From routine physical examinations, thyroid cancer is the most common malignant tumor detected, especially thyroid microcarcinomas of less than <1 cm. Nevertheless, the discovery of these microcarcinomas has caused panic in many people. The questions are:How do the so many thyroid microcarcinomas be treated? Should surgery be performed or should they just be observes? According to experience on thyroid microcarcinomas,the majority of them grew slowly therefore observations of the tumors have been recommended. For some cases with unsatisfactory locations or with obvious lymph node metastasis, surgery is highly recommended.

OBJECTIVE: To examine the effect of HMGB1 silencing on proliferation, survival ability, migration and invasion of human esophageal squamous cell carcinoma after X-ray radiation. METHODS: Using siRNA technique, sequences of HMGB1 mRNA were synthesized and transfected into cultured EC9706 cells as the HMGB1 shRNA group. A negative sequence was synthesized and used as negative control (NC). Both groups were irradiated or not with Xrays. Protein levels of HMGB1 and metastasis-related molecules were determined using Western blot. MTS method, clone formation, and wound healing, Transwell assay were employed to examine the proliferation, survival ability, migration and invasion of the cells. Flow cytometry assay was used to analyze cell apoptosis rates. Western blot was used to examine the expression of cell. RESULTS: Western blot results demonstrated that silencing HMGB1 gene significantly reduced the HMGB1 protein expression compared with NC group (P < 0.01). MTS data demonstrated that,after irradiation, HMGB1 silencing significantly inhibited the proliferation of esophageal squamous cell carcinoma cells compared with NC group (P < 0.01). The data from the clone formation assay revealed that the radiosensitivity was increased after down-regulation of HMGB1 expression compared with NC group (P < 0.01). The apoptosis rate of tumor cells in HMGB1 shRNA group after irradiation was markedly increased (P < 0.01). Wound-healing assays show that Wound-healing rate of HMGB1 silencing cells was significantly lower than that of NC group[(20.78±4.38)% and (39.02 ±3.25)%, respectively, P < 0.01]. Transwell assays with matrigel show the invasion capability of HMGB1 silencing cells at 3.5 h was deeply suppressed compared with NC group (67.00±16.56 and 194.00±19.74, respectively, P < 0.01). Our data show that apoptosis rate of HMGB1 silencing cells was significantly higher than that of NC group[(20.67 ±1.38)% and (13.64 ±1.24)%, respectively, P < 0.01]. In addition, expressions of N-cadherin,Vimentin,MMP-2 and MMP-9 protein were downregulated,and expressions of Ecadherin were upregulated after HMGB1 silencing compared with NC group (P < 0.01). CONCLUSION: siRNA interference inhibited expression of the HMGB1 gene, reduced the proliferation, migration and invasion of cells,induced cell apoptosis,and increased radiosensitivity,followed by regulating the expression of metastasisrelated molecules in oesophageal squamous cell carcinoma cells.

OBJECTIVE: To investigate associations between MTHFR rs1801131 polymorphisms and susceptibility to non-syndromic cleft lips with or without cleft palates (NSCL/P) in a Han population which was located in the Chaoshan area,China. METHODS: Peripheral blood samples from 357 NSCL/P children,199 fathers, 198 mothers and 354 healthy controls were collected. Genomic DNA samples were extracted and polymorphisms of the MTHFR rs1801131 was detected by matrix-assisted laser desorbed ionization time-offlight mass spectrometry (MALDI-TOF-MS). The chi-square test was used to compare the genotypes and allele frequencies between the case and the normal control groups. The transmission disequilibrium test (TDT) was conducted in the cleft case-parents trios. The family-based genetic analyses were completed by FBAT 2.0.2 software. The genotype frequency distributions of rs1801131 in both the case and control groups were in accordance with Hardy-Weinberg equilibrium. RESULTS: The case-control study showed that the genotype and allele distribution frequencies of the rs1801131 locus were not associated with risk of NSCL/P (P > 0.05), and there was no evidence of allele A or C transmission disequilibrium at rs1801131 in cleft case-parents trios (P > 0.05). Family-based association test (FBAT) analyses provided no evidence of NSCL/P risk with single allele A or C of rs1801131. CONCLUSION: Our study suggested that MTHFR rs1801131 polymorphisms were not associated with risk of NSCL/P in the study population.

OBJECTIVE: To explore possible mechanisms that mediated the translocation of p62 protein from nuclei into cytoplasm in esophageal squamous cell carcinoma. METHODS: After treatments with KPT330 in KYSE70,KYSE150 and KYSE510 ESCC cell lines,localizations of p62 were observed by immunofluorescence assay combined with confocal laser microscopy as compared with the untreated group. LC-MS/MS mass spectrometry was used to identify the phosphorylation sites of p62 in esophageal cancer cells. Site-directed mutagenesis was performed to construct mutants with different phosphorylation status of p62. Co-IP and PLA experiments were conducted to analyze interactions between p62 and GSK3β. p62 accumulated in the nucleus after treatment with nuclear export inhibitor. RESULTS: The results of mass spectrometry showed phosphorylation of p62 protein at four sites of T269/S272/S328/S332 in esophageal cancer cells. By transfecting different phosphorylated mutants of p62 into KYSE30 cells and KYSE150 cells with endogenous knockdown of p62,phosphorylation at T269/S272 and dephosphorylation of S328/S332 contributed to the transfer of p62 from cytoplasm to nucleus. Co-IP and PLA results showed direct interactions between p62 and GSK-3β. CONCLUSION: Our results indicates that nucleo-plasmic translocation of p62 in esophageal cancer cells was related to the phosphorylation status of S328/S332 and might be regulated by GSK-3β kinase.

OBJECTIVE: To analyze the trend of incidence, mortality and time of prostatic cancer in cancer registration area of Heilongjiang Province from 2013 to 2017. METHODS: The incidence (mortality) rate,age-standardized incidence (mortality) rate and cumulative rate (0-74 years old) of prostatic cancer in the tumor registration area of Heilongjiang Province from 2013 to 2017 were calculated. Chinese population census in 2000 and World Segi's standard population were used for age-standardized evaluation. Annual Percentage changes (APC) were calculated using Joinpoint software. RESULTS: From 2013 to 2017, the incidence of prostatic cancer in the tumor registration area was 6.50×10^-5, age-standardized incidence rate by Chinese standard population (ASR China) of 3.71×10-5, and the cumulative rate (0-74 years old) was 0.35%. The incidence of prostatic cancer in urban areas (8.05×10^-5) was higher than that in rural areas (2.83×10^-5). From 2013 to 2017,the mortality rate of prostatic cancer was 3.41×10^-5,ASR China mortality of 1.90×10^-5,and the cumulative rate (0-74 years old) was 0.16%. The mortality rate of prostatic cancer in urban areas (4.20×10^-5) was higher than that in rural areas (1.53×10^-5). The incidence and mortality rates increased with age. Although ASR China incidence and mortality showed an upward trend,the differences were not statistically significant (P < 0.05). CONCLUSION: The incidence rate and mortality rate of prostatic cancer in Heilongjiang cancer registry area increased with age. City areas were higher than that in rural area. It is suggested to strengthen the prevention and treatment of prostatic cancer in key populations,especially among the elderlies.

OBJECTIVE: To explore relationships between expression of metadherin (MTDH) and forkhead box M1 (FoxM1) in hepatocellular carcinomas (HCC) and the clinicopathological features and prognosis of patients. METHODS: Surgical specimens from 156 HCC patients were usedof. Expressions of MTDH and FoxM1 were detected by immunohistochemistry and to analyze the relationship between the expression of MTDH and FoxM1 and the clinical pathological characteristics and prognosis. RESULTS: Immunohistochemical results showed that positive rates of MTDH was 71.2% (111/156). Expressions of MTDH were significantly related to tumor numbers,TNM stages,microvascular invasions,satellite nodules (P < 0.05). Positive rates of FoxM1 was 64.7% (101/156). Expressions of FoxM1 were significantly related to cirrhosis, tumor numbers, TNM stages, microvascular invasions, satellite nodules (P < 0.05). RT-PCR showed that expression of MTDH and FoxM1 was significantly increased. Spearman correlation analyses showed that there was a significant correlation between MTDH and FoxM1. Results from the Kaplan-Meier survival analyses showed that survival times were significantly related to TNM stages, microvascular invasions, satellite nodules, expressions of MTDH and FoxM1 (P < 0.05). Results from the multivariate Cox regression analyses showed that TNM stages and MTDH expressions were the main factors which influenced survival times of the patients. CONCLUSION: For HCC patients,MTDH and FoxM1 expressions were related to many clinical factors. Particularly,MTDH expression was a prognostic factor and there was a significant correlation between MTDH and FoxM1 expressions.

OBJECTIVE: To establish and evaluate a modified local lymph node assay incorporated with bromodeoxyuridine and enzyme-linked immunosorbent assay. METHODS: Female BALB/c mice were organized into several groups of 4 mice per group:solvent control (AOO, ACE, DMSO or distilled water), positive control (25% hexyl cinnamaldehyde) and test substance (22 chemicals and 5 cosmetics were selected to form 1-3 different concentrations) groups. The dorsal sides of both ears of mice were treated with 25 μL/ear of test solutions at 9:00 am on 3 consecutive days with no treatment on day 4. BrdU solution was injected interperitoneally on day 5 with 0.5 mL per mouse. On day 6, ear punches were collected and weighted. The bilateral draining auricular lymph nodes were excised and made into single cell suspensions from which lymphocyte proliferations were measured using the BrdU ELISA kit. RESULTS: Ear punch weights from mice treated with 50% hexyl cinnamic aldehyde,5% cobalt dichloride,10%,25%,50% lactic acid and No.2 hair dye were increased significantly (P < 0.05 or P < 0.01),which might be considered as irritants. The others were negative. For LLNA, 15 chemicals (2,4-dinitrochloro-benzene, eugenol, isoeugenol, 3-aminophenol, transcinnamic aldehyde, hexyl cinnamic aldehyde, geraniol, hydroquinone, hexyl salicylate, formaldehyde, glutaraldehyde, nickel sulfate, cobalt dichloride, potassium dichromate, lactic acid) and No.2 hair dye were positive (SI>1.6),and the others were negative. The method was evaluated with positive prediction rate 93.3%, negative prediction rate 100%,false positive rate 14.3%,false negative rate 0,sensitivity 100%,specificity 85.7% and accuracy 95.2%. CONCLUSION: The LLNA:BrdU-ELISA assay using BALB/c mice appears to be useful for predicting sensitization and irritancy potential,and for safety evaluation of cometics.

OBJECTIVE: To explore effects of PM_2.5 samples from Shenzhen and Taiyuan on oxidative damage and apoptosis in human kidney 2 cells (HK-2 cells). METHODS: A medium-flow sampler was used to collect air samples at a university campus in Taiyuan and at the top of a building of the Shenzhen Center for Disease Control and Prevention. Membranes from the PM_2.5-adsorbed filters were eluted to prepare PM_2.5 suspensions. HK-2 cultures were divided into several groups:negative control,50 μg/mL Shenzhen PM_2.5,50 μg/mL Taiyuan PM_2.5 and positive control groups. Cells were treated for 24 h and levels of malondialdehyde (MDA),superoxide dismutase (SOD),reduced glutathione (GSH),glutathione peroxidase (GSH-PX) and rates of cell apoptosis were measured. RESULTS: Compared with the negative control group, MDA levels in the Shenzhen,Taiyuan and positive control groups were increased by 8.16%,34.51% and 72.23%,respectively; the SOD levels were reduced by 7.49%, 19.67% and 29.55%, respectively; GSH levels were reduced by 10.43%,16.39% and 37.43%,respectively. Among these data,differences between the Taiyuan PM_2.5 and the positive control groups were statistically significant (P < 0.05 or P < 0.01). In similar comparisons among the 4 groups, the GSH-PX levels were reduced by 42.70%, 61.62% and 60.98%, respectively; the differences between the positive control group,and the Shenzhen and Taiyuan groups were statistically significant (P < 0.01). The apoptosis rates were increased by 197.25%, 301.22% and 399.08%, respectively; and the differences were statistically significant (P < 0.05 or P < 0.01). CONCLUSION: Our data indicate that the PM_2.5 from Shenzhen and Taiyuan caused oxidative damage and increased apoptosis rates in HK-2 cells. In addition,the Taiyuan PM_2.5 were more toxic than that from Shenzhen.

OBJECTIVE: To study the protective effect of oleanolic acid (OA) on alcohol-induced gastric mucosal injury in rats. METHODS: Using the random number table method, 24 SD rats were randomly divided into 3 groups of 8 rats per group:control (isocaloric glucose solution),alcohol exposure (gavage 4 g/kg alcohol to construct alcoholic gastric wall injury model),OA intervention (perfusion to the stomach with an alcohol solution of 10 mg/kg OA). All treatments lasted 30 days. Rats were evaluated for general changes of the stomach wall and pathological changes of HE; content of malondialdehyde (MDA) and oxidized glutathione (GSSG) in the stomach wall; content of reduced glutathione (GSH) and ratio of GSH/GSSG,antioxidant enzyme transcription Factor Nrf-2 mRNA and protein expression; and TNF-α, IL-6 mRNA and protein expression levels. RESULTS: Compared with the control group,4 g/kg alcohol exposure for 30 days successfully induced oxidative damage and inflammatory reactions of the gastric wall in rats. Compared with the alcohol-exposed rats, pathological damage to the stomach wall in the OA intervention group was reduced, contents of MDA and GSSG in the stomach wall tissue were decreased (P < 0.05),GSH contents and the GSH/GSSG ratios were increased (P < 0.05),and Nrf-2 was increased. mRNA and protein expression levels were increased significantly (P< 0.05),while mRNA and protein expressions of TNF-α and IL-6 were decreased (P < 0.05). CONCLUSION: OA played a protective role on alcohol-induced gastric wall injury in rats through its antioxidant and antiinflammatory effects.

OBJECTIVE: To explore the use of tree shrews in the micronucleus test as an assay to evaluate medical devices. METHODS: 30 tree shrews were randomly divided into three groups:negative control, test sample (collagen implant) and positive control. Treated animals were each induced by two intraperitoneal injections of 10 mL/kg. Femoral bone marrow cells were removed from each animal to detect micronucleus rates, and to conduct other tests:blood biochemistry and electrolyte tests, spleen, liver and kidney organ coefficients and histopathological tests. At the same time,experiments were performed using mice as a reference for the system. RESULTS: The micronucleus rates from the negative control and the test sample groups of the tree shrews and that from mice were <5%. The difference in micronucleus rates between the test sample and the negative control groups were not significant (P > 0.05),but the difference between the positive control and the negative control groups were statistically significant (P < 0.01). The results of the tree shrew micronucleus test were consistent with those from the mice. Compared with the negative control group, the tree shrew blood routine, blood biochemistry, electrolyte and histopathological test results in the test sample group were not found to be abnormal. Compared with mice, the organ coefficient of tree shrews is closer to that of humans. CONCLUSION: Tree shrew can be used as an animal modal for micronucleus test evaluation,and tree shrew has the potential to be used as a biological evaluation of medical devices.

OBJECTIVE: To evaluate safety of MCPA-isooctyl and fluroxypyr-meptyl based on their digestion trends and residual levels on wheat. METHODS: The residue dynamics and final residue levels of 48% MCPA-isooctyl · fluroxypyr-meptyl · sulfentrazone suspension concentrates on wheat were carried out in three places for two years. The investigation involving three groups:wheat grain, plant and soil, and the digestion dynamics of MCPA-isooctyl and fluroxypyr-meptyl, were detected using high performance liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry. The three sets of samples were extracted with acetonitrile, the supernatant was purified by dispersive solid phase extraction, passed through the membrane,detected by gas chromatography-mass spectrometry and high-performance liquid chromatographytandem mass spectrometry,and also quantified by external standard method. RESULTS: The MCPA-isooctyl in wheat,plants and soil microorganisms was in the range of 0.005-0.2 mg/kg,and the fluroxypyr-meptyl in the range of 0.02-0.2 mg/kg. They showed a linear relationship with 0.05,0.1,0.2 mg/kg at 3 addition levels. The recovery rates were 77.0%-112.0%,the relative standard deviations were 1.1%-9.9%,and the limit of quantitative detection (LOQ) was 0.05 mg/kg. In 2 years (2016-2017) and in 3 places (Beijing, Henan, Ningxia), the digestion dynamic investigations show that the digestion half-life t_1/2 of fluroxypyr-meptyl on wheat plants in Beijing in 2016 was 5.92 d. In the soil samples in Beijing (2016) and Ningxia (2017),t_1/2 were 1.56 and 9.63 d,respectively. For comparison,the final residues of MCPA-isooctyl in wheat which were harvested at other times and other locations were <0.05 mg/kg. The final residues of fluroxypyr-meptyl were < 0.05 mg/kg which were lower than the national maximum residue limits for these two pesticides in wheat. CONCLUSION: MCPA-isooctyl and fluroxypyr-meptyl in winter wheat and soil had faster digestion rates. Therefore, 48% MCPA-isooctyl · fluroxypyr-meptyl · difluoro sulfentrazone suspension at the recommended dosage of 432 ga.i./hm² would probably not cause residual contamination of wheat and soil.

OBJECTIVE: The study investigated toxic effects of sub-chronic oral exposures for 90 days in rats to genetically modified (GM) corn with insect-resistant gene cry1Ab and cry3Bb, herbicide-resistant gene cp4epsps, high vitamin gene ZmHPT and ZmTMT. METHODS: 6-week-old Westar rats, half males and half females, were randomly divided into 7 groups with 20 per group according to weight:basic feed control group,non-GM corn groups (low-,medium- and high-doses),GM corn experimental groups (low-, medium- and high-doses). Rats were fed the different corns for 90 consecutive days. Then,they were observed for changes in body weight, food intake, blood biochemical indicators, organ coefficients and pathological features. RESULTS: Rats in each dose group grew normally during the test cycle. The body weight, food intake, blood biochemical indexes, organ coefficients of some GM corn and non-GM corn groups were significantly different from those in the control animals (P < 0.05). In addition, the differences were also significantly different between the GM corn compared with the non-GM corn groups (P < 0.05). However, no significant abnormalities were found in the pathology examinations of some main organ tissues. CONCLUSION: The experimental results show that sub-chronic exposure to the genetically modified corns did not lead to obvious pathological changes in rats.