Objective To clone and express the encoding sequence of glyceraldehydes-3-phosphate dehydrogenase(GAPDH)from periodic Brugia molayi(Bm).Methods Total RNA was extraeted from periodic Brugic malayi.The BmGAPDH gene was amplified by RT-PCR.The PCR product was cloned and then subeloned into pcDNA3.1(+)vector.The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification,and were transformed into COS-7 cell subsequently.The expressed protein was identified by SDS-PAGE.Results BmGAPDH mRNA was highiy expressed in transfected COS-7 cell.The deduced amino acid sequence was identical with that of BmGAPDH.The recombinant pnotein wag about Nr 43 000.Conclusion The recombinant plasmid peDNA3.1(+)-BmGAPDH has been constructed and the protein has been expressed correctly.

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.

OBJECTIVETo evaluate the overall diet quality and diet model of labor workers in Shenzhen using Chinese Diet Balance Index (DBI).

METHODSIn May 2009, 14 canteens from Baoan, Longgang and Nanshan districts were selected by stratified random sampling and 60 workers were randomly selected from each canteen by using random number method. Diet measurements were carried out among the 840 labor workers. Diet quality was evaluated by using DBI scoring and evaluating system.

RESULTSThe median values of labor workers' food intakes of cereal and meat & poultry were 483.8 and 121.7 g/d, which were more than the recommended amounts of their intakes of Chinese residents (cereal: 250 - 400 g/d, meat & poultry: 50 -70 g/d). The median values of the labor workers' intakes of fruit, dairy and eggs were 37.3, 20.6 and 23.5 g/d,which were less than recommended amounts in fruits (200 - 400 g/d), dairy (300 g/d) and eggs (25 - 50 g/d). The DBI-HBS scores of males and females in Shenzhen migrant workers were 24.4 +/- 6.1 and 22.6 +/- 6.3, respectively with a statistically significant difference (t = 4.21, P < 0.01). DBI-HBS scores of < 20 age group, 20 - 29 age group, 30 - 39 age group and > or = 40 age group in labor workers were 12.7 +/- 5.9, 11.3 +/- 6.3, 12.8 +/- 6.4 and 11.2 +/- 5.6 respectively (F = 3.67, P = 0.01). There were 7 dietary patterns among labor workers in this survey. Nearly 8.2% (68/830) of them belonged to Pattern A. Pattern B and E were the main dietary patterns, which accounted for 37.3% (310/830) and 31.0% (257/830) of the total population.

CONCLUSIONDBI can describe and evaluate the overall dietary quality and the major problem of the dietary patterns in labor workers. It is necessary to strength nutritional education to increase the intake of fruits, milk and eggs to improve nutritional status in labor workers in Shenzhen.

Adolescent ; Adult ; Dairy Products ; Diet ; statistics & numerical data ; Diet Surveys ; Eggs ; Feeding Behavior ; Female ; Fruit ; Humans ; Male ; Meat ; Middle Aged ; Nutritional Status ; Young Adult

The aim of this study was to investigate the ability of calcium ionophore (CI ) to induce the differentiation of CML cells into dendritic cells (DC), to analyze the P210 expression in DCs and to evaluate the stimulatory effect of CML-DC on production of cytotoxic activity against CML cells via activating the autologous T cells. The mononuclear cells were isolated from bone marrow of CML patients whose WBC counts were more than 30x10(9)/L when samples were collected, then the lymphocytes and monocytes were discarded by pouring out supernatant twice at different culture time point. Slightly adherent cells were cultured in RPMI 1640 containing 10% FCS, with or without CI (375 ng/ml) and GM-CSF (200 ng/ml) at 37 degrees C, 5% CO2, fully humidified atmosphere for 96 hours. The cell morphology was observed under the inverted microscope and electron microscope; the expression of CD antigens was analyzed with flow cytometry; the P210 expression was measured with Western blot. LDH assay was used to evaluate the effect of cultured CML cells (CML-DC) generating cytotoxic T lymphocyte (CTL) activity against CML cells. The results indicated that after treatment with calcium ionophore and GM-CSF for 96 hours, CML cells showed DC morphological characteristics under inverted microscope and electron microscope. The expression of CD83, CD86, CD40, CD80 and HLA-DR increased remarkably. P210 was expressed in the CML-DC, but the expression level was lower than that in CML cells without CI and GM-CSF treatment. LDH assay showed that the CTL activity against CML was found greater in autologous T cells activated by CML-DC than that by CML cells. It is concluded that the CML cells can be induced to quickly differentiate into DC when cultured with CI and GM-CSF. CML-DC expresses P210, but the expression level is lower than that in CML cells. CML-DC can stimulate autologous T cells to produce CTL against CML.
Antigens, CD ; metabolism ; Bone Marrow Cells ; cytology ; Calcium ; pharmacology ; Cell Differentiation ; Dendritic Cells ; cytology ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Ionophores ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; pathology ; Male ; Monocytes ; cytology ; Tumor Cells, Cultured

The aim of this study was to explore the effect of different conditions on two-dimensional gel electrophoresis of proteins from human acute promyelocytic leukemia cell line NB4. The 24 cm pH 3-10 linear immobilized pH gradient (IPG) strips were chosen, the isoelectric focusing was carried out by using IPGphor. Then, the second-dimensional SDS-PAGE was performed. After silver staining, the gel was analyzed by ImageMaster 2D Elite. The results showed that low ion intensity sample washing buffer improved the performance of isoelectric focusing. The lysis buffer containing 7 mol/L urea and 40 mmol/L DTT could solubilize the most proteins from NB4 cell line. The rehydration solution containing thiourea and urea increased the low molecular weight protein points to be resolved in the area of basic end. The reasonable sample load and Volt/hour of NB4 were about 100 micro g and 63 200 V/h for the 24 cm pH 3-10 IPG strips. It is concluded that the proteins from NB4 and similar cell line are complicated and affected by many factors, so that, it is very important to select the right methods for sample preparation and the conditions of two-dimensional gel electrophoresis.
Cell Line, Tumor ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Isoelectric Focusing ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Neoplasm Proteins ; analysis

BACKGROUNDAlthough antiangiogenesis therapy plays an important role in anti-neoplastic treatment with its recognized efficacy and slight adverse effect, there is no prospective clinical trial to define ideal markers for predicting efficacy of antiangiogenic therapy. This study was undertaken to investigate the changes of activated circulating endothelial cells (aCECs) and survivin after anti-angiogenesis therapy and their significance in predicting the efficacy of the therapy.

METHODSPatients of non-small cell lung cancer (NSCLC) treated with chemotherapy with or without Endostar were observed. The amount of activated CECs was detected by flow cytometry, and the expression of survivin mRNA was determined by real-time polymerase chain reaction (PCR).

RESULTSAfter treatment, the amount of activated CECs decreased significantly in clinical benefit cases (P = 0.021 in chemotherapy alone, P = 0.001 in chemotherapy plus Endostar), increased in disease progressive cases (P = 0.015 in chemotherapy alone, but P = 0.293 in chemotherapy with Endotatar). After therapy, the expression of survivin mRNA decreased in clinical benefit cases (P = 0.001) and increased in disease progressive cases (P = 0.018). A positive correlation was found between activated CECs and survivin in the chemotherapy group pre- and post-therapy (P = 0.001 and 0.021, respectively), but only in the chemotherapy with Endostar group pre-therapy (P = 0.030) rather than post-therapy. A positive correlation was found between the decreased activated CECs after therapy and time to progression (TTP) (r = 0.322, P = 0.012); a negative correlation was found between the amount of survivin mRNA in serum post-therapy and TTP (r = -0.291, P = 0.048).

CONCLUSIONSActivated CECs and survivin may be ideal markers forecasting efficacy and prognosis of NSCLC. The former can reflect more sensitively antiangiogenic efficacy and the latter is more sensitive to shrinkage or swelling of tumors. Their combination can evaluate more accurately the efficacy of antiangiogenic therapy of NSCLC.

Angiogenesis Inhibitors ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; metabolism ; Endostatins ; therapeutic use ; Endothelial Cells ; drug effects ; metabolism ; Female ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Lung Neoplasms ; drug therapy ; genetics ; metabolism ; Male ; Microtubule-Associated Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

This study was purposed to investigate the relationship between HLA-A, B allele polymorphisms and red blood cell parameters of patients with --(SEA/αα) subtype of α(0)-thalassemia in Han ethnic population of Wuzhou city. The HLA genetic polymorphisms were determined by polymerase chain reaction-sequence-based typing (PCR-SBT) in 57 patients with --(SEA/αα) subtype of α(0)-thalassemia of Han ethnic population in Wuzhou city, Guangxi province, China. Mean corpuscular volume (MCV), hemoglobin (Hb), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were detected by automatic blood analyzer system. HbA2 were detected by electrophoretic method. The statistical analysis was performed by ordinal polytomous logistic regression. The results showed that Hb and HbA2 levels were significantly lower in patients positive for HLA-A*33:03, B*15:01 or B*55:02, and were significantly higher in patients positive for B*15:02 (P < 0.05). It is concluded that several HLA alleles may be associated with Hb level of --(SEA/αα) subtype of α(0)-thalassemia of Han ethnic population in Wuzhou city. This result has the value for understanding phenotype-genotype relationships in thalassemia.
Adolescent ; Adult ; Alleles ; Child ; Child, Preschool ; China ; epidemiology ; Erythrocytes ; cytology ; Ethnic Groups ; genetics ; Female ; Genotype ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; Hemoglobins, Abnormal ; genetics ; Humans ; Male ; Middle Aged ; Young Adult ; alpha-Thalassemia ; blood ; classification ; epidemiology ; genetics

OBJECTIVEWe propose an image-based key frames gating method for intravascular ultrasound (IVUS) sequence based on manifold learning to reduce motion artifacts in IVUS longitudinal cuts.

METHODSWe achieved the gating with Laplacian eigenmaps, a manifold learning technique, to determine the low-dimensional manifold embedded in the high-dimensional image space. A distance function was constructed by the low-dimensional feature vectors to reflect the heart movement. The IVUS images were classified as end-diastolic and non-end-diastolic based on the distance function, and the IVUS images collected in end-diastolic stage constitutes the key frames gating sequences.

RESULTSWe tested the algorithm on 13 in vivo clinical IVUS sequences (images 915±142 frames, coronary segments length 15.24±2.37 mm) to calculate the vessel volume, lumen volume, and the mean plaque burden of the original and gated sequences. Statistical results showed that both the vessel volume and lumen volume measured from the gated sequences were significantly smaller than the original ones, indicating that the gated sequences were more stable; the mean plaque burden was comparable between the original and gated sequences to meet the need in clinical diagnosis and treatment. In the longitudinal views, the gated sequences had less saw tooth shape than the original ones with a similar trend and a good continuity. We also compared our method with an existing gating method.

CONCLUSIONThe proposed algorithm is simple and robust, and the gating sequences can effectively reduce motion artifacts in IVUS longitudinal cuts.

Algorithms ; Angiography ; methods ; Artifacts ; Electrocardiography ; Humans ; Motion ; Reproducibility of Results ; Sensitivity and Specificity

Intracranial atherosclerotic disease (ICAD) of a major intracranial artery, including middle cerebral artery (MCA),basilar artery, is the most common causes of stroke and is associated with a high risk of recurrent stroke in China. The difficulty to treatment these high-risk disease is to identify high-risk stroke subgroups and to develop more effective treatments (aggressive medical therapy/endovascular therapy). With the benefits, including non-invasive, in vivo, and no-ionizing radiation, 3.0 Tesla high-resolution magnetic resonance imaging (HR MRI) could be used to stratify high-risk patients, monitor progression of disease, and evaluate clinical efficacy, based on MCA wall structure and plaque characteristic. HR MRI has the latency of predicting high-risk patients benefit from endovascular therapy, having a broad application prospect during psot-SAMMPRIS era. The current research on MCA stenosis using HR MRI focuses on methodoiogy, diagnosis and differential diagnosis, etiology, and lacks of clinical efficiency evaluation and prognostic analysis of ICAD treatment, especially lacks the research on in-stent restenosis, which needs further investigation.
Brain ; China ; Constriction, Pathologic ; Diagnosis, Differential ; Humans ; Intracranial Arteriosclerosis ; Magnetic Resonance Imaging ; Middle Cerebral Artery ; pathology ; Plaque, Atherosclerotic ; Prognosis ; Stroke ; Treatment Outcome

OBJECTIVETo explore the relationship between mammaplasty and results after polyacrylamide hydrophilic gel (PAHG) removal from breast.

METHODSFrom Feb. 2003 to Aug. 2009, 130 patients with bilateral breast augmentation by PAHG injection were treated. Preoperative ultrasound examination and MRI were performed to know the distribution of PAHG and infiltration at the surrounding tissue. According to the conditions after removal, the patients were received implant augmentation immediately, or at the second stage, or no implant.

RESULTSThe patients were followed up for 3 months at the most with a very satisfactory rate of 63.84% (83/120), a satisfactory rate of 31.53% (41/120) and a unsatisfactory rate of 4.63% (6/120). Slight capsular contracture (Baker I) occurred in 5 cases with 6 breasts in satisfactory group. All the patients in unsatisfactory groups who selected unsuitable implants by themselves were re-operated to take out the implants. 3 cases with much residue PAHG insisted to receive breast implants. Among them, 2 cases achieved acceptable results even the surface of the breasts were not smooth. No other complication happened.

CONCLUSIONSThe breast reaugmentation after PAHG removal should be performed based on the deformity and condition of breast. Both cosmetic result and psychological relief could be obtained after mammaplasty.

Acrylic Resins ; adverse effects ; Adult ; Breast Implants ; adverse effects ; Device Removal ; Female ; Follow-Up Studies ; Humans ; Mammaplasty ; methods ; Middle Aged