The new drug of Chinese herbal prescription, which is the best carrier for the syndrome differentiation and treatment of Chinese medicine and is the main form of the new drug research and development, plays a very important role in the new drug research and development. Although there are many sources of the prescriptions, whether it can become a new drug, the necessity, rationality and science of the prescriptions are the key to develop the new drug. In this article, aiming at the key issues in prescriptions design, the source, classification, composition design of new drug of Chinese herbal prescriptions are discussed, and provide a useful reference for research and development of new drugs.
Drug Design ; Drug Prescriptions ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Humans

OBJECTIVETo discuss that pathogenesis evolvement regularity of Chinese miniature swine with phlege-stasis cementation syndrome of coronary heart disease.

METHODEighteen Chinese miniature swine were randomly divided to the normal control group, the model group and the Danlou tablet group, with six swine in each group. Except for the normal control group, all of the other groups were fed with high fat diet for two weeks. The coronary heart disease model with phlegm-stasis cementation syndrome was established by injuring left anterior descending artery with interventional balloons and continuously feeding with high fat diet for eight weeks. The levels of BMI, hemorheological parameters, lipids in serum and inflammatory cytokines were observed at the 0th (before the experiment), 2nd (before operation or drug administration), 6th (four weeks after drug administration) and 10th week (eight weeks after drug administration) of study. The levels of TG and TC in liver and the pathological changes in coronary artery tissues were also observed at the end of study.

RESULTCompared with the normal control group, the model group had showed significant increase in the levels of TC, TG, LDL-C and VLDL-C in serum (P < 0.01) from the second week to the end of the experiment, with notable rise in the whole blood viscosity under the shear rates of 5 s(-1) and 60 s(-1). At the 6th week, the levels of BMI and TG and TNF-alpha in serum significantly increased. At the 10th week, the levels of BMI and hs-CRP, IL-6 and TNF-alpha in serum significantly increased as well, with remarkable increase in coronary stenosis, intimal thickness and the ratio between intimal thickness and media thickness (P < 0.05 and P < 0.01), and significant rise in TC and TG in livers (P < 0.01). Compared with the model group, the Danlou tablet group showed obvious reduction in severity of coronary artery lesion, intimal thickness and lumen stenosis ratio and ratio between intimal thickness and media thickness (P < 0.01), BMI, TC, TG, LDL-C and VLDL-C in serum, TC and TG in liver, as well as hs-CRP, IL-6 and TNF-alpha levels in serum (P < 0.05 and P < 0.01), with notable decline in the whole blood viscosity under the shear rates of 5 s(-1) and 60 s(-1).

CONCLUSIONThe interaction of phlegm, blood stasis and toxin syndromes helps promote the progress and development of AS plaques, which is the key pathogenesis of phlegm-stasis cementation syndrome in coronary heart disease.

Animals ; Body Weight ; Coronary Disease ; blood ; physiopathology ; Female ; Hemodynamics ; Inflammation Mediators ; metabolism ; Lipids ; blood ; Liver ; metabolism ; Male ; Medicine, Chinese Traditional ; Swine ; Swine, Miniature

OBJECTIVETo establish the "clinical-mimetic" Chinese medicine symptom complex score method for the coronary heart disease Chinese miniature swine model with phlegm-stasis cementation syndrome, in order to observe the effect of formula of removing both phlegm and blood stasis (TYTZ) on the Chinese medicine symptom complex score for the coronary heart disease Chinese miniature swine model with phlegm-stasis cementation syndrome.

METHODTotally 36 Chinese miniature swine were randomly divided to the normal control group, the model group, the Shujiangzhi group, and TYTZ groups with doses of 2.0, 1.0 and 0.5 g x kg(-1), with six in each group. Except for the normal control group, all of the other groups were fed with high fat diet for two weeks. The coronary heart disease model with phlegm-stasis cementation syndrome was established by injuring left anterior descending artery with interventional balloons and continuously feeding with high fat diet for eight weeks. After the operation, the groups were administered with drugs for eight weeks. Their main symptoms, accompanied symptoms, tongue and pulse signs of the coronary heart disease Chinese miniature swine with phlengm-stasis cementation syndrome were observed according to the symptom-graded scoring method.

RESULTCompared with the model group, TYTZ in different doses could reduce the scores of main symptoms at the 6th and 10th week. Specifically, TYTZ in low dose could reduce the scores of tongue at the 6th week and the scores of accompanied symptoms, and tongue and pulse signs at the 10th week; And TYTZ in high dose could decrease all symptom scores at the 6th and 10th week (P < 0.05 or P < 0.01).

CONCLUSIONTYTZ can improve the scores of the main symptoms, accompanied symptoms, tongue and pulse signs in coronary heart disease Chinese miniature swine with phlegm-stasis cementation syndrome. It is suggested that the "clinical-mimetic" objective scoring for syndromes of Chinese miniature swine is of great significant to the development of new traditional Chinese medicine drugs.

Animals ; Chemistry, Pharmaceutical ; Coronary Disease ; blood ; diagnosis ; physiopathology ; therapy ; Female ; Male ; Medicine, Chinese Traditional ; methods ; Swine ; Swine, Miniature

OBJECTIVETo observe the effect of formula of removing both phlegm and blood stasis (TYTZ) in inhibiting the inflammatory reaction in Chinese mini-swine with coronary atherosclerosis.

METHODTotally 36 Chinese mini-swine were randomly divided to six groups: the normal control group, the model group, the Shujiangzhi group and TYTZ groups with does of 2.0, 1.0 and 0.5 g x kg(-1), and six each in every group. Except for the normal control group, all of other groups were fed with high-fat diet for 2 weeks. Interventional balloons are adopted to injure their left anterior descending artery endothelium. After the operation, they were fed with high-fat diet for 8 weeks to prepare the coronary atherosclerosis model. In the 8th week after the operation and administration, the intravascular ultrasound was adopted to observe the coronary artery plaque burden of each group and the pathological morphology of coronary artery. Such inflammatory factors as high-sensitivity C-reactive protein (hs-CRP), tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 were detected by ELISA. The expression of NF-kappaB p65 nuclear translocation was observed by the immunohistochemical method.

RESULTCompared with the normal control group, the model group showed significant increase in the coronary artery plaque burden at the end of the experiment (P < 0.01), notably abnormal structural changes in atherosclerotic vascular tissues, luminal stenosis, a large number of foam cells and inflammatory cell infiltration, remarkable growth of hs-CRP, TNF-alpha and IL-6 levels (P < 0.01). The immunohistochemical staining also showed the significant increase in the NF-kappaB p65 nuclear translocation of coronary artery of Chinese mini-swine in the model group. Compared with the model group, TYTZ could significantly attenuate atherosclerotic plaque burden (P < 0.01), inhibit the coronary luminal stenosis, reduce inflammatory cell infiltration, decrease such inflammatory cell factors as hs-CRP, TNF-alpha and IL-6 in serum, and inhibit the NF-kappaB p65 nuclear translocation of coronary artery (P < 0.05 or P < 0.01).

CONCLUSIONTYTZ can reduce the downstream inflammatory reaction by controlling NF-kappaB p65 nuclear translocation, so as to inhibit the occurrence and development of coronary atherosclerotic plaque in Chinese mini-swine.

Animals ; C-Reactive Protein ; metabolism ; Coronary Artery Disease ; blood ; complications ; drug therapy ; pathology ; Female ; Inflammation ; complications ; Interleukin-6 ; blood ; Male ; Medicine, Chinese Traditional ; methods ; Mucous Membrane ; drug effects ; secretion ; Swine ; Swine, Miniature ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo analyze and summarize changes of syndrome-related biological indices in acute lacuna encephalon infarction patients of upper hyperactivity of Gan yang syndrome (UHGYS), thus providing objective evidence for syndrome typing and disease identification.

METHODSRecruited were 50 patients at Department of Encephalopathy, Xiyuan Hospital, China Academy of Chinese Medical Sciences, who were in line with diagnostic criteria of UHGYS as the experimental group in this study. Another 40 healthy volunteers were recruited as the control group from May 2010 to July 2012. Blood routines (including WBC, RBC, Hb, NEUT%, and LY%), hepatic and renal functions tests (including ALT, AST, TBIL, TP, ALB, Cr, and BUN) were performed by automatic whole blood analyzer and colorimetric technique. The levels of fasting blood glucose, HbAlc, blood lipids (including TC, TG, HDL-C, LDL-C, and VLDL-C), and coagulation functions (including AT-III, PT, PTA, INR, TT, APTT, and FBG, reaction time), renin, angiotensin II, hs-CRP, and Hcy were also measured. The thyroid functions (including FT3, FT4, T3, T4, and TSH) were detected by electrochemiluminescence immunoassay. The levels of tumor necrosis factor alpha (TNF-alpha), IL-6 and IL-1 in serum were measured by ELISA and radioimmunoassay respectively.

RESULTSCompared with the control group, RBC, LY%, ALT, TP, ALB, HDL-C, AT-III activities, contents of PTA and FT4 obviously decreased, TBIL, BUN, Glu, HbAlc, TSH, hs-CRP, renin, Ang II, TNF-alpha, IL-1 and IL-6 significantly increased in the experimental group (P < 0.05, P < 0.01).

CONCLUSIONThe pathological process of acute lacuna encephalon infarction patients of UHGYS was closely correlated with thyroid functions, the renin-angiotensin-aldosterone system, the extrinsic and intrinsic coagulation systems, as well as inflammation reaction.

Acute Disease ; Adult ; Case-Control Studies ; Female ; Humans ; Infarction ; blood ; diagnosis ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Stroke, Lacunar ; blood ; diagnosis

OBJECTIVETo observe effect of formula of removing both phlegm and blood stasis (TYTZ) in improving hemorheology and blood fat of mini-swine with coronary heart disease of phlegm-stasis cementation syndrome.

METHODThirty-six Chinese mini-swine were randomly divided to six groups: the normal control group, the model group, the Shujiangzhi group and TYTZ groups with doses of 2.0, 1.0 and 0.5 g x kg(-1), with six mice in each group. Except for the normal control group, all of other groups were fed with high-fat diet for 2 weeks. Interventional balloons are adopted to injure their left anterior descending artery endothelium. After the operation, they were fed with high-fat diet for 8 weeks to prepare the coronary heart disease model of phlegm-stasis cementation syndrome. In the 8th week after the operation and administration, the changes in hemorheological parameters, serum lipid level, myocardial ischemia level and range were observed.

RESULTCompared with the normal control group, the model group showed significant increase in serum TC, TG, LDL-C and VLDL-C levels (P < 0.01), whole blood viscosity under the shear rate of 5 s (-1) and 60 s (-1) (P < 0.01), and myocardial ischemia degree and range (P < 0.01). Compared with the model group, TYTZ groups revealed significant decrease in myocardial ischemia degree and range (P < 0.01), serum TC, TG, LDL-C and VLDL-C levels (P < 0.05 or P < 0.01) and whole blood viscosity under the shear rate of 5 s(-1) and 60 s(-1) (P < 0.05).

CONCLUSIONTYTZ could improve the abnormal hemorheology in Chinese mini-swine with coronary heart disease of phlegm-stasis cementation syndrome, and regulate serum lipid, with a certain efficacy for coronary heart disease of phlegm-stasis cementation syndrome.

Animals ; Coronary Disease ; blood ; metabolism ; physiopathology ; therapy ; Female ; Hemorheology ; Lipids ; blood ; Male ; Medicine, Chinese Traditional ; methods ; Mucous Membrane ; secretion ; Swine ; Swine, Miniature

OBJECTIVETo observe the Chinese medicine constitution types and human leukocyte antigen (HLA)-DQA1 gene polymorphism in patients with hepatitis B (HB) virus infection in Chinese Han population of Zhejiang Province, for exploring the roles of constitution factor in pathogenesis of HB.

METHODSA total of 240 subjects, including 120 biopsy-proven chronic HB (CHB), 60 HB asymptomatic carrier (ASC) and 60 resolved from HBV infection spontaneously (RHBS) were studied. Their Chinese medicine constitution type was judged by Wangqi's classification, and their genotype of HLA-DQA1 was detected by polymerase chain reaction sequence specific primer for comparing the difference between groups in distribution frequency (DF) of constitution types and genes.

RESULTS(1) As compared with the RHBS group, DF of yin-deficiency constitution and phlegm-dampness constitution in the CHB group was significant higher (20.0% vs. 6.7% and 12.5% vs. 1.7%), and that of placid constitution was significant lower (11.7% vs. 31.7%), showing statistical significance between groups (OR = 3.5, 95% CI: 1.16-10.60; OR = 8.4, 95% CI: 1.09-65.42; OR = 0.161, 95% CI: 0.076-0.34; all P < 0.05). (2) As compared with the ASC group, DF of damp-heat constitution was significant higher (24.2% vs. 6.7%, P < 0.05, OR = 4.462, 95% CI: 1.49-13.36), and that of placid constitution was significant lower (11.7% vs. 45.0%, P < 0.01, OR = 0.285, 95% CI: 0.13-0.62) in the CHB group. (3) As compared with RHBS group, DF of HLA-DQA1 * 0201 allele in CHB group was significant higher (38.3% vs. 5.8%, P < 0.01, OR = 10.04, 95% CI: 4.48-22.48); and that of HLA-DQA1 * 0102 allele was significant lower (9.6% vs. 36.7%, P < 0.01, OR = 0.183, 95% CI: 0.10-0.32). (4) As compared with ASC group, DF of HLA-DQA1 * 0201 allele in CHB group was significant higher (38.3% vs. 7.5%, P < 0.01, OR = 7.667, 95% CI: 3.7-15.87), and that of HLA-DQA1 * 0102 allele was significant lower (20.0% vs. 9.6%, P < 0.01, OR = 0.424, 95% CI: 0.23-0.79).

CONCLUSIONBoth Chinese medicine constitution and HLA-DQA1 gene polymorphism show connection with the outcomes of HB virus infection in Chinese Han population, but the real association between them is required for further study.

Adolescent ; Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Body Constitution ; Carrier State ; virology ; Female ; Gene Frequency ; Genotype ; HLA-DQ alpha-Chains ; genetics ; Hepatitis B ; diagnosis ; genetics ; virology ; Hepatitis B virus ; Heterozygote ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Polymorphism, Genetic ; Young Adult

Objective To clne huma PAIl gene and prepare its monoclonal antibodies(McAbs) for determination of its expression in breast cancer cells.Methods Human PAI1 gene eDNA was amplified by RT-PCR from human breast cancer cell line MDA231 and inserted into the prokaryotic expression vector, which expressed fusion protein of MS2-PAI1 in E.coli.Fusion protein of MS2-PAI1 was purified and used for immunizing BALB/C mouse.Traditional hybridoma technology was used to produce hybridoma cells for preparation of monoclonal antibodies.Western blot and immunohistochemistry were used to detect PAI1 expression in breast cancer.Results The 1209 bp full PAI1 gene was cloned.The two hybridoma cell lines that secreted specific monoclonal antibodies against human PAI1 were identified by ELISA.The immunoglobulin subclasses of the McAbs were IgG1.The McAbs can specifically recognize PAI1 but not other proteins.Western blot showed that the McAbs against PAI1 can specifically react with MS2-PAI1 fusion protein and endogenous proteins in cells.The positive reaction was found in breast cancer cell line MDA231 and breast cancer tissues by immunochemical staining.Conclusions The McAbs against human PAI1 are successfully prepared by hybridoma technology with MS2-PAI1 fusion protein expressed in E.coll.It has been shown that PAI1 can be expressed in MDA231 and breast cancer tissues.The McAbs against PAI1 could be a useful tool for the further study of the human PAI1 functions and detection of clinical tumor samples.

Objective: To establish a multiplex PCR-microarray method for detecting important enteropahogens.Methods: Uniplex and multiplex PCR were performed to obtain the best primer sets for identifying the target bacteria at species and multi-species level.Fluorescent dyes were mixed into PCR reaction to determine whether it can affect the efficiency of amplification.To improve the efficiency of microarray,a 35 pairs primer-labeling system was optimized based on the hybridization results to find the best combination to avoid false negative results.Results: Specific PCR products were all obtained using species-specific primer sets.More preferential amplification may happen when more primer pairs were added to the reaction.The hybridization results showed a positive association between the efficiency of multiplex-PCR and signal intensity.Conventional PCR yielded more products than fluorescent dyes labeled PCR.Thirty-five primers were divided into three different combinations to label target respectively,hybridization results showed a high specificity.Conclusion: Mixing fluorescent dyes into PCR may reduce the efficiency of amplification and hybridization,but may have no effect on the analysis of hybridization results.The hybridization efficiency of microarray depends on the amplification efficiency of multiplex PCR.For microarray target labeling,three primer sets could be used to avoid negative hybridization led by preferential amplification of multiplex-PCR.It indicates that the multiplex PCR-microarray method is an attractive diagnosis tool for the high-throughput identification of enteropathogenic organisms especially for multiple causative agents and epidemiological investigations.

OBJECTIVETo investigate whole blood viscosity changes at different time points in rats model with lumbar vertebrae semidislocation, study Shi's theroy of qi and blood and "Gucuofeng and Jinchucao" [symbols: see text], also reveal pathological physiology characteristics of spinal disorder.

METHODSThirty-six SPF male rats weighted 350 to 450 g were randomly divided into rotatory fixation group (RF group), simple fixation group (SF group) and Sham group (Sham group), 12 rats in each group. Exterior vertebrae implanted through L4-L6 segments of lumbar vertebrae in RF and SF group were connected fixed device. In RF group, L5 spinous process were rotated to right, and caused L5 spinous process was non collinear with L4 and L6; in SF group, external fixed device were simple connected without rotation. At 1, 4, 8 and 12 weeks after fixation, whole blood viscosity changes were tested.

RESULTSAt 4 and 8 weeks after fixation, high (150/s), medium (60/s) and lower (10/s) shear rate in RF and SF group were higher than that of Sham group (P<0.05). At 1 and 12 weeks, there was no sigificant differences among three groups in whole blood viscosity (P>0.05).

CONCLUSION"Gucuofeng and Jinchucao" [symbols: see text] vertebrae could raise whole blood viscosity, increase degree of bloos stasis and induce or aggravate spinal disorder in further.

Animals ; Blood Viscosity ; Disease Models, Animal ; Joint Dislocations ; surgery ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Rats ; Rats, Sprague-Dawley ; Time Factors