Survival Outcome of Salivary Gland Carcinoma: A 50-Year Retrospective Study With Long-Term Follow-up.

PURPOSE: Salivary gland carcinomas (SGCs) can be classified into more than 20 subtypes with various clinical behaviors. The present study aimed to analyze the clinical and pathological features of SGCs and evaluate their long-term prognosis. METHODS: A retrospective cohort study was performed. This study investigated cases of histologically confirmed SGC at the authors' institution from January 1963 to December 2014. Data on sex, age, site, histopathological diagnosis, tumor-node-metastasis classification, postoperative radiotherapy and/or chemotherapy, local and regional recurrence, and distant metastasis (DM) were collected as covariates. The overall survival (OS) rate was analyzed as the outcome. Kaplan-Meier survival analysis and Cox multivariate analysis were used for survival analysis. The cohort was divided into 2 groups-before and after 1989. The clinicopathological characteristics of the 2 groups were compared using the χ2 test. RESULTS: The cohort included 1,637 patients who met the admission criteria and had a male-to-female ratio of 0.9:1. The median age was 47 years (range, 8 months to 86 years). The median follow-up time was 54 months (range, 1-432 months). The majority of the tumors occurred in the parotid gland (35.3%), followed by the palate gland (25.2%). Adenoid cystic carcinoma was the most common tumor type (34.3%), and mucoepidermoid carcinoma (29%) was the second most common type. In the 1,637 patients, the neck lymph node metastasis rate was 8.7% at the first surgery, and the overall DM rate was 14.1%. The 5-, 10-, and 15-year OS rates of the 1,637 cases were 93.1%, 87.2%, and 79.3%, respectively. Comparative analysis before and after 1989 showed statistically significant differences in sex, site, histologic subtype, T classification, local and regional recurrence rate, and radiotherapy (P < .05), while no significant differences were found in age, N classification, M staging, DM, or chemotherapy. CONCLUSIONS: The OS rates of SGC have improved significantly over the past 30 years. This is attributable to an increase in the proportion of patients diagnosed at the early stage and receiving radiotherapy, as this has led to a reduction in the local and regional recurrence rate and, consequently, an improvement in the survival rates.

ZnO nanotubes supported molecularly imprinted polymers arrays as sensing materials for electrochemical detection of dopamine.

In this study, ZnO nanotubes (ZNTs) were prepared onto fluorine-doped tin oxide (FTO) glass and used as supports for MIPs arrays fabrication. Due to the imprinted cavities are always located at both inner and outer surface of ZNTs, these ZNTs supported MIPs arrays have good accessibility towards template and can be used as sensing materials for chemical sensors with high sensitivity, excellent selectivity and fast response. Using K3[Fe(CN)6] as electron probe, the fabricated electrochemical sensor shows two linear dynamic ranges (0.02-5µM and 10-800µM) towards dopamine. This proposed electrochemical sensor has been applied for dopamine determination with satisfied recoveries and precision. More complex human urine samples also confirmed that the proposed method has good accuracy for dopamine determination in real biological samples. These results suggest potential applicability of the proposed method and sensor in important molecule analysis.

Identification of an RNA Polymerase III Regulator Linked to Disease-Associated Protein Aggregation.

Protein aggregation is associated with age-related neurodegenerative disorders, such as Alzheimer's and polyglutamine diseases. As a causal relationship between protein aggregation and neurodegeneration remains elusive, understanding the cellular mechanisms regulating protein aggregation will help develop future treatments. To identify such mechanisms, we conducted a forward genetic screen in a C. elegans model of polyglutamine aggregation and identified the protein MOAG-2/LIR-3 as a driver of protein aggregation. In the absence of polyglutamine, MOAG-2/LIR-3 regulates the RNA polymerase III-associated transcription of small non-coding RNAs. This regulation is lost in the presence of polyglutamine, which mislocalizes MOAG-2/LIR-3 from the nucleus to the cytosol. We then show biochemically that MOAG-2/LIR-3 can also catalyze the aggregation of polyglutamine-expanded huntingtin. These results suggest that polyglutamine can induce an aggregation-promoting activity of MOAG-2/LIR-3 in the cytosol. The concept that certain aggregation-prone proteins can convert other endogenous proteins into drivers of aggregation and toxicity adds to the understanding of how cellular homeostasis can be deteriorated in protein misfolding diseases.

Molecularly imprinted solid phase extraction method for simultaneous determination of seven nitroimidazoles from honey by HPLC-MS/MS.

In this work, a selective sample cleanup procedure that combined molecular imprinting technique with solid phase extraction was developed for the simultaneous extraction of the seven nitroimidazoles (NMZs) from honey samples. The molecular imprinting polymers for NMZs were prepared through bulk polymerization method using 2-methyl-5-nitroimidazole as template molecule, methacrylic acid as the functional monomer and ethylene glycol dimethacrylate as the cross linking agent. The obtained molecular imprinting polymers showed high affinity to template molecule and was used as selective sorbent for simultaneously selective extraction of the seven NMZs from honey matrix. An off-line molecularly imprinted solid phase extraction (MISPE) method followed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS-MS) for simultaneous determination of the seven NMZs from honey samples was also established. The proposed method was validated at 1.0, 2.0 and 10.0µg/kg, obtaining recoveries in the range of 79.7-110%, with repeatability and interday precision values (expressed as relative standard deviation) ≤11.4% and ≤15.2%, respectively. Limits of quantification for different NMZs were 1.0µg/kg, which were always below the minimum required performance limits established by the European Community Reference Laboratories (Commission Decision 2002/657/EC). It was demonstrated that this proposed MISPE-HPLC-MS-MS method could be applied to direct determination of NMZs from honey samples.

[Detection of the exogenous gene copy number of the transgenic tomato anti-caries vaccine].

PURPOSE: To detect the exogenous gene copy number of the transgenic tomato anti-caries vaccine by using the SYBR Green real-time PCR. METHODS: Recombinant plasmid pEAC10 and pEPC10 were used as standard to detect genome samples of exogenous gene pacA-ctxB and pacP-ctxB by SYBR green fluorescent quantitation, then the average value was calculated as gene copy number. RESULTS: The copy number of the transgenic tomato carrying pacA-ctxB was 1.3 and the pacP-ctxB was 3.2. CONCLUSIONS: The transgenic tomato plants which have high stability are low-copy transgenic plants. Supported by National Natural Science Foundation of China (30160086, 81260164), Science and Technical Fund of Guizhou Province (LKZ[2011]41), Project of Technology Innovation Team in Guizhou Province, Leading Academic Discipline Construction Project in Guizhou Province and Excellent Scientific Research Team Cultivation Project in Zunyi Medical College ([2012]12).

Structural shifts of gut flora in rat acute alcoholic liver injury and jianpi huoxue decoction's effect displayed by ERIC-PCR fingerprint.

OBJECTIVE: To study the structural shifts of gut flora in rats with acute alcoholic liver injury (AALI), and the effect of jianpi huoxue decoction (JPHXD) on the gut flora. METHODS: Thirty-six Sprague-Dawley rats were randomly allocated to the control, AALI and JPHXD groups equally. The rats in the control group were given water and those in AALI and JPHXD groups were given ethanol by intragastric gavage for 5 days, while rats in the JPHXD group were administered JPHXD simultaneously. The blood and liver tissue were collected at the end of the experiment. The activities of serum alkaline aminotransferase (ALT), aspartate aminotransferase (AST), hepatic γ-glutamyltranspetidase (γ-GT) and hepatic triglyceride (TG) levels were determined. Plasma endotoxin level in the portal vein was measured. Pathological changes of liver tissues were determined by hematoxylin and eosin (HE) staining and oil red O staining. The total DNA of gut flora were extracted from fecal samples by Bead-beating method and determined by ERIC-PCR fingerprint method. The similarity cluster analysis and principal component analysis were performed to analyze the ERIC-PCR fingerprint respectively. RESULTS: In the AALI group, the ratio of liver/body weight, activities of ALT, AST and hepatic γ-GT, amount of hepatic TG were elevated significantly compared with those in the control group (all P<0.01). JPHXD decreased the ratio, activities of ALT, AST, γ-GT and TG significantly compared with those in the AALI group (P<0.05 or P<0.01). HE and oil red O staining showed that fat deposited markedly in liver tissue, while JPHXD alleviated pathological changes markedly. Plasma LPS level in rat portal vein in the AALI group increased significantly (P<0.01), but it was decreased significantly in the JPHXD group (P<0.01). The cluster analysis and principal component analysis of ERIC-PCR fingerprint showed that gut flora in the AALI group changed markedly, and JPHXD could recover gut flora to some extent. CONCLUSIONS: The structure of gut flora shifted markedly during acute alcoholic liver injury, JPHXD had prevention effect through the modification of gut flora.

[Effect of jianpi huoxue recipe on gut flora in rats with alcoholic fatty liver induced by Lieber-DeCarli liquid diet].

OBJECTIVE: To study the effect of Jianpi Huoxue Recipe(JPHXR) on the gut flora in rats with alcoholic fatty liver (AFL) induced by Lieber-DeCarli liquid diet. METHODS: Forty Sprague-Dawley rats were divided into 4 groups: A: normal rats, B: rats fed with non-alcoholic liquid diet, C: rats fed with ethanol liquid diet to make AFL model, and D: AFL model rats intervened by gastrogavage of JPHXR 1.0 mL/100 g per day for 8 successive weeks, 10 rats in each group. Except those in Group D (to them an equal volume normal saline was given for Successive instead), JPXHR was administered to rats in other three groups. At the end of experiment, rats were sacrificed, their blood and liver tissue samples were collected for determining serum activities of alanine transaminase (ALT) and aspartate aminotransferase (AST), endotoxin level in portal vein (expressed by lipopolysacchrides content, abbr. as LPS), and pathological examination of liver with HE staining and oil O red staining. Moreover, total DNA of gut flora were extracted from fresh rat fecal samples by Bead-beating method for determining the ERIC-PCR fingerprint, and a cluster analysis on the fingerprint was performed. RESULTS: Compared with the levels of ALT and AST in Group A (31.15 +/- 7.04 U/L, and 53.23 +/- 10.28 U/L respectively) and Group B (26.96 +/- 8.12 and 52.09 +/- 8.62), the corresponding levels in Group C (92.72 +/- 25.83 and 72.60 +/- 23.31) significantly increased (P < 0.01), while the increments in Group D (65.28 +/- 20.36 and 59.11 +/- 10.32) were decreased (P < 0.01, P < 0.05). Pathological examination showed marked fat deposition in Group C, but which was significantly reduced in Group D. Endotoxin level in the portal vein was (0.033 +/- 0.010, EU/mL) in Group A and 0.043 +/- 0.018 in Group B, which was increased significantly in Group C (0.541 +/- 0.085, P < 0.01) and Group D (0.349 +/- 0.098 EU/mL, P < 0.01), but the increase in Group C was more significant (P < 0.01). The cluster analysis of ERIC-PCR fingerprint showed significant changes in gut flora of Group C and D, which was in Group D partially recovered. CONCLUSIONS: JPHXR had good preventive effect against alcoholic fatty liver in rats, and could modify the structure of gut flora to some extent.

High dose magnesium sulfate exposure induces apoptotic cell death in the developing neonatal mouse brain.

BACKGROUND: Magnesium sulfate (MgSO4) is often used as a treatment for pre-eclampsia/eclampsia and preterm labor, resulting in the exposure of a significant number of neonates to this drug despite a lack of evidence suggesting that it is safe, or effective as a tocolytic. While there is evidence that MgSO4 may be neuroprotective in perinatal brain injury, recent reviews have suggested that the effects are dependent upon dose, and that higher doses may actually increase neonatal morbidity and mortality. There is a lack of evidence investigating the neurotoxic effects of neonatal magnesium (Mg) exposure on the developing brain, specifically in terms of neurodevelopmental apoptosis, a cell-killing phenomenon known to be potentiated by other drugs with mechanisms of action at Mg-binding sites (i.e. NMDA receptor antagonists such as MK-801, ketamine, and PCP). OBJECTIVE: To investigate the effects of Mg exposure on the neonatal mouse brain at different postnatal ages to determine whether MgSO4 treatment causes significant cell death in the developing mouse brain. METHODS: C57Bl/6 mice were treated with four doses of MgSO4 (250 mg/kg) on postnatal days 3 (P3), 7 (P7) or 14 (P14). Caspase-3 immunohistochemistry, cupric silver staining, and electron microscopy techniques were used to examine Mg-treated brains for neurotoxic effects. RESULTS: Qualitative evaluation using cupric silver staining revealed widespread damage throughout the brain in P7 animals. Results of electron microscopy confirmed that the cell death process was apoptotic in nature. Quantitative evaluation of damage to the cortex, caudate-putamen, hippocampus, thalamus, and cerebellum showed that Mg treatment caused significant brain damage in animals treated on P3 and P7, but not P14. CONCLUSIONS: Administration of high doses of Mg may be detrimental to the fetal brain, particularly if exposure occurs during critical periods of neurodevelopment.

Detection of PERV by polymerase chain reaction and its safety in bioartificial liver support system.

AIM: To establish a method detecting porcine endogenous retrovirus (PERV) in China experimental minipigs and to evaluate the safety of PERV in three individuals treated with bioartificial liver support systems based on porcine hepatocytes. METHODS: Porcine hepatocytes were isolated with two-stage perfusion method, then cultured in the bioreactor, which is separated by a semipermeable membrane (0.2 microm) from the lumen through which the patients' blood plasma was circulated. After post-hemoperfusion, patients' blood was obtained for screening. Additionally, samples of medium collected from both intraluminal and extraluminal compartments of the laboratory bioreactor and culture supernate in vitro was analyzed. The presence of viral sequences was estimated by polymerase chain reaction (PCR) and reverse transcriptase-polymerase chain reaction (RT-PCR). Finally, the infection of virus in the supernate of common culture was ascertained by exposure to the fetal liver cells. RESULTS: PERV-specific gag sequences were found in the porcine hepatocytes using RT-PCR. and were detected in all samples from the intraluminal, extraluminal samples and culture supernate. However, culture supernatant from primary porcine hepatocytes (cleared of cellular debris) failed to infect human fetal liver cells. Finally, RT-PCR detected no PERV infection was found in the blood samples obtained from three patients at various times post-hemoperfusion. CONCLUSION: The assays used are specific and sensitive, identified by second PCR. PERVs could be released from hepatocytes cultured in bioreactor without the stimulation of mitogen and could not be prevented by the hollow fiber semipermeable membrane, indicating the existence of PERV safety in extracorporeal bioartificial liver support system (EBLSS).