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.

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.

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.

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.

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.

Objective To investigate epidemic characteristics of HCV and the distribution of naturally occurring resistance mutations of HCVNS5B in Hubei province,and provide the basis for individualized antiviral treatment for HCV infected pa-tients.Methods Collected HCV blood samples of 253 patients from March 2011 to May 2014 in Renming Hospital of Wu-han University,and determined the subtypes of HCVNS5B by first generation of sequencing technology,then compared the result with the nucleotide sequence of HCVNS5B online,located drug resistance loci and record.Results There were 4 gen-otypes and 6 different genetic subtypes in these 253 cases,namely 1a subtype 3,1b subtype 192,2a subtype 38,3a subtype 3, 3b subtypes 11 and 6a subtypes 6.Found 27 gene locus of resistance from 234 different places.The main drug resistance lo-ciare L159F (475-477)26.42%,followed by C316Y (946-948)16.98%,V321I (961-963)15.09% and S282T (844-846) 12.58%.Conclusion The epidemic characteristics of HCV in Hubei province were consistent with those of current in Chi-na;subtype 1b was the main types,followed by 2a subtype.The number of drug-resistant of HCVNS5B was more and com-plex,and presented some popular distributions.It was complex in mutations of 2a subtype,including 13 mutations of natural resistance.There were fewer mutations in subtype 1b,including sites of L159F and C316N.These results may provide the basis for antiviral therapy for HCV patients in this region.

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 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 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.

AIM: To observe the difference of actions among three traditional compl ex prescriptions with the same composition but the different dosage, including X iaochengqi Decoction, Houpudahuang Decoction and Houpusanwu Decoction, so as to provide evidence for their different clinical applications. METHODS: A series of experiments were performed, including carbon p rop ellance in small intestine, black stool excretion, cough induced by ammonia liqu or and phenol red secretion from trachea. RESULTS: Three prescriptions all enhanced carbon propellance in sma ll intestine at the high dosage, while at the low dosage only Houpusanwu Decoct ion showed the significant effect. They all shortened the latency of black stool a nd increased their total amounts in 6 hours at the high dosage, while at the low dosage Xiaochengqi Decoction and Houpudahuang Decoction had the significant p urgative action. Houpudahuang Decoction remarkably prolonged cough latency, dec reased cough times and enhanced phenol red secretion from trachea, while other t wo prescriptions had no obvious effects. CONCLUSION: Such results showed that Xiaochengqi Decoction had the better purgative action, while Houpusanwu Decoction regulated Qi strongerly, a nd Houpudanghuang Decoction showed the marked actions of relieving cough and re solving phlegm.