Identification and treatment of Hashimoto's hyperthyroidism / 中国实用内科杂志

Hashitoxicosis patients have the clinical characteristics of Graves' disease with hyperthyroidism and the pathological phenotype of Hashitoxicosis. The initial stage of hyperthyroidism is almost indistinguishable from Graves' disease in clinical manifestations, including increased uptake of radioactive iodine in the thyroid and positive antibodies to thyrotropin receptors, as well as high levels of thyroid peroxidase antibodies and/or thyroglobulin antibodies. Generally, the symptoms of hyperthyroidism are relatively mild, and the stage of hyperthyroidism is relatively short. Hashitoxicosis patients are sensitive to antithyroid drugs and are prone to hypothyroidism in the course of treatment. It will develop into permanent hypothyroidism after 3-24 months. Therefore, antithyroid drugs should be started at low doses, thyroid function should be detected frequently, and timely drug adjustment should be made.Radioactive iodine or surgical treatment should be avoided.

Expression of hepatitis B virus 1.3-fold genome plasmid in an SV40 T-antigen-immortalized mouse hepatic cell line.

AIM: To investigate the expression of the hepatitis B virus (HBV) 1.3-fold genome plasmid (pHBV1.3) in an immortalized mouse hepatic cell line induced by SV40 T-antigen (SV40T) expression. METHODS: Mouse hepatic cells were isolated from mouse liver tissue fragments from 3-5 d old Kunming mice by the direct collagenase digestion method and cultured in vitro. The pRSV-T plasmid was transfected into mouse hepatic cells to establish an SV40LT-immortalized mouse hepatic cell line. The SV40LT-immortalized mouse hepatic cells were identified and transfected with the pHBV1.3 plasmid. The levels of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in the supernatant were determined by an electrochemiluminescence immunoassay at 24, 48, 72 and 96 h after transfection. The expressions of HBsAg and hepatitis B c antigen (HBcAg) in the cells were investigated by indirect immunofluorescence analysis. The presence of HBV DNA replication intermediates in the transfected cells and viral particles in the supernatant of the transfected cell cultures was monitored using the Southern hybridization assay and transmission electronic microscopy, respectively. RESULTS: The pRSV-T plasmid was used to immortalize mouse hepatocytes and an SV40LT-immortalized mouse hepatic cell line was successfully established. SV40LT-immortalized mouse hepatic cells have the same morphology and growth characteristics as primary mouse hepatic cells can be subcultured and produce albumin and cytokeratin-18 in vitro. Immortalized mouse hepatic cells did not show the characteristics of tumor cells, as alpha-fetoprotein levels were comparable (0.58 ± 0.37 vs 0.61 ± 0.31, P = 0.37). SV40LT-immortalized mouse hepatic cells were then transfected with the pHBV1.3 plasmid, and it was found that the HBV genome replicated in SV40LT-immortalized mouse hepatic cells. The levels of HBsAg and HBeAg continuously increased in the supernatant after the transfection of pHBV1.3, and began to decrease 72 h after transfection. The expressions of HBsAg and HBcAg were observed in the pHBV1.3-transfected cells. HBV DNA replication intermediates were also observed at 72 h after transfection, including relaxed circular DNA, double-stranded DNA and single-stranded DNA. Furthermore, a few 42 nm Dane particles, as well as many 22 nm subviral particles with a spherical or filamentous shape, were detected in the supernatant. CONCLUSION: SV40T expression can immortalize mouse hepatic cells, and the pHBV1.3-transfected SV40T-immortalized mouse hepatic cell line can be a new in vitro cell model.

[Functional analysis of hepatitis B virus immune escape mutants with insertion mutations in the surface antigen].

OBJECTIVE: To evaluate the influence of insertion mutations occurring in the hydrophobic region, between amino acids 114 and 115, of the hepatitis B surface antigen (HBsAg) on viral antigenicity and replication. METHODS: Hepatitis B virus (HBV) DNA was obtained from patients with HBsAg-positive chronic hepatitis B (CHB) infection and subjected to sequence analysis and comparison to GenBank reference sequences for HBV genotype B (AB073826) and genotype C (AF286594). Insertion mutations detected in the HBsAg region were used to make recombinant expression plasmids via site-directed mutagenesis. After transfecting the recombinant HBsAg into Huh7 cells, the mutants' effects on viral antigenicity and replication were evaluated by chemiluminescence microparticle immunoassay (CMIA) and Southern blot hybridization, respectively. The viral antigenicity of each mutant was predicted by bioinformatic analysis, using the Jameson-Wolf method to predict the antigenic index, the Hopp-Woods method to predict hydrophilicity, the Emini method to predict the probability of a region lying of the protein's surface, and the Karplus-Schulz method to predict the flexibility of the protein backbone. RESULTS: Two CHB patients harbored HBV with insertion mutations in HBsAg: one with two (NT) and one with three (NTT) inserted amino acids between 114 and 115. The NTT recombinant HBsAg mutant showed no impact on viral replication and reacted weakly with anti-HBs in CMIA (P = 0.02). The antigen indices for the insertion of NTT were 1.00, -0.16, and 0.18, and insertion of the three amino acids affected the index values of five proximal amino acid sites (with an average increase of 0.13). The hydrophilic indices for the insertion of NTT were 0.2, -0.4, and -0.4, with no significant effect on the proximal amino acids. The insertion of the three amino acids changed both the surface probability (range: -0.55 to 2.97; affecting eight proximal amino acids) and the flexibility (range: -0.01 to 1.1; affecting five proximal amino acids). CONCLUSION: The NTT three amino acids insertion in the HBsAg open reading frame, between 114 and 115 of the wild-type sequence, detected in a CHB patient may play a role in HBV immune escape without influencing the viral replicative capacity.

[A retrospective follow-up study of prolonged nucleos(t)ide analogue and interferon combination therapy in HBeAg-positive chronic hepatitis B patients].

OBJECTIVE: To investigate the efficacy of a 96-week course of nucleos(t)ide analogue and interferon (IFN) combination therapy for achieving seroconversion at 24 weeks after completion in patients with chronic hepatitis B (CHB). METHODS: One-hundred-and-thirty-five CHB patients with positivity for hepatitis B e antigen (HBeAg) were recruited for study between January 2005 and December 2008. All patients were given a 96-week course of nucleos(t)ide analogue (lamivudine or adefovir dipivoxil) alone (monotherapy controls, n = 45) or in combination with IFN or Pegylated-IFN-alpha-2a (Peg-IFNa-2a) (n = 90). At treatment weeks 12, 24, 48, 72, and 96, and at 24 weeks after treatment completion, serum samples were collected from all patients for assessment of biochemical, virological and serological responses to treatment. The biochemical response was indicated by normalization of the alanine aminotransferase (ALT) level. The virologic response was indicated by a reduction in the hepatitis B virus (HBV) DNA level to less than 1000 copies/ml. The serological response was indicated by seroconversion of either HBeAg or hepatitis B surface antigen (HBsAg). Statistical analysis was performed with the Chi-squared test. RESULTS: Among the patients treated with nucleos(t)ide analogue and IFN combination therapy, 41.1% (37/90) achieved HBeAg seroconversion and 18.9% (17/90) achieved HBsAg seroconversion at the end of treatment. However, significantly less of the patients treated with nucleos(t)ide analogue monotherapy achieved HBeAg seroconversion and none achieved HBsAg seroconversion by end of treatment (33.3% and 0%, respectively; x2= 8.08, P less than 0.01 vs. the combination therapy group). Age stratification of the 17 HBsAg-seroconverted patients treated with combination therapy indicated that the HBsAg seroconversion rate was significantly higher in patients less than 30-years-old than those 30 and older (x2= 12.62 and 4.24, respectively, P less than 0.05). At post-treatment week 24, the 17 HBsAg-seroconverted patients treated with combination therapy showed HBsAg titers of less than 250 IU/ml; moreover, 11.8% (2/17) of these patients remained HBeAg-positive and 17.6% (3/17) showed abnormal ALT levels and elevated HBV DNA. CONCLUSION: Prolonged nucleos(t)ide analogue plus IFN combination therapy can significantly improve the rate of HBsAg seroconversion in HBeAg-positive CHB patients, and this treatment regimen is especially efficacious in patients under the age of 30.

Dynamic changes of sICAM-1 in patients suffering from hemorrhage fever with renal syndrome / 中国地方病学杂志

Objective To study the dynamic changes of soluble intracellular adhesion molecule-1 (sICAM-1), and to assess its role in the pathogenesis of hemorrhagic fever(HFRS) with renal syndrome. Methods A total of 30 patients with HFRS(case group) and 20 healthy subjects (control group) from Worker Hospital of Finance & Commerce of Qiqihar city were included in the study during 2006 - 2007. Double-antibody sandwich ELISA method was used to measure the level of sICAM-1 in serum samples. Results The serum levels of sICAM-1 in patients with HFRS in early stage, critical stage, convalescent stage were(47.56±6.51), (94.23±15.36), (54.19±8.42)ng/L,respectively. The serum level of sICAM-1 in control group was (22.63±3.40)ng/L. The above values were compared between any two groups, differences were statistically significant (all P < 0.01). The serum level of urea nitrogen in patients with HFRS in early stage, critical stage, convalescent stage were (5.16±0.12), (33.84±9.24),(8.20±1.30)mmol/L, respectively. The serum level of urea nitrogen in control group was (4.20±0.56)mmol/L.The above value were compared between any two groups, the differences were statistically significant (all P< 0.01). Conclusions sICAM-1 is involved in the pathogenesis of HFRS as protective factors. Cellular immune playes an important role in the pathogenesis of HFRS.

[Clinical implication of expression of perforin/granzyme in peripheral blood lymphocyte in severe hepatitis B].

OBJECTIVE: To explore role of perforin/granzyme expression of peripheral blood lymphocyte in injury of hepatocyte in severe hepatitis B, and evaluate relationship between perforin/granzyme expression levels and hepatitis B virus (HBV)-DNA load. METHODS: Thirty eight patients of severe hepatitis B were enrolled in the study. Fasting venous blood was collected on following morning of admission. Twenty adult healthy subjects served as healthy control group. Perforin/granzyme expression of peripheral blood lymphocyte was detected by flow cytometry, and serum HBV-DNA load was detected by fluorescence quantitative polymerase chain reaction (PCR). RESULTS: Positive rate of perforin/granzyme in peripheral blood lymphocyte in severe hepatitis B was higher than that of the healthy control group [perforin: (43.42+/-19.28)% vs. (19.65+/-9.27)%, granzyme: (40.35+/-12.26)% vs. (22.28+/-9.35)%, both P<0.01]. There was a significant negative correlation between the perforin/granzyme expression of peripheral blood lymphocyte and serum HBV-DNA load (r(perforin) =-0.92, r(granzyme) =-0.96, both P<0.01), the higher serum HBV-DNA load, the lower perforin/granzyme expression in severe hepatitis B. CONCLUSION: Perforin/granzyme overexpression in peripheral blood lymphocyte is an important factor in injury of hepatocyte in patients with severe hepatitis B, and the expression may be involved in HBV-DNA cleanup.

Detection of circulating antigen with a MAb-based sandwich-ELISA and its comparison with specific IgM detection in sera of patients with hemorrhagic fever with renal syndrome.

A monoclonal antibody (1H2) against nucleocapsid protein of Hantavirus was developed. A sandwich enzyme-linked immunosorbent assay (ELISA) with the monoclonal antibody (MAb) was established and evaluated for detecting circulating antigen (CAg) in serum of patients with hemorrhagic fever with renal syndrome (HFRS). Results were compared to that of an immunoglobulin M (IgM)-detecting ELISA. Of 143 patients with HFRS, 106 were positive for CAg of Hantavirus and 128 positive for specific IgM. Among the 15 HFRS patients in whom specific IgM was not detected, 10 were positive for CAg. Of 100 controls including 40 hepatitis B cases, 40 measles cases, and 20 healthy persons all were negative for both CAg and specific IgM. Detection of Hantavirus CAg with a MAb-based sandwich ELISA (MBS-ELISA) established in the present study adds a new diagnostic tool for HFRS, and it increases the diagnostic rate to conventional specific IgM detection, especially for patient in the early stage of HFRS.

Epidemiological study of the effects of smoking cigarette on thyroid gland / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the effect of cigarette smoking on thyroid gland volume, thyroid function and thyroid autoantibodies in the areas with different iodine intakes.</p><p><b>METHODS</b>A cross-sectional epidemiological study in Panshan (mild iodine-deficient area), Zhangwu (more than adequate iodine intake area) and Huanghua (iodine-excessive area) was conducted in 3761 subjects in 1999.80.2 % of them were followed up in 2004. Questionnaires, thyroid function, thyroid autoantibodies, urinary iodine concentration,and thyroid B ultrasound were performed.</p><p><b>RESULTS</b>The prevalence of goiter was higher in smokers than in non-smokers (15.1% vs. 11.5%, P< 0.05). The average thyroid volume was higher in smokers with phenomenon more obvious in Panshan and Huanghua areas. Data from logistic analysis showed that smoking cigarette was an independent risk factor of goiter. There was no difference in serum TSH and Tg level between smokers and non-smokers. The positive rate of TPOAb (>100 IU/ml) was higher in smokers than in non-smokers(10.8% vs. 9.0 % , P <0.05) and was especially obvious in Huanghua area. Smoking was a independent risk factor of increasing positive rate of TPOAb. During the prospective observation,it was found that the incidence of positive TPOAb(>,100 IU/ml) was 7.4% in the subjects that were from non-smokers turning to smokers and 2.9% in those whose smoking behavior did not change. Logistic analysis indicated that the shifting from non-smoking to smoking was independent risk factor for the increase on high incidence of positive TPOAb.</p><p><b>CONCLUSION</b>Smoking cigarette was a independent risk factor of goiter. Smoking was also a risk factor of increasing TPOAb positive rate. Shifting from not smoking to smoking was an independent risk factor of increasing high incidence of positive TPOAb.</p>

[Study on the cost-effectiveness, benefit and utility analysis on the infant inoculation hepatitis B vaccine in Shanghai].

OBJECTIVE: To study the cost-effectiveness, benefit and utility of infant hepatitis B vaccination in Shanghai from 1992 to 2001. METHODS: To calculate the cost of hepatitis B vaccination by cost analysis method. Both the numbers of persons with HBsAg positive and patient with hepatitis B, cirrhosis and liver cancer decreased as the index of direct effect. To study the sick-time and the cost of treating hepatitis B, cirrhosis and liver cancer patients, a face to face questionnaire was used and quasi method was adopted to understand the effect of cure and the course of hepatitis B. The cost benefit analysis method was also used to calculate the cost benefit of HBV vaccine. The disability adjusted life years (DALY) was regarded as an index of utility to measure the disease burden. RESULTS: Input of 501,129.49 Yuan might have the result of reducing one liver cancer patient, ten cirrhosis patients, one hundred chronic hepatitis B patients and one thousand HBsAg positive people. The cost of hepatitis B vaccination was 0.24 hundred million Yuan during the past ten years in Shanghai, which had obtained the total benefit value of 41.22 hundred million Yuan, with a cost benefit ratio of 1:172 Yuan. It was estimated that the total disease burden of hepatitis B, cirrhosis and liver cancer patients was 59,762.55 DALY in order to reduce one DALY loss cost of 402.50 Yuan. CONCLUSION: HBV vaccine inoculation in infants seemed to be a low-cost input and high-effect output strategy.

Determination of cut-off points of thyroid peroxidase antibody and thyroglobulin antibody and their clinical significance / 中华检验医学杂志

40 IU/ml were used as cut-off points.(4) TSH level was higher in subjects with positive thyroid autoantibodies than those without antibodies (P

Impact of iodine intake on thyroid diseases——A five-year prospective epidemiological study / 中华内分泌代谢杂志

Objective To evaluate the relationship between iodine intake level and the occurrence and development of thyroid diseases.Methods During the period of 1999-2004,a cross-sectional survey in 1999 and a 5-year follow-up survey were conducted in the 3 communities with different iodine intake levels in China:Panshan (mild deficiency),Zhangwu (more than adequate) and Huanghua (iodine excess).A total of 3 761 subjects were included in 1999 and 80.24% of them participated in the 5-year follow-up survey in 2004.The levels of serum thyroid hormones and thyroid autoantibodies were measured,and thyroid B ultrasound was performed.Results (1) The prevalence of overt hypothyroidism in 1999 in Zhangwu and Huanghua was 3.5 and 7.3 times,and that of subclinical hypothyroidism was 3.2 and 6.6 times as high as in Panshan,respectively.The 5-year cumulative incidence of subclinical hypothyroidism was 11.3 and 12.6 times as high as in Panshan,but no difference of incidence of overt hypothyroidism was found.(2) The prevalence of autoimmune thyroiditis (AIT) in 1999 in Zhangwu and Huanghua was 3.8 and 6.2 times,and the S-year cumulative incidence of AIT was 4.4 and 5.5 times as high as in Panshan.(3) During the 5-year follow-up period,the incidences of hypothyroidism in subjects with positive thyroid peroxidase antibody and euthyreidism in Zhangwu and Huanghua were 4.2 and 10.3 times as high as in Panshan.(4) No significant differences in both prevalence in 1999 and 5-year cumulative incidence of overt hyperthyroidism were found in the 3 communities.(5) The incidence of thyroid cancer was 19.37/100 000 per year in Huanghua,and all of the cases were papillary thyroid cancer,and no thyroid cancer was found in Zhangwu and Huanghua.Conclusion More than adequate or excessive dietary iodine intake may induce and promote the occurrence and development of hypothyroidism and AIT,therefore,median urinary iodine excretion (MUI)more than 200?g/L in the population appears not safe.The observation that a higher incidence of thyroid cancer occurred in the iodine excessive areas with MUI 600?g/L deserves further investigation.

Thyroid autoantibodies In a five-year follow-up survey of populations with different Iodine intakes / 中华内分泌代谢杂志

Objective To perform a follow-up survey about thyroid disorders in three rural communities with different iodine intakes in China,observe the incidences and natural outcomes of positive thyroid autoantibodies in euthyroid population,and also explore the influence of iodine intakes on these outcomes and autoantibodies.Methods In 1999,a cross-sectional study on thyroid disorders was performed in three rural communities of Pansan ( mild iodine deficient area),Zhangwu ( more than adequate iodine intake area) and Huanghua (excessive iodine intake area) in China.The 5-year follow-up study was performed in 2004.Both in 1999 and 2004,serum levels of TSH,thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TgAb) were measured with the same method.Iodine in urine and B ultrasound on thyroid were also examined.Results Among the euthyroid subjects with normal TSH level in 1999,an increased prevalence of positive TPOAb in Zhangwu and an increased prevalence of positive TgAb in Hnanghua were observed in 2004 (both P＜0.05 ).Most of euthyroid subjects with positive TPOAb or positive TgAb in 1999 remained positive thyroid antibodies in 2004. The percentage was even higher in those with high levels of antibodies.Thyroid dysfunction was more frequent in subjects with positive TPOAb and/or TgAh than in those without thyroid antibodies (14.44% vs 3.11%,P＜0.01).For those with positive antibodies in 1999,the incidence of hypothyroidism in 2004 was 1.32%,8.46% and 15.38% in Pansan,Zhangwu and Huanghua,respectively (P＜0.05).The 5-year cumulative incidences of positive TPOAb (≥50 U/ml) and TgAb (≥40 U/ml) were 2.81% and 3.82%,respectively.The incidence of positive TPOAb was the highest in Zhangwu ( 3.84% ).The incidence of positive TgAb was the highest in Huanghua (5.07%),significantly higher than those in other two areas (P＜0.05).Conclusion The incidence of thyroid dysfunction is higher in subjects with positive antibodies than that in those with negative antibodies.The high iodine intake is a risk factor of hypothyroidism in subjects with positive thyroid antibodies.Sustained excessive iodine intake increases the incidence of positive thyroid antibodies.

Incidences of hyperthyroidism in communities with different iodine intake levels:a five-year prospective and comparative epidemiological survey / 中华内分泌代谢杂志

Objective To analyze the relationship between different iodine intakes and epidemiological features of hyperthyroidism in 3 communities with different iodine intake levels,as well as to explore the risk factor for the pathogenesis of hyperthyroidism.Methods A cohort study was performed in 3 Chinese rural communities with long-term stable iodine nutrition status defined as mild deficient (Panshan),more than adequate (Zhangwu, previously mild iodine deficiency and salt iodization lasting 8 years until 2004) and excessive (Huanghua).A total of 3 761 (934 males,2 827 females) inhabitants aged above 13 participated in the original investigation in 1999 and 3 018(709 males,2 309 females) of them received identical examinations after 5 years.Results Median urinary iodine concentration of school-aged children was 87.61zg/L in Panshan,213.9?g/L in Zhangwu and 633.5?/L in Huanghua (P＜0.05) at fallowing up,and no statistic significance was found when comparing the corresponding data from original survey (83.5?g/L,242.9?g/L and 650.9?g/L).The accumulative incidence of hyperthyroidism was 13.6/1 000,9.4/1 000 and 8.1/1 000 (P＞0.05),being 16.4/1 000,11.2/1 000 and 9.1/1 000 (P＞0.05) in female,respectively in Panshan,Zhangwu and Huanghua.In original healthy subjects (without history of thyroid disease and overt thyroid dysfunction) merely with either positive thyroid peroxidase antibody (TPOAb＞50 IU/ml) or thyroglobulin antibody (TgAb＞40 IU/ml),the accumulative incidence of newly diagnosed hyperthyroidism was not significantly different among 3 communities in follow-up survey.TSH＜0.3 mIU/L,positive TPOAb or goiter in original healthy participants were associated with the occurrence of overt hyperthyroidism in 5 years [ Logistic Regression,OR = 5.7 (95% CI 2.1-15 ),OR = 3.8 (95% CI 1.7-8.7 ) and OR = 3.1 (95% CI 1.4-6.8) respectively].Conclusion Iodine supplementation in mild iodine deficient area does not increase the incidence of overt hyperthyroidism ultimately.Long-term excessive iodine intake does not affect the incidence and pattern of hyperthyroidism.TSH＜0.3 mIU/L,positive TPOAb and goiter in healthy individuals are the risk factors for hyperthyroidism.

A five-year follow-up study of hypothyroidism in areas with different iodine intakes / 中华内分泌代谢杂志

Objective To investigate epidemiologieal characteristics of hypothyroidism and factors influencing its outcome in the areas with different iodine intakes.Methods An epidemiologic follow-up study of thyroid diseases was performed in Panshan (iodine deficient),Zhangwu [mild iodine deficiency before universal salt iodination (USI) and more than adequate iodine intakes after USI ] and Huanghua ( iodine excessive) in China.A total of 3 761 subjects were investigated in 1999,and 3 018 (80.2% ) of them were followed up in 2004.FT_4,FT_3,TSH,thyroid autoantibodies and urinary iodine concentration were determined and thyroid B ultrasound was performed in all participants.Results Five-year cumulative incidence of overt hypothyroidism was 0.23%,0.47% and 0.35% in Panshan,Zhangwu and Huanghua,respectively.Twenty patients with autoimmune- induced overt hypothyroidism did not comply with the physician's orders to take thyroxine.Thyroid function spontaneously turned to normal in 55% of them,turned to subclinical hypothyroidism in 20% and maintained overt hypothyroidism in 25%.Five-year cumulative incidence of subclinical hypothyroidism in Zhangwu (2.60%) and Huanghua (2.89%) were significantly higher than that in Panshan(0.23% ) (both P＜0.01 ).One hundred patients with subclinical hypothyroidism were followed up,and 5% of them developed overt hypothyroidism,66% turned to euthyroidism and 29% maintained subclinical hypothyroidism.Raised serum TSH (＞6 mU/L) in the original survey ( OR = 3.4),positive thyroid autoantibodies in the follow-up study ( OR = 5.3 ) and more than adequate iodine supplementation in the mild iodine-deficient population ( OR = 8.0) were risk factors influencing outcome of subclinical hypothyroidism.Conclusion More than adequate iodine supplementation among a population in the mildly iodine-deficient area may increase the incidence of overt and subolinical hypothyroidism, and also influence outcome of subclinical hypothyroidism.