Objective: To study the factors affecting the content of magnolol which is one of the main active components in Magnolia Bark Three Agents Decoction. Methods: The contents of magnolol in extracts of Magnalia Bark Three Agents Decoction (medicinal materids were decocted alone, together or in different compatibleness, and residuals) were determined by HPLC. The effects of use amount of Radix et Rhizoma Rhei and Fructus Aurantii Immaturus, decoction time, and extraction times on magnolol content were studied by orthogonal design.Results: The content of magnolol in Magnolia Bark Three Agents Decoction of which medicinal materials were decocted together was over double that of which they were decocted alone. The dissolution sate of magnolol increased with the increasing of the use amount of Fructus Aurantii Immaturus and extraction times. Conclusion: Fructus Aurantii Immaturuns and extraction times are the main factors affecting magnolol content in Magnolia Bark Three Agents Decoction.

AIM: To isolate,identify and screen active compounds of simplitied formula of Danggui Shaoyao Power(poria,Rhizoma Atracty-lodis Macrocephalae,and Radix Angeliae Sinensis). METHODS: Compounds in the fraction of CO_2 supercritical fluid extraction(SFE-CO_2) were isolated by silica gel chromatography and detected the protective effect on KCl-induced PC12 cell damage. RESULTS: Twelve compounds were obtained,and nine of them were identified as atractylenolide Ⅰ,Ⅱ,Ⅲ,biatractylenolide,levistolid A,pachymic acid,ferulic acid,?-sitosterol and ?-daucosterol,of which only atractylenolide Ⅲ can improve the cell viability significantly. CONCLUSION: Atractylenolide Ⅲ may be one of the active compounds of FBD in curing vascular dementia.

Hydrolysate of Area subcrenata (HA) was found to be effective in lowering the elevated SGPT level induced by carbon tetrachloride, thioacetamide (TAA) and pred-nisolone. In addition, HA was shown to shorten sodium pentobarbital sleeping time in both normal and carbon tetrachloride intoxicated mice. The results indicated that HA possessesa significantl protective action on liver injuries in mice.

With the development of molecular diagnostic techniques, the study for the etiology of leukemia has been entering the era of the molecular biology.Molecular techniques for leukemia diagnosis, prognosis and individualized therapy are used widely, becoming one of the necessary routine tests for patients with leukemia. So far, molecular techniques for leukemiaincluding cytogenetic diagnosis, molecular genetics, molecular diagnostics based on PCR, mutation detection, as well as a variety of next-generation sequencing, have played an important role in the diagnosis of leukemia, minimal residual disease monitoring, prognosis and targeted therapy.

Objective:To investigate the effects of Origin-Preserving Decoction on the proliferation of marrow hematopoietic cells in aplastic anemia mice. Methods:Based on experimental AA models induced by Benzene and 60Co radiation in mice, the experiments including semi-solid colony culture (CFU-GM、CFU-E、BFU-E) and liquid culture (MTT Assay) were undertaken to investigate the effects of Origin-Preserving Decoction on the proliferation of marrow hematopoietic cells in AA mice. Results:Origin-Preserving Decoction (3g/kg) (ig.) could improve the outputs of CFU-GM, CFU-E and BFU-E in AA mice damaged by benzene and 60Co radiation: MTT assay showed that Origin-Preserving Decoction could stimulate the hematopoeisis of bone marrow of AA mice radiated by 60Co. Conclusions: Origin-Preserving Decoction has a good future for AA therapy.

Objective To explore the proper proportion of effective fractions in Shengmaisan(saponins of Radix ginseng,saponins of Radix ophiopogonis,Lignans of Fructus schisandrae)in different anoxia models.Methods Acute cerebral hypoxia was induced by sodium nitrite and decapitation in mice,and the orthogonal design was used in these two models to find the proper proportion of three effective fractions.The gasping time of the mice,which were decapitated was observed to compare the anti-anoxia effects of XZF with other clinical drugs,and cerebral ischemic reperfusion injury model of mice was also used to study the effect of XZF on related biochemical index.Results XZF(the proportion of saponins of Radix ginseng,saponins of Radix ophiopogonis,Lignans of Fructus schisandrae as 7 ∶ 2 ∶ 6)significantly prolonged the gasping time,and decreased brain nitrogen monoxidum(NO)content after reperfusion of the mice at dosages of 50 mg/kg and 150mg/kg.Meanwhile,XZF also reduced malondialdehyde(MDA)content and increased superoxide dismutase(SOD)activity at the higher dosage.Conclusion XZF obtained by experimental screening exerts a significant protective effect on cerebral ischemia injury in mice,which provide some pharmacological evidence for further development of new modern Chinese drug composed with effective fractions for cerebral vascular diseases.

Objective To explore the accuracy and clinical application of Pyrosequencing for detection of drug resistant relevant mutation in the polymerase gene of Hepatitis B virus (HBV).Methods Compared with Sanger sequencing,the accuracy and sensitivity of Pyrosequencing were assessed.Pyrosequencing was used to determine the serum of 1164 patients with chronic Hepatitis B and its results were analyzed.Results The sensitivity of Pyrosequencing was 1 × 103 KIU/L,the same as Sanger sequencing.But Pyrosequencing was more stable and specific than Sanger sequencing to detect low rate of mutation with over 10％ mutation.The mutations couldn't be detected in 248 patients with chronic Hepatitis B,but positive results occurred in 919 patients,among them,61.5 ％ without mutation,38.5 ％ with mutation (including 47.5％ of the single locus mutation and 52.5％ of the combined mutation).Other sites had different degree of mutations except the sites of rtI169T and rtA194T; the main mutation sites in patients with chronic Hepatitis B were rtM204V/I (32.1％),rtL180M (19.8％),rtA181V/T (6.7％) and rtN236T (5.3％),in which fractional mutation had high ratio in the sites of rtA181V/T (6.7％) and rtN236T (5.3％).Conclusions Pyrosequencing has good sensitivity,specificity and accuracy and can early detect the drug resistant relevant mutation in the polymerase gene of HBV,which is suitable for monitoring patients with chronic Hepatitis B for the treatment of nucleoside analogues (acid).The different sites and frequencies of mutations may be associated with different habits of doctors for using different anti-HBV drugs.

Objective To evaluate the value of detection of interleukin 28B (IL28B) rs12979860 by allele-specific PCR (AS-PCR) for the prediction of antiviral treatment hepatitis C patients.Methods One hundred seventy-four blood samples were random collected from hospitalized patients with hepatitis C,who came from department of infectious diseases,Renmin Hospital of Wuhan University from May 2011 to May 2012.Two pairs of specific primers were designed for rs12979860 gene polymorphisms,and one mutated base was introduced to the second or third site of the end of 3' with the reverse primer.rs12979860 gene polymorphism of 30 cases with hepatitis C was detected by AS-PCR,and gene sequencing was further used to verify the consistency of the two methods in parallel.Then,the frequency distribution of different rs12979860 genotypes with 174 cases were analyzed by the AS-PCR method in the population.Results The genotype CC,CT or TT of rs12979860 with 30 cases could be well identified by both AS-PCR and gene sequencing,and the coincidence rate was 100％ (x2 =60.0,P ＜ 0.01).Compared to gene sequencing,both of the sensitivity and specificity of AS-PCR were 100％.Compared to the control (CC genotype),TT genotype detection sensitivity by AS-PCR was 10-5,while sequencing sensitivity was 2 × 10-1.rs12979860 polymorphism in the TT,CC and CT genotype distribution in the Chinese population frequencies were 3.45％ (6/174),13.2％ (23/174) and 83.3％ (145/174),respectively.Conclusion AS-PCR can quickly,accurate,reliable,economic and efficiently detect IL28B rs12979860 gene polymorphism of hepatitis C in patients,which could predict the effect of antiviral therapy on patients with hepatitis C.

Objective: To observe the effect of DSP on three models of memory dysfunction,so as to provide evidence for its potential therapeuticaction for senile dementia.Methods: Three kinds of memory dysfunction are induced by scopolamine,reserpine and ovariectomy respectively,and memory ability is measured through step through and water maze tests.Meanwhile AchE activity in front cortex, hippocampus,striatum and limlic lobe of rats injected scopalamine is determined.Results: DSP can significantly prolong the step through latency,and decrease AchE activity in different parts of rats injeted scopalamine.And DSP can remarkably prolong latency,and increase uterus index of mice after ovariectomy.On the other hand,DSP can shorten water maze latency,enhance the correction percentage of mice injected reserpine.Conclusions:DSP can improve the memory dysfunction possibly through below pathways:(1)enhance the cholinergic system;(2)antagonize the decrease of monoamino transmitter;(3)has the possible estrogen like action.

Objective To study the effects of planting soil on active ingredients in different organs of Lonicera japonica Thunb.. Methods The contents of active ingredients in Lonicera japonica Thunb. from different origins, different varieties and different organs, such as chlorogenic acid, luteoloside, total flavonoids, total phenolic acid, and soil nutrient and inorganic elements, were detected. The data were analyzed by using SPSS 19.0 software. Results Lonicera japonica Thunb. from Pinyi area in Shandong Province had the highest content of active ingredients. In terms of content of active ingredients, PY-HBRD-1 was the most suitable variety in the Pingyi area. Analysis on the correlation between planting soil and active ingredients showed that the contents of K, Mn and other elements could be used as the index for quality control of Lonicera japonica Thunb.. Conclusion This study could provide guidance for selection of varieties of Lonicera japonica Thunb. in different planting areas, and enrich the materials about researches on producing area of Lonicera japonica.