Objective To investigate the trend of liver function changes in untreated adult patients with Graves’ disease in China, and to analyze the associated factors. Methods Patients with newly diagnosed as well as recurrent Graves’ disease from January 2006 to August 2014 were enrolled. They were over 18 years old and did not receive any treatments, Examination of liver function, thyroid function, and thyroid related antibodies as well as tests regarding virus hepatitis were performed. Results A total of 1 254 patients were enrolled. 33 patients with virus hepatitis were ruled out. Ultimately, 1 221 patients matched the criteria of our trial, with 347 males and 874 females [(39. 3 ± 9. 5) year old]. After inclusion, they were assigned to 2 groups according to their liver function results(605 in normal group and 616 in abnormal group). Compared to normal group, patients in the abnormal group were older [(40. 1 ± 9. 2 vs 38. 5 ± 8. 7) year old, P<0. 05] and with higher proportion of females(81. 8% vs 61. 2% , P<0. 05). Regarding the thyroid function and related antibody tests, some patients yielded results that were extremely high so as to exceed the upper limit of the normal range. These patients were more frequently seen in the group with abnormal liver function. The patients whose thyroid function parameters exceeded the upper limit had higher level of alanine aminotransferase[ALT,(37. 69 ± 7. 51 vs 31. 90 ± 5. 95) U/ L, P<0. 05], aspartate aminotransferase[AST, (31. 97 ± 5. 09 vs 27. 88 ± 3. 82) U/ L, P<0. 05], direct bilirubin[DBiL, (5. 58 ± 0. 77 vs 4. 54 ± 0. 71) μmol/ L, P<0. 05]than the group whose thyroid function on the detected range. In the patients with all results detected, patients in abnormal liver function group had higher level of triiodthyronine[T3 , (5. 42 ± 0. 29 vs 4. 94 ± 0. 33) nmol/ L, P<0. 05], thyroxin[T4 ,(217. 53 ±14. 32 vs 204. 22 ±13. 54) nmol/ L, P<0. 05], free triiodthyronine[FT3 ,(15. 88 ± 2. 86 vs 14. 48 ±4. 83) pmol/ L, P<0. 05], free thyroxin[FT4 ,(48. 91 ±8. 45 vs 42. 95 ±6. 14) pmol/ L, P<0. 05], thyroid peroxidase antibody[ TPOAb, (402. 75 ± 89. 99 vs 210. 70 ± 44. 63) IU/ ml, P < 0. 05] and thyrotrophin receptor antibody[TRAb,(14. 08 ± 5. 24 vs 9. 04 ± 2. 58) IU/ L, P<0. 05]. Multivariate logistic regression analysis revealed that patients’ age(OR=0. 98, 95% CI 0. 97-0. 99), gender(OR=0. 94, 95% CI 0. 91-0. 97), level of FT4 (OR=3. 08, 95% CI 2. 19-4. 32), TPOAb(OR = 0. 98, 95% CI 0. 97-0. 99), and TRAb(OR = 1. 07, 95% CI 1. 01-1. 12) were independent risk factors of their liver dysfunction. Conclusion Graves’ disease may lead to liver dysfunction, which is much more common and severe in elder and female patients, as well as patients who are suffering from hyperthyroidism and raised level of thyroid related antibodies.

ObjectiveTo investigate the mechanism of modified Shenhong Tongluo prescription on cell apoptosis in rats with myocardial ischemia-reperfusion injury （MIRI）. MethodSixty Sprague-Dawley （SD） rats were randomly divided into a blank group， a model group， low-， medium-， and high-dose groups of modified Shenhong Tongluo prescription， and a simvastatin group. Except for the blank group， a rat model of MIRI was prepared by ligating the left anterior descending coronary artery. Starting from the first day after successful modeling， the blank group （1.0 mL·kg^-1 physiological saline）， model group （1.0 mL·kg^-1 physiological saline）， low-， medium-， and high-dose groups of modified Shenhong Tongluo prescription （1.031， 2.063， and 4.126 g·kg^-1 Shenhong Tongluo prescriptiona standard concentrate）， and simvastatin group （0.71 mg·kg^-1 simvastatin） were orally administered once daily for 2 weeks. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of cardiomyocytes. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of serum creatine kinase isoenzyme （CK-MB）， interleukin-6 （IL-6）， and tumor necrosis factor-α （TNF-α）. TdT-mediated dUTP nick-end labeling（TUNEL） staining was used to detect the apoptosis rate of rat cardiomyocytes. Western blot was used to detect the expression levels of apoptosis-related proteins B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and caspase-3. ResultCompared with the blank group， in the model group， HE staining showed disturbed arrangement of cardiomyocytes， incomplete fibers， focal necrosis of cardiomyocytes， and inflammatory cell infiltration； serum CK-MB， IL-6， and TNF-α levels were significantly increased （P<0.05）； apoptosis rate of cardiomyocytes was significantly increased （P<0.01）， with significantly increased expression levels of Bax and Caspase-3 proteins， and significantly decreased Bcl-2 expression （P<0.05）. Compared with the model group， the low-， medium-， and high-dose groups of modified Shenhong Tongluo prescription significantly reduced CK-MB， IL-6， and TNF-α levels （P<0.05）， significantly downregulated cardiomyocyte apoptosis rate （P<0.05）， significantly decreased Bax and Caspase-3 proteins， and significantly increased Bcl-2 expression levels （P<0.01）. In the modified Shenhong Tongluo prescription groups， the expression levels of Bax and Caspase-3 proteins significantly decreased with increasing dosage， while the expression level of Bcl-2 significantly increased with increasing dosage of modified Shenhong Tongluo prescription （P<0.05）. ConclusionShenhong Tongluo prescription can alleviate myocardial tissue pathological damage and reduce myocardial cell apoptosis， possibly by inhibiting Caspase-3 and Bax expression and promoting Bcl-2 expression.

Rv2742 is a novel gene identified from Mycobacterium tuberculosis H37Rv by the proteogenomics strategy. The aim of this study was to establish a system of soluble expression and purification of the missing protein Rv2742 in M. tuberculosis H37Rv, to provide reference for further research on the biological function of Rv2742. The soluble protein was not successfully induced by prokaryotic expression vectors pGEX-4T-2-Rv2742, pET-32a-Rv2742, pET-28a-Rv2742 and pMAL-c2X-Rv2742. After the codon of novel gene Rv2742 was optimized according to E. coli codon usage frequency, only the recombinant strain containing plasmid pMAL-c2X-Rv2742 could produce soluble products of Rv2742 encoding gene. In addition, the expression effects of the desired fusion protein were also analyzed under different conditions including hosts, culture temperatures and IPTG concentrations. The optimum expression conditions were as follows: Rosetta (DE3) host, 16 °C culture temperature and 0.5 mmol/L IPTG. After being purified by affinity chromatography with amylose resin, the fusion protein sequence was confirmed by LC-MS/MS. These results indicated that the novel gene Rv2742 product could be successfully induced and expressed in a soluble form by the expression system pMAL-c2X with MBP tag. Our findings provide reference for studies on potential interaction and immunogenicity.
Chromatography, Liquid ; Cloning, Molecular ; Escherichia coli ; Mycobacterium tuberculosis ; genetics ; Recombinant Fusion Proteins ; Tandem Mass Spectrometry