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 explore the mutation spectrum of breast and ovarian cancer susceptibility gene 1/2 (BRCA1/2) which was related to breast cancer in female residents of Wuhan,and assess the relation of gene mutation and risk of suffering breast cancer.Methods 128 cases of female individuals,including 58 cases of breast cancer after surgery,70 cases of benign breast disease,and 50 femal hcalthy volunteers were selected by simple randon sampling from Department of Breast Surgery of Renmin Hospital of Wuhan University.BRCA1/2 gene was sequenced and compared with the standard template sequence to explore the possible mutations.Results In the breast cancer group,mutation emerged in 11 cases and the mutation rate was 19.0％ (11/58),including 8 cases of the BRCA1 gene mutations (3 cases of 185 del AG,5 case of 5382 ins C) and 3 cases of the BRCA2 gene mutations (2 cases of 6174 del T,1 case of C5773T) ; in the benign breast disease group,mutation emerged in 5 cases,the mutation rate was 7.1％(5/70),including 4 cases of the BRCA1 gene mutations (1 case of 185 del AG,3 cases of 5382 ins C),and 1 case of the BRCA2 gene mutation (6174 del T).There was no mutation detected in healthy control group.The mutation rate of the breast cancer group was significantly higher than that of benign breast disease group and healthy control group (x2 =4.05,10.56,P ＜ 0.05); However,there was no significant difference between benign breast disease group and healthy control group (x2 =3.73,P ＞0.05).Conclusions The mutation of BRCA1 gene (185 del AG,5382 ins C) and BRCA2 gene (6174 del T,C5773T) is in the presence of female residents in Wuhan.Furthermore,the mutation in BRCA1/2 gene increases the risk of breast cancer.

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 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 The aim of this study was to assess the relationship between the oral health habit and halitosis. It can provide an advisory basis for nurse giving individualized oral health instruction to patients of halitosis and can improve the result of prevention and treatment for halitosis accordingly. Methods 202 patients with periodontal disease were chosen to participate in this study. The study included the questionnaire of oral health habit and the halitosis diagnosis by organoleptic assessment.Comparisons of difference in the knowledge of oral health habit between the subjects with and without halitosis were conducted. Results Brushing times in subjects without halitosis were significantly more than that in subjects with halitosis. Brushing teeth in evening and interproximal brush usage were much better in subjects without halitosis than that in subjects with halitosis,respectively. Conclusions Good oral health habits could decrease the incidence of halitosis.

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.