Crystallization mechanism of the Pt50Au50 alloy with grain boundary segregation during the solidification process.

The crystalline mechanism of the Pt50Au50 alloy with grain boundary (GB) segregation during the rapid solidification process is investigated using molecular dynamics simulations. The cluster evolution and phase transformation processes during the GB segregation are analyzed by means of the energy temperature (E-T) curve, the pair distribution function (g(r)) curves, common neighborhood analysis (CNA), cluster-type index method (CTIM) and three-dimensional visualizing analyses. It is found that the GB segregation phenomenon of the Pt50Au50 alloy comes from various solidification temperatures of Pt- and Au-centered clusters. Four critical temperatures T1 (1153 K), T2 (1073 K), T3 (853 K) and T4 (753 K) are discovered during the liquid-solid transition, corresponding to the supercooled liquid, Pt-centered atom nucleation, Pt-centered cluster growth, Au-centered atom nucleation and grain growth process, respectively, which is observably different to the solidification process of other alloys. The Pt atoms begin to gather together in the high-temperature liquid before the liquid-solid transition. It is also found that the CTIM proposed by us would provide an effective tool to investigate the GB segregation process.

Evaluation of the semen swim-up method for bovine sperm RNA extraction.

Isolation of high-quality RNA is important for assessing sperm gene expression, and semen purification methods may affect the integrity of the isolated RNA. This study evaluated the effectiveness of the sperm swim-up method for seminal RNA isolation. Frozen semen samples in straws from three bulls of proven fertility were purified by the swim-up method. RNA extraction was carried out using the E.Z.N.A.(TM) Total RNA kit II, with non-swim-up sperm as a control. Total sperm RNA was analyzed by UV spectrophotometry, reverse transcription polymerase chain reaction (RT-PCR), and agarose gel electrophoresis, and expression of the sex-determining region on the Y chromosome (SRY), leptin (LEP), and ribosomal protein subunit 23 (RPS23) genes, were determined. 18S RNA was used as a positive control. Fewer somatic cells were found in sperm swim-up samples than in the non-swim-up counterparts (0 x 10(3) vs 17.33 ± 2.52 x 10(3) sperm, P < 0.05). In addition, high-quality RNA was obtained in about 2 h, with no significant difference between groups. Interestingly, the yields of RNA fragments containing ≥200 nucleotides were significantly reduced in sperm swim-up samples (0.92 ± 0.41 x 10(7) sperm) compared with the non-swim-up samples (1.36 ± 0.33 x 10(7) sperm, P < 0.05). After RT-PCR, clear bands representing SRY, LEP, and RPS23 in sperm cDNA were observed on agarose gel electrophoresis. Finally, no bands corresponding to 18S RNA were found in RNA samples from the sperm swim-up group. Our findings suggest that small amounts of sperm RNA can be efficiently extracted from frozen straw semen samples using the swim-up technique.

Expression patterns and association analysis of the porcine DHX58 gene.

RIG-1 signalling is responsible for the detection of cytoplasmic viral RNA molecules. DEXH(Asp-Glu-X-His) box polypeptide 58 (encoded by DHX58) is a negative regulator of the RIG-1 signalling pathway. In human, the DHX58 gene can be upregulated and can inhibit the RIG-1 signalling pathway during viral infection. In this study, porcine DHX58 gene expression patterns were studied. According to our results, the porcine DHX58 gene was upregulated not only by the stimulation of Poly I:C but also by the stimulation of lipopolysaccharides (LPS). One polymorphism (g.4919G>C), detected in the ninth intron,was significantly associated with some blood parameters including the red cell distribution width of 1-day-old pigs and white blood cell counts, lymphocyte absolute counts, and platelet distribution width of 17-day-old pigs (P < 0.05). Moreover, the individuals with the genotype GG have a significantly higher mean white blood cell count than individuals with genotype CC or GC (P < 0.05). Our study indicates that DHX58 is an important gene that is associated with the immune response in swine.

Finite element analysis of the effects of three preparation techniques on stresses within roots having curved canals.

AIM: To compare stress distribution within roots having curved canals prepared by three preparation techniques when subjected to occlusal loads and condensation loads as a consequence of different filling techniques. METHODOLOGY: Three preparation techniques (crown-down, step-back and reverse-flaring) were compared by finite element analysis (FEA). Based on an established FEA model within curved canal, three modified models prepared by different preparation techniques were established by replacing original canal with prepared ones. FEA was performed to investigate the stress distribution under occlusal forces, which were simulated by loads of 500 N in four directions (buccal, lingual, mesial and distal), at 0 (vertical), 30, 45 and 60 degrees to the longitudinal axis of the tooth. In addition, vertical and lateral condensation processes at the curvature were simulated to determine the influence of different canal filling techniques on stress distribution. RESULTS: When the occlusal and the filling loads were applied, stress distribution around the curvature and the orifice had little change on the three modified prepared models. The reverse-flaring technique resulted in the least stress with the lateral condensation process. In the case of vertical condensation, the maximum von Mises stress (46.205 MPa) occurred near the loading site. The model also revealed a tendency for stress concentration (30.635 MPa) just below the compacting level. CONCLUSIONS: The study confirms that appropriate canal preparation techniques in simulated curved canals have little influence on stress distribution around the curvature or the orifice. However, vertical compaction induced high stress in the region just below the loading site.

Computed tomography versus magnetic resonance imaging for diagnosing cervical lymph node metastasis of head and neck cancer: a systematic review and meta-analysis.

Computed tomography (CT) and magnetic resonance imaging (MRI) are common imaging methods to detect cervical lymph node metastasis of head and neck cancer. We aimed to assess the diagnostic efficacy of CT and MRI in detecting cervical lymph node metastasis, and to establish unified diagnostic criteria via systematic review and meta-analysis. A systematic literature search in five databases until January 2014 was carried out. All retrieved studies were reviewed and eligible studies were qualitatively summarized. Besides pooling the sensitivity (SEN) and specificity (SPE) data of CT and MRI, summary receiver operating characteristic curves were generated. A total of 63 studies including 3,029 participants were involved. The pooled results of meta-analysis showed that CT had a higher SEN (0.77 [95% confidence interval {CI} 0.73-0.87]) than MRI (0.72 [95% CI 0.70-0.74]) when node was considered as unit of analysis (P<0.05); MRI had a higher SPE (0.81 [95% CI 0.80-0.82]) than CT (0.72 [95% CI 0.69-0.74]) when neck level was considered as unit of analysis (P<0.05) and MRI had a higher area under concentration-time curve than CT when the patient was considered as unit of analysis (P<0.05). With regards to diagnostic criteria, for MRI, the results showed that the minimal axial diameter of 10 mm could be considered as the best size criterion, compared to 12 mm for CT. Overall, MRI conferred significantly higher SPE while CT demonstrated higher SEN. The diagnostic criteria for MRI and CT on size of metastatic lymph nodes were suggested as 10 and 12 mm, respectively.

[Short course radiotherapy in malignant lymphoma].

From 1979 to 1983, 32 cases of malignant lymphoma were treated by short course radiotherapy. 60Co teletherapy or conventional X-ray (H.V.L. 0.95 mm Cu) in a daily skin dose of 450 rad was given. In order to avoid radiation injury to the pharynx and spinal cord, lymph nodes on both sides of the neck were first irradiated by 60Co through tangential anterior fields, then by perpendicular conventional irradiation. Peak radiation reaction of skin or tumor was seen in 2-3 weeks. The larger the tumor, the slower was the complete regression. The 5 year survival rate was 52.2% and local recurrence rate was 6.2%. The short course radiotherapy could be used as a salvage treatment to residual tumor or resistant lesions after chemotherapy. It is used not only in advanced disease but also in early lesion. The short course irradiation, improving the treatment result of the advanced lymphomas and prolonging the survival, is capable of achieving more than the conventional course. The short course radiotherapy, being short in duration and low in cost, is advised.

Embryo transfer and sex determination following superovulated hinds inseminated with frozen-thawed sex-sorted Y sperm or unsorted semen in Wapiti (Cervus elaphus songaricus).

The aim of this study was to evaluate embryo production in superovulated wapiti hinds inseminated with either Y-sorted or unsorted semen. Eighteen hinds were allocated to three treatment groups: AI following multiple ovulation (CIDR/FSH) with 10×10(6) Y-sorted frozen-thawed semen (Y group, n=6), or 10×10(6) and 100×10(6) unsorted frozen-thawed semen for the unsorted (n=6) and the control group (n=6). The embryos from the sixth day following insemination were collected and classified. Fifteen embryos from the unsorted or the control group, and four embryos from the Y group were sex determinated based on DNA analysis of the amelogenin gene. Twenty-one embryos from the Y group and 42 embryos from the unsorted or the control group were transferred into 21 and 42 synchronized recipients via standard procedures on 6th day post estrus, respectively. There were no significant differences in the number of recovered eggs, transferable embryos, degenerated embryos or unfertilized oocytes per hind among the three groups of the control (9.2±3.6, 4.7±1.9, 3.0±2.0, 1.5±1.4), the unsorted (8.2±1.9, 4.8±0.7, 1.7±1.0, 1.7±1.0) and the Y group (8.8±4.2, 4.2±1.8, 2.2±1.2, 2.5±2.1), respectively (P>0.05). The sex ratio of embryos from the Y group (4M/0F) was significantly (P<0.05) distinct from that of the unsorted and control group (8M/7F). The sex ratio of the offspring from sexed embryos (8M/0F) was deviated significantly (P<0.05) from that of the non-sexed embryos (11M/9F). In conclusion, the results suggested that the male embryos of predicted sex can be achieved with AI of sex-sorted cryopreserved sperm. PCR amplification using the amelogenin gene primers can be applied to DNA analysis of micro samples from wapiti embryo biopsies for sex identification. The male offspring can be produced after transferred with the male embryos of predicted sex.

Successful low dose insemination of flow cytometrically sorted Sika (Cervus nippon) sperm in Wapiti (Cervus elaphus).

The purpose of this study was to determine a practical method in Wapiti (Cervus elaphus) of using predetermined sexed Sika (Cervus nippon) semen. Semen was collected by electro-ejaculation from one stag of proven fertility and transported to the laboratory where it was retained as unsorted (control) or was separated into X- and Y-chromosome-bearing sperm using a modified high-speed cell sorter. Wapiti hinds (n=81) were inseminated into the uterus by rectum manipulation with 1 x 10(6) (X1 and Y1 group, respectively) or 2 x 10(6) (X2 and Y2 group, respectively) of sorted frozen-thawed and 1 x 10(7) non-sorted frozen-thawed (a commercial dose control) Sika motile sperm 60-66h after removal of intra-vaginal progesterone-impregnated CIDR devices and administration of 700IU of PMSG at the time of CIDR removal. The percentage of hinds calving after insemination was similar for X1 (38.5%), X2 (41.7%), Y1 (44.4%), Y2 (38.9%) groups (P>0.05), but higher for control (75%) treatment (P<0.05). Ultimately 15 out of the 16 Sika and Wapiti-hybrid calves produced by Wapiti hinds inseminated with Y-sorted sperm were male (93.7%) and 10/10 (100%) Sika and Wapiti-hybrid calves from hinds inseminated with X-sorted sperm were female. The sex ratio of the Sika and Wapiti-hybrid calves born to hinds inseminated with sex-sorted sperm deviated significantly (P<0.05) from 50% and 50.0% in the control group. All Sika and Wapiti-hybrid calves were born between 237 and 250d of gestation. Male and female calves in the control group had similar birth weights and weaning weights as calves from hinds inseminated with X- or Y-sorted sperm. In conclusion it can be said that normal Sika and Wapiti-hybrid calves of predicted sex can be produced after artificial insemination of Wapiti does with low numbers of sex-sorted cryopreserved Sika sperm.

The alteration of Id-1 and TSP-1 expression in mucoepidermoid carcinoma associated with its clinical features and prognosis.

Expression of Id-1 (inhibitors of DNA binding/differentiation protein 1) and TSP-1 (thrombospondin-1) in mucoepidermoid carcinoma and their relationship to pathological features and prognosis was studied. Moderately and poorly differentiated groups had significantly higher Id-1 positive expression rate (p<0.05) than well differentiated carcinoma. Stages III-IV showed significant increase of Id-1 positive expression rate (p<0.05) compared with stages I and II. Id-1 positive expression was significantly higher in patients with cervical lymph node metastasis or relapse at 5 years (p<0.05). After that, patients with negative Id-1 expression had significantly higher tumor-free survival than patients with positive expression (p<0.05). Correlation between the expression of Id-1 and TSP-1 in mucoepidermoid carcinoma was negative (p<0.05). Poorly differentiated groups show significantly lower TSP-1 positive expression rate than well differentiated groups (p<0.05). No significant differences of TSP-1 positive expression were detected with clinical stage. TSP-1 positive expression was significantly lower in patients with cervical lymph node metastasis or relapse at 5 years (p<0.05). After 5 years, patients with positive TSP-1 expression had significantly higher tumor-free survival than patients with negative TSP-1. Positive Id-1 expression is associated with high malignancy/poor prognosis; positive TSP-1 expression is associated with low malignancy/good prognosis. Protein expression status may help assess tumor malignancy and patient prognosis.

Separation, cultivation and biological characteristics of oral carcinoma-associated fibroblasts.

OBJECTIVES: Carcinoma-associated fibroblasts (CAFs) have been suggested to regulate the initiation and progression of many types of solid tumors. The aim of the study was to separate, cultivate, identify oral CAFs, and to investigate their biological characteristics. MATERIALS AND METHODS: The primary CAFs and normal fibroblasts (NFs) of the tongue were obtained by tissue culture. Then cells were dissociated by 0.25% trypsin and purified by curettage method combining with trypsinization. The cells were verified according to morphological observation and immunohistochemical staining of certain proteins. Multiple proliferation indexes and karyotype of the cells were assayed. RESULTS: Third passage purified oral CAFs and NFs were attained successfully. The morphological characteristics of the CAFs changed significantly comparing to the NFs. The CAFs showed positive staining for vimentin, alpha-smooth muscle actin and matrix metalloproteinases-2. The proliferation and mitosis ability of the CAFs were significantly increased compared with the NFs (P < 0.05). No karyotypic abnormalities were found in the CAFs. CONCLUSIONS: There were obvious differences in the biological characteristics between oral CAFs and NFs. The results may provide us an experimental foundation for further studies on the roles of CAFs in the initiation and progression of oral cancer.