Comparative analysis of the genetic diversity of the neutral microsatellite loci and second exon of the goat MHC-DQB1 gene.

This study compared and analyzed the genetic diversity and population structure of exon 2 of the DQB1 gene and 13 autosomal neutral microsatellite markers from 14 Chinese goat breeds to explore the potential evolutionary mechanism of the major histocompatibility complex (MHC). A total of 287 haplotypes were constructed from MHC-DQB1 exon 2 from 14 populations, and 82 nucleotide polymorphic sites (SNPs, 31.78%) and 172 heterozygous individuals (79.12%) were identified. The FST values of the microsatellites and MHC-DQB ranged between 0.01831-0.26907 and 0.00892-0.38871, respectively. Furthermore, 14 goat populations showed rich genetic diversity in the microsatellite loci and MHC-DQB1 exon 2. However, the population structure and phylogenetic relationship represented by the two markers were different. Positive selection and Tajima's D test results showed the occurrence of a diversified selection mechanism, which was primarily based on a positive and balancing selection in goat DQB. This study also found that the DQB sequences of bovines exhibited trans-species polymorphism (TSP) among species and families. In brief, this study indicated that positive and balancing selection played a major role in maintaining the genetic diversity of DQB, and TSP of MHC in bovines was common, which enhanced the understanding of the MHC evolution.

Selection signatures of litter size in Dazu black goats based on a whole genome sequencing mixed pools strategy.

Litter size is considered to be the most important index for estimating domestic animal productivity. Due to its complexity, the molecular mechanism of litter size has not been elucidated, and it has restricted the use of marker-assisted selection to create high-yield populations in goats. A genome-wide selective sweep analysis was performed with 31 Dazu black goats to identify significant genomic regions and candidate genes related to litter size by a mixed pools strategy. A total of 96 candidate genes were identified, including NR6A1, STK3, IGF2BP2, AR, HMGA2, NPTX1, ANKRD17, DPYD, CLRB, PPP3CA, PLCB1, STK3 and HMGA2, using mixed pool analysis with ZHp and FST. We classified these candidate genes based on the functional classification and annotation of signaling pathways. According to the GO and KEGG analysis results, a total of 43 GO terms and 108 pathways were annotated from these genes. In particular, some novel candidate genes were enriched in reproduction-related pathways, such as the estrogen signaling pathway and oocyte meiosis. These findings provide insight into the influences of coding genes on the fecundity traits of goats.

[Recent research advances of 1-deoxynojirimycin and its derivatives].

The 1-DNJ named 1-deoxynojirimycinis (2R,3R,4R,5S)-2-(hydroxymethyl) piperidine-3,4,5-triol, which is the nature active components existingin mulberryresources including leaves, stems, roots and silkworm larva, silkworm chrysalis, etc.The 1-deoxynojirimycin is a polyhydroxylated piperidine alkaloid, which was first found in Streptomyces as an antibiotic. Then the Japanese researchers isolated it from the mulberry root. 1-DNJ can inhibit postprandial hyperglycemia by suppressing intestinal alpha glucosidase. Therefore, 1-DNJ is often used to treat treating diabetes and complicating disease and to prevent obesity and weight-related disorders. With the development of the researches, 1-deoxynojirimycin and its derivtiv was discovered to possess anti-hyperglycemic, anti-virus, anti-tumor functions and so on. Therefore,based on our current studythe existing knowledge on source, technique preparation process, pharmacokinetics, bioactivties,and in silico target fishing of 1-DNJ were summarized, so that the researchers may use it to explore future perspective of research on 1-DNJ.

A 1.1 Mb duplication CNV on chromosome 17 contributes to skeletal muscle development in Boer goats.

The Boer goat is one of the top meat breeds in modern animal husbandry and has attracted widespread attention for its unique growth performance. However, the genetic basis of muscle development in the Boer goat remains obscure. In this study, we identified specific structural variants in the Boer goat based on genome-wide selection signals and analyzed the basis of the molecular heredity of related candidate genes in muscle development. A total of 9 959 autosomal copy number variations (CNVs) were identified through selection signal analysis in 127 goat genomes. Specifically, we confirmed that the highest signal CNV (HSV) was a chromosomal arrangement containing an approximately 1.11 Mb (CHIR17: 60062304-61171840 bp) duplicated fragment inserted in reverse orientation and a 5 362 bp deleted region (CHIR17:60145940-60151302 bp) with overlapping genes (e.g., ARHGAP10, NR3C2, EDNRA, PRMT9, and TMEM184C). The homozygous duplicated HSV genotype (+/+) was found in 96% of Boer goats but was not detected in Eurasian goats and was only detected in 4% of indigenous African goats. The expression network of three candidate genes ( ARHGAP10, NR3C2, and EDNRA) regulating dose transcription was constructed by RNA sequencing. Results indicated that these genes were involved in the proliferation and differentiation of skeletal muscle satellite cells (SMSCs) and their overexpression significantly increased the expression of SAA3. The HSV of the Boer goat contributed to superior skeletal muscle growth via the dose effects of overlapping genes.

Clinical associations of the genetic variants of CTLA-4, Tg, TSHR, PTPN22, PTPN12 and FCRL3 in patients with Graves' disease.

OBJECTIVE: Graves' disease (GD) is an organ-specific autoimmune disorder. Both immune-modulating genes and thyroid-specific genes are involved in its genetic pathogenesis. It remains unclear, however, how the interactions of various susceptibility genes contribute to the pathogenesis and clinical severity of the disease. The purpose of this study was to investigate the relationships between GD and single nucleotide polymorphisms (SNPs) from CTLA-4, PTPN22, PTPN12, FCRL3 (general autoimmunity genes regulating T and B cells) and the TSHR and Tg genes (disease-specific genes). Furthermore, we evaluated the influences these SNPs have on the risk and severity of GD. DESIGN AND METHODS: This cross-sectional clinical study was performed in 436 GD patients and 316 healthy, gender-matched individuals. Twenty-eight SNPs from CTLA-4, PTPN22, PTPN12, FCRL3, TSHR and Tg genes were genotyped and their associations with the risk and severity of GD were analysed. RESULTS: The CTLA-4 rs231779, Tg rs2069550 and PTPN22 rs3789604 SNPs were associated with GD, with additive risk effects present in rs231779 and rs2069550. The ACACC and ACGCT haplotypes, composed of five SNPs in the CTLA-4 gene (rs4553808, rs5472909, rs231775, rs231777 and rs231779), were protective and risk haplotypes respectively. The AA genotype of PTPN22 rs3789604 and AA genotype of FCRL3 rs7528684 were correlated with a reduced risk of GD, while the CC genotype of TSHR rs2239610 was associated with higher serum concentrations of FT4 and TRAb. Logistic analysis confirmed the contribution of CTLA-4 rs231779 to the development of GD. CONCLUSIONS: These preliminary results demonstrate that the immune-regulatory gene CTLA-4 and the thyroid-specific gene Tg contribute to the risk of Graves' disease with additive effects, while PTPN22 rs3789604 and FCRL3 rs7528684 polymorphisms are protective against the disease. In addition, the TSHR rs2239610 SNP is related to the severity of Graves' disease.

Evolutionary relationship and population structure of domestic Bovidae animals based on MHC-linked and neutral autosomal microsatellite markers.

Major histocompatibility complex (MHC) genes are critical for disease resistance or susceptibility responsible for host-pathogen interactions determined mainly by extensive polymorphisms in the MHC genes. Here, we examined the diversity and phylogenetic pattern of MHC haplotypes reconstructed using three MHC-linked microsatellite markers in 55 populations of five Bovidae species and compared them with those based on neutral autosomal microsatellite markers (NAMs). Three-hundred-and-forty MHC haplotypes were identified in 1453 Bovidae individuals, suggesting significantly higher polymorphism and heterozygosity compared with those based on NAMs. The ambitious boundaries in population differentiation (phylogenetic network, pairwise FST and STRUCTURE analyses) within and between species assessed using the MHC haplotypes were different from those revealed by NAMs associated closely with speciation, geographical distribution, domestication and management histories. In addition, the mean FST was significantly correlated negatively with the number of observed alleles (NA), observed (HO) and expected (HE) heterozygosity and polymorphism information content (PIC) (P < 0.05) in the MHC haplotype dataset while there was no correction of the mean FST estimates (P> 0.05) between the MHC haplotype and NAMs datasets. Analysis of molecular variance (AMOVA) revealed a lower percentage of total variance (PTV) between species/groups based on the MHC-linked microsatellites than NAMs. Therefore, it was inferred that individuals within populations accumulated as many MHC variants as possible to increase their heterozygosity and thus the survival rate of their affiliated populations and species, which eventually reduced population differentiation and thereby complicated their classification and phylogenetic relationship inference. In summary, host-pathogen coevolution and heterozygote advantage, rather than demographic history, act as key driving forces shaping the MHC diversity within the populations and determining the interspecific MHC diversity.

Identification of a Goat Intersexuality-Associated Novel Variant Through Genome-Wide Resequencing and Hi-C.

Background: Polled intersex syndrome (PIS) leads to reproductive disorders in goats and exerts a heavy influence on goat breeding. Since 2001, the core variant of an 11.7 kb deletion at ~129 Mb on chromosome 1 (CHI1) has been widely used as a genetic diagnostic criterion. In 2020, a ~0.48 Mb insertion within the PIS deletion was identified by sequencing in XX intersex goats. However, the suitability of this variation for the diagnosis of intersex goats worldwide and its further molecular genetic mechanism need to be clarified. Results: The whole-genome selective sweep of intersex goats from China was performed with whole-genome next-generation sequencing technology for large sample populations and a case-control study on interbreeds. A series of candidate genes related to the goat intersexuality phenotype were found. We further confirmed that a ~0.48 Mb duplicated fragment (including ERG and KCNJ15) downstream of the ~20 Mb PIS region was reversely inserted into the PIS locus in intersex Chinese goats and was consistent with that in European Saanen and Valais black-necked goats. High-throughput chromosome conformation capture (Hi-C) technology was then used to compare the 3D structures of the PIS variant neighborhood in CHI1 between intersex and non-intersex goats. A newly found structure was validated as an intrachromosomal rearrangement. This inserted duplication changed the original spatial structure of goat CHI1 and caused the appearance of several specific loop structures in the adjacent ~20 kb downstream region of FOXL2. Conclusions: Results suggested that the novel complex PIS variant genome was sufficient as a broad-spectrum clinical diagnostic marker of XX intersexuality in goats from Europe and China. A series of private dense loop structures caused by segment insertion into the PIS deletion might affect the expression of FOXL2 or other neighboring novel candidate genes. However, these structures require further in-depth molecular biological experimental verification. In general, this study provided new insights for future research on the molecular genetic mechanism underlying female-to-male sex reversal in goats.

[Abdominal adipose area, serum adiponectin and leptin levels of nonalcoholic fatty liver disease in elderly males.].

OBJECTIVE: To study the abdominal adipose area, serum adiponectin and leptin levels of nonalcoholic fatty liver disease in elderly males. METHODS: A total of 238 elderly males (more than 60 years) were enrolled and divided into three groups: Nonalcoholic fatty liver disease (NAFLD) group (n = 76), matching group (age and body mass index matching with NAFLD group, n = 77), normal control group (n = 85). Serum levels of adiponectin and leptin were measured by RIA (radiological immunological assay). Abdominal adipose area was detected by computer tomography. RESULT: (1) body mass index (BMI), abdominal subcutaneous adipose area, visceral adipose area, total adipose area of NAFLD group and matching group were (26.87+/-2.62) kg/m2 and (26.63+/-1.97) kg/m2, (166.59+/-54.27) cm2 and (147.89+/-50.14) cm2, (148.94+/-53.72) cm2 and (150.06+/-45.47) cm2, (315.25+/-89.42) cm2 and (297.93+/-75.12) cm2, respectively; and were higher than those in control group (P less than 0.01). The abdominal subcutaneous adipose area is higher in NAFLD group than in matching group, however, the abdominal visceral adipose area and total adipose area were not significantly different between those two groups. (2) The serum leptin level in NAFLD group and matching group was significantly higher than that in control group, and serum leptin level was not significantly different between NAFLD group and matching group. The serum adiponectin of NAFLD group [(6.31+/-3.31)mug/ml] was significantly lower than that of matching group [(9.87+/-7.071)mug/ml, P less than 0.01] and control group (P less than 0.01). There was no difference in adiponectin level between matching group and control group. 3) AST, TG, abdominal subcutaneous adipose area, abdominal visceral adipose area were risk factors of NAFLD, while serum adiponectin was protective factor of NAFLD. CONCLUSION: These data indicate that elderly male NAFLD patients manifest abdominal obesity, high serum leptin, low serum adiponecin, and suggest that adiponectin may play a crucial role in the pathogenesis of NAFLD in elderly males.

Genetic diversity estimation of Yunnan indigenous goat breeds using microsatellite markers.

BACKGROUND: To assess the genetic diversity of seven Yunnan indigenous goat populations (Fengqing hornless goat, Mile red-bone goat, Longling goat, Ninglang black goat, Black-bone goat, Yunling black goat, and Zhaotong goat), their population structures were investigated using 20 microsatellite markers. RESULTS: The results indicated that the genetic diversity of these goats was rich. The observed heterozygosity ranged from 0.4667 ± 0.0243 to 0.5793 ± 0.0230, and the mean number of alleles ranged from 4.80 ± 1.61 and 4.80 ± 1.64 to 6.20 ± 2.93. The population structure analysis showed that these seven goat populations were separated into two clusters, consistent with the results from phylogenetic networks, pairwise differences, and STRUCTURE analyses. We speculate that this may have been caused by natural geographical isolation, human migration and economic and cultural exchanges. We suggest removing CSRD247 and ILSTS005, two loci identified to be under positive selection in the present study, from the microsatellite evaluation system of goats. CONCLUSIONS: The present study may provide a scientific basis for the conservation and utilization of Yunnan indigenous goats.

Genome-wide analysis of Chongqing native intersexual goats using next-generation sequencing.

Sex reversal has been studied extensively in vertebrate species, particularly in domestic goats, because polled intersex syndrome (PIS) has seriously affected their production efficiency. In the present study, we used histopathologically diagnosed cases of PIS to identify correlated genomic regions and variants using representative selection signatures and performed GWAS using Restriction-Site Associated Resequencing DNA. We identified 171 single-nucleotide polymorphisms (SNPs) that may have contributed to this phenotype, and 53 SNPs were determined to be located in coding regions using a general linear model. The transcriptome data sets of differentially expressed genes (DEGs) in the pituitary tissues of intersexual and nonintersexual goats were examined using high-throughput technology. A total of 10,063 DEGs and 337 long noncoding RNAs were identified. The DEGs were clustered into 56 GO categories and determined to be significantly enriched in 53 signaling pathways by KEGG analysis. In addition, according to qPCR results, PSPO2 and FSH were significantly more highly expressed in sexually mature pituitary tissues of intersexual goats compared to healthy controls (nonintersexual). These results demonstrate that certain novel potential genomic regions may be responsible for intersexual goats, and the transcriptome data indicate that the regulation of various physiological systems is involved in intersexual goat development. Therefore, these results provide helpful data for understanding the molecular mechanisms of intersex syndrome in goats.

Genome-wide selection signatures analysis of litter size in Dazu black goats using single-nucleotide polymorphism.

Litter size is considered to be the most important index for estimating domestic animal productivity. The number of indigenous goats in China with higher litter sizes than those of commercial breeds in other countries may be helpful for accelerating genetic improvements in goat breeding. We performed a genome-wide selective sweep analysis of 31 Dazu black goats with extreme standard deviation in litter size within the third fetus to identify significant genomic regions and candidate genes through different analyses. The analysis identified a total of 33,917,703 variants, including 32,262,179 SNPs and 1,655,524 indels. In addition, two novel candidate genes (LRP1B and GLRB), which are related to litter size, were obtained with π, Tajima's D, πA/πB, and F ST at the individual level with a 95% threshold for each parameter. These two genes were annotated in five GO terms (localization, binding, macromolecular complex, membrane part, and membrane) and two pathways (long-term depression and neuroactive ligand-receptor interaction pathway). Regarding the result of linkage disequilibrium (LD) analysis, in LRP1B and GRID2, the high-yield Dazu black goats exhibit significantly different LD patterns from low-yield goats. Litter size variability has low heritability and is related to multiple complex factors found in domestic animals. Obtaining a clear explanation and significant signal by genome-wide selective sweep analysis with a small sample size is difficult. However, we investigated some candidate genes, particularly LRP1B and GLRB, which may provide useful information for further research.

Association of the interleukin (IL)-16 gene polymorphisms with Graves' disease.

Interleukin (IL)-16 was one of the cytokines with the function of T helper cell recruitment, whose expression in the thyrocyte and orbital fibroblast of Graves' disease (GD) patients was increased. Recently association of IL-16 gene polymorphisms with autoimmune diseases had been reported. However, there was little known about the impact of IL-16 gene polymorphisms on GD. In this study, we performed a case-control association study of three tagSNPs (rs4778889-rs1131445-rs4778641) within the IL-16 gene on 258 patients with GD and 208 healthy subjects in the Chinese population. Our data showed that common IL-16 variants were associated with GD (P=0.013-0.0186) and Graves' disease associated ophthalmopathy (GO) (P=0.0033-0.041). A novel protective haplotype containing the three tagSNPs (C-T-C) was observed in association with GO (P=0.013). In conclusion, IL-16 gene was significantly associated with susceptibility to Graves' disease and Graves' disease associated ophthalmopathy in the Chinese population.

Transgenic expression of cytotoxic T-lymphocyte-associated antigen 4-immunoglobulin prolongs xenogeneic skin graft survival without extensive immunosuppression in rat burn wounds.

BACKGROUND: We sought to establish a transgenic animal line skin-specifically overexpressing cytotoxic T-lymphocyte-associated antigen 4-immunoglobulin (CTLA4Ig) as a reproducible source of xenogeneic skin grafts with extended survival for wound coverage. We tested this strategy in mice based on a previously established transgenic mouse line that stably and skin-specifically expresses CTLA4Ig for lifetimes and generations. METHODS: CTLA4Ig expression was examined by immunohistochemical assay, and its bio-activity was tested by mixed lymphocyte reaction. The survival of transgenic mouse skin grafted onto rat burn wounds was observed. The impact of transgenic skin grafting on recipient immunity was evaluated by inspecting the survival of the wild-type skin grafted along with transgenic skin onto a separate wound on the same rat. The circulatory CTLA4Ig protein in recipient was detected by sandwich enzyme-linked immunosorbent assay, and its impact on recipient lymphocyte response against donor antigen was tested by mixed lymphocyte reaction. RESULTS: The transgenic CTLA4Ig protein suppressed lymphocyte proliferation in vitro, and the transgenic skin graft survival was remarkably prolonged compared with the wild-type skin derived from the same mouse strain. The survival of the wild-type skin grafted along with transgenic skin exhibited no significant difference from that grafted alone. Circulatory CTLA4Ig protein was detected in recipients, however, no significantly reduced recipient lymphocyte response against donor antigen was observed. CONCLUSION: transgenic expression of CTLA4Ig may be a potential and safe method to prolong xenogenic skin graft survival in burn wounds, and transgenic animal lines can be established as a reproducible source of skin grafts with extended survival for wound coverage.

[Prognostic factors in the relapse of Graves disease].

OBJECTIVE: To evaluate the variables which can be used as prognostic factors in predicting the outcome of Graves disease (GD) after treatment with antithyroid drugs. METHODS: We performed a retrospective audit of 204 patients with newly diagnosed Graves disease consecutively at the Ruijin Hospital. RESULTS: Overall, 110 patients (53.9%) were considered to be treatment failures. Age at the time of diagnosis was (31.0 +/- 12.2)years in the successful group and (36.3 +/- 14.0) years in the failure group. Free T3 (FT3) was (25.60 +/- 9.52) pmol/L and (19.16 +/- 6.38) pmol/L in the failure and the successful group (P = 0.001). FT3 to FT4 ratio and thyrotrophin receptor antibody (TRAb) levels were higher in the failure group (P = 0.001). Logistic regression analysis showed that thyroid size, FT3 to FT4 ratio and TRAb at the time of diagnosis were associated with failure outcome. The patients reached euthyroid state at 3, 6, 9 and 12 months respectively and in the failure group the patients with continued thyrotropin suppression were more than those in the successful group (P = 0.001). CONCLUSIONS: Graves disease patients with large thyroid size, high levels of TRAb and FT3 to FT4 ratio before drug treatment are more likely to fail to respond to antithyroid drug treatment. We also found that patients with continuing thyrotropin suppression and attainment of euthyroid state in the course of treatment had low remission rate and prolonged therapy.

Sheep mitochondrial heteroplasmy arises from tandem motifs and unspecific PCR amplification.

The mitochondrial DNA control region (D-loop) is a widely used molecular marker in evolutionary and phylogeographic research. However, the occurrence of heteroplasmy of the D-loop region within individuals has rarely been investigated. In this study, a total of 85 Chinese sheep were used to amplify a partial D-loop region, and 15 heteroplasmic animals (17.64%) were identified. A comparative analysis of the PCR amplification and cloning of the D-loop sequences from the heteroplasmic samples revealed most of the sequencing profile from the heteroplasmic regions started at the beginning of a 75-bp random repeat motif. In addition, a total of 22 nonsyngeneic sequences with a D-loop were found in 61 of the clones obtained from the 4 random heteroplasmic and 3 homozygote animals, and their genomic locations were compared for homology. In summary, the D-Loop sequencing profiles appear to be heteroplasmic and could arise from tandem repeat motifs and unspecific replication during PCR amplification; however, they are not likely due to the presence of multiple mitochondrial genomes within an individual.

Genetic diversity of the Chinese goat in the littoral zone of the Yangtze River as assessed by microsatellite and mtDNA.

The objective of this study was to assess the genetic diversity and population structure of goats in the Yangtze River region using microsatellite and mtDNA to better understand the current status of those goat genetic diversity and the effects of natural landscape in fashion of domestic animal genetic diversity. The genetic variability of 16 goat populations in the littoral zone of the Yangtze River was estimated using 21 autosomal microsatellites, which revealed high diversity and genetic population clustering with a dispersed geographical distribution. A phylogenetic analysis of the mitochondrial D-loop region (482 bp) was conducted in 494 goats from the Yangtze River region. In total, 117 SNPs were reconstructed, and 173 haplotypes were identified, 94.5% of which belonged to lineages A and B. Lineages C, D, and G had lower frequencies (5.2%), and lineage F haplotypes were undetected. Several high-frequency haplotypes were shared by different ecogeographically distributed populations, and the close phylogenetic relationships among certain low-frequency haplotypes indicated the historical exchange of genetic material among these populations. In particular, the lineage G haplotype suggests that some west Asian goat genetic material may have been transferred to China via Muslim migration.

MEN1 mutation analysis in Chinese patients with multiple endocrine neoplasia type 1.

Multiple endocrine neoplasia type 1 (MEN1) is an inherited tumour syndrome characterized by the development of tumours of the parathyroid, anterior pituitary and pancreatic islets, etc. Heterozygous germ line mutations of MEN1 gene are responsible for the onset of MEN1. We investigated the probands and 31 family members from eight unrelated Chinese families associated with MEN1 and identified four novel mutations, namely 373_374ins18, 822delT, 259delT and 1092delC, as well as three previously reported mutations, such as 357_360delCTGT, 427_428delTA and R108X (CGA>TGA) of MEN1 gene. Furthermore, we detected a loss of heterozygosity (LOH) at chromosome 11q in the removed tumours, including gastrinoma, insulinoma and parathyroid adenoma from two probands of MEN1 families. RT-PCR and direct sequencing showed that mutant MEN1 transcripts remained in the MEN1-associated endocrine tumours, whereas normal menin proteins could not be detected in those tumours by either immunohistochemistry or immunoblotting. In conclusion, MEN1 heterozygous mutations are associated with LOH and menin absence, which are present in MEN1-associated endocrine tumours.

[The clinical patterns and RET proto-oncogene in fifteen multiple endocrine neoplasia type 2A pedigrees].

OBJECTIVE: To detect the phenotype and rearranged mutations during transfection of RET proto-oncogene in fifteen multiple endocrine neoplasia type 2A (MEN2A) probands and their family members. METHODS: Totally 119 family members of the fifteen MEN2A pedigrees were recruited. Total genomic DNA was extracted from peripheral blood for PCR. PCR products of exon 8, 10, 11, 13, 14, 15 and exon16 of the RET proto-oncogene were purified and direct gene sequencing was performed. RESULTS: The germline mutations of RET proto-oncogene were detected in 49 members of the fifteen MEN2A pedigrees. Among them, 37 patients had MEN2A phenotype and the remaining 12 were gene carriers. The mean age at which the members of the former group were diagnosed as MEN2A was significantly later than those of the latter group [(43.0 +/- 13.9) yr vs (9.8 +/- 7.4) yr, P < 0.01]; The incidences of medullary thyroid carcinoma (MTC), pheochromocytoma (PCC) and hyperparathyroidism (HPT) in 37 MEN2A patients were 91.9%, 56.8% and 10.8%; Five germline mutations which all located in codon 634 of exon11 in RET proto-oncogene were detected in the fifteen MEN2A pedigrees. They were C634W (13.3%), C634Y (46.7%), C634R (26.7%), C634F (6.7%) and C634S (6.7%); Through the analysis of the genotype-phenotype of the 37 MEN2A patients, no statistic significance was found in the age of diagnose for MEN2A, the incidence of PCC, the percentage of cervical lymph node metastasis and the five different genotypes. CONCLUSION: The mean age at which the 37 patients of the fifteen MEN2A pedigrees were diagnosed was older than that reported in the overseas literatures. The occurrence of MTC in MEN2A patients is earlier than that of PCC and HPT. The incidences of MTC and PCC are basically consistent with those in the overseas archives, while the incidence of HPT is lower than that in the corresponding report. The detected mutations of RET proto-oncogene in MEN2A patients were all located in codon 634, being simpler than that in foreign reports.

[Freezing-thawing damages plasma membrane integrity of goat spermatozoa].

OBJECTIVE: To evaluate the plasma membrane integrity and morphology of fresh and frozen goat spermatozoa. METHODS: The ejaculates of three male goats were obtained by the artificial vagina method of collection and the rates of sperm abnormality and acrosome integrity were detected after freezing-thawing processing. The plasma membrane integrity of the fresh and frozen-thawed goat spermatozoa was evaluated with a combination of fluorescent probes, carboxyfluorescein diacetate and propidium iodide. RESULTS: The freezing-thawing process significantly influenced the viability and integrity of the spermatozoa ([74.43 +/- 13.78]% vs. [46.25 +/- 2.69]%; [64.26 +/- 7.03]% vs. [6.27 +/- 2.90]%, P < 0.01). The results showed differences in acrosome integrity rate between the fresh and frozen samples ([80.77 +/- 10.70]% vs. [58.42 +/- 18.05]% , P < 0.05). CONCLUSION: The freezing-thawing process significantly reduces sperm viability and acrosome integrity and seriously damages the plasma membrane integrity.

[Thyroid-associated ophthalmopathy: a clinical study of 381 cases].

OBJECTIVE: To find the clue of making accurate diagnosis for thyroid-associated ophthalmopathy (TAO) by analyzing 381 cases. METHODS: We analyzed 381 cases with TAO diagnosed in Ruijing Hospital to summarize the characteristics of clinical symptoms and signs, the type and duration of thyroid disorders, the image study of computerized tomography and to know the effect of some influencing factors on the course and severity of TAO. RESULTS: (1) In the 381 cases of TAO, only 86.4% had hyperthyroidism, as TAO could be associated with hypothyroidism or euthyroidism. Ophthalmopathy occurred simultaneously with the development of hyperthyroidism or followed it 18.8 months later in the majority of the patients. (2) Upper eyelid retraction was the commonest sign of the lids. 120 cases complained of double vision. 89 had eye movement restriction, especially to upper direction. Mean extent of proptosis was 17.9 mm. 14.5% had high intraocular pressure. The extraocular muscles involvement was characteristically bilateral and multiple. The inferior rectus was the most frequently involved with enlargement. (3) Only 19.2% cases received corticosteroids, retrobulbar irradiation or surgery because of the severity of ocular symptoms and signs. CONCLUSIONS: TAO way occur with the development of hyperthyroidism simultaneously or follow it later. The disease has its own clinical characteristics and natural course. Making accurate diagnosis earlier is essential for instituting treatment and predicting prognosis.