Simultaneous Determination of 7 Active Constituents in Tangshen Qingdu Granule by HPLC-DAD / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of 7 active constituents in Tangshen qing-du granule. METHODS:HPLC was performed on the column of SHIMADZU Inert Sustain C18 with mobile phase of acetonitrile-0.1%phosphoric acid at a flow rate of 1.0 mL/min,detection wavelength was 327 nm for chlorogenic acid and caffeic acid,280 nm for baicalin,228 nm for arctiin and 276 nm for wogonoside,baicalein and wogonin,column temperature was 35℃,and injection volume was 10 μL. RESULTS:The linear range was 4.830-154.6 μg/mL for chlorogenic acid and(r=0.9998),0.750-24.1 μg/mL for caffeic acid(r=0.9997),22.859-731.5 μg/mL for baicalin(r=0.9997),8.491-271.7 μg/mL for arctiin(r=0.9993),2.471-79.0μg/mL for wogonoside(r=0.9996),6.656-213.0 μg/mL for baicalein(r=0.9994) and 2.756-88.2 μg/mL for wogonin (r=0.9998);RSDs of precision,stability and reproducibility tests were lower than 2.0%,recoveries were 96.86%-100.82%(RSD=1.46%,n=6),98.79%-101.09%(RSD=0.93%,n=6),97.57%-101.51%(RSD=1.37%,n=6),97.76%-99.63%(RSD=0.77%,n=6),97.99%-100.12%(RSD=0.76%,n=6),96.54%-101.07%(RSD=1.87%,n=6) and 96.60%-99.59%(RSD=1.14%,n=6). CONCLUSIONS:The method is simple with good precision,stability and reproducibilty,and can be used for the simultaneous determination of 7 active constituents in Tangshen qingdu granule.

Study on Water Extraction Technology of Yinju Jiedu Oral Liquid / 中国药房

OBJECTIVE:To optimize the water extraction technology of Yinju jiedu oral liquid,and provide reference for the industrial production of the preparation. METHODS:According to the investigation of extraction time-extraction rate curves of chlo-rohenic acid of Yinju jiedu formula and extraction rate of chlorohenic acid in Lonicera japonica and other combined medicinal mate-rials in the formula,decoction methods and time of L. japonica were determined. Using the comprehensive scores of linarin,harpa-goside,(R,S)-epigoitrin,psoralen+angelicin contents and dry extraction yield as indexes,L9(34)orthogonal test was designed to detect the effects of adding water amount,decoction time times and optimize the extraction technology of the residues and other me-dicinal materials. Verification test was conducted. RESULTS:The optimal technology was L. japonica decocted first for 30 min with 8-fold water;the residues and other medicinal materials were decocted with 8-fold water for 3 times,1 h each time;combin-ing all the syrups. In verification test,the average contents of chlorohenic acid,linarin,harpagoside,(R,S)-epigoitrin,psoralen+angelicin were respectively 34.51,10.31,1.97,0.21,9.79 mg/g(RSD=1.24％,1.19％,1.40％,1.71％,1.28％,n=3);aver-age dry extraction yield was 25.4％(RSD=1.64％,n=3);average extraction rate of chlorohenic acid was 78.95％(RSD=1.24％,n=3). CONCLUSIONS:In the optimized water extraction technology,both the extraction rate of chlorohenic acid and contents of other ingredients are relatively high. The technology is stable and feasible.

The frequency and function of FoxP3~+ regulatory T cell in patients with acute hepatitis B / 细胞与分子免疫学杂志

AIM: To investigate the dynamic variety of frequency and function of FoxP3~+ regulatory T cells in patients with acute hepatitis B (AHB). METHODS: Peripheral blood mononuclear cells (PBMCs) from 15 AHB patients at acute phase (week 1 of illness), convalescent phase (primary occurrence of both ALT level normalization and HBsAg negative conversion), resolved phase (at least 8 weeks after both ALT normalization and HBsAg seroconversion, and 15 health subjects were analyzed for FoxP3 (Forkhead/winged helix transcription factor) mRNA expression in MACS magnetic beads-purified CD4~+ T cells by real-time RT-PCR assay. The effects of Treg cells on the proliferation of CD4~+CD25~- T cells were examined by a ~3[H]-thymidine incorporation assay. RESULTS: AHB patients presented a significantly higher FoxP3 mRNA expression at convalescent phase than acute phase (t=-6.04, P<0.01) and resolved phase (t=4.45, P<0.01), and healthy controls (t=3.44, P<0.01). We also observed that the suppression efficiency of Treg cells on proliferation of CD4~+CD25~- T cells was lower at acute phase than convalescent phase (t=-5.30, P<0.01) and resolved phase (t=-3.20, P<0.05), but there was no significant difference between healthy controls and any phase of AHB. CONCLUSION: AHB patients presented lower circulating Treg frequency and suppression function at acute phase, and both of them are increased at convalescent phase, and then return to normal level along with disease resolved. This follow-up study furthers our understanding of Treg' s role in immunopathogenesis of hepatitis B.

EFFECT OF CADMIUM ON ULTRASTRUCTURE OF HEPATOPANCREAS CELLS OF FRESHWATER CRAB(SINOPOTAMON YANGTSEKIENSE) / 解剖学报

Objective After an experimental exposure to low dosage of Cd 2+(39*!?mol/L) for 30 days,the changes of Cd 2+ on ultrastructure of the hepatopancreas cells of the freshwater crab (Sinopotamon yangtsekiense♂)at three different times were studied. Methods Sixty freshwater crabs(♂)were divided into two groups:control(n=30) and test(n=30)groups.The test(n=30) group was exposed to low dosage of Cd 2+(39*!?mol/L)while the control crabs were in water without cadmium.During the 5*!d,15*!d,30*!d,the ultrastructure of the hepatopancreas cells was observed by using transmission electro-microscopy(TEM). Results In the course of the experiment,the nuclear membrane changed from spreading all over the place to disintegrated,and the penetration of the mitochondrial outer membrane and inner membrane was changed.The mitochondria showed swelling and vacuolating.Mitochondrial cristae partly disintegrated,then fully disintegrated.Besides,the Cd 2+ exposure affected the gross form and the structure of the rough endoplasmic reticulum(RER).At first,the RER became the different sized vesicles,then,the ribosomes on the little vesicles began to fall,and finally,the RER turned to the concentric round lamellar structures.But the smooth endoplasmic reticulum(SER)increased markedly.In addition,the numbers and types of lysosome were increased in the mean time and some lysosomes became void vesicles,the autophagosomes or the myeloid bodies(MD).The microvilli showed partly falling and vacuolating.Conclusion After the cadmium treatment the detoxification mechanism of the freshwater crab's hepatopancreas cells lost,the structural and functional integrality of the mostly organelles,such as the nucleous,the mitochondria and the rough endoplasmic reticulum,were destroyed,so that the normal physiological function of hepatopancreas cells was affected.