Iodine Nutrition, Thyroid-stimulating Hormone, and Related Factors of Postpartum Women from three Different Areas in China: A Cross-sectional Survey

Objective Studies on the relationship between iodine,vitamin A(VA),and vitamin D(VD)and thyroid function are limited.This study aimed to analyze iodine and thyroid-stimulating hormone(TSH)status and their possible relationships with VA,VD,and other factors in postpartum women.Methods A total of 1,311 mothers(896 lactating and 415 non-lactating)from Hebei,Zhejiang,and Guangxi provinces were included in this study.The urinary iodine concentration(UIC),TSH,VA,and VD were measured.Results The median UIC of total and lactating participants were 142.00µg/L and 139.95µg/L,respectively.The median TSH,VA,and VD levels in all the participants were 1.89 mIU/L,0.44μg/mL,and 24.04 ng/mL,respectively.No differences in the UIC were found between lactating and non-lactating mothers.UIC and TSH levels were significantly different among the three provinces.The rural UIC was higher than the urban UIC.Obese mothers had a higher UIC and a higher prevalence of excessive TSH.Higher UICs and TSHs levels were observed in both the VD deficiency and insufficiency groups than in the VD-sufficient group.After adjustment,no linear correlation was observed between UIC and VA/VD.No interaction was found between vitamins A/D and UIC on TSH levels.Conclusion The mothers in the present study had no iodine deficiency.Region,area type,BMI,and VD may be related to the iodine status or TSH levels.

Analysis of the value and safety of thyroid-stimulating hormone in the clinical efficacy of patients with thyroid cancer

BACKGROUND Thyroid cancer(TC)is a common malignant tumor in the endocrine system.In recent years,the incidence and recurrence rates of TC have been raising due to increasing work pressure and irregular lifestyles.Thyroid-stimulating hormone(TSH)is a specific parameter for thyroid function screening.This study aims to explore the clinical value of TSH in regulating the progression of TC,so as to find a breakthrough for the early diagnosis and treatment of TC.AIM To explore the value and safety of TSH in the clinical efficacy of patients with TC.METHODS 75 patients with TC admitted to the Department of Thyroid and Breast Surgery of our hospital from September 2019 to September 2021 were selected as the observation group,and 50 healthy subjects were selected as the control group during the same period.The control group was treated with conventional thyroid replacement therapy,and the observation group was treated with TSH suppression therapy.The soluble interleukin(IL)-2 receptor(sIL-2R),IL-17,IL-35levels,free triiodothyronine(FT3),free tetraiodothyronine(FT4),CD3+,CD4+,CD8+,CD44V6,and tumor supplied group of factor(TSGF)levels were observed in the two groups.The occurrence of adverse reactions was compared between the two groups.RESULTS After treatment with different therapies,the levels of FT3,FT4,CD3+,and CD4+in the observation group and the control group were higher than those before treatment,while the levels of CD8+,CD44V6,and TSGF were lower than those before treatment,and the differences were statistically significant(P<0.05).More importantly,the levels of sIL-2R and IL-17 in the observation group were lower than those in the control group after 4 wk of treatment,while the levels of IL-35 were higher than those in the control group,and the differences were statistically significant(P<0.05).The levels of FT3,FT4,CD3+,and CD4+in the observation group were higher than those in the control group,and the levels of CD8+,CD44V6,and TSGF were lower than those in the control group.There was no significant difference in the overall incidence rate of adverse reactions between the two groups(P>0.05).CONCLUSION TSH suppression therapy can improve the immune function of patients with TC,lower the CD44V6 and TSGF levels,and improve serum FT3 and FT4 levels.It demonstrated excellent clinical efficacy and a good safety profile.

Correlation analysis of serum thyroglobulin,thyroid-stimulating hormone levels,and thyroid-cancer risk in thyroid nodule surgery

BACKGROUND Thyroglobulin(Tg)is one of the markers of thyroid cancer,and its concentration may be elevated in patients with malignant thyroid tumors.Thyroid-stimulating hormone(TSH)is secreted by the pituitary gland,which has a significant impact on thyroid gland function.Excessively high or low TSH levels may be associated with an increased risk of thyroid cancer.Thus,in-depth studies on the association of serum Tg and TSH levels with thyroid cancer risk in patients with thyroid nodules are warranted.This can help determine whether Tg and TSH levels can predict the degree of malignancy of thyroid nodules,which can in turn guide doctors in making accurate diagnoses and treatment decisions.Furthermore,such studies can provide more accurate diagnostic methods for thyroid nodules and help patients become aware of the presence of thyroid cancer as early as possible,thereby improving the success rate of treatment and prognosis.AIM To investigate the association of serum Tg and TSH levels with the risk of thyroid cancer in patients undergoing thyroid nodule surgery.METHODS The clinical data and laboratory examination results of 130 patients who underwent thyroid nodule surgery were retrospectively analyzed.Furthermore,their preoperative serum Tg and TSH levels were recorded.Histopathological examination conducted during follow-up revealed the presence of thyroid cancer.Correlation analysis were used to analyze the association of Tg and TSH levels with the risk of thyroid cancer.RESULTS Of the 130 patients,60 were diagnosed with thyroid cancer.Statistical analysis revealed a significant positive correlation between serum Tg levels and the risk of thyroid cancer(P<0.05).This suggests that high serum Tg levels are associated with an increased risk of thyroid cancer.However,no significant correlation was observed between serum TSH levels and the risk of thyroid cancer(P>0.05).CONCLUSION In patients who underwent thyroid nodule surgery,serum Tg levels exhibited a significant correlation with the risk of thyroid cancer but serum TSH levels did not.These findings suggest that serum Tg can serve as an important biomarker for assessing the risk of thyroid cancer in these patients.

Comprehensive treatment of thyroid-stimulating hormone(TSH)-secreting pituitary adenoma:one case report

Thyroid-stimulating hormone(TSH)-secreting pituitary adenoma is a rare type in all pituitary tumors.Recently we treated a TSH-secreting pituitary tumor in our hospital.The patient had been treated for hyperthyroidism,in which methimazole had been prescribed for 10 years,but the symptoms had not been alleviated.MRI imaging demonstrated the typical features of a sellar tumor,and the diameter was approximately 2.7 cm.Based on the laboratory studies:T3 at 6.27 nmol/L,T4 at 260.10 nmol/L,FT3 at 17.22 pmol/L,FT4 at 76.06 pmol/L,TSH at 9.93 Mu/L,the patient was diagnosed with a TSH-secreting pituitary tumor and central hyperthyroidism.After the patient was given octreotide for one week,he received resection of tumor via single-nostril transsphenoidal approach.After discharge,the patient received the radiation therapy two courses about 20 days.Through the comprehensive treatment of surgery,radiotherapy and drugs,the patient received a satisfactory result.

Thyroid-stimulating hormone-thyroid hormone signaling contributes to circadian regulation through repressing clock2/npas2 in zebrafish

Thyroid-stimulating hormone(TSH)is important for the thyroid gland,development,growth,and metabolism.Defects in TSH production or the thyrotrope cells within the pituitary gland cause congenital hypothyroidism(CH),resulting in growth retardation and neurocognitive impairment.While human TSH is known to display rhythmicity,the molecular mechanisms underlying the circadian regulation of TSH and the effects of TSH-thyroid hormone(TH)signaling on the circadian clock remain elusive.Here we show that TSH,thyroxine(T4),triiodothyronine(T3),and tshba display rhythmicity in both larval and adult zebrafish and tshba is regulated directly by the circadian clock via both E′-box and D-box.Zebrafish tshba^(−/−)mutants manifest congenital hypothyroidism,with the characteristics of low levels of T_(4)and T_(3)and growth retardation.Loss or overexpression of tshba alters the rhythmicity of locomotor activities and expression of core circadian clock genes and hypothalamic-pituitary-thyroid(HPT)axis-related genes.Furthermore,TSH-TH signaling regulates clock2/npas2 via the thyroid response element(TRE)in its promoter,and transcriptome analysis reveals extensive functions of Tshba in zebrafish.Together,our results demonstrate that zebrafish tshba is a direct target of the circadian clock and in turn plays critical roles in circadian regulation along with other functions.

拮抗剂方案双扳机次日血清β-hCG水平对新鲜胚胎移植结局的影响

目的:探讨拮抗剂方案双扳机次日血清β-人绒毛膜促性腺激素(β-human chorionic gonadotropin,β-hCG)水平对体外受精/卵胞质内单精子注射(in vitro fertilization/intracytoplasmic sperm injection,IVF/ICSI)新鲜胚胎移植周期妊娠结局的预测价值。方法:回顾性分析2017年1月-2024年1月于南京医科大学附属妇产医院生殖中心行拮抗剂方案促排卵患者共717个周期的临床资料,所有周期均采用双扳机(hCG+促性腺激素释放激素激动剂)诱导排卵,按照扳机次日血清β-hCG水平分为5组,A组(42个周期)β-hCG≤50 U/L,B组(282个周期)50 U/L<β-hCG≤100 U/L,C组(250个周期)100 U/L<β-hCG≤150 U/L,D组(92个周期)150 U/L<β-hCG≤200 U/L,E组(51个周期)β-hCG>200 U/L。比较各组的促排卵实验室结局及妊娠结局。结果:5组间体质量指数、基础卵泡刺激素、促性腺激素天数及总量、hCG日雌孕激素水平及高评分囊胚数的差异均有统计学意义(均P<0.05),其中体质量指数越高,扳机次日β-hCG水平越低。5组可利用胚胎数、高评分囊胚率、临床妊娠率、流产率及活产率比较,差异均无统计学意义(均P>0.05)。结论:在拮抗剂方案新鲜胚胎移植周期中,双扳机次日血清β-hCG水平并不能预测IVF/ICSI治疗周期的妊娠结局。

Efficacy of the long-acting octreotide formulation in patients with thyroid-stimulating hormone-secreting pituitary adenomas after incomplete surgery and octreotide treatment failure

Background Little information about the current management of patients with thyroid-stimulating hormone （TSH）-secreting pituitary adenomas or about the usefulness of the somatostatin analogue octreotide was contained in the literature. This study aimed to report the efficacy and safety of the long-acting octreotide formulation in patients with TSH-secreting pituitary adenomas after incomplete surgery and octreotide treatment failure. Methods Fifteen patients with TSH-secreting pituitary adenomas （8 men and 7 women）, who previously underwent incomplete surgical resection and/or adjuvant radiotherapy （n=12） and failure of octreotide treatment （n=15）, followed between 2007 and 2010 in Beijing Tiantan Hospital were included in this study. All patients received 1- to 2-months of the long-acting octreotide formulation treatment after the above combination of treatment. Paired samples t-test was used to analysis the variables. Results After two-month duration of the long-acting octreotide formulation treatment, the mean serum free or unbound thyroxine （FT4） （（16.02±1.72） pmol/L） and free triiodothyronine （FT3） （（2.87±0.43） pmol/L） levels of 15 patients significantly decreased compared with those after octreotide-treatment （FT4, （35.36±7.42） pmol/L, P 〈0.001; FT3, （17.85±7.22） pmol/L, P 〈0.001）. Mean TSH levels stayed in the normal range after the long-acting octreotide formulation treatment （（0.72±0.21） mUlL） and were significantly lower than the pretreatment value （（5.27±1.04） mUlL, P 〈0.001）, post-surgery value （（3.37±0.31） mU/L, P 〈0.001） and post-octreotide-treatment value （（4.52±0.41） mU/L, P 〈0.001）. In these patients with TSH-secreting pituitary adenomas there was no evidence of tachyphylaxis. Conclusion The long-acting octreotide formulation may be a useful and safe therapeutic tool to facilitate the medical treatment of TSH-secreting pituitary adenomas in patients who underwent incomplete surgery or need long-term somatostatin analog therapy.

Enzyme-loaded liposome with biocatalytic precipitation for potentiometric immunoassay of thyroid-stimulating hormone in thyroid carcinoma

A simple and feasible potentiometric immunosensing platform based on enzymatic biocatalytic precipitation technique was designed for the sensitive detection of thyroid-stimulating hormone(TSH;a typical kind of biomarkers for thyroid carcinoma),using horseradish peroxidase(HRP)-loaded liposome for the signal amplification.To construct such an assay system,a sandwich-type immunoreaction was readily carried out on monoclonal anti-TSH capture antibody-coated electrode by using polyclonal antiTSH secondary antibody-conjugated HRP-loaded liposome.Accompanying the formation of sandwichtype immunocomplex,the carried liposome was lysed through the added Triton X-100 to release the entrapped HRP molecules,which catalyzed the oxidation of 4-chloro-1-naphthol to produce an insoluble and uncharged organic precipitation on the electrode surface,thereby causing the change of the local electrical potential.Two labeling protocols with and without the liposome were investigated for detection of target TSH,improved analytical features were achieved with HRP-entrapped liposome.Under optimal conditions,the potentiometric immunosensor had good responses for TSH detection within the linear range of 0.01-30 p IU/mL at a detection limit of 0.0067 p IU/mL.Good reproducibility,high specificity and long-time stability were acquired during the assay procedure.Importantly,a wellmatched accuracy between the potentiometric immunosensor and commercial human TSH ELISA kit was gave for the analysis of human serum samples.

The effects of thyroid-stimulating hormone,estradiol and prolactin on sodium/iodide symporter mRNA expression in mouse lactating mammary gland cells under different iodine levels

Objective The present study investigated the sodium/iodide symporter mRNA expression in mouse lactating mammary gland cells under different iodine levels and the effects of thyroid-stimulating hormone(TSH),estradiol(E2)and prolactin(PRL)on NIS mRNA expression in mouse lactating mammary gland cells.

人促甲状腺激素受体A亚单位蛋白在昆虫细胞体系中的分泌性表达

目的拟构建昆虫细胞分泌表达体系,在草地夜蛾卵巢细胞分泌性促甲状腺激素受体(thyroid stimulating hormone receptor,TSHR)A亚单位蛋白。方法将TSHR A亚单位蛋白(22-289)和绿色荧光蛋白(green fluorescence protein,GFP)基因连接到质粒,并通过转化、蓝白斑筛选获得重组杆粒,将其转染入sf9昆虫细胞收获重组杆状病毒,扩增并测定滴度。最终通过蛋白印迹实验鉴定重组蛋白的表达并优化表达条件。结果在蛋白表达体系的构建过程中,PCR鉴定和测序均证实了重组质粒和重组杆粒序列的正确性。将重组杆粒转染入细胞后观察到病毒出芽迹象,空斑实验鉴定P1代病毒的滴度为2×10^(7) pfu/m。蛋白印迹显示重组蛋白分子质量为55 ku,主要在培养基中。蛋白表达的最佳感染复数(multiplicity of infection,MOI)为1,最佳感染时程为72 h。结论本研究构建了TSHR A亚单位的昆虫细胞杆状病毒表达体系,该体系能够外泌性表达分子质量为55 ku左右的蛋白TSHR 22-289,且蛋白能够成功的糖基化修饰。该系统为其工业化平台的建设及生产提供前期基础,也针对日后TSHR蛋白的研究和Graves病的病因预防提供了有用的工具。

Atypical pituitary hormone–target tissue axis

A long-held belief is that pituitary hormones bind to their cognate receptors in classical target glands to actuate their manifold functions.However,a number of studies have shown that multiple types of pituitary hormone receptors are widely expressed in non-classical target organs.Each pituitary gland-derived hormone exhibits a wide range of nonconventional biological effects in these non-classical target organs.Herein,the extra biological functions of pituitary hormones,thyroid-stimulating hormone,follicle-stimulating hormone,luteinizing hormone,adrenocorticotrophic hormone,and prolactin when they act on non-classical organs were summarized,defined by the novel concept of an“atypical pituitary hormone–target tissue axis.”This novel proposal explains the pathomechanisms of abnormal glucose and lipid metabolism,obesity,hypertension,fatty liver,and atherosclerosis while offering a more comprehensive and systematic insights into the coordinated regulation of environmental factors,genetic factors,and neuroendocrine hormones on human biological functions.The continued exploration of the physiology of the“atypical pituitary hormone–target tissue axis”could enable the identification of novel therapeutic targets for metabolic diseases.

IGFALS基因多态性与特发性矮小症患儿rHGH治疗效果及血清IGF-1水平的关系

目的探究胰岛素样生长因子不稳定亚单位(IGFALS)基因多态性与特发性矮小症(ISS)患儿重组人生长激素(rHGH)治疗效果及血清胰岛素样生长因子-1(IGF-1)水平的关系。方法选择2021年1月至2022年8月大理白族自治州人民医院收治的120例ISS患儿作为ISS组,另选健康儿童120例作为对照组,提取两组外周血全基因组DNA,扩增IGFALS基因rs767089671位点基因片段并进行测序。采用rHGH治疗ISS患儿12个月,比较不同IGFALS基因分型患儿生长速率(GV)、年龄对应身高标准差积分(HtSDS)、血清IGF-1、胰岛素样生长因子结合蛋白3(IGFBP-3)水平变化,分析IGFALS基因多态性与rHGH治疗效果及血清IGF-1水平的关系。结果ISS组和对照组IGFALS基因rs767089671位点的基因分型分布符合Hardy-Weinberg平衡定律(P>0.05);ISS组IGFALS基因rs767089671(c.1310C>T)位点基因分型及C/T等位基因分布频率与对照组比较差异有统计学意义(P<0.05);治疗前不同基因分型患儿性别、年龄、体质量、身高、遗传靶身高、骨龄比较差异无统计学意义(P>0.05);治疗后不同基因分型患儿GV、HtSDS、血清IGFBP-3、IGF-1水平均升高,且CC型患儿GV、HtSDS、血清IGFBP-3、IGF-1水平变化更明显(P<0.05);Spearman相关性分析显示,ISS患儿治疗前血清IGF-1、IGFALS基因多态性与治疗前后ΔHtSDS呈正相关(r=0.932、0.576,P<0.05)。结论ISS患儿IGFALS基因rs767089671位点多态性与rHGH促生长效果有关,且GV增长不明显可能与CT和TT基因分型患儿血清IGF-1水平增加不足有关。

FSHβ基因PCR-SSCP多态性及其与济宁青山羊高繁殖力关系的研究

采用PCR-SSCP技术检测促卵泡素β亚基(follicle-stimulatinghormoneβ,FSHβ)基因5′调控区、外显子1和外显子2在高繁殖力山羊品种(济宁青山羊)和低繁殖力山羊品种(辽宁绒山羊、波尔山羊、安哥拉山羊)中的单核苷酸多态性,同时研究该基因对济宁青山羊高繁殖力的影响。结果表明:山羊与绵羊的FSH基因该段核苷酸序列同源性为98%。9对引物中,只有P9的扩增片段存在多态性。P9的扩增片段在济宁青山羊和辽宁绒山羊中检测到AA、AB和AC3种基因型;在波尔山羊中检测到AA、CC和AC3种基因型;在安哥拉山羊中检测到AA、BB、CC、AB、AC和BC共6种基因型。测序分析发现BB型与AA型相比在外显子2的第94bp处有G→A突变,并引起氨基酸改变(丙氨酸→苏氨酸);CC型与AA型相比在外显子2的第174bp有一处C→T沉默突变。济宁青山羊AA、AB和AC基因型频率分别为0.686、0.137和0.177。AA基因型济宁青山羊产羔数最小二乘均值比AB基因型的多0.78只(P<0.05),比AC基因型的多0.64只(P<0.05)。

牛促卵泡激素α亚基cDNA的克隆与序列分析

从牛脑垂体中提取总ＲＮＡ，分离ｍＲＮＡ，反转录获得ｃＤＮＡ，ＰＣＲ扩增获得长为３８０ｂｐ的牛促卵泡激素α亚基ｃＤＮＡ片段，将它克隆于ｐＵＣ１９中，进行序列分析，结果表明：所克隆的α亚基基因编码区序列与Ｅｒｗｉｎ所发表的序列基本相同，仅编码第２４位赖氨酸的密码有差异，同时将所获基因的核苷酸序列及相应氨基酸序列与人、啮齿类的同类基因相比较，具有很高的同源性．

免疫抑制素和卵泡抑素阻止夏季热应激导致的猪精液品质下降的研究

【目的】通过免疫抑制素和卵泡抑素促进精子发生,在夏季热应激状态下保持公猪精液品质。【方法】在夏季热应激状态下对精液品质出现下降的成年种公猪单独免疫抑制素、或联合免疫抑制素α亚基和卵泡抑素重组蛋白(n=6)3次,每次间隔21 d;然后在9周试验期间定期采血测定血液抗体效价、FSH和睾酮浓度,每周采精观察精子活力和精子密度作为精液品质指标。【结果】抑制素单独免疫、抑制素和卵泡抑素联合免疫都使猪血液中抗抑制素的抗体效价显著高于对照组值(P<0.01),联合免疫也使猪血液中抗卵泡抑素的抗体效价显著高于对照组值(P<0.01)。单独免疫或联合免疫都显著升高了血液FSH浓度;联合免疫还升高了血液睾酮水平。在试验期间对照组中猪的精子活力持续下降,而两免疫组的精子活力都上升,其中联合免疫猪的精子活力显著高于对照组猪。试验开始时对照组猪精子密度显著高于两免疫组,但在试验期间持续下降;而免疫组中猪精子密度在试验期间未下降,特辑是在联合免疫组于试验后期还显著高于对照组猪。【结论】免疫抑制素和卵泡抑素可以促进公猪FSH和睾酮分泌,克服夏季热应激的不良影响,促进猪精子发生,并提高精液精子活力和密度,有望用于热应激状态下保持种用公猪的繁殖性能。

ESR、FSHβ及OPN基因多态性与母猪繁殖性状的关联分析

采用PCR-RFLP法对大白猪和长白猪猪群进行了雌激素受体基因(ESR)、卵泡促激素β亚基基因(FSHβ)和骨桥蛋白基因(OPN)等3个与繁殖性能相关的基因多态性检测,并对不同基因型的总产仔数、产活仔数及出生窝重进行了相关分析。结果表明:①在长白猪中,ESR和OPN基因的优势基因是A等位基因,FSHβ基因的优势基因是B等位基因;在初产母猪中,ESR基因BB型个体比AA型个体总产仔数、产活仔数及出生窝重高,FSHβ基因AA型个体比BB型个体总产仔数、产活仔数及出生窝重上高,OPN基因BB基因型个体比AA基因型个体总产仔数少1.20头,差异显著(P<0.05);在经产母猪中,OPN和FSHβ基因AA型个体比BB型个体总产仔数和产活仔数高,ESR基因总产仔数、产活仔数规律与初产母猪相反。②在大白猪群体中,ESR基因的优势基因是A等位基因,FSHβ和OPN基因的优势基因是B等位基因;在初产母猪中,ESR基因AA型比BB型个体产活仔数和出生窝重高,OPN和FSHβ基因AA型个体比BB型个体总产仔数和产活仔数高;在经产母猪中,ESR、OPN及FSHβ基因全都表现出BB型比AA型个体总产仔数和产活仔数高的趋势。

山羊卵泡刺激素α亚基cDNA的分子克隆与序列分析

从新屠宰的雌山羊脑垂体中提取总RNA ,反转录获得cDNA .以此cDNA为模板用PCR法扩增目的片段 ,获得长为 380bp的山羊卵泡刺激素α亚基cDNA片段 .将它克隆至pMD 18 T Verctor.随机挑选 3个阳性重组子进行测序 ,将测序结果与绵羊、牛、猪等多种哺乳动物该基因的核苷酸序列及相应氨基酸序列进行比较 .结果表明 ,山羊卵泡刺激素α亚基基因氨基酸序列与绵羊、水牛的同源性最高 ,达 96 % ,与牛的同源性达 95 % ,与人的同源性较低 ,为 74 % .山羊卵泡刺激素α亚基基因编码区的核苷酸与绵羊的同源性最高 ,达 95 % ,与水牛、牛的同源性达 94 % ,与马和大鼠的同源性较低 ,为 85 % .总体来看 ,在哺乳类动物中FSHα亚基基因同源性还是很高的 .

中国大鲵脑cDNA文库构建及促甲状腺激素β亚基基因cDNA的克隆和序列分析

以大鲵脑组织为材料,提取总RNA,应用CloneminerTM文库构建技术构建了cDNA文库。经检测,文库初始滴度为5.0×105cfu.mL-1,重组率为91.70%,插入片断大小在0.35～3.9kb,平均插入片段大小为0.791kb,文库扩增后滴度为4.0×109cfu.mL-1。随机挑选200个阳性克隆测序,经生物信息学分析,发现20个片段与已报道的基因有较高同源性,包括促甲状腺激素基因、生长激素基因及催乳素基因等,其中三个克隆为促甲状腺激素β亚基(TSHβ)基因。序列分析表明,TSHβcDNA全长序列为705bp,包含417bp开放读码框,编码138个氨基酸序列,其中前19个氨基酸为大鲵TSHβ亚基信号肽部分,后接119个氨基酸组成的TSHβ亚基成熟肽。同源性分析显示,大鲵TSHβ亚基与两栖类牛蛙、蟾蜍及爪蟾同一蛋白的同源性分别为53.6%、52.2%和60.3%。采用邻接法和最大简约法对部分脊椎动物TSHβ氨基酸序列构建分子系统树,结果显示有尾目大鲵为两栖动物中较早分化的一支,表明其进化地位较原始。RT-PCR组织分布分析发现,TSHβ基因除垂体有表达外,性腺也有少量表达。

籽鹅促卵泡激素α亚基基因的克隆、序列分析及其原核表达载体的构建

以雌性籽鹅脑垂体的总RNA为模板,利用特异性引物通过RT-PCR扩增获得长为363 bp的籽鹅促卵泡激素α亚基的cDNA片段,将扩增的促卵泡激素α亚基基因片段克隆至pMD18-T载体后进行测序。将测序结果与鸡、鹌鹑、火鸡、鼠、羊、牛等多种禽类和哺乳动物的该基因核苷酸序列及推导的氨基酸序列进行同源性分析。结果表明,这些物种促卵泡激素α亚基基因序列具有较高的保守性,其中与鸡、鹌鹑、火鸡的核苷酸同源性最高,均为95.9%,推导的氨基酸序列与鸡、鹌鹑、火鸡的同源性最高,均为97.5%。为了对克隆的籽鹅促卵泡激素α亚基基因功能研究提供基础,将籽鹅促卵泡激素α亚基基因克隆至pET-32a(+)原核表达载体。

促黄体素β基因多态性及其与济宁青山羊产羔数的关系

为了解促黄体素β基因多态性与济宁青山羊产羔数的关系,采用PCR-SSCP技术检测促黄体素β亚基(Luteinizing hormone beta-subunit,LHβ)基因5′调控区和3个外显子在高繁殖力(济宁青山羊)、中等繁殖力(波尔山羊)和低繁殖力山羊品种(辽宁绒山羊和安哥拉山羊)中的单核苷酸多态性,同时研究该基因对济宁青山羊高繁殖力的影响。结果表明:山羊与绵羊的LHβ基因核苷酸序列相似性为98%。6对引物中,仅引物P1和P5扩增片段存在多态性。对于P1扩增片段,在4个山羊品种中均检测到AA、AB和BB 3种基因型;测序分析发现BB与AA基因型相比在5′调控区存在2处突变202C→A和210C→T;济宁青山羊3种基因型之间产羔数差异均不显著(P>0.05)。对于P5扩增片段,在济宁青山羊中检测到CC、CD和DD 3种基因型,辽宁绒山羊中检测到CC和CD2种基因型,波尔山羊和安哥拉山羊中都只检测到CC基因型;测序分析发现DD与CC基因型相比在外显子2存在单碱基突变1124C→T;济宁青山羊CC、CD和DD基因型频率分别为0.38、0.44和0.18;DD和CD基因型济宁青山羊平均产羔数分别比CC基因型的多0.99(P<0.01)和0.87只(P<0.01)。济宁青山羊在P1、P5两个座位上都处于哈迪-温伯格平衡状态(P1座位:χ2=1.66,P=0.437;P5座位:χ2=1.22,P=0.544)。