Hepatitis B is a serious infectious disease worldwide, and hepatitis B virus (HBV) is the direct cause of this disease. In recent years, as an essential part of its evolutionary process, HBV mutation has been extensively studied domestically and globally. However, the study on the conserved sequences in HBV sequences is still in its infancy. In this study, we applied multiple EM for motif elicitation (MEME) algorithm to discover HBV motif and proposed a new metric, conservative index (CI), to carry out phylogenetic analysis based on HBV sequences. Then, the constructed phylogenetic tree was subjected to reliability assessment. The results demonstrated that the new metric CI combined with the MEME algorithm can effectively help to discover motifs in HBV sequences and construct a phylogenetic tree based on them and to analyze the evolutionary relationship between HBV sequences; in addition, the possible ancestral sequences of samples may be obtained by conservative analysis. The proposed method is valuable for the exploratory study on large HBV sequence data sets.
Computational Biology ; Conserved Sequence ; Evolution, Molecular ; Hepatitis B virus ; genetics ; Humans ; Nucleotide Motifs ; Phylogeny ; Reproducibility of Results

OBJECTIVETo observe the clinical effects of acupuncture method of "Huoxue Sanfeng, Shugan Jianpi" (activating blood and eliminating wind, soothing liver and strengthening spleen) on morning blood pressure in patients with cerebral infraction combined with essential hypertension.

METHODSSixty-eight patients were randomly divided into an observation group and a control group, 34 cases in each one. The patients in the two groups were treated with acupuncture method of "Xingnao Kaiqiao" (consciousness-restoring resuscitation) and oral administration of nifedipine. In addition, patients in the observation group were treated with acupuncture method of "Huoxue Sanfeng, Shugan Jianpi" that met the criteria of standard manipulation, in which bilateral Renying (ST 9), Quchi (LI 11), Hegu (LI 4), Zusanli (ST 36) and Taichong (LR 3) were selected. The treatment was given once a day, five times a week, for totally six weeks. The improvement and control rate of morning blood pressure in the two groups were observed.

RESULTS(1) After treatment, the morning blood pressures were decreased significantly in the two groups (all P < 0.05); after 15 treatments, the reduction of systolic pressure and diastolic pressure in the observation group was superior to that in the control group, but the difference was not significant (both P > 0.05); after 30 treatments, the reduction of systolic pressure and diastolic pressure in the observation group was significantly superior to that in the control group (both P < 0.05). (2) After 30 treatments, the control rate of morning blood pressure in the observation group was significantly higher than that in the control group [82.4% (28/34) vs 58.8% (20/34), P < 0.05].

CONCLUSIONAcupuncture method of "Huoxue Sanfeng, Shugan Jianpi", characterized with standard manipulation criteria, can effectively control morning blood pressure in patients with cerebral infraction combined with essential hypertension.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Blood Pressure ; Cerebral Infarction ; physiopathology ; therapy ; Essential Hypertension ; Female ; Humans ; Hypertension ; physiopathology ; therapy ; Liver ; physiopathology ; Male ; Middle Aged ; Spleen ; physiopathology ; Treatment Outcome

Objective To detect serum protein spectrum in gastric cancer, screen the serum proteins related to gastric carcinoma and build auxiliary diagnosis model with SELDI-TOF-MS, Methods The serum proteomic fingerprints were detected by SELDI-TOF-MS and CM10 proteinchip. The sera came from 50 first visit gastric cancer patients who were diagnosed by pathological method and 16 healthy peo- ple. The data were analyzed by Biomarker Wizard Software and the proteomic fingerprint of gastric carcinoma model was developed by BPS (biomarker pattern software). Results Compared with the healthy people group, there were 34 significant different protein peaks in the gas- tric carcinoma group. And the diagnosis model composed with 6 proteins (M/Z values were 6016, 6744, 2822, 4474, 7892 and 3242) could classify the 2 groups correctly. In the test group, the sensitivity and specificity were 96% and 93.75% respectively, the accuracy was 95.45%. Conclusion SELDI-TOF-MS method show features such as microcontent, fast and high- resolution etc. It could be utilized to screen significant proteins and develop an auxiliary diagnosis model in gastric carcinoma, which may be used to diagnose this disease.

Objective: To establish an HPLC method for the simultaneous determination of five active components (paeoniflorin, liquiritin, ammonium glycyrrhizinate, edomin and osthole)in Pifukang lotion with multiple UV wavelengths.Methods: A Diamonsil C18 (200 mm×4.6 mm,5 μm) column was used with the mobile phase consisting of 0.1% hydrochloric acid and acetonitrile with gradient elution.The flow rate was 1.5 ml·min-1, and the column temperature was 30℃.Paeoniflorin was detected at 232 nm(0-6 min), liquiritin was detected at 277 nm(6-10 min),and ammonium glycyrrhizinate, edomin and osthole were detected at 254 nm(after 10 min).Results: The linear range of paeoniflorin,liquiritin,ammonium glycyrrhizinate, edomin and osthole was 28.200-282.000 μg·mL-1(r=0.999 7),12.150-121.500 μg·ml-1(r=0.998 8),13.420-134.200 μg·ml-1(r=0.999 5),0.047-0.466 μg·ml-1(r=0.999 9) and 2.38-23.8 μg·ml-1(r=0.999 9), respectively.The average recovery (RSD) was 98.49%(0.71%), 99.00%(0.62%), 98.38%(0.85%), 97.36%(0.92%) and 97.70%(0.78%), respectively (n=6).Conclusion: The established method is simple, accurate, highly sensitive and well reproducible, which can be used for the determination of the five active ingredients in Pifukang lotion.

Objective To explore the influence of Akt inhibitor MK2206 in the proliferation and apoptosis of tongue squamous cell carcinoma TCA-8113 cells,and to clarify the possible mechanism.Methods The tongue squamous carcinoma TCA-8113 cells at the logarithmic phase were randomly divided into control group and 1,5,25,125, 250 nmol·L-1 MK2206 groups.The inhibitory rate of proliferation of TCA-8113 cells was detected with MTT method,and the apoptotic rate of TCA-8113 cells was determined with flow cytometry(FCM),and the expressions of caspase-9,Bad,GSK-3β,p-Akt and T-Akt proteins in the TCA-8113 cells were detected with Western blotting method.Results The IC50 of tongue squamous cell carcinoma TCA-8113 cells after treated with MK2206 for 12, 24,and 36 h were (112.54±1.67),(79.67±2.01),and (33.33±1.98)nmol·L-1 .The FCM results showed that the apoptotic rates of TCA-8113 cells after treated with 1,5,25,125,and 250 nmol·L-1 MK2206 for 12 h were (14.2±0.74)%,(19.3±0.45)%,(35.1±0.45)%,(39.6±0.48)% and (52.1±0.19)%;there were significant differences compared with control group(P<0.01).The Western blotting method results showed that the expressions of p-Akt, Bad and GSK-3βwere decreased with the increasing of dose and time of MK2206;compared with theβ-actin in control group,the bands got darken;the expression level of caspase-9 was increased, compared with theβ-actin in control group, the bands got darken;the T-Akt protein expression did not change significantly;compared with the β-actin in control group, the color of bands had no significant difference.Conclusion Akt inhibitor MK2206 can inhibit the proliferation of tongue squamous cell carcinoma TCA-8113 cells and induce apoptosis.

Objective To explore the changes of QRS combined data and myocardial enzymes during ischemia-reperfusion in rabbits,and observe the changes of ST segment and T wave of lead Ⅱ,and arrhythmias.Methods The models of ischemia-reperfusion(A,B,C and D group)were made by ligating the left circumflex territory for 10,20,30,40 min and then reperfusion for 60 min.The sham-operated control was designed.The changes of the QRS combined data,ST segment and T wave and rises of arrhythmias about electrocardiogram of lead Ⅱ were observed.Meanwhile,the activity of serum myocardial enzymes was measured.Results The QRS combined data of the group D was higher significantly than the sham-operated control,group A and group B(P<0.05).T waves of different groups were not significantly different(P>0.05).The J spot elevation of lead Ⅲ in 19 rabbits of the ligating group and 5 rabbits of the sham-operated control was observed before operation.The changes of ST segment were clearly observed in the ligating group after operation:apparent mono-phasic curve elevation in 21 rabbits,level type depression in 2 and no change in 1.ST segment of other animals was declined more than 0.2 mV in 60 min reperfusion,except ST segment of 2 rabbits elevated again more than 0.1 mV,2 rabbits were normal and 1 rabbit was depressed.Two rabbits of the group D respectively died of sinus bradycardia in reperfusion 40 min and 43 min,and two rabbits appeared premature ventricalar contraction during reperfusion 3-5 min.The activity of serum aspartate aminotransferase(AST)and lactate dehydrogenase(LDH)obviously increased in the group D(P<0.05).Creatine kinase(CK)was high significantly except the sham-operated control(P<0.05).The activity of serum CK-MB of the groups C and group D was significantly higher than that of the sham-operated control(P<0.05).Conclusion The rising of LDH,CK-MB and QRS combined date in the group D is related with ischemia-reperfusion injury.

The study is aimed to distinguish morphological characteristics of Dalbergiae Lignum collected from crude drug's markets and establish a identification methods and the quality standard for Dalbergiae Lignum. The macroscopic and microscopic features of Dalbergiae Lignum from crude drug's market were observed, analyzed and compared according to Hongmu specification issued by the People's Republic of China in 2000, and by the characteristics recorded in domestic monograph of Mucai Shibie (wood identification). The redwood of Dalbergiae Lignum cut into small pieces as medicinal material are dry heart wood of mahogany (trees from Dalbergia sp.), which characteristics of the small pieces as crude drug are different. There are differences in macroscopic and microscopic features about texture of wood and color, odor, taste, transverse section, radial section, tangential section. The results can provide basis for identification, application and improment of the quality standard of Dalbergiae Lignum as medicinal material.
China ; Dalbergia ; anatomy & histology ; chemistry ; classification ; Herbal Medicine ; economics ; Plants, Medicinal ; chemistry ; classification ; Quality Control ; Xylem ; anatomy & histology ; chemistry

Hepatitis B is one of the most serious global threats to human health. Phylogenetic analysis of hepatitis B virus (HBV) can reveal the evolutionary relationship between HBV sequences and thus provide a basis for the prediction and treatment of hepatitis B and other aspects. In this study, we performed sequence analyses on the HBV sequences of five clinical HBV samples and the HBV sequences retrieved from the GenBank, EMBL, and DDBJ to construct a phylogenetic tree and analyze sequence structures. The experimental results revealed that the C gene of one cloned sequence had a recombinant structure of HBV B/ C subtype. Moreover, the phylogenetic results proved the existence of a newly found subtype HBV/B6 in Xishuangbanna of Yunnan Province, China. The experimental conclusion represents certain value for phylogenetic studies of HBV in Yunnan ethnic minority groups.
DNA, Recombinant ; genetics ; Genes, Viral ; genetics ; Genotyping Techniques ; Hepatitis B virus ; classification ; genetics ; Humans ; Phylogeny

Objective To investigate the effects of curcumine treatment on the expression of TNF-α in the peritoneum of acute peritonitis rats. Methods The rats were intrapertoneally injected Staphylococcus epidemidis (S.epidermidis) for the mixlelization of acute peritonitis. After a survival time of 3,6,12,24 and 48 hours,TNF-α expression in the peritoneum was detected by Western blot and RT-PCR in the rats of curcumin treatment group and non-curcumin treatment group. Results TNF-a expression showed time-dependent increase at 6,12,24 hours after the infection in the non-curcumin treatment group. In curcumin treatment group,TNF-α expression decreased at 6,12,24 hours after infection compared to non-curcumin treatment group. Conclusion Curcumin may be a protective agent in acute peri-tonitis induced by S.epidermidis.

Objective To investigate the prevalence of latent tuberculosis infection(LTBI),and to identify the risk factors in primary schoolchildren from Shanghai through the population-based field investigation combined with the tuberculosis infection enzyme-linked immunospot assay(T-SPOT.TB)assay.Methods The children in grade 4 and 5 were enrolled from four primary schools in Pudong new district and Yangpu district of Shanghai.Questionnaire interview was applied to investigate the soeiodemographic and clinical information related to LTBI.The T-SPOT.TB assay was used to detect LTBI in the enrolled subjects.Univaitate and multivariate analyses were used to identify the risk factors associated with LTBI among the primary schoolchildren.Results Totally 472 schoolchildren were enrolled in the present study,with 439(93.0%)being vaccinated with bacillus calmette-guerin (BCG) and ten (2.1%) having contact history with tuberculosis (TB) patients.Among the 472 eligible subjects,16(3.4%) children were T-SPOT.TB positive,who had no clinical symptoms andsigns relevant to TB and were defined as LTBI.The LTBI prevalence in BCG vaccinated and unvaccinated children were 2.7% and 12.1%,respectively (OR:6.972;95%CI:1.834-26.500);those in TB contacts and children without TB contact history were 30.0% and 2.8%, respectively (OR: 16. 38; 95% CI: 3. 692-72. 700). Conclusions The prevalence of LTBI among senior schoolchildren in Shanghai is 3.4%. BCG vaccination is protective for children from LTBI, while daily contacts with TB patients increases the risk of LTBI in schoolchildren.