Retraction Notice to: A Novel Mechanism of Doxorubicin Resistance and Tumorigenesis Mediated by MicroRNA-501-5p-Suppressed BLID.

[This retracts the article DOI: 10.1016/j.omtn.2018.06.011.].

Evaluating the effects of microparticle addition on mycelial morphology, natural yellow pigments productivity, and key genes regulation in submerged fermentation of Monascus purpureus.

Morphology plays an important role in fungal fermentation and secondary metabolites biosynthesis. One novel technique, microparticle-enhanced cultivation was successfully utilized to control the morphology of Monascus purpureus precisely and enhance the yield of yellow pigments. The production of yellow pigments increased to 554.2 U/ml when 4 g/L 5000 mesh talc added at 24 h. Field emission scanning electron microscope observation indicated that the actual effect depends on the properties of microparticle. Sharp-edged microparticles showed better stimulatory effects than smooth, round-shaped ones. Particle size analysis, scanning electron microscope, and cell integrity evaluation proved obvious morphological changes were induced by talc addition, including smaller mycelial size, rougher hyphae, and decreased cell integrity. Furthermore, the expression levels of MrpigG, MrpigD, MrpigE, and MrpigH were significantly upregulated by the addition of talc. It indicated that the microparticle could not only affect the mycelial morphology, but also influence the expression levels of key genes in biosynthetic pathway of Monascus yellow pigments.

A standardized pathological proposal for evaluating microvascular invasion of hepatocellular carcinoma: a multicenter study by LCPGC.

BACKGROUND AND AIMS: Microvascular invasion (MVI) is a key pathological factor that severely affects the postoperative prognosis of patients with hepatocellular carcinoma (HCC). However, no MVI classification schemes based on standardized gross sampling protocols of HCC are available at present. METHODS: 119 HCC specimens were sampled at multiple sites (3-, 7-, and 13 points) for the optimum MVI detection rate. 16,144 resected HCCs were graded as M0, M1 or M2 by adopting three-tiered MVI grading (MVI-TTG) scheme based on the seven-point sampling protocol (SPSP). Survival analyses were performed on 2573 patients to explore the advantages of MVI-TTG. RESULTS: The MVI detection rate determined by SPSP was significantly higher than that determined by the 3-point sampling method (34.5% vs. 47.1%, p = 0.048), but was similar to that determined by the 13-point sampling method (47.1% vs. 51.3%, p = 0.517). Among 16,144 resected HCCs, the proportions of M0, M1 and M2 specimens according to SPSP were 53.4%, 26.2% and 20.4%, respectively. Postoperative survival analysis in 2573 HCC patients showed that the 3-year recurrence rates in M0, M1 and M2 MVI groups were 62.5%, 71.6% and 86.1%, respectively (p < 0.001), and the corresponding 3-year overall survival (OS) rates were 94.1%, 87.5% and 67.0%, respectively (p < 0.001). M1 grade was associated with early recurrence, while M2 grade was associated with both early and late recurrence. MVI-TTG had a larger area under the curve and net benefit rate than the two-tiered MVI grading scheme for predicting time to recurrence and OS. CONCLUSIONS: SPSP is a practical method to balance the efficacy of sampling numbers and MVI detection rates. MVI-TTG based on SPSP is a better prognostic predictor than the two-tiered MVI scheme. The combined use of SPSP and MVI-TTG is recommended for the routine pathological diagnosis of HCC.

A Novel Mechanism of Doxorubicin Resistance and Tumorigenesis Mediated by MicroRNA-501-5p-Suppressed BLID.

Doxorubicin is a widely used anthracycline-based anti-tumor agent for both solid and liquid tumors. Mounting evidence has demonstrated that microRNAs (miRNAs) are involved in chemoresistance and tumorigenesis. However, the roles of microRNA-501-5p (miR-501) in doxorubicin resistance and gastric cancer cell proliferation and invasion are still not fully understood. In this study, we identified that BLID (BH3-like motif-containing protein, cell death inducer) was directly regulated by miR-501 at the post-transcriptional level in multiple gastric cancer cell lines. Endogenous miR-501 was higher, whereas BLID was lower, in doxorubicin-resistant gastric cancer SGC7901/ADR cells compared with their parental SGC7901 cells. miR-501 suppressed gastric cancer cell apoptosis, induced resistance to doxorubicin, and enhanced cell proliferation, migration, and invasion. Subcutaneous injection of miR-501 lentivirus-infected SGC7901 cells resulted in rapid growth of xenograft tumors and resistance to doxorubicin treatment, unlike injection of negative miRNA lentivirus-infected SGC7901 cells. This is achieved at least partially by directly targeting BLID and subsequent inactivation of caspase-9 and caspase-3 and phosphorylation of Akt. Taken together, miR-501 induces doxorubicin resistance and enhances the tumorigenesis of gastric cancer cells by suppressing BLID. miR-501 might be a potential target for doxorubicin resistance and gastric cancer therapy.

[Lab-scale ANAMMOX Process in a Wastewater Treatment Plant].

A lab-scale, completely anaerobic ammonium oxidation (ANAMMOX) process was operated in a municipal wastewater treatment plant (WWTP). Sewage effluent treated by an A/O process and nitrification process was input as the substance to start up the up-flow ANAMMOX filter reactor. After the 109th day, the ammonia removal rate and nitrite removal rate were greater than 90% for 15 successive days and the nitrogen removal rate was higher than 70%. The ANAMMOX filter reactor successfully started up. From days 245 to 333, the reactor was running during the winter. The weight of biomass reached 12.24 mg·g-1, and the average nitrogen removal rate was 54.3%. Backwash was adopted at day 461, and the weight of biomass decreased to 8.01 mg·g-1. From days 605 to 693, the reactor was running in the winter again. The weight of biomass was 10.41 mg·g-1, and the average nitrogen removal rate was sustained at 69.7%. Compared with the previous winter, the weight of biomass was lighter but the total nitrogen removal loading was 23% greater. For the entire operation, the ANAMMOX rate at high temperature was stable but that at low temperature increased from 1.5 kg·(kg·d)-1 to 3.6 kg·(kg·d)-1. The results show:Long-term domestication at low temperature was in favor of improving treatment efficiency of ANAMMOX process in cold environment and realized ANAMMOX process operated efficiently in winter.

[Effect of Carbon Source on Lab-scale SAD Process in a Wastewater Treatment Plant].

Lab-scale anaerobic ammonia oxidation and denitrification (SAD) processes were operated simultaneously in a municipal waste water treatment plant (WWTP). Sewage treated by the A/O and nitrification process was used as the substance to start up an anaerobic ammonia oxidation filter reactor. Adding glucose and sodium propionate to influent was used as the substance to start up the SAD filter reactor after the successful start-up of the ANAMMOX reactor. The SAD process performed well with an average total nitrogen concentration in the effluent of 6.41 mg·L-1 when 30 mg·L-1 glucose was added to the effluent sewage at ambient temperature. Compared with the ANAMMOX process, the total nitrogen concentration in the effluent from the SAD process decreased 42%. The stability of the SAD process was destroyed and the SAD process turned into a denitrification process when 30 mg·L-1 glucose was added in the influent sewage in a low temperature environment. In normal and low temperature environments, the SAD process functioned well, and the average total nitrogen concentration of the effluent was 6.54 mg·L-1 when 30 mg·L-1 sodium propionate was added in the influent sewage. Compared with glucose, sodium propionate had little influence on the SAD process.

[Laboratory-scale CANON Processes Applied to Wastewater Treatment Plants].

A laboratory-scale completely autotrophic nitrogen removal over nitrite (CANON) process was operated in a municipal wastewater treatment plant (WWTP). Sewage effluent treated by the anaerobic/oxic (A/O) process and was used to operate a WWTP to obtain the initial substance for the start-up of a CANON filter reactor. On the 48th day, the ammonia removal rate was measured at greater than 90% in successive 10 d samples and the nitrogen removal rate was greater than 70%. The CANON filter was successful at start up. From the 49th to the 129th day, the dissolved oxygen in the reactor was maintained at fairly low concentration of 0.2-0.5 mg·L-1. The effluent contained nearly no ammonia and the maximum total nitrogen (TN) concentration was 15.6 mg·L-1, which exceeded the national Class 1A Discharge Standards for pollutants from municipal wastewater treatment plants. Nitrite oxidizing bacteria (NOB) proliferated excessively in the reactor. Backwash was implemented on 129th, 169th and 213th days. The nitrogen removal rate was more than 70% for a long time and TN concentration in effluent was below 12 mg·L-1. The nitrogen concentration in effluent fitted the national Class 1A Discharge Standards and the NOB were effectively inhibited. These results show that backwash has negligible on the structure of filter and its impact on the thickness of the bio-membrane and its functional bacteria was small, however, it is capable of effectively inhibiting the activity of the NOB. Periodically backwashing can be utilized as an engineering application to maintain stable operation of the CANON process.

[Application of reverse molecular docking technology in target prediction, active ingredient screening and action mechanism exploration of traditional Chinese medicine].

Traditional Chinese medicine(TCM) modernization has gradually become a worldwide trend. Reverse docking technology has also gradually become a useful tool for TCM modernization. It involves docking a small-molecule drug in the potential binding cavities of a set of clinically relevant macromolecular targets. Detailed analysis of the binding characteristics was used for the ranking of the targets according to the tightness of binding. This process can be used to potentially identify the novel molecular targets for the drug which may be relevant to its mechanism of action or side effect. In order to explore the action mechanism, screen the active ingredients and seek the treating target of TCM, reverse molecular docking technology has been widely used and has achieved remarkable results in recent years. In this review, we summarized the application of reverse molecular docking technology in the target seeking, active ingredients screening and potential mechanism exploration of TCM, which may provide more scientific basis for the clinical research and development of new herbal drugs.

Value of multidetector computed tomography in the diagnosis of mucosa-associated lymphoid tissue-lymphomas in the parotid gland: A case report and review of the literature.

The present study aimed to review the multidetector computed tomography (MDCT) imaging features of eight mucosa-associated lymphoid tissue (MALT)-lymphoma cases of the parotid gland and to explore the diagnostic value of MDCT. A total of eight patients with pathologically confirmed MALT-lymphomas of the parotid gland underwent pre-operative MDCT plain and dual-phase scans. The changes in the CT values and enhancement patterns of the tumors were assessed. Quantitative analysis was performed to determine the CT value changes of the tumors in the various enhanced phases compared with the plain scan. The MALT-lymphomas of the parotid gland exhibited even density isodense or hyperdense nodules, with occasional calcification and necrosis. The dual-phase scan of the MALT-lymphomas revealed a pattern of lower or moderate enhancement, circumambient enhancement or delayed enhancement. The MALT-lymphomas were closely associated with Sjögre's syndrome and demonstrated malignant features and isodense or hyperdense nodules and lower or moderate enhancement on the CT scans.

Expression patterns of prion protein gene in differential genotypes sheep: quantification using molecular beacon real-time RT-PCR.

Determination of the transcription level of cellular prion protein (PrP(C)) is essential for understanding its role in organisms and revealing mechanism of susceptibility and resistance to scrapie. However, the expression of prion protein (PrP) mRNA in sheep has not been quantified in great detail in digestive tract which is important during scrapie spread through oral route. Herein, we report on measurement of sheep PrP mRNA using absolute quantitative real-time RT-PCR. Total RNA was isolated from five different regions of the central nervous system (CNS), four regions of lymphoid system, eleven regions of digestive tract, and two reproductive organ tissues of eight sheep of two different genotypes (ARR/ARQ and ARH/ARQ) and PrP mRNA was quantified by real-time RT-PCR using molecular beacon. The results showed that highest levels of PrP mRNA were expressed in thalamus and cerebrum (P < 0.01) of CNS examined, followed by cerebellum, spinal cord, and brain stem. In peripheral organs examined, lymph tissue showed moderate level of PrP expression similar to that in digestive tract and reproduction organs. PrP expression levels in the same tissue of different genotype sheep had significant variation. Our study provided the first detail, tissue-specific and genotype-specific data of PrP mRNA expression in sheep for further studies of pathogenesis of prion diseases.

Significance of margin in nephron sparing surgery for renal cell carcinoma of 4 cm or less.

BACKGROUND: Current surgical practice for nephron sparing surgery allows at least 1 cm margin of normal tissue around the tumour. However, recent studies show that the width of the margin is not important, even simple enucleation is as effective as partial nephrectomy. We explored whether margin size has significant impacts on clinical outcomes in nephron sparing surgery for renal cell carcinoma of 4 cm or less. METHODS: Between 1998 and 2006, 115 patients with sporadic, pathologically confirmed, renal cell carcinoma 4 cm or less (T1a) and normal contralateral kidney were treated by nephron sparing surgery using a margin less than 5 mm. The surgical margin status was evaluated from frozen and permanent paraffin sections. RESULTS: Mean and median tumour diameter were 3.3 cm and 3.5 cm (range 1.0-4.0). The mean margin width was 2.2 mm (median 2.0, range 0-6). In addition, 114 cases had margins 5 mm or less (99.1%), 97 cases (84.3%) had margin 3 mm or less, and 26 cases had margin zero (22.6%). None of the patients had positive surgical margins. No patients died during follow-up (mean 65 months). There were no any major surgical complications and no distant metastasis was detected. Local recurrence was detected in one case (0.9%) at a different site of the kidney. CONCLUSIONS: For early localized renal cell carcinoma of 4 cm or less, as long as tumour is completely excised, the size of margin in nephron sparing surgery is not important. Nephron sparing surgery with 5 mm margin is enough for tumour control. It provides excellent renal function preservation, favourable long term progression free survival and is not associated with an increased risk of local recurrence.

[Long-term outcomes of mini-margin nephron sparing surgery for renal cell carcinoma].

OBJECTIVE: To study the safety and effects of mini-margin nephron sparing surgery (NSS) for renal cell carcinoma (RCC). METHODS: From January 1998 to December 2006, 115 cases of RCC with diameter of 4 cm or less and stage of T1aN0M0 were treated with NSS using a margin of 5 mm or more. The mean diameter of the tumors was 3.3 cm (range 1.0-4.0 cm). Of the cases, 3 were with synchronous bilateral cancer while 112 cases were with normal opposite kidneys. The clinical results were followed and analyzed. RESULTS: All of the operations were technically successful. The mean duration of surgical procedures was 90 min (ranged 80-120 min). The blood loss was 50 -200 ml. No patient needed blood transfusion. Renal arteries were occluded in 98 cases under hypothermic technique for a mean duration of 22 min (20-25 min). While in 17 cases, renal parenchyma squeezing was used for bleeding control. All of the 115 cases were of negative margin by weather frozen or routine pathologic study. The mean follow-up was 62 months (6-96 months). Local recurrence was found in 1 case during follow-up, with a local recurrence rate of 0.9%, while no distant metastasis was detected. All the patients were alive with no evidence of tumor bearing until last evaluation. Secondary gross hematuria occurred in 3 cases during hospital stay and cured by bed limitation. There were no major complications such as bleeding and urinary leakage or urinoma requiring re-operation. CONCLUSIONS: Mini-margin nephron sparing surgery is likewise safe and effective in treating early localized renal cell carcinoma 4 cm or less. It provides excellent renal function preservation, favorable long-term progression-free survival, and is not associated with an increased risk of local recurrence.

Safety and efficacy of mini-margin nephron-sparing surgery for renal cell carcinoma 4-cm or less.

OBJECTIVES: To explore whether mini-margin nephron-sparing surgery (NSS) is clinically safe and effective for renal cell carcinoma (RCC) of 4 cm or less with a normal contralateral kidney. METHODS: A total of 115 patients with sporadic, pathologically confirmed RCC of 4 cm or less (Stage T1a), with a normal contralateral kidney, were treated by NSS using a mini-margin of less than 5 mm from 1998 to 2006. The surgical margin status was evaluated by both frozen and permanent paraffin section studies. The patients were followed up, and the data were analyzed. RESULTS: The mean and median tumor diameter was 3.3 and 3.5 cm (range 1.0 to 4.0). None of the patients had positive surgical margins detected at either frozen section or final paraffin section analysis. The mean margin width was 2.2 mm (median 2.0, range 0 to 6). Of the 115 patients, 114 had margins of 5 mm or less (99.1%), 97 (84.3%) had margins of 3 mm or less, and 26 had margins of 0 mm (22.6%). At a mean follow-up of 65 months (median 66, range 9 to 105), all patients were alive. No distant metastasis was detected. Local recurrence was detected in 1 patient (0.9%) at a different site in the kidney. No major surgical complications, such as hemorrhage or urinary leakage/urinoma requiring reoperation, occurred. Considering only the 97 patients with follow-up of more than 3 years in the analysis, the mean and median follow-up time was 73 and 69 months (range 37 to 105), respectively. All 97 patients were alive with no evidence of disease at the last visit. CONCLUSIONS: The results of our study have shown that mini-margin NSS is a safe and effective approach for treating early localized RCC of 4 cm or less.

Infrequent COX-2 expression due to promoter hypermethylation in gastric cancers in Dalian, China.

Cyclooxygenase-2 (COX-2) has been shown to play oncogenic roles during stepwise gastrocarcinogenesis, and its expression is correlated with Helicobacter pylori infection, tumor necrosis factor alpha-mediated nuclear factor (NF)-kappaB activation, and Wnt signaling. To examine COX-2 expression and the status of its regulatory factors, we examined 49 gastric cancers (GCs), 21 premalignant tissues, and 10 noncancerous gastric mucosa from residents of Dalian, China. Unexpectedly, it was found that COX-2 expression was infrequent in the gastric samples (18.8%, 15/80) regardless of the type of lesion or morphological phenotype. H pylori infection was detected in 19 of 35 tested GC cases. Tumor necrosis factor alpha expression, NF-kappaB nuclear translocation, or Wnt2 overexpression was observed in 56 (82.3%) of 68, 40 (50.0%) of 80, and 62 (77.5%) of 80 of the gastric tissue samples, respectively. Methylation-sensitive restriction enzyme digestion followed by polymerase chain reaction of COX-2 promoter regions revealed a remarkably high hypermethylation rate (100%, 20/20) among the COX-2-negative GCs, which was associated with the overexpression of DNA methyltransferase (DNMT) 1 (r = 0.587, P < .01). These results indicate that (1) in contrast to previous findings using other GC sources, our results show that COX-2 activity may not be a critical molecular event during GC formation, (2) the tumor-promoting effects of H pylori infection and Wnt and NF-kappaB activities may be mediated by COX-2-independent pathways, and (3) promoter hypermethylation is the major cause of COX-2 silencing in Dalian GCs, apparently because of increased expression of DNMTs (especially DNMT1). Consequently, a COX-2-oriented preventive or therapeutic strategy is not practical for Dalian GCs. The frequent COX-2 hypermethylation observed in Dalian GCs could have insightful epigenetic and epidemiologic implications.

Frequent loss of membranous E-cadherin in gastric cancers: A cross-talk with Wnt in determining the fate of beta-catenin.

The potential correlation of E-cadherin reduction and Wnt2 up-regulation in determining the intracellular distribution of beta-catenin in gastric cancers was investigated by the methods of frozen tissue array-based immunohistochemistry, Western blot and RT-PCR analysis. It was revealed that membranous E-cadherin was reduced frequently in the two major subtypes of gastric cancer (intestinal gastric cancer, i-GC and diffuse gastric cancer, d-GC) and closely correlated with the risk of lymphoid node metastasis (P < 0.05). The reduction of membranous E-cadherin was paralleled with cytosolic and nuclear accumulation of beta-catenin and the increased Wnt2 expression. These results indicate that the reduced E-cadherin is a common genetic phenotype of GCs and plays beneficial roles in tumor metastasis. Altered beta-catenin distribution may result from the imbalance of E-cadherin production and Wnt expression, which confers on gastric cancer cells more aggressive behaviors.

Optimal margin in nephron-sparing surgery for renal cell carcinoma 4 cm or less.

OBJECTIVE: Renal cell carcinoma (RCC) of 4 cm or less is with a low incidence of multicentricity and metastasis and is usually considered suitable for nephron-sparing surgery (NSS). This study was designed to investigate the distance between extra-pseudocapsule cancer lesions and primary tumors, and to suggest the optimal margin of normal parenchyma in NSS for RCC 4 cm or less. METHODS: We prospectively studied 82 kidneys in which RCCs of 4 cm or less were resected by radical nephrectomy. According to UICC TNM classification (1997), all tumors were staged as T1 and classified as conventional RCC in 76 cases and papillary RCC in 6 cases. The kidney samples were first step sectioned at 3mm intervals and examined for multicentricity. Then, on each layer of tissue sectioned, parenchyma margins of 15 mm beyond pseudocapsule were continuously sectioned and examined microscopically to investigate completeness of pseudocapsule and possible presence of extra-pseudocapsule cancer lesions. The greatest distance between extra-pseudocapsule lesions and primary tumors was measured. RESULTS: The diameter of 82 primary tumors was 3.4+/-0.7 mm (range 1.5-4.0 cm). Of them, 31.7% (26/82) were found without intact pseudocapsule. Of the 82 cases, 19.5% (16/82) were with positive cancer lesions beyond pseudocapsule, with invasion into normal parenchyma in 12.2% (10/82), into venule in 2.4% (2/82) and satellite tumors in 4.9%(4/82). The average distance between extra-pseudocapsule cancer lesions and primary tumors was 0.5+/-1.3mm (range 0-5.0mm), with a 95% confidential interval (CI) (0.11, 0.94). No significant difference was found in the incidence of extra-pseudocapsule cancer lesions between the tumors 2.5 cm or less and that greater than 2.5 cm. CONCLUSIONS: These data suggest that when partial nephrectomy is performed in RCC 4 cm or less, a 10mm margin may be too large and go against renal function maintaining. Enucleation alone was associated with a significant risk of incomplete excision, and therefore liable for local recurrence. Thorough inspection of the whole kidney before and during operation may help to avoid leaving over large and distant multifocal lesions.

[Optimal margin in nephron-sparing surgery for renal cell carcinoma 4 cm or less in diameter].

OBJECTIVE: To investigate the optimal margin in nephron-sparing surgery (NSS) for renal cell carcinoma (RCC) 4 cm or less in diameter. METHODS: Eighty-two kidneys with RCC 4 cm or less in diameter resected by radical nephrectomy were prospectively studied. The kidney samples were sectioned at 3 mm interval and examined for multicentricity. On each layer of tissue sectioned, parenchyma margin of 15 mm beyond pseudocapsule was continuously sectioned and examined for completeness of pseudocapsule and extra-pseudocapsule cancer lesion. The farthest distance between extra-pseudocapsule lesion and primary tumor was measured. PCNA expression was detected in 41 patients by using standard SP immunohistochemistry technique. RESULTS: The diameter of 82 primary tumors was 3.4 +/- 0.8 cm (range 1.5 - 4.0 cm). Of these, 31.7% (26/82) were found without intact pseudocapsule and 17.1% (14/82) with positive cancer lesions beyond pseudocapsule. The average distance between extra-pseudocapsule cancer lesion and primary tumor was 0.5 +/- 1.3 mm (range 0 - 5.0 mm), with a confidential interval (CI) of 95% (0.11, 0.94). Statistically, the one side percentile P(95) was 4.9 mm, P(97.5) was 5.0 mm and P(100) was 5.0 mm. The mean PCNA index in the 41 patients with RCC was (29.5 +/- 17.6)%, which was (49.6 +/- 21.5)% in the group with extra-pseudocapsule cancer lesions and (24.6 +/- 12.7)% in the group without (t = 3.162, P = 0.013). The ratio of strong expression was 5/8 in the group with extra-pseudocapsule cancer lesions, and 18.2% (6/33) in the group without the lesions (chi(2) = 6.442, P = 0.011). Logistic regression analysis showed that completeness of pseudocapsule and PCNA index were significant predictors of extra-pseudocapsule cancer lesions (P = 0.019). CONCLUSIONS: These data suggest that when NSS is performed in RCC 4 cm or less in diameter, a margin of more than 5 mm of adjacent parenchyma should be excised with the tumor. Enucleation alone was associated with a significant risk of incomplete excision, and therefore liable for local recurrence. Tumors with incomplete pseudocapsule and(or) high PCNA indices are more likely to have extra-pseudocapsule cancer lesions, so intensive follow-up is necessary after NSS.