Using transvaginal ultrasonography and MRI to evaluate ovarian volume and follicle count of infertile women: a comparative study.

AIM: To compare two-dimensional (2D) transvaginal ultrasonography (TVUS) and 2D/three-dimensional (3D) magnetic resonance imaging (MRI) in estimating ovarian volume and follicle count. MATERIALS AND METHODS: The ovarian volume (OV) and follicle count (FC) of 84 women with infertility were evaluated by 2D TVUS and 2D/3D MRI. Bland-Altman analysis was used for comparison. RESULTS: The OV from 3D MRI was 0.50 ml (95% confidence interval [CI], 0.25-0.74, p<0.001) smaller than that by 2D TVUS. OV from 2D MRI was 2.65 ml (95% CI, 2.36-2.95, p<0.001) and 3.15 ml (95% CI, 2.77-3.53, p<0.001) smaller than that from 3D MRI and 2D TVUS, respectively. The FC1-9 mm and total follicle count (tFC) estimated by 2D TVUS were 7.81 (95% CI, 6.96-8.66, p<0.001) and 7.82 (95% CI, 6.97-8.67) smaller than those from 2D MRI, respectively. Further analysis showed that 2D TVUS detected lower FC1-3 mm but higher FC4-6 mm than 2D MRI. No significant difference was shown in the results of FC7-9 mm and FC ≥ 10 mm. CONCLUSION: In women with infertility, 2D MRI underestimated OV as compared with 2D TVUS. OV from 3D MRI was lower but very close to that from 2D TVUS. For patients unsuitable for TVUS, 3D MRI is recommended for OV evaluation. 2D TVUS underestimated FC1-9 mm and tFC compared with 2D MRI. In fertility counselling and research, 2D MRI is a useful alternative to TVUS when an accurate FC is needed.

17ß-Estradiol improves osteoblastic cell function through the Sirt1/NF-κB/MMP-8 pathway.

Objective: This study aims to investigate the beneficial effects of 17ß-estradiol supplementation on the function of osteoblastic cells through the Sirtuin-1/nuclear transcription factor-κB/matrix metalloproteinase-8 (Sirt1/NF-κB/MMP-8) pathway.Methods: Mouse primary osteoblasts were obtained from neonatal mouse calvaria, and the cells were treated with or without 17ß-estradiol. We first detected the effect of 17ß-estradiol on the function of osteoblastic cells. Then, the changes in estrogen receptor-α (ERα), Sirt1, NF-κB, and MMP-8 were determined after the osteoblasts were treated with 17ß-estradiol. During supplementation with 17ß-estradiol, knockdown of Sirt1 in osteoblasts was used to further measure the changes of NF-κB and MMP-8 and observe the cell function.Results: In primary osteoblastic cells, exposure to 17ß-estradiol improved cell viability and increased the levels of bone formation biomarkers, including osteocalcin, osteoprotegerin (OPG), procollagen type 1 N-terminal propeptide (P1NP), and alkaline phosphatase (ALP). In addition, 17ß-estradiol supplement activated ERα and Sirt1 expression and inhibited NF-κB and MMP-8 expression. Moreover, these effects induced by 17ß-estradiol were reversed by knockdown of Sirt1 in mouse primary osteoblasts.Conclusion: 17ß-Estradiol replacement therapy may treat postmenopausal osteoporosis by improving osteoblastic cell function via the Sirt1/NF-κB/MMP-8 pathway.

Immune efficacy of a porcine circovirus type 2 vaccine purified using Gram-positive enhancer matrix surface display technology.

AIMS: Purification of porcine circovirus type 2 (PCV2) using Gram-positive enhancer matrix (GEM) surface display technology and immunogenicity evaluation of the purified antigen. METHODS AND RESULTS: A recombinant bifunctional protein containing a protein anchor domain and a 'virus anchor' domain was designed as a protein linker (PL) between PCV2 and GEM particles. By incubating with PL and GEM particles sequentially, PCV2 could be purified and enriched through a simple centrifugation process with GEM surface display technology. Our data showed that one unit (2·5 × 109 particles) of GEM particles with 80 µg PL could purify 100 ml of PCV2-containing culture supernatant (viral titre: 106·5 TCID50 per ml-1 ) with a recovery rate up to 99·6%. The impurity removal efficiency of this method, calculated according to decreased total protein content during purification, was approximately 98%. Furthermore, in vivo experimentation showed that piglets immunized with purified PCV2 could elicit strong immune responses to prevent against PCV2 infection. CONCLUSION: Porcine circovirus type 2 could be efficiently purified and enriched with GEM display technology via a crucial PL, and the purified PCV2 could elicit effective immune responses against PCV2 infection. SIGNIFICANCE AND IMPACT OF THE STUDY: The GEM-based purification method established here is cost-efficient and high-throughput, and may represent a promising large-scale purification method for PCV2 vaccine production.

Tempo and mode of recurrent polyploidization in the Carassius auratus species complex (Cypriniformes, Cyprinidae).

Polyploidization is an evolutionarily rare but important mechanism in both plants and animals because it increases genetic diversity. Goldfish of the Carassius auratus species complex can be tetraploids, hexaploids and octaploids. Polyploidization events have occurred repeatedly in goldfish, yet the extent of this phenomenon and its phyletic history are poorly understood. We explore the origin, tempo and frequency of polyploidization in Chinese and Japanese goldfish using both mitochondrial (mtDNA) and nuclear DNA sequences from up to 1202 individuals including the outgroup taxon, Cyprinus carpio. Analyses of de novo nuclear gene data resolve two clusters of alleles and the pattern supports the prior hypothesis of an ancient allotetraploidization for Carassius. Alleles shared by tetraploid and hexaploid individuals indicate recent autoploidizations within the C. auratus complex. Sympatric tetraploids and hexaploids share mtDNA haplotypes and these frequently occur independently within six well-supported lineages and sublineages on a small spatial scale. Gene flow estimates (Fst values) indicate that hexaploids differ only slightly from sympatric tetraploids, if at all. In contrast, allopatric populations of tetraploids and hexaploids differ from one another to a far greater extent. Gene flow between sampled localities appears to be limited. Coalescence-based time estimations for hexaploids reveal that the oldest lineage within any sampled locality is around one million years old, which is very young. Sympatric, recurrent autoploidization occurs in all sampled populations of the C. auratus complex. Goldfish experience polyploidization events more frequently than any other vertebrate.

A new high-frequency allele of the BM2113 locus in the Yunnan mithun population.

The BM2113 locus was amplified in Yunnan mithun (Bos frontalis) from the southwest mountains of China. It showed a high degree of polymorphism with a total of 12 alleles. The 121-bp polymorphic allele of the BM2113 locus that accounted for 37.1% of homozygotes was the predominant allele with a frequency of 58.57%, identified as mithun-specific for Bos species in Yunnan mithun. The polymorphism information content value was high with a mean of 0.6170, the expected and observed heterozygosity was moderate with values of 0.6427 and 0.6000, respectively, and the BM2113 locus was under Hardy-Weinberg equilibrium (P = 0.2897) in the Yunnan mithun population. This study elucidated the genetic diversity, multi-origin, specific alleles, and characterization of mithun.

Formation of and countermeasures for subacute coronary stent thrombosis in elderly diabetic patients.

The aim of this study was to evaluate and investigate the pathogenetic mechanism and countermeasures of subacute thrombosis (SAT) after coronary stenting in elderly diabetic patients. The clinical characteristics and pathogenetic mechanisms in 3 cases of SAT after stent implantations in elderly diabetic patients were retrospectively examined to determine the treatment strategies for SAT. Among 98 patients with diabetes who had coronary stents implanted or were >60 years of age, three (3.06%) had SAT. One case of SAT was diagnosed by angiography; coronary balloon dilatation, thrombolysis, and re-perfusion resulted in full recovery in this case. The second case involved potential SAT, and in the third case, SAT was not ruled out. Two cases were characteristic of ST-segment elevation myocardial infarction, and one case, in which SAT was not ruled out, resulted in sudden death. SAT within a stent may be related to intraoperative stent malapposition caused by a grade C lesion, age, diabetes, chronic total occlusion, or postoperative irregular administration of medication.

Chicken domestication: an updated perspective based on mitochondrial genomes.

Domestic chickens (Gallus gallus domesticus) fulfill various roles ranging from food and entertainment to religion and ornamentation. To survey its genetic diversity and trace the history of domestication, we investigated a total of 4938 mitochondrial DNA (mtDNA) fragments including 2843 previously published and 2095 de novo units from 2044 domestic chickens and 51 red junglefowl (Gallus gallus). To obtain the highest possible level of molecular resolution, 50 representative samples were further selected for total mtDNA genome sequencing. A fine-gained mtDNA phylogeny was investigated by defining haplogroups A-I and W-Z. Common haplogroups A-G were shared by domestic chickens and red junglefowl. Rare haplogroups H-I and W-Z were specific to domestic chickens and red junglefowl, respectively. We re-evaluated the global mtDNA profiles of chickens. The geographic distribution for each of major haplogroups was examined. Our results revealed new complexities of history in chicken domestication because in the phylogeny lineages from the red junglefowl were mingled with those of the domestic chickens. Several local domestication events in South Asia, Southwest China and Southeast Asia were identified. The assessment of chicken mtDNA data also facilitated our understanding about the Austronesian settlement in the Pacific.

Two-year caries clinical study of the efficacy of novel dentifrices containing 1.5% arginine, an insoluble calcium compound and 1,450 ppm fluoride.

A 2-year double-blind randomized three-treatment controlled parallel-group clinical study compared the anti-caries efficacy of two dentifrices containing 1.5% arginine, an insoluble calcium compound (di-calcium phosphate or calcium carbonate) and 1,450 ppm fluoride (F), as sodium monofluorophosphate, to a control dentifrice containing 1,450 ppm F, as sodium fluoride, in a silica base. The 6,000 participants were from Bangkok, Thailand and aged 6-12 years initially. They were instructed to brush twice daily, in the morning and evening, with their randomly assigned dentifrice. Three trained and calibrated dentists examined the children at baseline and after 1 and 2 years using the National Institute of Dental Research Diagnostic Procedures and Criteria. The number of decayed, missing and filled teeth (DMFT) and surfaces (DMFS) for the three study groups were very similar at baseline, with no statistically significant differences among groups. After 1 year, there were no statistically significant differences in caries increments among the three groups. After 2 years, the two groups using the dentifrices containing 1.5% arginine, an insoluble calcium compound and 1,450 ppm F had statistically significantly (p < 0.02) lower DMFT increments (21.0 and 17.7% reductions, respectively) and DMFS increments (16.5 and 16.5%) compared to the control dentifrice. The differences between the two groups using the new dentifrices were not statistically significant. The results of this pivotal clinical study support the conclusion that dentifrices containing 1.5% arginine, an insoluble calcium compound and 1,450 ppm F provide significantly greater protection against caries lesion cavitation, in a low to moderate caries risk population, than dentifrices containing 1,450 ppm F alone.

Origins of domestic dog in southern East Asia is supported by analysis of Y-chromosome DNA.

Global mitochondrial DNA (mtDNA) data indicates that the dog originates from domestication of wolf in Asia South of Yangtze River (ASY), with minor genetic contributions from dog-wolf hybridisation elsewhere. Archaeological data and autosomal single nucleotide polymorphism data have instead suggested that dogs originate from Europe and/or South West Asia but, because these datasets lack data from ASY, evidence pointing to ASY may have been overlooked. Analyses of additional markers for global datasets, including ASY, are therefore necessary to test if mtDNA phylogeography reflects the actual dog history and not merely stochastic events or selection. Here, we analyse 14,437 bp of Y-chromosome DNA sequence in 151 dogs sampled worldwide. We found 28 haplotypes distributed in five haplogroups. Two haplogroups were universally shared and included three haplotypes carried by 46% of all dogs, but two other haplogroups were primarily restricted to East Asia. Highest genetic diversity and virtually complete phylogenetic coverage was found within ASY. The 151 dogs were estimated to originate from 13-24 wolf founders, but there was no indication of post-domestication dog-wolf hybridisations. Thus, Y-chromosome and mtDNA data give strikingly similar pictures of dog phylogeography, most importantly that roughly 50% of the gene pools are shared universally but only ASY has nearly the full range of genetic diversity, such that the gene pools in all other regions may derive from ASY. This corroborates that ASY was the principal, and possibly sole region of wolf domestication, that a large number of wolves were domesticated, and that subsequent dog-wolf hybridisation contributed modestly to the dog gene pool.

Karyotype analysis of mithun (Bos frontalis) and mithun bull x Brahman cow hybrids.

We examined the cytogenetics of mithun (Bos frontalis), a domesticated version of the Asian gaur, and hybrids (F(1) generation) produced by artificial insemination of Brahman cows (Bos indicus) with mithun semen. Reproductive potential was also examined in the F(1) generation and a backcrossed heifer for utilization of heterosis. Metaphase chromosome spreads were examined by conventional staining and fluorescence in situ hybridization hybridized with the entire chromosome 1 of mithun as a specific probe. Chromosome 1 of mithun was found to be equivalent to Bos taurus chromosomes 2 and 28. The karyotype of the female mithun (N = 4) comprised 58 chromosomes, including 54 acrocentric and four large submetacentric chromosomes, without the four acrocentric chromosomes found in the domesticated species B. indicus. However, one of the four female mithuns with a normal mithun phenotype had an abnormal karyotype (2n = 59), indicating introgression from B. taurus or B. indicus. The F(1) karyotypes (N = 6, 3♂3♀) of the mithun bull × Brahman cow cross had 2n = 59, intermediate between their parents; they were consistent heterozygous carriers with a centric fusion involving rob(2;28), as expected. Two pronounced red signals were seen in the mithun karyotypes, three red signals in the mithun × Brahman hybrids, and four red signals in the Brahman cattle, in good agreement with centric fusion of bovine rob(2;28). The female backcross hybrid (N = 1) with 2n = 59 had a similar chromosome configuration to the F(1) karyotypes and had rob(2;28). Such female backcross hybrids normally reproduce; however, the F(1) bulls (N = 3) had not yet generated normal sperm at 24 months.

Clinicopathologic and prognostic significance of long non-coding RNA myocardial infarction-associated transcript in multiple cancers.

OBJECTIVE: The aim of the meta-analysis was to explore the clinicopathological and prognostic significance of long non-coding RNA (lncRNA) myocardial infarction-associated transcript (MIAT) in various cancers. MATERIALS AND METHODS: We searched multiple databases, including PubMed, China National Knowledge 53 Infrastructure (CNKI), Springer, Web of Science, and Cochrane, for articles on the prognostic value of lncRNA MIAT in various cancers before 25 March 2021. The odds ratio (OR) and 95% confidence interval (CI) were adopted to evaluate the clinicopathological features and outcomes of cancers. The Cancer Genome Atlas dataset was used to identify the differential expression and prognostic significance of lncRNA MIAT. RESULTS: We enrolled 14 publications, including 1,573 cancer patients. Higher lncRNA MIAT expression was significantly related to worse overall survival (OR=3.13, 95% CI: 2.47-3.96, p<0.05), regardless of cancer types, sample size, and follow-up time of the included studies. Additionally, higher lncRNA MIAT expression was associated with larger tumour sizes (OR=1.67, 95% CI: 1.24-2.26, p<0.05), advanced clinical stage (OR=4.79, 95% CI: 3.38-6.79, p<0.05), lymph nodes metastasis (OR=7.33, 95% CI: 4.61-11.67, p<0.05), and distant metastasis (OR=2.62, 95% CI: 1.88-3.66, p<0.05), but not associated with age and gender. We found no publication bias, and sensitivity analysis indicated that the results were reliable. CONCLUSIONS: Higher lncRNA MIAT expression may predict larger tumour sizes, advanced clinical stage, metastasis of cancers, and lower overall survival rate. LncRNA MIAT may serve as a useful clinicopathological and prognostic biomarker for cancers.

Replication study of candidate genes/loci associated with osteoporosis based on genome-wide screening.

UNLABELLED: Osteoporosis is a major public health problem characterized by low bone mineral density (BMD). This replication study confirmed 38 single-nucleotide polymorphisms (SNPs) out of 139 SNPs previously reported in three recent genome-wide association studies (GWASs) in an independent US white sample. Ten SNPs achieved combined p < 3.6 x 10(-4). INTRODUCTION: BMD is under strong genetic control. This study aims to verify the potential associations between BMD and candidate genes/loci reported by GWAS of FHS100K, Icelandic deCODE, and UK-NL. METHODS: Eight promising (at the genome-wide significant level after Bonferroni correction) and 131 available sub-promising (at the most stringent p value, p < 5.5 x 10(-5) in the three GWASs reports) SNPs were selected. By using genotypic information from Affymetrix 500 K SNP arrays, we tested their associations with BMD in 1,000 unrelated US whites. Fisher's combined probability method was used to quantify the overall evidence of association. BMD was measured by dual energy X-ray absorptiometry. RESULTS: Two promising SNPs, rs3762397 and rs3736228, were replicated in the current study with p < 0.05. Besides, 36 sub-promising SNPs were replicated at the same significant level. Ten SNPs achieved significant combined p < 3.6 x 10(-4) (0.05/139 SNPs, corrected for multiple testing). CONCLUSIONS: Osteoporosis susceptibility of 38 SNPs was replicated in 1,000 unrelated US whites. This study showed promise for replication of some initial genome-wide association signals.

Genome-wide association analyses suggested a novel mechanism for smoking behavior regulated by IL15.

Cigarette smoking is the leading preventable cause of death in the United States. Although smoking behavior has a significant genetic determination, the specific genes and associated mechanisms underlying the smoking behavior are largely unknown. Here, we carried out a genome-wide association study on smoking behavior in 840 Caucasians, including 417 males and 423 females, in which we examined approximately 380,000 single nucleotide polymorphisms (SNPs). We found that a cluster of nine SNPs upstream from the IL15 gene were associated with smoking status in males, with the most significant SNP, rs4956302, achieving a P-value (8.80 x 10(-8)) of genome-wide significance. Another SNP, rs17354547 that is highly conserved across multiple species achieved a P-value of 5.65 x 10(-5). These two SNPs, together with two additional SNPs (rs1402812 and rs4956396) were selected from the above nine SNPs for replication in an African-American sample containing 1251 subjects, including 412 males and 839 females. The SNP rs17354547 was replicated successfully in the male subgroup of the replication sample; it was associated with smoking quantity (SQ), the Heaviness of Smoking Index (HSI) and the Fagerstrom Test for Nicotine Dependence (FTND), with P-values of 0.031, 0.0046 and 0.019, respectively. In addition, a haplotype formed by rs17354547, rs1402812 and rs4956396 was also associated with SQ, HSI and FTND, achieving P-values of 0.039, 0.0093 and 0.0093, respectively. To further confirm our findings, we carried out an in silico replication study of the nine SNPs in a Framingham Heart Study sample containing 7623 Caucasians from 1731 families, among which, 3491 subjects were males and 4132 were females. Again, the male-specific association with smoking status was observed, for which seven of the nine SNPs achieved significant P-values (P<0.05) and two achieved marginally significant P-values (P<0.10) in males. Several of the nine SNPs, including the highly conserved one across species, rs17354547, are located at potential transcription factor binding sites, suggesting transcription regulation as a possible function for these SNPs. Through this function, the SNPs may modulate the gene expression of IL15, a key cytokine regulating immune function. As the immune system has long been recognized to influence drug addiction behavior, our association findings suggest a novel mechanism for smoking addiction involving immune modulation through the IL15 pathway.

Artificial selection of the melanocortin receptor 1 gene in Chinese domestic pigs during domestication.

Black coat colour is common in Chinese indigenous domestic pigs, but not among their wild ancestors, and it is thus presumed to be a 'domestication trait.' To determine whether artificial interference contributes to morphological diversification, we examined nucleotide variation from 157 Chinese domestic pigs and 40 wild boars in the melanocortin receptor 1 (MC1R) gene, which has a key role in the coat pigmentation of Sus scrofa. Compared with a pseudogene GPIP, our results showed that the joint effects of demography and selection have resulted in markedly low genetic diversity of MC1R in Chinese domestic pigs. Coalescent simulation and selection tests further suggest that the fixation of two non-synonymous substitutions associated with black colour is the result of artificial selection. In contrast, a much higher genetic diversity and only a single non-synonymous substitution were found among the wild boars, suggesting a strong functional constraint. Moreover, our conclusion is consistent with the preference for black colour in the ancient Chinese sacrificial culture. This case provides an interesting example of a molecular evaluation of artificial livestock selection and its associated cultural impact in ancient China.

Gene polymorphisms of SOCS1 and SOCS2 and acute lymphoblastic leukemia.

OBJECTIVE: Acute lymphoblastic leukemia (ALL) causes the dysfunction of the systemic blood system and immune system. The etiology and predisposing factors of ALL are unknown. The suppressor of cytokine signaling 1 (SOCS1) and SOCS2 are inhibitors of cytokine signal transduction. Gene polymorphisms of SOCS1 and SOCS2 and their expressions may be related to ALL. PATIENTS AND METHODS: A total of 200 ALL patients in our hospital and 200 healthy people were enrolled in ALL group and control group, respectively. Genomic deoxyribonucleic acids (DNAs) and total RNAs were extracted from the peripheral blood of each subject. Gene polymorphisms of SOCS1 at rs33977706, rs243327, and rs33932899 and those of SOCS2 at rs3816997 were amplified by polymerase chain reaction (PCR) and sequenced. Besides, the expression levels of SOCS1 and SOCS2 in ALL patients were detected by real-time fluorescence quantitative PCR. RESULTS: The frequency of the allele C of SOCS1 rs33977706 in ALL group was lower than that in the control group, displaying a significant difference between the two groups (p=0.015). The frequency of allele A of SOCS2 rs3816997 was notably higher in ALL group than that of the control group (p=0.000). In addition, the frequency of CA genotype of SOCS1 rs33977706 in ALL group was markedly lower than that in the control group, showing a significant difference (p=0.000). ALL group had remarkably higher frequencies of AA genotype of SOCS2 rs3816997 (p=0.000) and ACC haplotype of SOCS gene (p=0.000), and lower frequencies of ATG (p=0.026) and CCC (p=0.006) haplotypes. The two loci, SOCS1 rs33932899 and SOCS1 rs243327, were linked to each other (D'=0.781). Moreover, the expression level of SOCS1 in ALL group was lower than that in the control group, in which the expression of the CT genotype of SOCS1 rs243327 was relatively higher (p=0.021). SOCS2 level was lower in ALL group. Particularly, SOCS2 level in ALL patients carrying AC genotype was lower than those carrying AA and CC genotypes (p=0.000). ALL patients carrying CT genotype of SOCS1 rs243327 had shorter period of agranulocytosis (p=0.000), a lower ratio of bone marrow primitive/immature cells (p=0.001), and a higher hemoglobin (Hb) level in blood (p=0.000). The ratio of bone marrow primordial/immature cells was lower in ALL patients with AC genotype of SOCS2 rs3816997 (p=0.038). CONCLUSIONS: The expression levels of SOCS1 and SOCS2 are prominently related to ALL, and their polymorphisms are associated with the susceptibility to ALL.

MiR-127-3p inhibits proliferation of ovarian cancer in rats through down-regulating MAPK4.

OBJECTIVE: To reveal the anti-tumor effect of micro ribonucleic acid (miR)-127-3p on epithelial ovarian cancer (EOC). PATIENTS AND METHODS: The expression of miR-127-3p in 7 kinds of EOC cell lines and 10 cases of clinical samples of EOC patients was detected via quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). OVCAR-3 and Caov-3 cell lines were transfected with lentiviruses to overexpress endogenous miR-127-3p. Then, the anti-tumor effect of miR-127-3p on EOC cells was explored through the in vitro cell proliferation assay, bufalin sensitivity assay, wound healing assay, and invasion assay. In addition, whether the mitogen-activated protein kinase 4 (MAPK4) gene is a downstream target of miR-127-3p in EOC was verified via Dual-Luciferase reporter assay and qRT-PCR. The involvement of MAPK4 in regulating phenotypes of OVCAR-3 and Caov-3 cells was finally explored. RESULTS: MiR-127-3p was downregulated in both EOC cell lines and EOC tissues (p<0.05). After lentivirus-mediated overexpression of miR-127-3p, in vitro proliferation and invasion of EOC cells were inhibited, and the sensitivity to bufalin was enhanced (p<0.05). MiR-127-3p directly regulated MAPK4 gene in EOC. Moreover, the upregulation of MAPK4 inhibited the anti-tumor effect of miR-127-3p on EOC, manifested as the remarkably enhanced cell proliferation and migration (p<0.05), and the weakened sensitivity to bufalin (p<0.01). CONCLUSIONS: MiR-127-3p exerts an inhibitory effect on EOC cells via regulating MAPK4 level.

Genetic analysis of first-trimester miscarriages with a combination of cytogenetic karyotyping, microsatellite genotyping and arrayCGH.

Miscarriage is the spontaneous loss of an embryo or fetus before the 20th week of pregnancy. Most miscarriages occur before the end of the first trimester (<13 weeks). Although many risk factors relate to this occurrence, genetic factors play the most important role. Chromosomal abnormalities, including both numerical and structural anomalies, underlie the majority of miscarriages. In this study, we employed a comprehensive approach using cytogenetic karyotyping, polymerase chain reaction (PCR)-based genotyping, and microarray-based comparative genomic hybridization (arrayCGH) in combination to analyze chromosomal profiles of 115 first-trimester miscarriages of Chinese women. Seventy cases (61%) were found to have chromosomal anomalies, of which 90% were numerical and 10% were structural. Cytogenetic karyotyping identified 78.6% (55/70), PCR assays 2.9% (2 triploids), and arrayCGH 18.6% (13/70) of the anomalies. In this study, a microdeletion of 108 kb and four microduplications sizing from 300 to 1460 kb were observed. An advantage of using this combination approach is that microsatellite genotyping and arrayCGH can be accomplished in spite of culture failure and maternal cell contamination. In addition, arrayCGH can detect submicroscopic chromosomal anomalies and gene dosage alterations.

Function of a TGF-beta inducible nuclear protein in the silk gland in Bombyx mori.

A TGF-beta inducible nuclear protein 1 (BmTINP1) was cloned from silkworm, Bombyx mori. Polyclonal antibodies against BmTINP1 were produced and subsequently used in immunoblotting and immunohistochemistry analyses. The immunoblotting analyses demonstrated that BmTINP1 was specifically expressed in the anterior silk gland (ASG) and the middle silk gland (MSG) but not in the posterior silk gland (PSG). There were two bands that suggested the existence of an isoform of BmTINP1. The expression profiles of BmTINP1 in ASGs and MSGs were similar, and they manifested a high level of expression throughout the period during which silk gland grew exponentially. Immunohistochemistry results revealed that BmTINP1 was translocated from the nucleus into the cytoplasm when larvae developed from the 4th-HCS into the 5th instar. 20-hydroxyecdysone (20E) promotes the translocation, while the methoprene [a juvenile hormone (JH) analog] restrains the process. Our findings indicate that BmTINP1 is involved in silk produce along with the rapid growth of ASGs and MSGs during the last instar larvae, and the process could be regulated by hormones via control of BmTINP1 translocation from the nucleus to the cytoplasm.

Polymorphisms in the C-type lectin genes cluster in chromosome 19 and predisposition to severe acute respiratory syndrome coronavirus (SARS-CoV) infection.

BACKGROUND: Polymorphisms of CLEC4M have been associated with predisposition for infection by the severe acute respiratory syndrome coronavirus (SARS-CoV). DC-SIGNR, a C-type lectin encoded by CLEC4M, is a receptor for the virus. A variable number tandem repeat (VNTR) polymorphism in its neck region was recently associated with susceptibility to SARS infection. However, this association was controversial and was not supported by subsequent studies. Two explanations may account for this discrepancy: (1) there may be an unknown predisposition polymorphism located in the proximity which is linked to the VNTR; or (2) it was a spurious association due to unrecognised population structure in the VNTR. METHODS: We performed a comprehensively genetic association study on this C-type lectin gene cluster (FCER2, CLEC4G, CD209, and CLEC4M) at 19p13.3 by a tagging single nucleotide polymorphisms (SNPs) approach. RESULTS: 23 tagSNPs were genotyped in 181 SARS patients and 172 population controls. No significant association with disease predisposition was detected. Genetic variations in this cluster also did not predict disease prognosis. However, we detected a population stratification of the VNTR alleles in a sample of 1145 Han Chinese collected from different parts of China. CONCLUSION: The results indicated that the genetic predisposition allele was not found in this lectin gene cluster and population stratification might cause the previous positive association.

Peripheral Blood Leukocyte Telomere Length Is Associated with Age but Not Renal Function: A Cross-Sectional Follow-Up Study.

OBJECTIVES: We aimed to evaluate the relationship between baseline renal function and changes in telomere length in Han Chinese. METHODS: The telomere restriction fragment (TRF) length of leukocytes in the peripheral blood was measured in healthy volunteers recruited in 2014. The estimated glomerular filtration rate (eGFR) was calculated based on serum creatinine (Scr) and serum cystatin C (CysC)-eGFRcys and eGFRScr-cys through the Cockcroft-Gault formula (eGFRC-G) or the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI / eGFRCKD-EPI) equation. The correlation between telomere length changes over time and renal function was analyzed. RESULTS: Leukocyte TRF lengths were negatively correlated to age (r = -0.393, p < 0.001) and serum CysC (r = -0.180, p < 0.01), while positively associated with eGFRCKD-EPI, eGFRC-G, eGFRcys, and eGFRScr-cys (r = 0.182, 0.122, 0.290, and 0.254 respectively, p < 0.01). The 3-year change of telomere length was 46 bp/years. When adjusted for age, the associations between telomere length changes and baseline, subsequent TRF lengths, and serum CysC were no longer present. No association was observed between TRF length changes and renal function. CONCLUSION: The rate of telomere length changes was affected by age and baseline telomere length. The telomere length changes might be important markers for aging.