Heavy metal ions exchange driven protein phosphorylation cascade functions in genomic instability in spermatocytes and male infertility.

DNA double-strand breaks (DSBs) are functionally linked to genomic instability in spermatocytes and to male infertility. The heavy metal cadmium (Cd) is known to induce DNA damage in spermatocytes by unknown mechanisms. Here, we showed that Cd ions impaired the canonical non-homologous end-joining (NHEJ) repair pathway, but not the homologous recombination (HR) repair pathway, through stimulation of Ser2056 and Thr2609 phosphorylation of DNA-PKcs at DSB sites. Hyper-phosphorylation of DNA-PKcs led to its premature dissociation from DNA ends and the Ku complex, preventing recruitment of processing enzymes and further ligation of DNA ends. Specifically, this cascade was initiated by the loss of PP5 phosphatase activity, which results from the dissociation of PP5 from its activating ions (Mn), that is antagonized by Cd ions through a competitive mechanism. In accordance, in a mouse model Cd-induced genomic instability and consequential male reproductive dysfunction were effectively reversed by a high dosage of Mn ions. Together, our findings corroborate a protein phosphorylation-mediated genomic instability pathway in spermatocytes that is triggered by exchange of heavy metal ions.

SNP rs12794714 of CYP2R1 is associated with serum vitamin D levels and recurrent spontaneous abortion (RSA): a case-control study.

PURPOSE: Vitamin D (VD) deficiency seems to be associated with the risk of recurrent spontaneous abortion (RSA). Vitamin D receptor (VDR) and cytochrome P450 family 2 subfamily R member 1 (CYP2R1) are two genes which are vital for VD metabolism and actions. However, whether single-nucleotide polymorphisms (SNPs) in these genes are correlated with the risk of RSA are poorly understood. Therefore, we aimed to characterize the relationships among VDR SNPs, CYP2R1 SNPs and RSA. METHODS: This case-control study enrolled 75 RSA patients and 83 controls. Serum VD and some cytokines were detected with LC-MS/MS and flow cytometry, respectively. Genotyping for three SNPs of CYP2R1 (rs10741657, rs10766197 and rs12794714) and five SNPs of VDR (rs7975232, rs1544410, rs2189480, rs2228570 and rs2239179) was done with polymerase chain reaction (PCR) and high-throughput sequencing. All the data were analyzed with appropriate methods and in different models. RESULTS: The results revealed a significant correlation between the AG genotype of CYP2R1 rs12794714 and VD levels (OR 0.686; 95% CI 0.49-0.96; p = 0.028). Besides, the AG and GG genotypes of CYP2R1 rs12794714 were markedly related to the risk of RSA (OR 52.394, 59.497; 95% CI 2.683-1023.265, 3.110-1138.367; p = 0.009, 0.007, respectively). CONCLUSION: Our results indicate that CYP2R1 rs12794714 might be a risk factor for RSA. Hence, early screening of pregnant women for CYP2R1 rs12794714 is necessary to warrant proactive counseling and treatment against RSA.

Extraction and Characterization of Flaxseed Oil Obtained with Subcritical n-Butane.

In this study, subcritical n-butane was adopted to extract oil from flaxseed. The extraction conditions i.e. extraction temperature, extraction time, and liquid-solid ratio were investigated and optimized by response surface methodology. The flaxseed oil obtained by subcritical n-butane were characterized and compared with those prepared by n-hexane and cold pressing. Results indicated that the optimal combination of parameters was 53.93℃, 56.82 min, and 19.98:1 mL/g. Subcritical n-butane had higher yield (28.75%) than n-hexane and cold pressing. GC analysis indicated that subcritical n-butane extraction had no obvious influence on the fatty acid composition. Nevertheless, the oil obtained by subcritical n-butane with higher contents of phytosterols (2.93 mg/g) and carotenoids (46.56 mg/kg), and presented a higher oxidation stability (9.27 h). Thus, it was suggested that subcritical n-butane extraction is a promising alternative to extract high quality flaxseed oil.

[An analysis on chromosome X, Y and 18 in the spermatozoa of asthenospermia patients by triple-color fluorescence in situ hybridization].

OBJECTIVE: To analyze the numerical aberration of chromosome X, Y and 18 in the spermatozoa of asthenospermia patients by triple-color fluorescence in situ hybridization. METHODS: The experiment included 10 asthenospermia patients and 5 healthy men with normal semen quality as controls. Fluorescence in situ hybridization (FISH) and probes for chromosomes including X, Y and 18 were used to determine the frequency of the aneuploid of the chromosomes in spermatozoa. RESULTS: Of the 45,547 spermatozoa counted from the semen samples, the hybridization rate was 99.18%. The frequencies of the chromosome disomies including XX18, XY18, YY18, X1818 and Y1818 were (0.124 +/- -0.086)%, (0.360 +/- 0.380)%, (0.109 +/- 0.195)%, (0.342 +/- 0.746)% and (0.299 +/- 0.564)% in the case group and (0.014 +/- 0.019)%, (0.090 +/- 0.080)%, (0.030 +/- 0.031)%, (0.068 +/- 0.103)% and (0.075 +/- 0.083)% in the control. The sperm aneuploid rate was 9.25% in the former and 2.70% in the latter, with significant difference in between (P< 0.01). CONCLUSION: Asthenospermia patients have a higher aneuploid rate of sperm chromosome than normal fertile men. However, larger samples are yet to be studied to obtain more scientific evidence.

[Role of cerebral computed tomography in the evaluation of brain injury following hypoxia in neonates].

OBJECTIVE: To investigate the role of cerebral computed tomography (CT) in the evaluation of the severity of brain injury following hypoxia in neonates. METHODS: A total of 114 full-term newborns who had perinatal hypoxia, including 25 cases of hypoxic-ischemic encephalopathy (HIE), 36 cases of neonatal asphyxia and 53 cases of simple intrauterine fetal distress, were enrolled in this study. Twenty normal newborns served as the Control group. All had cerebral CT scan at 2-7 days of age. Neonatal behavior neurological assessment (NBNA) was performed at 5 days of age. RESULTS: The average NBNA scores were significantly lower and the abnormality rate of NBNA was significantly higher in the HIE group than in the other three groups (P < 0.05). The Asphyxia and the Distress groups had also lower NBNA scores and higher abnormality rate of NBNA than the Control group (P < 0.05). Twenty-two patients were found to have cerebral CT abnormality in the HIE group, but there was only 1 case in the Control group (P < 0.01). The abnormality rate of cerebral CT in the Asphyxia and the Distress groups was not statistically different from that of the Control group. Twenty-five cases of HIE were divided into mild (n=15), medium (n=6) and severe (n=4) by clinical grading but were divided into normal (n=3), mild (n=10), medium (n=7) and severe (n=5) by CT grading. CT and clinical grading on HIE was not consistent. The sensitivity of CT in the diagnosis of mild, moderate and severe HIE was 47%, 33% and 50% respectively, the specificity was 70%, 74% and 86% respectively and the accuracy was 48%, 64% and 80% respectively. CONCLUSIONS: CT evaluation on mild brain injury induced by asphyxia or intrauterine fetal distress is not of any value and the role of CT evaluation on the HIE grade is uncertain and doubtful.

[Archival tags and geolocation methods for marine animals: A review].

Archival tags, a group of data storable electronic tags, are widely used as strong tools for obtaining long term and large scale activity information of marine animals, specifically highly migratory oceanic fishes, and corresponding environmental data. Though retrieving tags is an indispensable step for obtaining data, which is a shortage of archival tags, a series of achievements have been made on marine animals by using archival tags since the 1990s. With the appearance of pop-up satellite tag, which solved the problem of data retrieving and was fully independent of the fishing, both breadth and depth of marine animals' studies are extended by the end of the 1990s. Geolocation based on light intensity is the key to estimate marine animals' movement and has achieved some progress in the past 20 years. However, the accuracy of geolocation for latitude is not high enough, and there is still much room for improvement. To date, most geolocation methods that use ambient daylight involve identifying the times when the sun is at a precisely known zenith angle (e.g., sunrise and sunset). The problem of estimating longitude has been proved easy to solve, but accurate latitude estimates remain elusive. This paper mainly introduced two tags, i. e., archival tags and pop-up tags, and three geolocation methods, i.e. , 1) the "fixed reference" method, 2) the "variable reference" method, and 3) the "reflection" method. We also presented a prospect analysis on archival tags and possible research direction of geolocation methods. We believed that miniaturization and multi-sensor integration are the trends for electronic tags while more environmental factors such as depth, SST (sea surface temperature) or magnetic field intensity, instead of single factor, as auxiliary parameters would be used for improving the geolocation accuracy in the future.