[Expression of HMGB1 protein in breast cancer and its clinicopathological significance].

Objective: To investigate the expression and clinicopathological significance of high mobility group box protein B1 (HMGB1) protein in breast cancer. Methods: The expression of HMGB1 protein in 26 normal breast tissues and 417 invasive breast cancer tissues diagnosed at Dongyang People's Hospital, Zhejiang Province from 2016 to 2018 were detected by immunohistochemical EnVision method. The relationship between nuclear and cytoplasmic HMGB1 protein expression and clinicopathologic features of breast cancer patients were analyzed. Results: The nuclear and cytoplasmic expression of HMGB1 protein was 80.8% (337/417) and 16.8% (70/417) respectively in breast cancer, and was 46.2%(12/26) and 0(0/26) respectively in normal breast tissue. Both nuclear and cytoplasmic expression of HMGB1 protein in breast cancer were significantly higher than normal breast tissue (P<0.001, P=0.046, respectively). The nuclear expression of HMGB1 protein was also higher in high grade, estrogen receptor (ER) negative, progesterone receptor (PR) negative (P=0.006, P=0.004, P<0.001, respectively); whereas the cytoplasmic expression of HMGB1 protein was also higher in high grade, estrogen receptor (ER) negative, progesterone receptor (PR) negative (P<0.001 in all) breast cancers. Multivariate logistic regression model showed that nuclear HMGB1 expression correlated with histologic grade (OR=2.188, 95%CI=1.078-4.443, P=0.030), while cytoplasmic HMGB1 expression correlated with histologic grade (OR=3.031, 95%CI=1.600-5.742, P=0.001), ER (OR=0.129, 95%CI=0.034-0.494, P=0.003) and TNM staging (OR=3.820, 95%CI=1.042-14.001, P=0.043). Multivariate analysis of Cox proportional hazard model showed that nuclear HMGB1 expression was an independent risk factor for the overall survival of breast cancer patients (HR=0.366, 95%CI=0.138-0.972, P=0.044). Conclusion: Nuclear and cytoplasmic HMGB1 proteins are related to multiple poor prognostic factors in breast cancer, and may be a potential biomarker for breast cancer treatment.

Isolation, sequence analysis, and characterization of androgen receptor in Southern catfish, Silurus meridionalis.

Androgen receptor (AR), the mediator of androgen, plays important roles in the androgen signal pathway. In the present study, we isolated and analyzed the cDNA sequence and tissue distribution of androgen receptor in Southern catfish (scAR). The full-length cDNA of scAR contains 3,116 bp with an open reading frame (ORF) of 2,415 bp, encoding a protein of 804 amino acids (aa). Tissue distribution analysis of scAR revealed that it was expressed in all tissues examined, with no sexual dimorphism in the ovary and testis. Phylogenetic analysis and multiple amino acids sequence alignment indicated the close relationship and high similarity of scAR with ARs from cypriniform species. In addition, partial sequences of ARs from 7 other siluriform species were also isolated. Comparison of catfish ARs with those from other vertebrates revealed that an extra C-terminal tail of about 20aa exists in all the ARs from siluriform fishes investigated, but not in other ARs. The extra sequence was resulted from a 4-bp insertion before the stop codon of other vertebrate ARs, and it was identical in ARs from siluriform species of the same genus but different among ARs from species of different genera. We report here for the first time that the ARs from siluriform species are longer in C-terminal than those from other vertebrates and it might be useful in reconstruction of the phylogenetic relationship among siluriform fishes. The significance of the extra C-terminal tail for AR function remains elusive.

Prevalence and genotype of Giardia duodenalis from faecal samples of stray dogs in Hualien city of eastern Taiwan.

Giardia duodenalis is a zoonotic protozoan parasite that causes diarrhea through waterborne transmission or fecal-oral infection. The cysts are chlorine-resistant and, therefore, can pollute drinking water and induce a pandemic disease. In this study, we aimed to detect G. duodenalis infection in stray dogs in Hualien, Taiwan. We collected faecal samples from 118 dogs and amplified DNA sequences of the ß-giardin gene by nested polymerase chain reactions (nested PCR). Eleven of the 118 faecal samples tested positive for the parasite. The genotype analysis of the 11 samples indicated that 7 samples belonged to assemblage C and four samples belonged to assemblage D. Our study provided a better understanding of the infection rate and genotypes of G. duodenalis in dogs from Hualien City, and human infection could not be induced by this zoonotic infection pathway in Hualien City.

Study of chloropropanols in soy sauce by gas chromatography-triple quadrupole mass spectrometry with coupled column separation without derivatisation.

A qualitative and quantitative determination method for chloropropanols in soy sauce was developed by GC-MS/MS with coupled column separation without derivatisation. The target compounds were 1,3-dichloropropan-2-ol (1,3-DCP), 2,3-dichloropropan-1-ol (2,3-DCP), 3-monochloropropane-1,2-diol (3-MCPD) and 2-monochloropropane-1,3-diol (2-MCPD). 3-MCPD-d(5) was used as an isotope internal standard for MCPDs and 1,3-DCP-d(5) for DCPs. After spiking with internal standards and mixed with 1 g of Extrelut™ NT, about 1 g of the sample was washed by 4 ml of hexane and the analytes were eluted with 15 ml of 95% (v/v) ethyl ether/hexane mixture. The concentrated extract was directly injected without derivatisation, separated by a coupled column - a 3 m Innowax (polyethylene glycol) combined with a 30 m DB-5 ms ((5%-phenyl)-methylpolysiloxane) by a quartz capillary column connector - and detected by GC-MS/MS. The limits of detection (LODs) in the sample matrix were 1.0, 2.5, 5.0 and 5.0 µg kg(-1) for the above chloropropanols, respectively. The relative proportions of 2-MCPD/3-MCPD ranged from 0.19 to 3.74 in soy sauce samples. 2,3-DCP and 2-MCPD were more stable than 1,3-DCP and 3-MCPD under alkaline condition. The levels of chloropropanols can be decreased by an alkaline treatment process.

RNA synthesis in germinating embryonic axes of soybean and wheat.

The rate of synthesis of RNA during early germination of wheat and soybean embryos was investigated by ascertaining the incorporation of radioactive uridine into RNA. In wheat embryos, where the lag period preceding rapid growth is 5.5 hours, there is a 2-fold increase in RNA synthesis between 1.5 and 5.5 hours, with half of the increase occurring by 3.5 hours. In soybean axes, where the lag period is 9.5 hours, the increased rate of RNA synthesis is 5.5-fold between 1.5 and 9.5 hours, with three fourths of this increase occurring after 4 hours.Analysis of the ratio of radioactivity incorporated into the 18S and 26S rRNAs of the germinating embryos provided a further measure of the increased rates of RNA synthesis. With wheat embryos, the 26S/18S ratio increased from 1.0 at 1.5 hours to 1.5 at 3.5 hours, while with the soybean axes, distinct ribosomal patterns were obtained only after 4 hours and the 26S/18S rRNA ratio increased from 0.4 at 4 hours to 1.0 at 9 hours. The extent of methylation of the rRNA synthesized at 4 and 9 hours in the soybean axes was similar, indicating that the methylating capacity of the axes is probably not rate limiting to rRNA synthesis. In both seed embryo systems the level of UTP increased 2 to 3-fold during the lag phase of germination. With wheat embryos, the time course of the increase in UTP correlated approximately with the change in the rate of RNA synthesis. With the soybean axes, however, the increase in the rate of RNA synthesis occurred predominantly after the rise in the level of UTP.