemical characteristics of related products and information from the literature, indicated possible TNT conversion pathways. Sulfide can initiate both nitroreduction and denitration of TNT simultaneously. Nitroreduction led to the accumulation of primary intermediates 4-hydroxylaminodinitrotoluene and 4-aminodinitrotoluene, whereas denitration resulted in the production of unidentified substances with molecular weight less than that of TNT. Also, polyreaction between the above intermediates formed many unidentified substances. Humification was concluded to be the best choice for remediation of TNT-contaminated soil and water due to the formation of intermediates with stable, intact aromatic systems. However, the denitration pathway of TNT offered the possibility of mineralization.

Two new 7, 20-epoxy-ent-kauranes from the aerial parts of Isodon eriocalyx.

Two previously undescribed 7, 20-epoxy-ent-kauranes along with six known ent-kauranoids, were isolated from the aerial parts of Isodon eriocalyx. The structures of new compounds were established on the basis of extensive spectroscopic analyses. Compound 2 could inhibit the production of interleukin - 1ß (IL - 1ß) in monosodium urate (MSU) and lipopolysaccharide (LPS) induced macrophages.

Semaphorin 4D promotes the proliferation and metastasis of bladder cancer by activating the PI3K/AKT pathway.

The present study aimed to investigate the role of semaphorin 4D (Sema4D) in bladder cancer cell proliferation and metastasis in vivo and in vitro. Effects of Sema4D modulation on cancer cell viability and clonogenic abilities were assessed by MTT assay and colony formation assay. Cell apoptosis, cell cycle analysis, transwell assays, and wound-healing assays were also assayed. A mouse model of bladder cancer was established to observe the tumorigenesis in vivo. Our data showed that Sema4D was 4-fold upregulated in clinical bladder cancer tissues relative to noncancerous ones and differentially expressed in bladder cancer cell lines. Knockdown of Sema4D in bladder cancer T24 and 5637 cells significantly decreased cell proliferation, clonogenic potential, and motility. On the contrary, overexpression of Sema4D in bladder cancer SV-HUC-1 cells significantly increased cell viability and motility. Concordantly, knockdown of Sema4D impaired while overexpression of Sema4D promoted bladder cancer cell growth rates in xenotransplanted mice. Cell cycle was arrested by modulation of Sema4D. Cell apoptotic rates and the mitochondrial membrane potentials were consistently increased upon knockdown of Sema4D in T24 cells and 5637 cells. Western blotting revealed that epithelial-mesenchymal transition was promoted by Sema4D. The PI3K/AKT pathway was activated upon Sema4D overexpression in SV-HUC-1 cells, while it was inactivated by knockdown of Sema4D in T24 cells. All these data suggest that Sema4D promotes cell proliferation and metastasis in bladder cancer in vivo and in vitro. The oncogenic behavior of Sema4D is achieved by activating the PI3K/AKT pathway.

Knockdown of lncRNA XIST suppresses osteosarcoma progression by inactivating AKT/mTOR signaling pathway by sponging miR-375-3p.

BACKGROUND: Osteosarcoma (OS) is one of the most common bone tumors in adolescents and young adults. Emerging evidence suggested ncRNA (lncRNA and miRNA) are closely associated with cell progression, apoptosis and autophagy. However, the role of regulatory network between ncRNA and mRNA in OS has not been fully verified. METHODS: lncRNA XIST, miRNA expression were detected by qRT-PCR. The protein expression of LC3, p62, AKT, p-AKT, mTOR and p-mTOR was measured by western blot. MTT assay and flow cytometry were applied to measure cell proliferation and apoptosis. Luciferase assay was used to ensure the relationship between lncRNA, miRNA and mRNA. GFP-LC3 cells were observed using fluorescence microscope. RESULTS: XIST expression was up-regulated but miR-375-3p was down-regulated in OS tissues and cells. Luciferase assay results demonstrated that miR-375-3p was a target of XIST and mTOR was a target mRNA of miR-375-3p. In addition, knockdown of XIST and mTOR inhibited OS cell proliferation and autophagy, but induced apoptosis. Knockdown of XIST could reverse the effect of miR-375-3p inhibitor on OS cells. The effects of si-mTOR of OS cells could be reversed by silencing miR-375-3p. Moreover, knockdown of XIST inhibited AKT/mTOR signaling pathway via sponging miR-375-3p. CONCLUSION: Knockdown of XIST inhibited cell growth and autophagy but induced cell apoptosis by suppressing the AKT/mTOR signaling pathway by sponging miR-375-3p.

[Cystadenoma of the seminal vesicle: 1 case report].

OBJECTIVE: To evaluate the clinical presentation pathologic characteristics, diagnosis and treatment of cystadenoma of the seminal vesicle. METHODS: One case of cystadenoma of the seminal vesicle was analyzed and relevant literature reviewed. RESULT: The patient underwent surgical removal of the right seminal vesicle along with the tumor, which was pathologically confirmed to be cystadenoma of the seminal vesicle. Ultrasonography and CT scanning revealed no recurrence 3 months later. CONCLUSION: Cystadenoma of the seminal vesicle is extremely rare and easily missed in diagnosis. Routine digital rectal examination (DRE) is necessary and ultrasonography and CT scan are helpful to diagnosis. Seminal vesiculectomy with tumor removal is an ideal option with good prognosis.

Inhibition of WWP2 suppresses proliferation, and induces G1 cell cycle arrest and apoptosis in liver cancer cells.

Primary liver cancer is one of the most common and aggressive human malignancies worldwide. As numerous studies have revealed that WW domain containing E3 Ub­protein ligase 2 (WWP2) exerts cancer­specific functions, the present study assessed the role of WWP2 in liver cancer. WWP2 was revealed to be significantly overexpressed in liver cancer tissues compared with paired normal tissues at the mRNA as well as at the protein level. Furthermore, small interfering RNA-mediated WWP2 knockdown in liver cancer cell lines was demonstrated to inhibit cell proliferation, cause cell cycle arrested in G1 phase and to induce apoptosis as revealed by a Cell Counting Kit-8 assay and flow cytometric analysis. In addition, western blot analysis revealed that WWP2 knockdown significantly increased the expression of apoptosis-associated markers caspase­7, caspase­8 and B-cell lymphoma 2 (Bcl-2)-associated X in liver cancer cell lines, while Bcl­2 was significantly decreased. In conclusion, the present study suggested that WWP2 may exert important functions in the over­proliferation and evasion of apoptosis of liver cancer, likely through regulating the expression of apoptosis-associated markers. Furthermore, WWP2 may represent a novel diagnostic marker and molecular therapeutic target for liver cancer.

Primary pleomorphic rhabdomyosarcoma of the adrenal gland in an adult: A case report.

A 61-year-old female was referred to The First Affiliated Hospital, College of Medicine, Zhejiang University (Hangzhou, China) due to a right adrenal tumor. A pre-operative transcutaneous fine-needle aspiration biopsy and right adrenalectomy were performed, and pathological analysis resulted in the diagnosis of pleomorphic rhabdomyosarcoma (RMS). Primary pleomorphic RMS of the adrenal gland in an adult is a rare condition. To the best of our knowledge, this is the first case of pleomorphic RMS of the adrenal gland in an adult diagnosed by light microscopy and immunohistochemical stains.

Ureteral obstruction and urinary fistula due to fibrin glue after partial nephrectomy: A case report and review of the literature.

In the present study, we report the case of a 69-year-old female who developed urinary leakage following partial nephrectomy (PN) to remove left renal masses. The results of CT and MR urography revealed left proximal ureteral obstruction and urinary fistula. Reoperation was performed on the 16th postoperative day to explore the left kidney and ureter in order to relieve the obstruction. The left proximal ureter was found to be enfolded by fibrin glue and showed marked stiffness and adhesion during the reoperation. The lesion of the ureter was resected and the ureter was anastomosed with the routine double-J stent. Pathological examination of surgical specimens revealed fat fibrous scar tissue hyperplasia with inflammatory cell infiltration. The patient recovered completely without exudate. Our experience suggests that care should be taken to avoid touching the ureter with fibrin glue during PN surgery.

Hand-assisted-drawing-needle running suture with a curved needle: an effective technique to create a W-ileal neobladder.

BACKGROUND: An important milestone in the area of urinary diversion was the advent of a series of orthotopic bladder substitution (OBS). However, reconstruction of OBS by the traditional hand suture method (THSM) is a time-consuming process. Stapling techniques are considered to be inferior to hand-sewn methods. We report our experience and functional results in patients with W-ileal neobladder by a hand-assisted-drawing-needle running suture (HADNRS). METHODS: Between April 1993 and December 2011, 347 patients (338 men and 9 women) aged 28 - 77 years (median age: 59 years) underwent radical cystectomy, followed by the creation of a modified W-ileal neobladder by HADNRS with a curved needle. A total of 347 (20 patients in 2003) were evaluated by urodynamic tests. RESULTS: The operative time ranged from 110 to 310 minutes (mean 148 minutes), and the mean time of reconstruction by HADNRS, excluding ureterointestinal and ileouretral anastomosis, was (20.2 ± 4.3) minutes. Histopathological analysis of removed specimens showed that 317 patients had transitional cell bladder carcinoma. Of these 317 patients, 19 also had squamous carcinoma and 13 had adenocarcinoma. Glandularis and prostate cancer occurred in 16 and 14 patients, respectively. Three patients (0.8%) had neobladder abdominal fistula. No other early complications or injury to the surgeon's hands occurred due to HADNRS. Of the 20 cases with urodynamic examinations in 2003, two suffered from daytime incontinence and six had nocturnal incontinence. The maximum capacity of the neobladder was (492.9 ± 177.8) ml, and the maximum pressure within the reservoir at the end of filling was (32.1 ± 8.6) cmH2O. CONCLUSION: Reconstruction of W-ileal neobladder by HADNRS is effective and economical.