Omitting the lower neck and sparing the glottic larynx in node-negative nasopharyngeal carcinoma was safe and feasible, and improved patient-reported voice outcomes.

BACKGROUND: Worsening voice and speech quality was frequently reported in head-and-neck patients after radiotherapy to the neck; omitting the lower neck and sparing the glottic larynx in node-negative nasopharyngeal carcinoma (NPC) patients might be safe and feasible, and improve voice and speech outcomes. METHODS: From January 2009 to January 2013, 71 patients were analyzed. All patients received bilateral neck irradiation. Upper group (UG) patients spared the glottic larynx while lower group (LG) patients did not. Voice and speech quality were evaluated at two time-points (T1 and T2) using the Communication Domain of the Head and Neck Quality of Life (HNQOL) instrument and the Speech question of the University of Washington Quality of Life instrument. RESULTS: At a median follow-up time of 32 months (T1),71.6% of patients reported worsened voice and speech quality. UG patients resulted in significant decreases in glottic larynx dose. With a median follow-up time of 71 months (T2), no patients experienced out-of-field nodal recurrence;there was no difference in the 5-year overall survival and nodal recurrence-free survival between two groups (P = 0.235 and 0.750, respectively). At T1, in patients who without concurrent chemotherapy (CCT), UG patients showed significantly better patient-reported voice quality, (P = 0.022). UG patients without CCT also showed higher scores in the HNQOL communication domain and pain domain (P = 0.012 and P = 0.019). CONCLUSIONS: For node-negative NPC patients, omitting the lower neck and sparing the glottic larynx was safe and feasible, and better voice outcomes were achieved in patients without CCT. Further prospective longitudinal studies to investigate whether this approach would be beneficial to node-negative patients are warranted.

hBMP-7 induces the differentiation of adipose-derived mesenchymal stem cells into osteoblast-like cells.

The aim of this study was to investigate the differentiation potential of adipose-derived mesenchymal stem cells (ADMSCs) into osteoblasts by human bone morphogenetic protein-7 (hBMP-7) induction. ADMSCs were isolated from the subcutaneous adipose tissue of a rabbit, and then transfected with the pcDNA3.1 vector alone and pcDNA3.1-hBMP-7 (hBMP-7), respectively. Untransfected ADMSCs were used as the control group. After transfection, the morphology and green fluorescent protein (GFP) fluorescence intensity of ADMSCs were observed by fluorescent microscopy. The 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide assay was performed to detect the growth of ADMSCs at 1, 3, and 5 days, respectively. Transmission electron microscopy was performed to observe the ultrastructural morphology of ADMSCs. In addition, ADMSCs were stained with quinalizarin and toluidine blue to reflect the content of osteoblasts and chondrocytes, respectively. Finally, the expression of collagen I and osteocalcin in ADMSCs was detected by western blot. ADMSCs were successfully isolated. Obvious GFP fluorescence and high expression of hBMP-7 demonstrated the successful transfection of hBMP-7. Specific morphological characters with a metabolically active ultrastructure were exhibited on the ADMSCs transfected with hBMP- 7. In addition, the growth rate of ADMSCs transfected with hBMP-7 was significantly higher than that of the cells in the vector and control groups. Successfully induced osteoblast-like cells were identified by an obvious erythrine area and high expression of collagen I and osteocalcin in ADMSCs transfected with hBMP-7. Thus, ADMSCs can be successfully differentiated into osteoblast-like cells by hBMP-7 induction in vitro.

Genetic effect of an A/G polymorphism in the HSP70 gene on thermotolerance in chicken.

Studying thermotolerance is important for the prevention of thermostress in chickens. This study aimed to analyze the effect of mutations in the heat shock protein 70 (HSP70) gene on chicken thermotolerance. The C.-69A>G SNP in the 5'-flanking region of the HSP70 gene was genotyped in Lingshan and White Recessive Rock (WRR) chickens. Association of this SNP with thermotolerance traits revealed it to be significantly associated with CD4+/CD8+, and potentially associated with heterophil-to-lymphocyte ratio in WRR chickens exposed to thermoneutral temperature (15°C). Online prediction detected a putative myeloid zinc finger protein 1 binding factor in the C.-69A>G mutation. Under acute thermostress, mRNA levels of HSP70 in individuals with different C.-69A>G genotypes varied in the heart, leg muscle, and liver tissues. The HSP70 protein was expressed at higher levels in individuals with the GG genotype than in those with the AA genotype. In heart and liver, protein expression of HSP70 in individuals with the GG genotype was significantly higher than in those with the AA genotype. In leg muscle, protein expression was higher in birds with the GG genotype than in those with the AA and AG genotypes. Luciferase activity of the GG genotype was significantly higher than that of the AA genotype, suggesting that the C.-69A>G SNP regulates HSP70 gene expression. These results indicate that the C.-69A>G SNP in the 5'-flanking region of the HSP70 gene might affect chicken thermotolerance and that the GG genotype might be advantageous for the prevention of thermostress.

Molecular characterization of the gene checkpoint homolog 1 in Daphnia carinata during different reproductive phases.

Full-length cDNA of the gene checkpoint homolog 1 (Chk1) was cloned from Daphnia carinata and designated DcarChk1. DcarChk1 cDNA was 1817 bp in length and encoded a 497-amino acid polypeptide. Phylogenetic analyses revealed that DcarChk1 was most closely related to Chk1 of Daphnia pulex, followed by homologous genes of insects. Expression of DcarChk1 was higher in adult Daphnia than in larvae, and significantly higher in males than females, as determined by real-time polymerase chain reaction analysis. Using whole-mount in situ hybridization techniques, DcarChk1 in parthenogenetic females was found to be expressed mainly on the head surface, capillus, and carapace valve edge. In contrast, in sexual females, DcarChk1 was expressed mainly in the joint of the second antenna, and in the thoracic limbs and capillus. These results suggest that DcarChk1 plays a significant role in both the growth and development, as well as in regulating reproductive plasticity, in D. carinata.

Protective effect of thymoquinone on cholestatic rats with liver injury.

The aim of this study was to investigate the protective effects of thymoquinone treatment on cholestatic rats with liver injury. Thirty-two Sprague-Dawley rats were divided randomly into four groups: normal control, bile duct ligation model control, low-dose thymoquinone (25 mg/kg), and high-dose thymoquinone (50 mg/kg). Thymoquinone gavage was administered continuously 3 days before bile duct ligation, and saline, at the same volume, was administered to the control group. The rats were sacrificed after 2 weeks of treatment, and the liver tissues were obtained and frozen. The contents of hydroxyproline (HP), malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in the homogenate of the liver tissues were determined to evaluate the changes in hepatic tissue pathology by fibrosis scoring. The HP and MDA levels were significantly lower and the SOD and GPx levels were significantly higher in the thymoquinone-treatment group than the corresponding levels in the model control group, and the differences were statistically significant (P < 0.05) and dose-dependent. The hepatic necrosis areas and hepatic fibrosis scores of the thymoquinone-treatment groups were significantly lower than those of the model group (P < 0.05). Thymoquinone increased the antioxidative capacity of liver and reduced the oxidative stress damage to the liver. Thymoquinone can be used as a liver protectant in patients with cholestasis.

Characterization of HSP70 and its expression in tissue: correlation with physiological and immune indices in goose (Anser cygnoides) serum.

We cloned the goose heat shock protein 70 gene (HSP70), to determine its sequence variation and elucidate its mRNA expression. We designed primers to amplify the entire goose HSP70 sequence. We used 10 commercial Wuzong goslings in a heat-stress experiment. We collected tissue samples for RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR). We analyzed the variation in expression of goose HSP70 before and after heat stress. We constructed a DNA pool from six different species, for single nucleotide polymorphism (SNP) screening. We detected 18 SNPs and selected three of these SNPs for correlation analysis with biological and immune traits in 200 Wuzong geese. We showed that T+237C was significantly correlated with the serum corticosterone level, whereas T+1122C was significantly correlated with the heterophil to lymphocyte ratio. Goose HSP70 contained no introns. The results of qRT-PCR analysis revealed significant gender differences in the expression of goose HSP70 at 40°C but not at 25°C; moreover, in general, expression was significantly higher at 40°C than at 25°C. With the exception of the leg muscle and cerebellum, HSP70 expression was significantly higher in male geese than in female geese. Our results indicate that goose HSP70 plays an important role in response to severe heat stress.

Association analysis between SNPs in the 5'-flanking region of the chicken GRP78 gene, thermotolerance parameters, and tissue mRNA expression.

Glucose-regulated protein 78 (GRP78) is a molecular chaperone in the endoplasmic reticulum and can be induced by different kinds of environmental and physiological stress. Thus far, the role of the GRP78 gene in thermotolerance in chickens has not been investigated. In the present study, we detected sequence variations in the 5ꞌ-flanking region of the GRP78 gene and evaluated several thermotolerance parameters, such as T3, corticosterone, H/L ratio, and levels of CD3(+), CD4(+), and CD8(+) T cells, to further determine its associations at 35° and 15°C. The sequencing results revealed 10 SNPs in the 5'-flanking region of the GRP78 gene, and seven mutations were chosen for further genotyping in a White Recessive Rock (WRR) chicken population. The SNP C.-744C>G in WRR chickens was significantly correlated with heat tolerance parameters under both conditions; it may therefore exert a potential hereditary effect on heat tolerance, and the genotype GG may be advantageous for thermotolerance. The heart, liver, brain, and leg muscle tissues of 8-day-old WRR chickens were sampled from heat stress groups, which were defined by exposure to 1, 2, 3, and 6 h of persistent thermal stress, and a control group, which was not exposed to thermal stress. Quantitative real-time polymerase chain reaction assay indicated that the mRNA expression level of the GRP78 gene increased gradually under heat stress, peaked at 3 h, and then decreased. We conclude that the mRNA expression of the GRP78 gene is time- and tissue-dependent.

Analysis of genetic diversity of the heat shock protein 70 gene on the basis of abundant sequence polymorphisms in chicken breeds.

This study was designed to detect the sequence variation of the chicken heat shock protein 70 (HSP70) gene. A total of 102 individuals from 8 native Chinese breeds together with Dwarf White Chicken and Red Junglefowl were used to detect sequence variations. The coding regions of the chicken HSP70 gene from 102 individuals were cloned and sequenced. Thirty-six variations were identified, which included 34 single nucleotide polymorphisms and 2 indel mutations. Fifty-seven haplotypes were observed, of which, 43 were breed-specific and 14 were shared. There were 7 Red Junglefowl-specific haplotypes, while Haidong and Silkie only had 2 specific haplotypes. Eleven and 3 haplotypes were shared between and within species, respectively. The variation in nucleotide diversity (Pi) and average number of nucleotide differences (K) among species were consistent. The total Pi of HSP70 was 0.0016, and the total K was 4.1998. The Pi value of Red Junglefowl was the highest (0.0018) and K was 4.8000, while the Pi of Silkie was the lowest (0.0010) and K was 2.5000. These results demonstrated that variation in chicken HSP70 was abundant between and within species.

Alteration of HSF3 and HSP70 mRNA expression in the tissues of two chicken breeds during acute heat stress.

This study aimed to estimate changes in HSF3 and HSP70 mRNA expression in stress-sensitive tissues of 2 chicken breeds during acute heat stress. Lingshan chickens (LSC) and White Recessive Rock (WRR) (24 chickens of each breed) were randomly divided into 4 groups (0, 2, 3, and 6 h of heat treatment). With increasing heat treatment time, both HSF3 and HSP70 expression first declined and then showed a significant increase in both breeds. However, HSP70 expression decreased in the heart following 6 h of heat treatment, whereas HSF3 expression continued to increase. After 2 h of heat treatment, HSF3 expression was significantly higher in the brain and leg muscle of LSC compared to WRR (P < 0.05, P < 0.01). In comparison, HSP70 expression was significantly higher in the liver and leg muscle of WRR compared to LSC (P < 0.01, P < 0.05). After 3 h of heat treatment, HSF3 expression was significantly higher in the brain and leg muscle of LSC compared to WRR (P < 0.01). In comparison, HSP70 expression was significantly higher in the liver and heart of LSC compared to WRR (P < 0.01). These results indicate that the expression of HSF3 and HSP70 mRNA in LSC and WRR exhibit species-specific and tissue-specific differences during heat treatment.

Role of inflammatory parameters in the susceptibility of cerebral thrombosis.

We aimed to investigate the association of inflammation-related genes such as IL-10, IL-6 and IL-1B with risk of ischemic stroke. We included 426 cases with ischemic stroke and 426 health controls from Xinxiang, China. Genomic DNA was extracted from the buffy coat layer of collected blood with the TIANamp blood DNA kit. Diabetes, hypertension, obesity, and smoking habits were associated with risk of ischemic stroke. We found that individuals carrying the CC genotype of IL-1B rs1864169 had a higher risk of ischemic stroke when compared with the TT genotype (OR = 1.80, 95%CI = 1.16-2.80). The IL-6 rs1800796 TT genotype was associated with increased risk of ischemic stroke. We found that IL-1B rs1864169 and IL-6 rs1800796 polymorphisms may interact with diabetes, hypertension and obesity. Our study suggests that IL-6 rs1800796 and IL-1B rs1864169 polymorphisms are associated with ischemic stroke risk in the Chinese population.

CD4 mononuclear cell infiltrates and Fas/Fas ligand positive mammary gland cells in breast tissue from a patient with Sjögren's syndrome.

We describe a 49-year-old patient with lip biopsy proven Sjögren's syndrome (SS) and keratoconjunctivitis sicca, who had dental caries, xerostomia, recurrent upper respiratory tract infections, arthritis in her hands, elbows and knees, and recurrent parotid inflammation. She developed bilateral breast nodules in 1988. Right breast nodules were excised in 1993 and 1995, but reappeared in 1996, requiring 2 more excisions. Breast tissue samples showed remarkable intralobular and perilobular mononuclear cell infiltrates that were predominantly CD4+ T cells and expressed bcl-2. A few cells stained CD20+ and CD8+. SS breast glandular epithelial cells stained more intensely for Fas compared to normal cells. CD4+ T cells and Fas mediated cell death may be involved in the mammary gland lesions in SS.