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Objective:To study the best extraction technology of Qishan sugar-free granule. Methods:The effects of boiling dura-tion, boiling times and water addition ratio on the extraction process were investigated by using L9 (34 ) orthogonal design, and the con-tents of total flavonoids, baicalin and berberine were taken as the indices. Results:The optimum conditions were as follows:10-fold a-mount of water for 2 cycles, and every extracting time was 1 hour. Conclusion: The best extraction technology of Qishan sugar-free granule is obtained, and the process is stable and feasible, and the contents of active components are high.
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Objective To analyze the relationship between thyroid function and grades of diabetic nephropa-thy (DN).Methods One hundred and twenty patients with definite DN were classified into the four groups,and forty patients with non-DN diabetic was NDN group.Their serum albumin,blood hemoglobin,serum potassium,functional parameters of thyroid and dependability to DN were analyzed.Results The plasma albumin ofⅢ,Ⅳ and Ⅴ group were (41.3 ±3.6)g/L,(30.5 ±4.8)g/L and (28.3 ±5.9)g/L,which were lower than those of NDN group (F=11.36,P<0.05).24h urine protein ofⅢ,Ⅳ and Ⅴ were (0.48 ±0.29)g,(1.86 ±0.54)g and (1.69 ±0.67)g, which were higher than those of group NDN (F=7.12,P<0.05).Triiodothyronine(T3)ofⅢ,ⅣandⅤgroup were (0.98 ±0.38)nmol/L,(0.75 ±0.41)nmol/L and (0.60 ±0.28)nmol/L,which were lower than those of NDN group (F=7.64,P<0.05).Tetraiodothyronine(T4)of III,IV and V group were (6.06 ±1.76)nmol/L,(5.31 ±1.98)nmol/L and (4.65 ±1.87)nmol/L,which were lower than those of NDN group (F=6.83,P<0.05 ).Free Triiodothyronine (FT3)ofⅢ,Ⅳ and Ⅴ group were (4.37 ±2.12)pmol/L,(3.33 ±2.30)pmol/L and (2.91 ±1.82)pmol/L, which were lower than those of NDN group (F=7.14,P<0.05 ).Correlation analysis indicated that FT3 was an independent predictor for the severity of diabetic nephropathy.Conclusion In the patients with DN Ⅳ and Ⅴ,DN has attained a very severe status and the treatment of DN should be intensive.
ABSTRACT
In the present study, through a functional strategy, a metagenome library of the marine microbes from the surface water of the South China Sea was screened for beta-glucosidase and six positive clones were obtained. One of these clones, pSB47B2, was subcloned and further analysed in sequence. The result showed that there was an open reading frame for a novel beta-glucosidase, which was nominated as bgl1B. Using pET22b(+) as vector and Escherichia coli BL21(DE3) as host, Bgl1B was overexpressed recombinantly with high yield obtained and substantial enzymatic activity detected. The recombinant protein (rBgllB) was purified by Ni-NTA affinity chromatography and further biochemically characterized. The results indicated that, with pNPG as substrate, the optimum pH and temperature for the hydrolytic activity of rBgl1B were about 6.5 and 40 degrees C respectively. Under the optimum conditions, rBgl1B hydrolyzed pNPG with an activity up to 39.7 U/mg, Km and Vmax being 0.288 mmol/L and 36.9 micromol/min respectively. In addition, rBgl1B could also hydrolyze cellobiose, with a Km of 0.173 mmol/L and a Vmax of 35 micromol/min. However, we did not detect evident hydrolytic activity of rBgl1B to lactose, maltose, sucrose, and CMC. The enzymatic activity of rBgl1B to pNPG was stimulated to certain degrees by low concentration of Ca2+ or Mn2+, whereas it exhibited significant tolerance against high Na+. Distinguished from most of the beta-glucosidases derived from fungi, which display the highest activities under acidic conditions, rBgl1B exhibited relatively higher activity and stability at pH between 7.0 and 9.0.