OBJECTIVE:To determine plasma concentration of irbesartan by HPLC with fluorescence detection.METHO-DS:Samples were separated on Diamonsil C18 with mobile phase consisted of 0.02mol?L-1 KH2PO4-acetonitrile(50∶50) at a flow rate of 1.0mL?min-1.The fluorescence detector was set at an excitation wavelength of 250nm and an emission waveleng-th of 375nm.The column temperature was 40℃.RESULTS:The linear range for irbesartan was from 1.0 to 1000ng?mL-1 (r=0.999 8).The mean relative recoveries of irbesartan at low,middle and high concentrations(2.0,50.0,and 500.00ng? mL-1) were 97.4%,106.1% and 101.6%,respectively.Both intra-day and inter-day RSD were below 10%.CONCLUSION:The method is simple,rapid,accurate and specific,and it can be used for the plasma concentration determination and pharmacokinetic study of irbesartan.

Protein kinase D (PKD) is a novel family of serine/threonine kinases and diacylglycerol (DAG) receptors and has been documented in a variety of cellular processes. To get high purity catalytic domain of PKD1 (PKD1-cat) for crystallography study, the GST-tagged PKD1-cat gene was cloned into a baculovirus transfer vector pFastBac1 (donor plasmid). When the recombinant plasmid was transformed into DH10Bac competent Escherichia coli, which contains a baculovirus shuttle vector (bacmid), transposition occurs to generate a recombinant bacmid containing the gene of interest (GST-PKD1-cat). The recombinant bacmid DNA was transfected into Spodoptera frugiperda Sf9 insect cells to generate recombinant baculovirus, which was then amplified through multiple rounds of infection in Sf9 cells. After that, Trichoplusia ni insect cells in suspension culture were infected with baculoviral stock at a multiplicity of infection (MOI) of 5 PFU/cell. SDS-PAGE and Western blotting analysis confirmed the detection of a 68 kDa protein by the glutathione S-transferase (GST) monoclonal antibody. The recombination protein was purified by Glutathione sepharose affinity chromatography and cleaved by PreScission Protease to remove GST tag, and a highly pure 42 kDa protein which was consistent with the molecular weight of the expected PKD1-cat protein was detected on SDS-PAGE. The activity of purified PKD1-cat protein was determined by in vitro PKD kinase assay. Our data showed that the kinase activity increased with the concentration of purified PKD1-cat protein. These results showed that the truncated recombinant PKD1-cat protein was highly active and pure, and could potentially be used for solving 3D structure of this protein by Nuclear Magnetic Resonance (NMR) or crystallography.
Animals ; Baculoviridae ; Blotting, Western ; Catalytic Domain ; Cell Line ; Electrophoresis, Polyacrylamide Gel ; Genetic Vectors ; Plasmids ; Protein Kinase C ; chemistry ; Recombinant Proteins ; chemistry ; Spodoptera

Objective To introduce a proposal that can standardize the clinical information of urine manifestation in Chinese medicine.Methods A hypothesis of symptomatic units was proposed in this paper,herein,the attribute,name and definition of the symptomatic units of urine manifestations in the clinical records of Chinese medicine in the past dynasties were summarized according to the stipulation of logic classification and denomination of a scientific term.Results The 23 symptomatic units,2 bounds and their assembly can express 3867 descriptions on the clinical manifestation of urine.Conclusion The symptomatic unit is a feasible hypothesis that can simplify the clinical information of Chinese medicine.The attribute extraction,denomination and definition may become one of the effective procedures to standardize the clinical information of Chinese medicine.

Objective Investigate the postmortem stability of the six immunohistochemical markers of fibronectin(Fn),fibrinogen(Fg),C5 complement(C5), myoglobin(Mb), actin(HHF35)and desmin(Dm)for the postmortem diagnosis of myocardial infarction.Method The areas of depletion ofMb、HHF35 and Dm,and the positive reaction areas of Fn、Fg and C5 in the ischemic myocardium were studied with immunohistochemistry,image analysis technique and statistical system.The postmortem stability of the six immunohistochemical markers was compared.Results The specimens of normal myocardium kept at 4℃ for 1 to 2 days showed homogenous brown reactions for Dm, HHF35 and Mb; depletion of Dm, HHF35 and Mb became evident when these specimens were kept at 4℃ for 3 days postmortem, and the depletion area increased with the lapse of postmortem interval; the depletion area of Dm, HHF35 and Mb in ischemic myocardial tissues also increased with the lapse of postmortem interval;the positive reaction areas of Fg, C5 and Fn in ischemic myocardial tissues decreased with the lapse of postmortem interval.Fg became negative when the ischemic myocardium were kept at 4℃ for 2 weeks postmortem,C5 became negative when kept at 4℃ for 3 weeks postmortem,but Fn remained positive when kept at 4℃ for 4 weeks postmortem.No positive reactions for Fg,C5 and Fn could be found in normal myocardium when kept at 4℃ for different time intervals. The image analysis result showed that the positive reaction areas decreased with the lapse of postmortem interval. Conclusion The Dm and HHF35, Mb showed least postmortem stability, easily influenced by autolysis, only suitable for detection in fresh corpses(1to 2dayspostmortem) ;Fgislittlebitbetter,suitableforcorpsesat4℃ 1weekpostmortem ;C5isbet ter,suitableforcorpsesat 4℃ 2weekspostmortem ;Fnisthebestmarker ,suitableforcorpsesat 4℃ 4 weekspostmortem .

0.05) while other indicators did not. All the data were simply related to each other. The primary influencing factors to NDS score 1, in decreasing order of impact were volume of infarction, position of infarction, hs-CRP and FBS. While the primary influencing factors to NDS score 3 at the end of 1 month treatment, in decreasing order of impact, were NDS score 1, size of infarction, FG, hs-CRP and TC. Conclusions Volume of infarction and hs-CRP are very important determining factors in both NDS score 1 at the beginning of the treatment and NDS score 3 at the end of 1 month treatment. However, in determining NDS score 3 at the end of 1 month treatment, the impact of FG is greater than hs-CRP. The other influencing factors include FBS, TC and so on. Therefore we conclude that it is helpful to recover from neurologic deficit through lowing these influencing factors directly by medication.

Objective　To evaluate the treatment of in-stent restenosis with rotational atherectomy and balloon angioplasty. Methods　The rotational atherectomy and 4～6 atm low pressure balloon angioplasty was performed in 3 patients with in-stent restenosis and follow up after treatment. Results　All cases were succeeded. The bradycardia occurred in one patient was quickly disappeared without treatment, two other patients were found no effect on heart rate, hemodynamic performance, global LV function, or regional wall motion. No complications, angina, death or other coronary event occurred during the follow up for 6～12 months. Two of them was performed coronary angiography after 6 months and showed the diameter of target vessel was less than 30% as compared with that on coronary angiography which performed immedately after operation. Conclusion　The management of in-stent restenosis in target vessels using a combination of rotational atherectomy and balloon angioplasty is safe and efficient.

Objective To explore clinical effects of laparoscopy applied in the female patients with infertility. Methods 468 cases of female infertility patients were diagnosed and treated by laparoscopy and 168 cases received follow-up survey. Results Among those infertility patients, the patients of normal pelvic cavity and the abnormal one were 49(10.47% ) and 419( 89.53% ). Among those patients of abnormal pelvic cavity .fallopian tube disease in 276 cases(65.87% ) was at the first place. The constituent ratio of endometriosis,ovary tumor,polycystic ovary,uterine myoma,female genital tract malformation and pelvic tuberculosis was 14. 80% ,5. 97% ,4. 30% ,3. 82% , 3. 34% and 1.91% .respectively. The patients of primary infertility and the secondary ones were 208 and 260 cases. In secondary infertility patients 69.23% (180/260) of patients were caused due to fallopian tube diseases,and the constituent ratio was higher than that in primary infertility 46.15% (96/208) (x2 = 14.05 ,P <0.05),while in primary infertility,the constituent ratio of endometriosis and normal pelvic cavity were 20.19% ,15.38% ,which were higher than that in secondary infertility respectively(x2 = 11. 78,11. 16, all P < 0.05). The pregnancy rate of 168 cases followed-up was 45.24% (76/168).47.37% ,84.21% pregnancy occurred within 6,12 months with laparoscopy. The shortest and longest time of pregnancy was 2 and 30 months, respectively. The median was 8months. Conclusion Laparoscopy playd an important role in diagnosis of infertility and improvement of pregnancy rate.

Objective Determine the common syndrome elements of viral hepatitis.Methods Refering to the syndrome elements hypothesis proposed by Zhang Zhibin,syndrome elements from the collection of 19,341 cases of Viral Hepatitis were extracted which were diagnosed by National Science and Technology Major Project of AIDS and Viral Hepatitis and Other Major Infectious Diseases Prevention and Control.And the results were compared with industry standards.Results There were 27 involved syndrome elements of viral hepatitis.Common syndrome elements were Qi stagnation,internal dampness,internal fire,blood stasis,yin deficiency,Qi deficiency,toxin,pathogenic water,and yang deficiency; The main excessive syndromes were Qi stagnation,internal dampness,internal fire,and blood stasis.The main deficiency syndromes were yin deficiency and Qi deficiency.Conclusion Excessive syndromes were the main syndromes of viral hepatitis.Dampness,heat,blood stasis,and toxin played key roles in the development of viral hepatitis.In addition to this,Qi stagnation,pathogenic water and phlegm were also important syndrome elements.

Objective:To standardize the clinical information of the nose.Methods:Based on the 1034 descriptions on the clinical manifestation of nose in the database of the clinical records of Chinese medicine in the past dynasties,according to the hypothesis of symptomatic unit,the symptomatic units,the second-level clinical characteristics and the limiting factors were summarized in the paper.Results:All the descriptions on the clinical manifestation of nose include 11 symptomatic units,7 second-level clinical characteristics and 1 limiting factor.Conclusion:Comparing with the symptoms in the authoritative works of Chinese medicine,cold-feeling of the nose while breathing was the new fi nding,nares flaring and ozena recorded in the authoritative works were the second-level clinical characteristics of polypnea and rhinorrhoea respectively.Nose diseases,such as nasal eczema,were usually diagnosed by several symptomatic units,so they were not symptomatic units.