Effects of excessive iodide intake during pregnancy and lactation on lipid metabolism in offspring male rats / 中华地方病学杂志

Objective:To study the impact of excessive iodide intake during pregnancy and lactation on lipid metabolism in offspring male rats.Methods:Forty-eight six-week-old Wistar rats (half male and half female) were fed adaptively for one week. The cage was closed according to the ratio of male and female 1∶1. The pregnant rats were divided into two groups according to their weight (220-240 g) by random number table. (1) 10 times high iodine (10 HI) intake during pregnancy and lactation until the postnatal day 21 (PN21) of their offspring: pregnant rats were divided into normal iodine group (NI group, drinking deionized water), 10 HI group (drinking potassium iodide solution with iodine content of 2 250 μg/L). Breast milk was used to feed the offspring rats to PN21, and the offspring male rats were taken as the research subjects, with 6 rats in each group. (2) 100 times high iodine (100 HI) intake during pregnancy and lactation to the offspring postnatal day 120 (PN120): pregnant rats were divided into NI group (drinking deionized water) and 100 HI group (drinking potassium iodide solution with iodine content of 24 750 μg/L). After feeding the offspring rats with breast milk until PN21, the offspring were continued to drink potassium iodide solution with the same iodine content as the mother's to PN120. The offspring male rats were taken as the research subjects, with 6 rats in each group. The levels of free triiodothyronine (FT 3), free thyroxine (FT 4), thyrotropin (TSH) in serum, low-density lipoprotein cholesterol (LDL-C), triglyceride (TG) and total cholesterol (TC) in serum and liver tissue homogenates were measured by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of cholesterol 7α-hydroxylase (CYP7A), low-density lipoprotein receptors (LDLR), sterol regulatory element-binding protein (SREBP)-1c, malic enzyme (ME) and thyroid hormone receptor β (TRβ) in the liver tissue were measured by real-time quantitative PCR. Results:(1) Effects of 10 HI intake during pregnancy and lactation on PN21 offspring male rats: compared with NI and 10 HI groups, the serum FT 3 [(7.53 ± 0.74), (8.88 ± 0.99) pmol/L], FT 4 [(5.58 ± 0.56), (7.68 ± 0.30) pmol/L], TSH levels [(16.69 ± 1.05), (14.49 ± 0.16) ng/ml] of offspring male rats were statistically significant ( t=- 2.91,-8.76, 3.59, P < 0.05). The levels of LDL-C, TG, TC in serum and liver of offspring male rats of 10 HI group were significantly lower than those of NI group ( t=3.28, 8.71, 3.44, 3.70, 3.49, 2.74, P < 0.05). The differences of mRNA expression levels of LDLR, ME, SREBP-1c in the liver of PN21 offspring male rats of 10 HI and NI groups were statistically significant ( t=- 3.50,-3.92, 5.58, P < 0.05). Among them, the levels of LDLR and ME in 10 HI group were higher than those in NI group, while the level of SREBP-1c in 10 HI group was lower than that in NI group. There no significant difference in CYP7A and TRβ mRNA levels between the two groups ( t=- 2.44, 3.20, P > 0.05). (2) Effects of 100 HI intake during pregnancy and lactation on PN120 offspring male rats: there were significant differences in serum FT 3, FT 4 and TSH levels of offspring male rats between 100 HI and NI groups ( t=- 4.39,-3.19, 4.72, P < 0.05). The levels of serum FT 3 and FT 4 in 100 HI group were lower than those in NI group, and the level of TSH in 100 HI group was higher than that in NI group ( P < 0.05). Compared with NI group, the serum and liver LDL-C, TG and TC levels in the offspring male rats of 100 HI group were significantly higher ( t=4.49, 12.85, 16.62, 4.35, 11.04, 16.01, P < 0.05). The differences of CYP7A, LDLR, ME, TRβ and SREBP-1c mRNA levels in liver of PN120 offspring male rats of 100 HI and NI groups were statistically significant ( t=26.40, 54.85,-10.98, 32.52, 10.50, P < 0.05). Among them, the CYP7A, LDLR, ME and TRβ mRNA levels in 100 HI group were lower than those of NI group, while the SREBP-1c mRNA level was higher than that of NI group ( P < 0.05). Conclusions:10 HI intake during pregnancy and lactation to the offspring male rats PN21 showed the serological changes of hyperthyroidism, the levels of blood lipids and liver lipids decreased, the levels of LDLR and ME mRNA increased, and SREBP-1c mRNA decreased in liver. However, 100 HI intake during pregnancy and lactation to the offspring male rats PN120 showed serological changes of hypothyroidism, the levels of blood lipids and liver lipids increased, the levels of CYP7A, LDLR, ME mRNA decreased, and SREBP-1c mRNA increased in liver.

Effects of high iodine given during pregnancy and lactation period of maternal rats and their offspring on thyroid function, memory / 中华地方病学杂志

Objective:The aim of this study was to investigate the alteration of thyroid function, learning and memory induced by high iodine exposure of both maternal rats during pregnancy and lactation period and their offspring and the possible mechanisms.Methods:Wistar rats(6 - 8 weeks old, 220 - 240 g), after one week of adaptation, female rats were mated with male rats (1 ∶ 1). Pregnant rats were divided into normal iodine intake group (NI), 10 times high iodine intake group (10 HI) and 100 times high iodine intake group (100 HI), iodine uptake was 7.5, 75.0, 750.0 μg/d through drinking water and feed, 8 pregnant rats in each group. Their offspring were continuously exposed to different doses of iodine until 180 d after weaning; learning and memory ability was assessed by Morris water maze. Blood sample was taken from the common carotid artery of the offspring and serum was separated. Serum free triiodothyronine (FT 3), free thyroxine (FT 4), and thyroid stimulating hormone (TSH) levels were measured by enzyme linked immunosorbent assay (ELISA). Total RNA was extracted from the hippocampus of the brain and the mRNA expressions of 10-11 translocation methylcytosine dioxygenase (TET 1, TET 2 and TET 3) were detected by Real-time qPCR. The co-localized expressions of thyroid hormone receptor beta 1 (TRβ1) and TET 3 in brain tissue were detected by immunofluorescence staining. Results:Compared with NI group [(4.95 ± 0.53), (12.62 ± 1.81) pmol/L, (7.70 ± 1.04) mU/L], in 10 HI and 100 HI groups, the levels of serum FT 3 and FT 4 [(4.60 ± 0.49), (4.62 ± 0.54), (11.67 ± 1.54), (10.85 ± 1.56) pmol/L] were significantly decreased ( P < 0.05), and serum TSH levels [(9.62 ± 0.93), (9.77 ± 1.28) mU/L] were significantly increased ( P < 0.05). Compared with NI group, the average escape latency of 10 HI and 100 HI groups in the first five days was significantly increased ( P < 0.05), and the space exploration experiment in the sixth day showed that the number of crossing platforms was significantly decreased ( P < 0.05); the mRNA expressions of TET 3 in the hippocampus of the brain were significantly decreased in 10 HI and 100 HI groups ( P < 0.05). Immunofluorescence staining showed that TRβ1 and TET 3 were co-localized in the cerebrovascular intima, choroid plexus and neurons in rats' brain in the 100 HI group. Conclusions:Under the condition of high iodine intake during pregnancy and lactation of the maternal rats, and continued exposure to their offspring until 180 d after weaning, the decreased ability of spatial learning and memory may be related to the alteration of thyroid hormone and the interaction of TET 3 with TRβ1 in the brain.

Iodinated thyroglobulin and autoimmune thyroiditis / 中华地方病学杂志

Iodinated thyroglobulin (TG) can reflect the storage of thyroid hormones and iodine in the thyroid, which has an important influence on thyroid function. However, the study on iodinated TG and autoimmune thyroiditis has yet to be further conducted. This paper reviews the formation of iodinated TG, the measuring method of TG, and the molecular mechanism of iodinated TG function, including the relationships between iodinated TG and thyroid peroxidase adjustment, the expression of thyrotropin receptor, thyroid transcription factor 1, paired box gene 8, and iodine and sodium transporter, last but not least, we aim to investigate the immune and autophagy mechanisms of high iodinated TG-induced autoimmune thyroiditis.

Propylthiouracil, Perchlorate, and Thyroid-Stimulating Hormone Modulate High Concentrations of Iodide Instigated Mitochondrial Superoxide Production in the Thyroids of Metallothionein I/II Knockout Mice

BACKGROUND: Increased oxidative stress has been suggested as one of the underlying mechanisms in iodide excess-induced thyroid disease. Metallothioneins (MTs) are regarded as scavengers of reactive oxygen species (ROS) in oxidative stress. Our aim is to investigate the effects of propylthiouracil (PTU), a thyroid peroxidase inhibitor, perchlorate (KClO4), a competitive inhibitor of iodide transport, and thyroid stimulating hormone (TSH) on mitochondrial superoxide production instigated by high concentrations of iodide in the thyroids of MT-I/II knockout (MT-I/II KO) mice. METHODS: Eight-week-old 129S7/SvEvBrd-Mt1(tm1Bri) Mt2(tm1Bri)/J (MT-I/II KO) mice and background-matched wild type (WT) mice were used. RESULTS: By using a mitochondrial superoxide indicator (MitoSOX Red), lactate dehydrogenase (LDH) release, and methyl thiazolyl tetrazolium (MTT) assay, we demonstrated that the decreased relative viability and increased LDH release and mitochondrial superoxide production induced by potassium iodide (100 µM) can be relieved by 300 µM PTU, 30 µM KClO4, or 10 U/L TSH in the thyroid cell suspensions of both MT-I/II KO and WT mice (P<0.05). Compared to the WT mice, a significant decrease in the relative viability along with a significant increase in LDH release and mitochondrial superoxide production were detected in MT-I/II KO mice(P<0.05). CONCLUSION: We concluded that PTU, KClO4, or TSH relieved the mitochondrial oxidative stress induced by high concentrations of iodide in the thyroids of both MT-I/II KO and WT mice. MT-I/II showed antioxidant effects against high concentrations of iodide-induced mitochondrial superoxide production in the thyroid.

Early effects of iodine excess on spleen cells of methallothionein Ⅰ/Ⅱ knockout mice / 中华地方病学杂志

Objective To investigate the effects of iodine excess on spleen cell viability,lactate dehydrogenase (LDH) leakage,mitochondrial superoxide production and peroxiredoxin (Prx)3 expression in methallothionein Ⅰ / Ⅱ knockout (MT-Ⅰ / Ⅱ KO)mice.Methods Spleen cell suspensions were prepared from six to eight-week old and healthy male MT-Ⅰ / Ⅱ KO mice and wild type (WT) mice; the cell number was adjusted to 5 × 107/L and the cells were plated in 96-well plates (100 μl each well); the cells were exposed to various concentrations of KI (0,10-4,10-3,10-2 mol/L) and 10-3 mol/L H2O2,respectively,for two hours,and control group did not give KI nor H2O2.Cell viability was assayed by methyl thiazolyl tetrazolium (MTT) colorimetric method.Cell damage was detected by chemical colorimetric method.Mitochondrial superoxide production in the spleen cells was measured by flow cytometry.Western blotting technology was used to investigate the expression of Prx3.Results In both MT-Ⅰ/Ⅱ KO and WT mice,the differences of cell viability,LDH leakage,mitochondrial superoxideproduction and the expression of Prx3 of spleen cells among the treatment groups were statistically significant (F =357.92,71.03,130.36,10.36,179.58,26.92,187.43,and 7.16,all P ＜ 0.05).Compared to the control group [(100.00 ± 2.00)％,(100.00 ± 1.63)％,(3 202.22 ± 85.63),(3 161.51 ± 144.49)U/L,43.82 ± 1.56,38.60 ± 2.81,0.61 ± 0.09,0.50 ± 0.08],cell viability of 10-4,10-3,10-2 mol/L KI treatment and 10-3 mol/L H2O2 groups [(80.77 ± 1.86)％,(89.89 ± 2.90)％,(76.08 ± 1.92)％,(87.66 ± 1.74),(73.26 ± 1.86)％,(84.30 ± 2.23)％,(66.22 ± 1.71)％,(70.80 ± 1.49)％] was decreased (all P ＜ 0.05); LDH leakage [(3 880.00 ± 190.62),(3 431.17 ± 170.45),(4 178.33 ± 170.43),(3 598.63 ± 189.09),(4 388.61 ± 123.79),(3 863.72 ± 195.64),(4 615.28 ± 196.17),(4 148.12 ± 195.81)U/L] was increased significantly (all P＜ 0.05); and mitochondrial superoxide production in the spleen cells (53.83 ± 3.22,47.03 ± 1.60,58.92 ± 4.00,50.48 ± 2.59,72.72 ± 2.14,68.53 ± 2.97,80.76 ± 4.11,75.26 ± 3.41) was increased significantly (all P ＜ 0.05); Prx3 expressions in 10-3、10-2 mol/LKI and 10-3 mol/L H2O2 treatment groups (0.82 ± 0.12,0.65 ± 0.12,0.96 ± 0.15,0.73 ± 0.16,1.04 ± 0.13,0.85 ± 0.16) significantly increased (all P ＜ 0.05),the differences of Prx3 expressions between 104 mol/L KI groups (0.73 ± 0.15,0.55 ± 0.09),and control groups were not statistically significant (all P ＞ 0.05).In 104,10-3,10-2 mol/L KI and 10-3 mol/L H2O2 treatment groups,cell viability of MT-Ⅰ/Ⅱ KO mice spleen was lower than that of WT mice (t =6.47,10.93,9.30 and 4.96,all P ＜ 0.05); LDH leakage was higher than that of WT mice (t =4.30,5.58,5.56 and 4.13,all P ＜ 0.05); mitochondria superoxide production was higher than that of WT mice (t =4.64,4.33,2.80 and 2.52,all P ＜ 0.05); Prx3 expression was higher than that of WT mice (t =2.54,2.37,2.59 and 2.27,all P ＜ 0.05).Conclusions KI may decline the cell viability,increase the leakage of LDH and increase the production of mitochondrial superoxide production and Prx 3 expression,which are much more significant in MT-Ⅰ /Ⅱ KO mice,suggesting that MT Ⅰ /Ⅱ has some antioxidative effect in high concentration of iodide induced oxidative stress in the spleen.

Peroxide effects of iodide excess on mitochondria in Fischer rat thyroid cell line in the early period / 中华地方病学杂志

Objective To investigate the peroxide effects of iodide excess on mitochondria in Fischer rat thyroid cell line(FRTL) in the early period.Methods After treatment with 0.0 mmol/L(control group) or 0.1 mmol/L potassium iodide(KI) for 2,4 and 24 h,respectively,changes of mitochondrial superoxide formation were assayed by flow cytometry and fluorescence microscopy using mitochondria-targeted hydroethidine(MitoSOX).The cytochrome c (cyt c) released from mitochondria to cytoplasm was detected by immunocytochemistry.The lactate dehydrogenase (LDH) released into supernatant was measured by a LDH kit using colorimetry.The percent of dead cells was assayed by flow cytometry using propidium iodide (PI).DNA with loading buffer was separated in 1％ agarose gel.Results Mitochondrial superoxide production in FRTL cells treated with 0.1 mmol/L KI was increased at 2; 4 and 24 h,especially at 2 h.The rates of cyc c protein immunity positive cells at 2,4 and 24 h in 0.1 mmol/L KI group were (35.3 ± 3.6)％,(45.8 ± 5.5)％ and (30.3 ± 6.1)％,respectively,which were significantly higher than that in control group[(14.8 ± 1.2)％,all P ＜ 0.05].The LDHs released into supernatant at 2,4 and 24 h in 0.1 mmol/L KI group were (1.85 ± 0.32),(6.63 ± 0.42) and (8.35 ± 0.34)U/mg,and these values at 4 and 24 h in 0.1 mmol/L KI group were significantly higher than that of control group[(0.89 ± 0.04)U/mg,all P ＜ 0.05].After incubation with 0.1 mmol/L KI for 2,4 and 24 h,the percentages of dead FRTL cells were 7.52％,9.29％ and 13.71％,respectively,while that of control group was 4.66％.After FRTL cells were treated with 0.1 mmol/L KI for 24 h,DNA ladder appeared.Conclusion Iodide excess (0.1 mmol/L) can cause mitochondrial peroxide injury in FRTL cells at 2 h and cell apoptosis at 24 h.

Effects of high concentrations of iodide exposure on mitochondrial superoxide production in the thyroid of metallothionein Ⅰ/Ⅱ knockout mice / 中华地方病学杂志

Objective To investigate the effects of high concentrations of iodide exposure on mitochondrial superoxide production,cell viability and cell damage in the thyroid of metallothionein Ⅰ/Ⅱ knockout (MT-Ⅰ/Ⅱ KO) mice and corresponding wild type (WT) mice.Methods Thyroid cell suspension of six to eight weeks old healthy male MT-Ⅰ/Ⅱ KO mice and WT mice were prepared.The thyroid cells were treated with high concentrations (10-4,10-3,10-2 mol/L) of potassium iodide(KI),or 10-3 mol/L hydrogen peroxide(H2O2) for 2 hours,respectively.Cell viability was evaluated with methyl thiazolyl tetrazolium(MTT) assay.Lactate dehydrogenase (LDH) level in cell culture medium was detected by enzyme-linked immunosorbent assay(ELISA).Mitochondrial superoxide production in the thyroid cells was measured by flow cytometry using a fluorescent probe,mitochondrial superoxide(MitoSOX).Results Compared to the control group[(100.00 ± 0.00)％,(100.00 ± 0.00)％],the cell viability of 10-4,10-3,10-2 mol/L KI and 10-3 mol/L H2O2 exposure groups were significantly decreased in the thyroid cells of both WT [(73.63 ± 2.05)％,(72.41 ± 2.26)％,(69.63 ± 2.29)％,(44.90 ± 2.93)％] and MT-Ⅰ/Ⅱ KO mice[(65.40 ± 2.39)％,(64.51 ± 2.27)％,(61.48 ± 2.33)％,(40.80 ± 2.76)％,all P＜ 0.05].Compared to the control group [(1 995.28 ± 30.52),(2 004.96 ± 19.71)U/L],significantly increased LDH activities were detected in the thyroid cells of WT [(2 809.22 ± 156.53),(2 850.80 ± 137.83),(2 920.45 ± 152.92),(4 487.49 ± 130.67)U/L] and MT-Ⅰ / Ⅱ KO mice [(3 261.06 ± 120.44),(3 474.19 ± 142.15),(3 597.08 ± 150.86),(4 706.64 ± 148.57)U/L,all P ＜ 0.05].Compared to the control group (26.49 ± 7.66,37.11 ± 8.48),the MitoSOX red fluorescence intensities of 10-2 mol/L KI and 10-3 mol/L H2O2 groups were significantly increased in WT mice(58.96 ± 5.11,87.95 ± 4.25) and MT-Ⅰ/ⅡKO mice(71.21 ± 5.55,99.76 ± 4.42) by flow cytometry (all P ＜ 0.05).Compared to the thyroid cells in WT mice,significantly decreased cell viability (all P ＜ 0.05),significantly increased LDH activity(all P ＜ 0.05) and significantly increased MitoSOX red fluorescence intensity by flow cytometry (all P ＜ 0.05) were detected in the thyroid cells of MT-Ⅰ/Ⅱ KO mice following treatment with KI or H2O2.Conclusions High concentrations of iodide (10-2 mol/L) and 10-3 mol/L H2O2 may lead to significant increase of mitochondrial superoxide production and LDH activity,decrease of cell viability in both WT and MT-Ⅰ / Ⅱ KO mice.More significant increase of superoxide production is detected in MT-Ⅰ / Ⅱ KO mice,indicating the potential protective role of metallothionein in the thyroid cells of WT mice.

Influence of Hydrogen Peroxide on Mitochondrial Membrane Potential and Superoxide Production in FRTL Cell / 天津医药

Objective:To investigate the effects of exogenous hydrogen peroxide(H_2O_2)on mitochondrial superoxide production and mitochondrial membrane potential changes(△ψ)in fisher rat thyroid cell line(FRTL).Methods:Following 1 mmol/L H_2O_2 exposure in FRTL cells for 10 min,30 min and 24 h,mitochondrial superoxide production was measured by living cell imaging and flow cytometry using MitoSOX.The mitochondrial membrane potential was assayed by spectrofluorimeter and fluorescent microscopy using rhodamine 123(rh123).The cell viability was detected by MTT colorimetric method.Morphological changes were observed by invert microscope.Apoptosis assay was performed by acridine orange staining.Results:Quantitative measurements of the mean intensities of MitoSOX demonstrated significant increase with 1 mmol/L H_2O_2 following 10 min,30 min and 24 h treatment in FRTL cells compared with that of control.Fluorescence intensity of rh123 and optical density of MTr were significantly decreased in FRTL ceils with 1 mmol/L H_2O_2 following 30 min and 24 h treatment(P ＜ 0.01).Under light microscope and fluorescence microscope the characteristic morphological features of programmed cell death,pickuosis,karyorrhexis,and cell shrinkage were observed after acridine orange staining.Conclusion:Acute and chronic exogenous H_2O_2 exposure cause oxide stress in FRTL cells,which result in the increase of mitochondrial superoxide production,△ψdecline,cell necrosis and apoptosis.

Application of Small Dose of Ketamine in Painless Gastroscopy / 中国康复理论与实践

@#Objective To observe the effect of intravenous small dose of ketamine combined with continuous infusion of propofol and sufentanyl in painless gastroscopy.Methods 62 patients undergoing painless gastroscopy under intravenous anesthesia were randomly divided into the control group and ketamine group with 31 cases in each group. The cases of the control group were treated with continuous infusion of propofol (target controlled infusion rate: 3.0 μg/ml) and sufentanyl (loading dose: 0.15 μg/kg, basal rate 0.15 μg/kg/h); those of the ketamine group were treated with intravenous small dose of ketamine 0.5 mg/kg combined with continuous infusion of propofol (target controlled infusion rate: 3.0 μg/ml) and sufentanyl (loading dose: 0.1 μg/kg, basal rate 0.1 μg/kg/h). The procedure of gastroscopy was started when patients were in unconsciousness and their vital signs were stable, and the infusion of medicine was stopped when the gastroscopy was finished. The systolic blood pressure (SBP) and heart rate (HR) at the time of before anesthesia, 1 minute after anesthesia, inserting the gastroscope, 10 minutes after inserting the gastroscope, and recovering from the anesthesia were recorded. The cases needing special respiratory management (SRM), displaying body movement (BM) in gastroscopy and post-operative nausea and vomiting (PONV) were recorded. The recovery time (RT) after gastroscopy was also recorded.Results No significant difference was found in SBP, HR, BM, PONV and RT between two groups ( P>0.05). Eight cases in the control group and one case in the ketamine group needed SRM before inserting the gastroscope due to transient respiratory depression ( P<0.01).Conclusion The application of intravenous small dose of ketamine combined with continuous infusion of propofol and sufentanyl in painless gastroscopy is effective and safe.

Leptin and chronic renal failure / 中国实用内科杂志

Leptin,a polypeptide hormone coded by Ob-gene,has the functions of depressing appetite,reducing caloric intake and enhancing caloric consumption,etc.Leptin has a close relationship with kidney,for it is metabolized and eliminated via kidney.Meanwhile,leptin can also directly affect kidney functions,influencing the process of chronic renal failure.Furthermore,hyper-leptinemia is associated with anorexia,malnutrition and cardiovascular diseases.Different dialysis patterns may also play different roles in cleaning leptin.For example,common dialysis could not clear leptin,but other dialysis modes such as high permeability dialysis,hemodiafiltration,and mastic absorption hemoperfusion can alleviate hyper-leptinemia through cleaning leptin partly.For kidney transplantation patients,hyper-leptinemia may affect the immune balance which has been maintained for a long time.

Concentrations of Morphine in Colostrum and Plasma During Postcesarean Epidural Anesthesia / 中华围产医学杂志

Objective To study the effect of morphine used in epidural anesthesia on puerpera after cesarean section and the safety of morphine to neonate. Method One hundred puerpera undergone cesarean section were randomly divided into test group and control group with continuous 2% lidocaine epidural anesthesia, as soon as the operation were finished, 2 mg morphine was injected into vacum epidurale for test group, nothing for control group. Colostrum and plasma, urine samples of puerpera and neonate were collected, morphine and metabolite level were tested by GC-MS and FPIA. Result Morphine concentrations ranged from 0.05). Conclusion As the epidural analgesia medicine after cesarean section, morphine has no side-effect to neonate and is safe to neonate.

Characterization of thyrotropin receptor molecule and its active peptide as immunogens in BALB/c mice / 中国病理生理杂志

AIM: To study the characterization of thyrotropin receptor (TSHR) and its active fragment TSHR aa352-366 as immunogens in BALB/c mice. METHODS: BALB/c mice were injected peritoneally with TSHR aa352-366-KLH (hemocyanin from keyhole limpets) and the mixture of TSHR aa352-366-KLH and guinea pig TSHR every 15 days, respectively. The levels of thyroid hormones and TSHR antibodies and TSHR mRNA were measured, and the pathological changes of thyroid tissue were observed. RESULTS: In the group injected with TSHR aa352-366-KLH, the serum levels of TT_3 and TT_4 decreased (P