To construct, express and purify fusion protein containing HBcAg and HBsAg preS1 epitope peptide for the purpose of investigating a novel HBV vaccine with both prophylactic and therapeutic functions. Using DNA recombinant technology, prokaryotic expression plasmid pBTcs1 expressing HBcAg and HBsAg pre-S1 epitope peptide fusion protein was constructed. After expressed in E.coli. HB101, the production BTcs1 was purified by sucrose density gradient ultracentrifugation and identified by SDS-PAGE, SEC, Western-blot and electron microscope. The results indicated that expression plasmid pBTcs1 was constructed successfully, and 20~25 mg purified BTcs1 fusion protein was obtained from 1L LB culture. Result of DOT-BLOT indicated that the distribution of BTcs1 was mainly in 30~50% sucrose, the purity of BTcs1 was greater than 95% by SDS-PAGE and SEC analysis. BTcs1 could probe with specific antibodies at 28 kDa by Western-blot, BTcs1 could also self assemble VLP by electron microscope analysis, its diameter was 30~34 nm approximately. The present study lay a foundation for further research functions and applications of BTcs1.

Nowadays,small peptides are always expressed in the form of fusion protein.The expression product contains many superfluous amino acids which can affect the biological functions of small peptides even expressed by GST fusion protein expression system.SUMO protease can cut SUMO fusion protein expressed by fusion expression system without any amino acid residues left on target protein thus become a hot topic in this field.Recombinant His-UlP1/pET3c/BL21(DE3)engineering strain was constructed by genetic engineering technology and the expression conditions were optimized in shake flaks.The process of high density fermentation was explored and different purification conditions were detected by chromatography.The results showed that SUMO protease could be expressed well after inducing the engineering strain by IPTG of 1.0mmol/L at 30℃ for 6 hours.The expression level of the strain in fermentation pots could reach 24.3% analyzed by SDS-PAGE.The purity of SUMO protease was more than 98% after further purification by cation exchange chromatography.The yield was 355mg SUMO protease per liter fermentation liquid.Western blot analysis demonstrated that there were immune reactions between IlP1 and 6?His antibodies,so it has established a good foundation for large-scale industralazation in the future.

Objective To explore the best vibration frequency for activating human muscles.Methods Nineteen healthy college students accepted vibration stimulation at frequencies between 10 and 50 Hz.Surface electromyograms (sEMG) were recorded.The subjects were sitting,standing,squatting (knee flexion 30°) and recumbent.Their left anterior tibial muscles and the medial heads of the gastrocnemius were targeted as test muscles.The vibration stimulation point was on the surface of the left distal tibia.The sEMG characteristics of the calf muscles were analyzed under vibration stimulation at different frequencies. ResultsThe leg muscles were activated significantly at all vibration frequencies,but the sEMG values of the anterior tibialis were significantly different at different frequencies,except for among 30 Hz,40 Hz and 50 Hz in any position.The gastrocnemius sEMG values were not significantly different at different frequencies. ConclusionVibration at 30 to 50Hz may be the normal human muscle activation frequency.

Macrophages are involved in many important biological processes and membrane proteins are the key effector molecules for their functions. However, membrane proteins are difficult to analyze by 2-DE based method because of their intrinsic tendency to self-aggregate during the first dimension separation (IEF). To circumvent the obstacle hampering membrane protein analysis, we combined one-dimensional SDS-PAGE with capillary liquid chromatography-tandem mass spectrometry (LC-MS/MS). Using this technique, we identified 458 GO annotated membrane proteins with extremely high confidence, including most known markers of peritoneal macrophages (e.g., CD11b, F4/80, CD14, CD18, CD86, CD44, CD16 and Toll-like receptor). Thirteen other CD antigens and 18 Ras-related small GTPase were also identified. In addition to those known macrophage membrane proteins, a significant number of novel proteins have also been identified. This research provides a valuable data set of macrophage membrane proteins, thus allowing for more comprehensive study of membrane proteins and a better understanding of the function mechanisms of macrophages in many biological processes.

OBJECTIVETo observe the effects of blood activating wind dissipating acupuncture (BAWDA) on blood pressure (BP) of prehypertension (PHT) patients.

METHODSTotally 60 PHT patients were assigned to the control group and the acupuncture group according to random digit table, 30 in each group. All patients were intervened by life style. BAWDA was additionally performed in patients in the acupuncture group for 6 weeks (30 times). The improvement of BP after intervened by acupuncture was observed. BP success rates and the proportion of PHT progressing to hypertension (HT) were also observed after 6-week intervention of acupuncture and at 1-year follow-up.

RESULTSSystolic blood pressure (SBP) and diastolic blood pressure (DBP) decreased after 6-week intervention in the acupuncture. The BP control rate was 56.7% (17/30 cases) in the acupuncture group vs.10.0% (3/30 cases) in the control group with statistical difference (chi2 = 14.70, P < 0.01). At 1-year follow-up BP success rate was 36.7% (11/30 cases) in the acupuncture group, remarkably higher than that of the control group [13.3%, (4/30 cases)] (chi2 = 4.36, P < 0.05).

CONCLUSIONSBAWDA showed BP regulating roles in a gradually stable decreasing tendency. It also could elevate BP success rate of PHT, and reduce the risk of PHT progressing to HT.

Acupuncture ; methods ; Acupuncture Therapy ; methods ; Blood Pressure ; Humans ; Hypertension ; Prehypertension ; therapy ; Wind

OBJECTIVETo estimate the prevalence of subcretins in 8 minorities of Yunnan province and to provide scientific basis for public health policy-making, as well as for prevention and control of iodine deficiency disorders.

METHODSFour thousand two hundred and twenty-two minority school children aged 8-12 years selected from 29 schools in 8 minority counties were measured by Combined Raven's Test in China (CRT-C(2)) and Jinyi Psychomotor test Battery (JPB).

RESULTSAverage of intelligence quotient (IQ) on 4,222 children was 91 +/- 19. Among 277 pupils with IQ between 55 - 69, 119 of them showed abnormal on JPB test and 5.4% of the children was found to have Goiter under ultrasonography. Median level of urinary iodine was 466.9 microg/L, and the qualified rate of iodized salt was 94.9%.

CONCLUSIONThe estimated prevalence rate of subcretin was 2.8%.

Child ; Child Development ; China ; epidemiology ; Congenital Hypothyroidism ; epidemiology ; Ethnic Groups ; Female ; Humans ; Intelligence Tests ; Male ; Prevalence

BACKGROUNDIn vitro fertilization (IVF) researches have suggested that cystathionine beta synthase (CBS) is involved in oocyte development. However, little is known about the regional and cellular expression patterns of CBS in the ovary. The purpose of this study was to analyze the localization of CBS in mice ovaries and to investigate the expression profile during follicular development.

METHODSWe used in situ hybridization and immunohistochemical analysis to determine CBS expression in the ovaries of female Balb/c mice. Then the follicles were collected from F1 (C57BL x Balb/c) mice and cultured in vitro. With the method of semi-quantitative RT-PCR, we also investigated the expression profile of CBS during follicular development.

RESULTSCBS was absent in the oocytes, although it was ubiquitously expressed in the ovary with the strongest expression in follicular cells at all stages. In late antral follicles, CBS expression was markedly higher in granulosa cells located close to the antrum and in cumulus cells around the oocyte. The semi-quantitative RT-PCR showed that CBS mRNA was detected in follicles at all stages in vitro. In cumulus-oocyte complexes superovulated, CBS expression also increased rapidly.

CONCLUSIONSCBS was located mainly in the follicular cells in the ovaries. The level of CBS expression is high in follicles during folliculogenesis in mice. Differences in the CBS expression profile between oocyte and follicular cells suggest a role for CBS as a mediator in interactions between oocyte and granulosa cells.

Animals ; Cystathionine beta-Synthase ; analysis ; genetics ; Female ; Gene Expression Profiling ; Immunohistochemistry ; In Situ Hybridization ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Ovarian Follicle ; physiology ; Ovary ; enzymology ; RNA, Messenger ; analysis

Objective To contrast HPV genotyping and clinical characteristics of women in Yan'an and provide data support for cervical cancer screening and prevention.Methods The genotyping of HPV infection was carried out by the method of PCR amplification and reverse hybridization.To statistical analysis clinical infection distribution data.Results Found 16.48％ (332 of 2014) were positive for HPV DNA in these women.HPV-16,52 and 53 were in the top three HPV high-risk types.HPV-81,6 and 44 were in the top three HPV low-risk types.There were 219 cases (65.96％) with pure high-risk type infection,74 cases (22.29％) with pure low-risk type infection and 39 cases (11.75％) with mixed infection in positive specimens.The HPV infection rate had showed first increasing then decreasing with age,and there were significant differences among the each groups (x2=34.238,P＜0.01).There were also differences in HPV positive detection rates in different occupations and there were significant differences among the each groups (x2=50.35,P＜0.01).There were also differences in HPV positive detection rates in different regions,and there were significant differences among the each groups (x2 =12.084,P＜0.05).Single infection was highest in HPV infection.Conclusion HPV infection was different in different populations,regions and ages,and it is of great significance to carry out HPV screening for early prevention and treatment of cervical cancer.

OBJECTIVETo study the influence of anastomoses sequence on the hemodynamics in off-pump coronary artery bypass grafting (CABG), and to investigate the strategy to stabilize the hemodynamics.

METHODSFrom March 2005 to March 2007, 67 patients with serious triple-vessel coronary artery lesions (male 45, female 22) with a age range from 44 to 81-years-old were enrolled for off-pump CABG. All the patients underwent left internal mammary artery-left anterior descending branch (LIMA-LAD) anastomose firstly, followed by the foreword anastomose of the other two vessels. According to the anastomose sequence of posterior descending branch (PDA) and obtuse marginal branch (OM), the patients were divided into three groups. Group I (n = 22) did the sequence of PDA-OM-aortic root (Ao). Group II (n = 14) did the sequence of Ao-PDA-OM. Group III (n = 31) did the sequence of Ao-OM-PDA. The hemodynamics markers, including heart rate (HR), central venous pressure (CVP), mean artery pressure (MAP), cardiac index (CI), left ventricular-stroke work index (LVSWI) and right ventricular stroke work index (RVSWI), were analyzed before LIMA-LAD anastomose to serve as baseline. And these markers were re-measured at LIMA-LAD anastomose, immediately after LIMA-LAD anastomose, at PDA anastomose and at OM anastomose.

RESULTSAll the patients survived after off-pump CABG. Compared with the baseline, there were no significant hemodynamic changes when doing LIMA-LAD anastomose. When the anastomose finished, the hemodynamic indices improved significantly. When performing OM anastomose, HR and CVP increased significantly, and MAP, CI, LVSWI and RVSWI decreased remarkably in group I and group III, while there were no significant changes of MAP, CI and RVSWI in group II . When performing PDA anastomose, HR and CVP increased significantly, and MAP, CI, LVSWI and RVSWI decreased remarkably in group I and group II, while there were no significant hemodynamic changes in group III except HR.

CONCLUSIONProximal anastomose first when performing off-pump CABG, followed by distal anastomoses of target vessels is beneficial to those who have hard exposure and difficult anastomose of OM and PDA.

Adult ; Aged ; Aged, 80 and over ; Anastomosis, Surgical ; methods ; Coronary Artery Bypass, Off-Pump ; methods ; Coronary Artery Disease ; physiopathology ; surgery ; Coronary Vessels ; surgery ; Female ; Hemodynamics ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; Retrospective Studies

Objective To analyze the clinical characteristics and prognosis of Chinese patients with apical hypertrophic cardiomyopathy (AHCM). Methods A total of 188 patients with AHCM diagnosed at Fuwai Hospital were included in this retrospective study. Clinical characteristics, mortality and cardiovascular morbidity were analyzed. A multiple logistic regression was performed to adjust for potential confounding factors. Results Males predominated with a number of 139 (73.9%) in this cohort. Patient's age ranged from 15 to 81 (51.9 ± 12. 6) years. There were 120 patients (63. 8% ) with "pure" type and 68 patients (36.2%) with "mixed" type of AHCM, 171 patients were followed up for (5.0 ± 3.0)years,cardiovascular mortality was 1.2%, 28 patients (16.4%) experienced one or more cardiovascular events. Conclusion The prevalence of AHCM is high in Chinese HCM patients, pure type AHCM is more common, and AHCM patients have a benign clinical course.