[Establishment of a patient-derived xenograft humanized mouse model for hepatoblastoma in children].

Objective: To establish a patient-derived xenograft (PDX) humanized mouse model for hepatoblastoma in children. In addition, compare the biological consistency between successfully modeled PDX tumors and primary tumors in children while comparing and analyzing the influence of PDX model modeling success as a key factor. Methods: A PDX tumor model was constructed from fresh tumor tissue samples from 39 children with hepatoblastoma. The tumor growth time and volume size were recorded in detail. Simultaneously, 39 children's data were collected for experimental and clinical analysis. The difference in tumorigenesis rate between different parameters was analyzed by χ (2) test (categorical variable). Continuous variables with a normal distribution were compared using the t-test. Results: After cell passage and pathological diagnosis, 21 cases of hepatoblastoma PDX models were successfully constructed, with a success rate of 53.8% (21/39). Tumor samples from each generation of successfully modeled PDX models had pathology results that were consistent with those of the corresponding primary tumors. The analysis of the key factors affecting the tumor formation rate of PDX revealed that the metastasis rate was more successful in primary tumors than in liver in situ tumors (7/8 vs. 14/31, P = 0.049). However, there was no significant difference between tumor formation rates and pathological subtypes. According to the PDX tumor formation group comparison between the primary tumor and the metastatic tumor, there was no statistically significant difference between the two groups in terms of tumor formation time and tumor volume. Hematoxylin-eosin staining in hepatoblastoma's PDX mouse was consistent with the primary tumor. Immunohistochemistry positivity rates of four proteins, namely hepatocyte antigen (Hepatocyte), phosphatidylinositol glycan 3, ß-catenin, and alpha-fetoprotein, in primary tumor tissues and PDX mouse models were 100% vs. 100%, 100% vs. 95.24%, 100% vs. 100%, and 95.24% vs. 85.71%, respectively. Conclusion: A PDX mouse model for hepatoblastoma has been successfully established in children. The tumor formation rate is high, with metastatic tumors having a higher tumor formation rate than primary tumors and transplanted tumors retaining the biological characteristics of primary tumors.

[Effects of microRNA-106b on migration and invasion of human malignant pleural mesothelioma cell NCI-H2452].

Objective: To investigate the inhibitory effect of microRNA-106b in the process of migration and invasion of human malignant pleural mesothelioma cell NCI-H2452. Methods: In April 2017, the expression level of miRNA-106b in malignant pleural mesothelioma cells (NCI-H2452, MSTO-211H, NCI-H2052) and normal mesothelial cells MeT-5A was detected and analyzed. Using NCI-H2452 cells as a model, the NCI-H2452 cell model with miRNA-106b overexpression was established by transfecting miRNA-106b mimics. The expression level of miRNA-106b in the cells was detected by real-time fluorescent quantitative PCR. The effect of miRNA-106b on the migration and invasion ability of NCI-H2452 cells was analyzed. The gene expression data of malignant mesothelioma and the downstream target gene data of miRNA-106b in public databases were analyzed to screen the downstream target genes of miRNA-106b in mesothelioma cells that affect cell migration and invasion ability, and to verify the expression of this gene in NCI-H2452 cells with miRNA-106b overexpression. Results: The expression of miRNA-106b in three MPM cells was decreased compared with MeT-5A cells (P<0.001) . The expression level of miRNA-106b was significantly increased after transfection of miRNA-106b mimics (P<0.001) . The scratch migration levels of the experimental group were 28.45%±4.37%, 38.12%±4.82% and 50.06%±8.92% at 24h, 31h and 48h, respectively. Compared with the control group, the migration level decreased by 37.48%±2.65%, 49.21%±3.45% and 68.14%±3.81% (P<0.01) . The number of cell migration and invasion decreased in the experimental group compared with the control group (P<0.001) . Public databases were used to screen and analyze the possibility that TCF21 gene, as a downstream target gene, could affect the migration and invasion ability of MPM cells. The expression level of TCF21 gene was increased after transfection of miRNA-106b mimics in NCI-H2452 cells (P=0.009) . Conclusion: MiRNA-106b can inhibit the migration and invasion of NCI-H2452 cells and increase the expression of TCF21 gene.

Niraparib maintenance therapy in patients with platinum-sensitive recurrent ovarian cancer using an individualized starting dose (NORA): a randomized, double-blind, placebo-controlled phase III trial☆.

BACKGROUND: This study evaluated maintenance treatment with niraparib, a potent inhibitor of poly(ADP-ribose) polymerase 1/2, in patients with platinum-sensitive recurrent ovarian cancer. PATIENTS AND METHODS: In this phase III, double-blind, placebo-controlled study conducted at 30 centers in China, adults with platinum-sensitive recurrent ovarian cancer who had responded to their most recent platinum-containing chemotherapy were randomized 2 : 1 to receive oral niraparib (300 mg/day) or matched placebo until disease progression or unacceptable toxicity (NCT03705156). Following a protocol amendment, patients with a bodyweight <77 kg or a platelet count <150 × 103/µl received 200 mg/day, and all other patients 300 mg/day, as an individualized starting dose (ISD). Randomization was carried out by an interactive web response system and stratified by BRCA mutation, time to recurrence following penultimate chemotherapy, and response to most recent chemotherapy. The primary endpoint was progression-free survival (PFS) assessed by blinded independent central review. RESULTS: Between 26 September 2017 and 2 February 2019, 265 patients were randomized to receive niraparib (n = 177) or placebo (n = 88); 249 patients received an ISD (300 mg, n = 14; 200 mg, n = 235) as per protocol. In the intention-to-treat population, median PFS was significantly longer for patients receiving niraparib versus placebo: 18.3 [95% confidence interval (CI), 10.9-not evaluable] versus 5.4 (95% CI, 3.7-5.7) months [hazard ratio (HR) = 0.32; 95% CI, 0.23-0.45; P < 0.0001], and a similar PFS benefit was observed in patients receiving an ISD, regardless of BRCA mutation status. Grade ≥3 treatment-emergent adverse events occurred in 50.8% and 19.3% of patients who received niraparib and placebo, respectively; the most common events were neutrophil count decreased (20.3% versus 8.0%) and anemia (14.7% versus 2.3%). CONCLUSIONS: Niraparib maintenance treatment reduced the risk of disease progression or death by 68% and prolonged PFS compared to placebo in patients with platinum-sensitive recurrent ovarian cancer. Individualized niraparib dosing is effective and safe and should be considered standard practice in this setting.

[microRNA-30d can inhibit the proliferation, migration and invasion of human mesothelial cell MSTO-211H].

Objective: To investigate the inhibitory effect and molecular mechanism of microRNA-30d (miR-30d) in the process of proliferation, migration and invasion of malignant mesothelioma cell line MSTO-211H. Methods: In April 2017, the human MSTO-211H cells was used to establish miR-30d overexpressed MSTO-211H cell model by transfection of miR-30d mimics. The qRT-PCR was performed to detect the expression level of miR-30d in the cells transfected miR-30d mimics. The effects of miR-30d on the proliferation, apoptosis, migration and invasion of MSTO-211H cells were analyzed by CCK-8 experiment, flow cytometry, cell scratch experiment and Transwell method. Results: After transfection of miR-30d, the expression level of miR-30d in the MSTO-211H+miR-30d cells group was significantly higher than MSTO-211H+miR NC cells group (P<0.01) . The cell activity of MSTO-211H+miR-30d group (105.13%±2.35%) was significantly lower than MSTO-211H+miR NC cells group (115.40%±1.35%) , and the level of apoptosis (3.97%±0.36%) was significantly higher than MSTO-211H+miR NC cells group (1.47%±0.10%) (P<0.01) . The relative migration areas at 12 and 24 h of MSTO-211H+miR-30d cells group (9.35±3.16 µm(2) and 58.19±1.82 µm(2)) were significantly lower than MSTO-211H+miR NC cells group (54.42±5.26 µm(2) and 88.32±1.96 µm(2)) (P<0.01) . Compared with the MSTO-211H+miR NC cells group, the numbers of cell migration and cell invasion were reduced in the MSTO-211H+miR-30d cells group (P<0.01) . Conclusion: miR-30d can regulate the progression of malignant pleural mesothelioma by inhibiting the proliferation, apoptosis, migration and invasion of MSTO-211H cells.

[Study on malignant transformation of MeT-5A cells induced by chrysotile].

Objective: To study the cytotoxicity and malignant transformation ability of chrysotile on MeT-5A cells. Methods: In June 2016, lactate dehydrogenase (LDH) method was used to detect the cytotoxicity of chrysotile to MeT-5A cells. MeT-5A cells were treated with 5 µg/cm(2) chrysotile intermittently for 24 h a time, once a week and a total of 28 times. After the cells showed anchorage independent growth, the cell features of malignant transformation were identified by colony forming frequency in soft agar, and the soft agar colony formation rates were calculated. The activities of key speed limiting enzymes of glycolysis metabolism including hexokinase (HK) , phosphofructokinase (PFK) and pyruvate kinase (PK) were determined by UV colorimetry. Results: Chrysotile was cytotoxic to MeT-5A cells in a concentration-dependent decline. Compared with the control group, the relative survival rates of MeT-5A cells were significantly decreased after exposed to chrysotile at 10, 20, 40 and 80 µg/cm(2) (P<0.05) . After 28 times of exposure, the growth rate of the cells in chrysotile transformed MeT-5A cells was accelerated, the arrangement was disordered, the contact inhibition was lost, and the double layer growth appeared, which could grow on soft agar. The colony forming rate of the chrysotile transformed MeT-5A cells was 18.33‰±2.49‰. Compared with the control group (0) , the difference was statistically significant (P<0.01) . The activities of glycolysis related kinase including PK [ (19.51±1.52) U/L], PFK[ (0.12±0.02) U/10(4) cell] and HK[ (0.26±0.01) U/10(4) cell] were increased in the chrysotile transformed MeT-5A cells compared with control group [ (25.00±1.04) U/L、(0.15±0.01) U/10(4) cell and (0.33±0.01) U/10(4) cell] (P<0.01) . Conclusion: Chrysotile can induce malignant transformation of MeT-5A cells and increase the activities of glycolysis related kinases including PK, PFK and HK.

[Pathological diagnosis of lung cancer based on deep transfer learning].

Objective: To establish an artificial intelligence (AI)-assisted diagnostic system for lung cancer via deep transfer learning. Methods: The researchers collected 519 lung pathologic slides from 2016 to 2019, covering various lung tissues, including normal tissues, adenocarcinoma, squamous cell carcinoma and small cell carcinoma, from the Beijing Chest Hospital, the Capital Medical University. The slides were digitized by scanner, and 316 slides were used as training set and 203 as the internal test set. The researchers labeled all the training slides by pathologists and establish a semantic segmentation model based on DeepLab v3 with ResNet-50 to detect lung cancers at the pixel level. To perform transfer learning, the researchers utilized the gastric cancer detection model to initialize the deep neural network parameters. The lung cancer detection convolutional neural network was further trained by fine-tuning of the labeled data. The deep learning model was tested by 203 slides in the internal test set and 1 081 slides obtained from TCIA database, named as the external test set. Results: The model trained with transfer learning showed substantial accuracy advantage against the one trained from scratch for the internal test set [area under curve (AUC) 0.988 vs. 0.971, Kappa 0.852 vs. 0.832]. For the external test set, the transferred model achieved an AUC of 0.968 and Kappa of 0.828, indicating superior generalization ability. By studying the predictions made by the model, the researchers obtained deeper understandings of the deep learning model. Conclusions: The lung cancer histopathological diagnostic system achieves higher accuracy and superior generalization ability. With the development of histopathological AI, the transfer learning can effectively train diagnosis models and shorten the learning period, and improve the model performance.

Reduction of soy isoflavones by use of Escherichia coli whole-cell biocatalyst expressing isoflavone reductase under aerobic conditions.

UNLABELLED: Soy isoflavone metabolites are currently receiving much attention due to the stronger and wider bioactivities than that of isoflavones. Therefore, biosynthesis of isoflavone metabolites by isolated isoflavone biotransforming bacteria is important. However, the biosynthesis process must be under obligate anaerobic conditions due to the reduction reactions catalysed by isoflavone biotransforming bacteria. In this study, we cloned the daidzein and genistein reductase gene (dgr) from Slackia sp. AUH-JLC159. The recombinant Escherichia coli (E. coli) whole-cell was used for the first time as the biocatalyst for aerobic biosynthesis of dihydrodaidzein (DHD) and dihydrogenistein (DHG) from soy isoflavones daidzein and genistein. Our results indicated that the recombinant E. coli whole-cell was able to reduce daidzein and genistein to DHD and DHG under aerobic conditions, while the maximal concentration of the substrate daidzein or genistein that the E. coli whole-cell was able to convert efficiently was only 0·4 mmol l(-1) . Under the optimized conditions, the maximal concentration of daidzein or genistein that the E. coli whole-cell was able to convert efficiently was increased to 1·4 mmol l(-1) . Our results demonstrated that E. coli whole-cell is an efficient biocatalyst for biosynthesis of isoflavone metabolites under aerobic conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Soy isoflavone metabolites, which are more biologically active than their precursor isoflavones, are currently receiving much more attention. However, the non-natural isoflavone metabolites are synthesized or biosynthesized under obligate anaerobic conditions. Here, we describe a new approach to the reduction of soy isoflavones daidzein and genistein under aerobic conditions by use of the recombinant Escherichia coli whole-cell expressing isoflavone reductase. Our study provides the first evidence that isoflavone metabolites, such as dihydrodaidzein and dihydrogenistein, are able to be produced efficiently under aerobic conditions.

[Up-regulation of DLK1 in non-small cell lung cancer and the relevant molecular mechanism].

OBJECTIVE: To explore the expression of delta-like 1 homolog (DLK1) gene in non-small cell lung cancer (NSCLC) and its regulatory mechanism. METHODS: The expression levels of DLK1 protein in 204 NSCLC tissues were examined by immunohistochemical (IHC) staining, and the correlation between DLK1expression and clinicopathological features was analyzed. Bisulfate sequencing PCR (BSP) of DNA samples from the tumor tissues of 18 NSCLC patients was performed to evaluate the DNA methylation status of CpG island in the DLK1 promoter region, and also compared with the corresponding IHC staining of DLK1 protein in the same samples. RESULTS: Among the 102 squamous cell carcinoma (SCC) tissue specimens and their adjacent normal bronchial epithelia, DLK1 was up-regulated in 72 and 37 samples, respectively (P=0.001), and among 102 adenocarcinomas (ADC) tissues and their adjacent alveolar tissues, DLK1 was up-regulated in 77 and 7 samples, respectively (P<0.001). In addition, overexpression of DLK1 was significantly associated with histological type, clinical stage and tumor size of NSCLC (P<0.05 for all). The expression of DLK1protein was inversely correlated with its promoter methylation (P<0.05). CONCLUSION: DLK1 expression is up-regulated in NSCLCs, which may be due, at least in part, to the DNA hypomethylation in the promoter region of theDLK1 gene.

[Application of temporary balloon occlusion of the abdominal aorta in the treatment of complete placenta previa complicated with placenta accreta].

Objective: To investigate the value of temporary balloon occlusion of the abdominal aorta in the treatment of complete placenta previa with placenta accreta. Methods: From January 2015 to February 2016, 24 cases of complete placenta previa with placenta accreta were treated with temporary balloon occlusion of the abdominal aorta(the study group)before cesarean, and 24 cases of complete placenta previa with placenta accreta did not receive balloon occlusion(the control group). The operation time, intraoperative blood loss, intraoperative blood transfusion volume, the perioperative hemoglobin level, the hysterectomy rate and the related complications were compared retrospectively.Also, the hospitalization time, the blood coagulation parameters after operation, including activated partial thromboplastin time(APTT), fibrinogen(FIB), D-Dimer and reperfusion injury parameters including creatine phosphokinase(CK), creatine phosphokinase isoenzyme(CK-MB), lactate dehydrogenase(LDH)and serum creatinine were compared between the 2 groups. Results: The blood loss[750 ml(400- 2 000 ml)vs 2 000 ml(1 500- 2 375 ml); Z=-3.214, P=0.001]and blood transfusion volume[200 ml(0-800 ml)vs 800 ml(0-1 200 ml); Z=- 2.173, P=0.030]in the study group were lower than in the control group. The hemoglobin difference between before and after operation in the study group was lower than the control group[(12.8±13.4)g/L vs(22.9±20.1)g/L; t=-2.041, P=0.047]. In the study group, there were still bleeding in 13 cases after releasing the balloon, 5 of them received uterine artery embolization, 5 cases received uterine artery ligation, and 3 cases received uterine packing. One case had venous thrombosis in the right lower limb. Two cases(8%,2/24)in the control group had hysterectomy, while none in the study group, there was no statistical significance(P= 0.489). Conclusions: Temporary balloon occlusion of the abdominal aorta can effectively reduce blood loss and blood transfusion in the treatment of complete placenta previa with placenta accreta, but there is still the risk of continuing bleeding after releasing the balloon. Other methods of hemostasis might be needed.

[Research of prognostic immunophenotypes in 163 patients of diffuse large B-cell lymphoma].

Objective: To screen and analyze the prognostic protein biomarkers of DLBCL, and to explore their value in the prognostic evaluation. Methods: 163 cases of confirmed DLBCLs from January 2011 to December 2016 were collected with their clinical, pathological and follow-up data, which were all from our hospital. The expression of protein markers were tested using immunohistochemical staining (IHC) . The immune phenotypes independent of the International Prognostic Index (IPI) that affect overall survival (OS) and progression-free survival (PFS) of DLBCL were explored by COX regression model, and the effect of their co-expression on the prognosis were also analyzed. Result: BCL6 negative (PFS: HR=1.652, 95%CI 1.030-2.649, P=0.037) , P53 positive (OS: HR=1.842, 95%CI 1.008-3.367, P=0.047) , and BCL2 strong positive expressions (S+) (OS: HR=2.102, 95%CI 1.249-3.537, P=0.005; PFS: HR=2.126, 95%CI 1.312-3.443, P=0.002) are adverse prognostic factors of DLBCL that are independent of IPI. BCL6(-) (PFS: HR=2.042, 95%CI 1.021-4.081, P=0.043) , P53(+) (OS: HR=3.069, 95%CI 1.244-7.569, P=0.015) and BCL2(S+) (OS: HR=2.433, 95%CI 1.165-5.082, P=0.018; PFS: HR=3.209, 95%CI 1.606-6.410, P=0.001) are adverse prognostic factors in the group of age≤60-year-old; in the group of IPI score 0-2, cases with BCL6(-) (OS: HR=2.467, 95%CI 1.322-4.604, P=0.005; PFS: HR=2.248, 95%CI 1.275-3.965, P=0.005) and BCL2(S+) (PFS: HR=2.045, 95%CI 1.119-3.735, P=0.020) have worse prognosis. The co-expression of BCL6(-) and BCL2(S+) has significant influence on prognosis of DLBCL (P=0.005 and P<0.001) , in which BCL6(+)/non-BCL2(S+) (n=86) has the best prognosis[3-year-OS (71.6±4.9) %, 3-year-PFS (67.0±5.1) %], and BCL6(-)/BCL2(S+) (n=10) has the worst prognosis[3-year-OS (20.0±12.6) %, 3-year-PFS (10.0±9.5) %]; the co-expression of BCL6(-) and P53(+) has no significant influence on prognosis (P=0.061 and P=0.089) , however, those cases with BCL6(+)/P53(-) (n=98) often get better prognosis[3-year-OS (70.6±4.7) %, 3-year-PFS (64.6±4.9) %] than others; the co-expression of P53(+) and BCL2(S+) has significant influence on prognosis of DLBCL (P<0.001 and P<0.001) , and P53(+)/BCL2(S+) (n=5) has the worst prognosis (3-year-OS and 3-year-PFS are both 0) ; BCL2(S+) cases get shorter OS and PFS, regardless of the expression of BCL6 and P53. Conclusion: The expression and co-expression of BCL6 negative, P53 positive and BCL2(S+) have certain value in the prognostic evaluation of DLBCL, especially in the group of age≤60-year-old and IPI score 0-2.

Monitoring of CaSO(4) deposition behaviors on biofilm during nanofiltration via ultrasonic time-domain reflectometry (UTDR).

The ultrasonic time-domain reflectometry (UTDR) as a non-destructive real-time method was employed to monitor the CaSO(4) deposition behaviors on biofilm during nanofiltration (NF). Two parallel experiments were performed to compare the different behaviors of CaSO(4) deposition with and without biofilm on the membrane. Results showed that the flux decline during combined fouling was slower than that in case of CaSO(4) fouling alone. The Ca(2 + ) rejection obtained with biofilm was higher than that without. A larger acoustic differential signal obtained by UTDR in the combined fouling revealed a denser and thicker layer formed on the membrane surface. Furthermore, the amount of CaSO(4) deposition on the biofouled membrane was more than that on non-biofouled membrane as a result of microorganisms as crystal nucleus to induce CaSO(4) crystallization and deposition. SEM images indicate that the CaSO(4) crystals deposited in order on the non-biofouled membrane, whereas on the biofouled membrane they were embedded in the biofilm. The denser and thicker fouling layer formed with biofilm was impermeable, resulting in a high Ca(2 + ) rejection. The complexation of Ca with polysaccharide in biofilm would eliminate the cake-enhanced osmotic pressure effect leading to a slow flux decline. To sum up, the independent measurements corroborate the ultrasonic measurements.

[Clinical analysis of selective laryngeal reinnervation using upper root of phrenic nerve and hypoglossal nerve branch in the treatment of bilateral vocal fold paralysis].

Objective: To evaluate the airway and voice quality improvement in patients with bilateral vocal fold paralysis (BVFP) who underwent selective laryngeal reinnervation surgery. Methods: From January 2012 to December 2016, a retrospective study was conducted in 39 patients with BVFP who underwent selective laryngeal reinnervation surgery in Department of Otorhinolaryngology Head and Neck Surgery, the First Affiliated Hospital of Navy Medical University. All patients were examined by videostroboscopy, vocal function assessment, laryngeal electromyography and pulmonary function test before and after the surgery, and followed up for at least 2 years to evaluate the efficacy and safety of the surgery.Wilcoxon signed rank test was used to analyze the G score and VHI-10 score data. Paired t-test was used to analyze acoustic parameters, MPT values and pulmonary function parameters. Results: Postoperative infection and hemorrhage occurred in one patient separately.Videostroboscopic videos showed that at 4-8 months postoperatively, vocal folds in 35 patients achieved moderate or severe abduction during inspiration, 2 patients only achieved mild abduction, 2 patients showed no abduction,while all patients achieved adduction in bilateral vocal cords during phonation. The recovery rate of moderate-to-severe abduction was 89.7% (35/39), and these patients were decannulated successfully. At 12 months after operation, G score and VHI-10 score were significantly lower than those before operation (P<0.05), and the acoustic parameters jitter, shimmer, HNR and MPT were significantly improved (P<0.05). Most of the parameters of the pulmonary function test at 3 months postoperatively returned to the normal reference level, while the maximum inspiratory pressure (PImax) at 12 months after operation was still slightly lower than the normal level, but it was significantly improved compared with preoperative value (P<0.05). The EMG data at 12 months postoperatively showed full interference potentials in 37 patients in bilateral posterior cricoarytenoid muscles during inspiration, and full interference potentials in bilateralthyroarytenoid muscles during phonation. Obvious misdirected regeneration electric activitieswere found in two of them. Potentials in posterior cricoarytenoid muscle were weak in 2 cases with poor abduction. During long-term follow-up, only one case showed decreased abduction, but did not affect respiratory function. Conclusions: The selective laryngeal reinnervation procedure applied in the present study can restore physiological motion of vocal cords. The success rate was high, the curative effect was stable, and the complications were rare. It is worth of promotion.

Transcriptome analysis revealed that aflatoxin M1 could cause cell cycle arrest in differentiated Caco-2 cells.

Being a hydroxylated metabolite of aflatoxin B1 (AFB1) and the most threatening aspect of AFB1 contamination, aflatoxin M1 (AFM1) can lead to hepatotoxicity and hepato-carcinogenicity, and possess intestinal cytotoxicity. However, little is known about the potential mechanisms of the extrahepatic effect. The aim of this study was to investigate intestinal dysfunction induced by AFM1 via transcriptome analysis. Gene expression profiling was analyzed to comparatively characterize the differentially expressed genes (DEGs) after differentiated Caco-2 cells were exposed to different concentrations of AFM1 for 48 h. A total of 165 DEGs were significantly clustered into two down-regulated patterns. Protein-protein interaction (PPI) network analysis based on Search Tool for Retrieval of Interacting Genes (STRING)suggested that 23 key enzymes mainly participated in the regulation of the cell cycle. Q-PCR analysis was performed to validate that key 12 genes (BUB1, BUB1B, MAD2L1, CCNA2, RB1, CDK1, ANAPC4, ATM, KITLG, PRKAA2, SIRT1, and SOS1) were involved. This study firstly revealed that the toxicity of AFM1 to intestinal functions may be partly due to the occurrence of cell cycle arrest, which is linked to changes in CDK1, SOS1/Akt, and AMPK signaling molecules.

Individual and combined cytotoxicity assessment of zearalenone with ochratoxin A or α-zearalenol by full factorial design.

The combined mycotoxins zearalenone (ZEA) with ochratoxin A (OTA) or α-zearalenol (α-ZOL) are frequently found together in milk. Toxicological data concerning the combined effects of these mycotoxins are sparse. In present study, individual and combined ZEA, OTA and α-ZOL caused cytotoxicity and oxidative damage, including reductions in intracellular superoxide dismutase and glutathione peroxidase activities and glutathione content, along with increases in malonaldehyde content on human Hep G2 cells after 48 h of exposure. Among individual mycotoxins, OTA had the greatest cytotoxic effect followed by α-ZOL. Compared with individual mycotoxins, combinations produced more serious negative effects, more importantly, ZEA + OTA was antagonistic for these effects, whereas ZEA + α-ZOL was antagonistic at low concentrations, but synergistic at high concentrations of ZEA, which were evaluated by 3 × 3 full factorial analysis and estimated marginal means plots. Our results also demonstrated a significant correlation between cytotoxicity and oxidative damage in response to these combinations.

[The efficacy of ansa cervicalis anterior root for unilateral recurrent laryngeal nerve injury].

Objective: To discuss the long-term efficacy of laryngeal reinnervation using the anterior root of the ansa cervicalis in the treatment of unilateral vocal fold paralysis (UVFP) caused by thyroid surgery. Method: From January 2010 to January 2016, a total of 39 UVFP patients who underwent ansa cervicalis anterior root-to-recurrent laryngeal nerve (RLN) anastomosis and who had suffered nerve disfunction for 6 to 24 months were enrolled as UVFP group.Another 39 age and gender matched normal subjects served as control group. Videostroboscopy, vocal function assessment (acoustic analysis, perceptual evaluation and maximum phonation time), and laryngeal electromyography were performed preoperatively and postoperatively for assessing surgery efficacy. Paired sample t test was used for statistical analysis. Result: Videostroboscopic reports indicated that the glottic closure, vocal fold edge, vocal fold position, phase symmetry and regularity were significantly improved in the UVFP group (P<0.01, respectively, postoperative vs. preoperative)and showed no statistical differences compared to the control group (P>0.05, respectively). Both the postoperative GRBAS assessment and acoustic parameters were also significantly improved in the UVFP group, Pre-operative acoustic parameters/Post-operative acoustic parameters were 1.68±0.82/0.39±0.27, 10.08±2.56/4.58±2.96, 0.203±0.216/0.018±0.038, 5.96±1.92/17.42±4.11(P<0.01, respectively) and Pre-operative acoustic parameters/Post-operative acoustic parameters were 0.39±0.27/0.32±0.19, 4.58±2.96/3.32±1.27, 0.018±0.038/0.014±0.027, 17.42±4.11/18.76±5.29, which showed no statistical differences compared to the control group (P>0.05, respectively). Conclusion: Delayed laryngeal reinnervation with the anterior root of ansa cervicalis, it can restore the physiological laryngeal phonatory function to the normal or a nearly normal voice quality, which is a feasible and effective approach for the treatment of thyroid surgery-related UVFP.

Enhanced toxicity to the cyanobacterium Microcystis aeruginosa by low-dosage repeated exposure to the allelochemical N-phenyl-1-naphthylamine.

It has been puzzling whether and how a plant could exert a strong allelopathic inhibition to the target organisms by releasing low concentrations of allelochemicals. Plant allelochemicals have been proposed to be released continuously, however, direct evidence from specific allelochemicals is urgently required. In the present study, the toxicity of allelochemical N-phenyl-1-naphthylamine (NPN) towards the cyanobacterium Microcystis aeruginosa by two different exposure patterns was compared. One was low-dosage repeated exposure (LRE), in which 50  µg L-1 NPN was repeatedly dosed to simulate the continual release of allelochemicals, and the other one was high-dosage single exposure (HSE) as per the routine toxicity assay. The results showed a significant growth inhibition to M. aeruginosa in the LRE group, where the inhibition rate reached above 90% from day 6 to day 9. The cell-membrane damage ratio increased from 64.05% on day 5 up to 96.60% on day 9. PSII photosynthesis activity expressed as Fv/Fm, ΦPSII, NPQ and ETRmax was also thoroughly inhibited in this group. Whereas the growth and PSII photosynthesis activity of M. aeruginosa in the HSE group were inhibited initially, but recovered gradually from day 4 or 5, which was accompanied by a continuous reduction of NPN content in culture solutions. Although NPN content in the LRE group was relatively lower, it remained at a more stable level throughout the experiment. These results indicate that continual release of low-dosage allelochemicals by aquatic plants plays crucial roles in their potent inhibition against cyanobacteria. Low-dosage continual exposure pattern needs to be investigated further.

Aflatoxin M1 cytotoxicity against human intestinal Caco-2 cells is enhanced in the presence of other mycotoxins.

Aflatoxin M1 (AFM1), a class 2B human carcinogen, is the only mycotoxin with established maximum residue limits (MRLs) in milk. Toxicological data for other mycotoxins in baby food, containing cereals and milk, either in isolation or in combination with AFM1, are sparse. The aim of this study was to investigate the cytotoxicity of AFM1, ochratoxin A (OTA), zearalenone (ZEA), and α-zearalenol (α-ZOL), individually and in combinations, in human Caco-2 cells. The tetrazolium salt (MTT) assay demonstrated that (i) OTA and AFM1 had similar cytotoxicity, which was higher than that of ZEA and α-ZOL, after a 72 h exposure; and (ii) the quaternary combination had the highest cytotoxicity, followed by tertiary and binary combinations and individual mycotoxins. Isobologram analysis indicated that the presence of OTA, ZEA, and/or α-ZOL with AFM1 led to additive and synergistic cytotoxicity in most combinations. The cytotoxicity of OTA was similar to that of AFM1, suggesting that OTA in food poses a health risk to consumers. Furthermore, AFM1 cytotoxicity increased dramatically in the presence of OTA, ZEA, and/or α-ZOL (p < 0.01), indicating that the established MRLs for AFM1 should be re-evaluated considering its frequent co-occurrence with other mycotoxins in baby food which contains milk and cereals.

[Comparing the free fasciocutaneous flap with free jejunium in reconstruction for hypopharyngeal and cervical esophageal defects].

Objective:To investigate the application of the reconstruction methods for hypopharyngeal and cervical esophageal defects due to the resection of hypopharyngeal cancer and advanced laryngeal cancer between free fasciocutaneous flaps and free jejunium transfer.We compared the superiorities and inferiorities of these two reconstruction methods. Method:Retrospective review of the archives of 56 patients from 2000 to 2010 who underwent pharyngoesophageal reconstruction with free flaps (n=32) or free jejunal transfer(n=24),comparison of indications,complications, hospitalization duration, swallowing function recovery and postoperative survival time. Result:The overall 3 year survival rate of free flap group and free jejunal transfer group was 59.3%,55.7% respectively; the overall 5 year survival rate was 38.5%,37.1% respectively. The overall rate of complication rate was 18.8%, 16.7% respectively. The patients with free flaps had higher incidence rate of fistula and scarring in the donor site and lower incidence rate of hues and stricture than the ones with free jejunal transfers. The mean hospitalization duration was (15.00±7.06) days and(13.00±6.75) days. The mean time of first oral food intake was(13.00±5.83)days and (11.00±6.67) days. The differences between two groups had no statistical significance(P>0.05). Conclusion:Free flaps and free jejunium transfer are the two most common reconstruction methods for the hypopharyngeal and cervical esophageal defects. Each has its own advantages and disadvantages respectively. We should choose reconstruction method according to the site and extent of the hypopharyngeal and cervical esophageal defects, preoperative and postoperative radiotherapy requirement.

[Bacteria condition of low genital tract of pregnant women and neonatal infection].

The swab from low genital tract of 74 cases of pregnant women, which were selected at random, were taken in labour and delivery room before delivery. A piece of placenta tissue and the swab of neonatal respiratory tract were obtained after delivery and were cultured in anaerobic and aerobic condition within 24 hours. Thirty-eight percent of bacteria that cultured in neonatal respiratory tract were the same species that obtained from maternal low genital tract and placentae. There were 15 infected neonates from 74 neonates, with positive culture in 9 neonates. Staphylococcus aureus played an important part in neonatal infection. Sixty seven percent of bacteria from the infected neonates were similar to the bacteria detected in low genital tract and placentae. Vertical transmission may play an important role in neonatal infection.