The cloning of promoter is important for studying the genetic engineering and the regulation of gene expression in plants. Two promoters Os772 and Os359, which are predicted to be highly expressed in the endosperm of rice from the EST database were cloned. After construction of the Os772∶∶GUS and Os359∶∶GUS expression vectors, they were transformed into rice. X-Gluc staining of transgenic plants showed that Os772 and Os359 can promote GUS gene expression in matured endosperm but not in root, stem, leaf and flower. This result indicates Os772 and Os359 are two rice endosperm-specific promoters.

Objective To analyze the epidemiological characteristics of Mycobacterium tuberculosis by enterbaeterial repetitive intergenic consensus-polymerase chain reaction(ERIC-PCR)DNA fingerprint. Methods Mycobacterium tuberculosis positive sputum samples between September 2003 to May 2006 were collected and cultured.Chromosomal DNA were extracted and ERIC-PCR DNA fingerprinting was analyzed by software,such as RAPD PHYLIP and Treeview.Results A total of 42 different fingerprints were detected.Phylogenetic analysis showed that they could be classified into three clusters,the clustering rate was 72.6%.The characteristics of ERIC-PCR fingerprint patterns were related to age,drug resistance,and type of resistance.Conclusions ERIC-PCR DNA fingerprinting technique used in this study is good for epidemiological studies with its strong discrimination,simplicity and rapidness.A high level of recent transmission is found in our city.

Background Stromal cell-derived factor-1_?(SDF-1_?)has been demonstrated to be essential for stern cell mobilization/homing.Recent evidence indicates that SDF-1_? has been expressed in injured carotid arter- ies.Besides,high SDF-1_? plasma levels are clinically associated with stable coronary artery disease.Objective To investigate whether SDF 1 involves in mobilization of endothelial progenitor cells(EPC)and reendothelialization after vascular injury.Methods SDF-1_? was detected by RT-PCR and Western blot in carotid arteries of mice at different time points after wire-induced injury.SDF-1_? determination in peripheral blood samples and BM was per- formed by SDF-1_? enzyme-linked immunosorbent assay(ELISA)kit.EPC in peripheral blood collected at different time points after vascular injury were quantified by flow cytornetry.In subgroup,blocking SDF-1 rnonoclonal anti- body was injected,peripheral blood EPC were quantified after vascular injury and reendothelialization of injured ar- teries was determined 14 days later.Results Expression of SDF-1_? was evident at day 1,and peaked at day 3 after arterial injury.A rise in plasmatic concentration of SDF-1_? and a significant reduction of SDF-1_? in bone marrow concentration was noticed at all time points following injury.The amount of circulating EPC was increased shortly after induction of vascular injury and persisted up to 7 days(P

The high performance liquid chromatography-fluorescence micelle assay (HPLC-FMA) method for the content determination of polysorbate 80 in monoclonal antibody drugs was validated to study its applicability and transferability between various laboratories, and the feasibility to be included in the Chinese Pharmacopoeia. Both J.T. Baker and Nanjing Well-sourced polysorbate 80 was used in the collaborative validation of polysorbate 80 content analysis in seven different laboratories. The results show that when the protein concentration was no more than 20 mg·mL^-1 and the concentration of polysorbate 80 ranged from 0.05 to 0.5 mg·mL^-1, the method had good specificity. The recovery rates of the spiked samples ranged from 92.20% to 117.70% for J.T.Baker and from 93.90% to 117.20% for Nanjing Well. The intra-laboratory precision (%RSD) was less than 4.30% for J.T. Baker, and less than 2.60% for Nanjing Well, while the overall precision was less than 5.45% for J.T. Baker, and less than 6.70% for Nanjing Well. The linear correlation coefficient was more than 0.98 for J.T. Baker and more than 0.99 for Nanjing Well. The results of the collaborative validation prove that the HPLC-FMA method has good accuracy, precision, linearity, and specificity, and could be used for release control analysis of polysorbate 80 content in monoclonal antibodies across different laboratories.

The expression of plant gene is controlled by its promoter. The isolation and the function analysis of promoter are important for studying the genetic engineering and the regulation expression of plant genes. In this paper, we cloned a promoter, 0s252, which was predicted to be highly expressed in the stem of rice from the EST database. After the construction of the Os252::GUS expression vector, it was transformed into rice. The integration of transgenes into transgenic rice genome was confirmed through PCR analysis. X-Gluc staining showed that Os252 can promote GUS gene expression in leaf, stem and matured seed. GUS enzyme activities driven by Os252 promoter in leaf and seed are about 190% and 250% of that driven by the 35S promoter. Thus, the Os252 promoter can be applied for rice genetic engineering.
Gene Expression Regulation, Plant ; Oryza ; genetics ; metabolism ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Promoter Regions, Genetic ; genetics

OBJECTIVETo observe the effects of C reactive protein (CRP) on endothelial progenitor cell (EPCs) function.

METHODSMononuclear cells (MNCs), isolated from bone marrow by density gradient centrifugation combined with adherent cell filtration, were plated on fibronectin coated culture dishes. After 7 days, adherent cells were cultured with different concentrations of CRP (0, 5, 10, 15, 20 microg/ml) for 48 hours. EPCs proliferation and migration ability were observed and adhesion assay was performed. The eNOS mRNA expression of EPCs were measured by RT-PCR.

RESULTSThe number of EPCs in CRP groups (10, 15, 20microg/ml) was obviously lower than that in control group (54 +/- 3, 47 +/- 3, 39 +/- 5 vs.60 +/- 3, P < 0.01). EPCs proliferation capacity was inhibited in CRP groups (10, 15, 20 microg/ml) compared with that in control group (0.297 +/- 0.036, 0.273 +/- 0.013, 0.259 +/- 0.035 vs. 0.345 +/- 0.014, P < 0.01). EPCs migration capacity was inhibited significantly in CRP groups (5, 10, 15, 20 microg/ml) than that in control group (28 +/- 2, 22 +/- 3, 19 +/- 3, 16 +/- 2 vs. 30 +/- 2, P < 0.05). EPCs adhensive number was lower in CRP groups than that in control group (11 +/- 2, 9 +/- 2, 6 +/- 2, 5 +/- 1 vs. 12 +/- 2, P < 0.05). The mRNA expressions of eNOS in CRP groups were significantly lower in control group. And compared with control group, NOS activity decreased significantly in CRP groups (10, 15, 20 microg/ml) (57.44 +/- 3.25, 48.37 +/- 3.86, 36.82 +/- 4.89 vs. 68.56 +/- 2.82, P < 0.01).

CONCLUSIONCRP could both reduce EPCs number and inhibit EPCs functions.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; C-Reactive Protein ; pharmacology ; Cells, Cultured ; Endothelial Cells ; drug effects ; metabolism ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; drug effects

To analyze and discuss placebo-related information in clinical research literatures in the past 30 years, including placebo's dosage form, ingredients, preparation process and quality control. Effort were made to research the CNKI. full-text database to preliminary find 700 placebo-related clinical research literature, screen out 301 eligible articles by hand, read the literatures to extract placebo-related information and make statistics and discussions. According to the results, Chinese randomized placebo-controlled clinical studies were characterized by diverse dosage forms of placebo with lack of reports for components, as evidenced by the only 17 literatures describing placebo's preparation or specific composition among the 301 literatures. Placebo-controlled clinical trials covered a wide range of disease spectra, but with a specific tropism of diseases in terms of system classification. Although placebo plays a key role in blinded clinical studies, researchers made less records of placebo, perhaps because they paid less attention to placebo or more attention to the research process or restricted by other objective conditions. Moreover, placebo production, quality control and quality evaluation also need to be further standardized.
Biomedical Research ; history ; standards ; China ; History, 20th Century ; History, 21st Century ; Humans ; Placebo Effect ; Quality Control ; Randomized Controlled Trials as Topic ; history ; standards

OBJECTIVETo explore the relationship between the genetic polymorphism of fibronectin (FN) (4 genetic locus) and pneumoconiosis.

METHODS128 male I-period pneumoconiosis were selected as cases who were examined with radiography and diagnosed by the Pneumoconiosis Diagnosis Expert Panel, based on the Chinese National Diagnosis Criteria of Pneumoconiosis (GBZ 70 - 2002). According to 1:1 paired matching method, 128 dust exposure workers were selected as control who were exposed to same dust as cases. The difference of age and cumulative length of service between case and control was not over five years and two years, respectively. 5 ml peripheral venous blood was drawn and anticoagulated with 2% EDTA. The polymorphisms of FN (MspI, TaqIb, HindIII, HaeIIIb) were detected, using the method of polymerase chain restriction fragment length polymorphism (PCR-RFLP) techniques and PCR.

RESULTSThe frequencies of FN Msp I (CC) in cases and control groups were 10.9% and 3.9%, respectively. The difference was significant (P < 0.05). The frequencies of FN (MspI) C allele were 41.8% and 31.2% in case and control, and the difference between cases and controls was significant (P < 0.05). The frequencies of FN HaeIIIb (AA) genotype in cases (24.2%) was higher than that in control groups (17.9%), OR = 5.0 (95% CI: 4.840 approximately 24.210). The frequencies of FN (HaeIIIb) A allele were 51.9% and 42.2% in case and control, and the difference was significant (P < 0.05). The difference of TaqIb and HindIII genotype between cases and controls were not significant (P > 0.05).

CONCLUSIONThe risk of suffering from pneumoconiosis increases in workers carrying FN (MspICC or HaeIIIb AA) genotype after exposure to dust. Workers both carrying FN (HaeIIIb AA) and (MspICC) genotypes are more susceptible to pneumoconiosis. The relationship between genetic polymorphism of FN (TaqIIb, HindIII) and pneumoconiosis has not been found.

Adult ; Aged ; Aged, 80 and over ; Fibronectins ; genetics ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; genetics ; Polymorphism, Restriction Fragment Length

The purpose of this study was to detect the minimal residual disease (MRD) in peripheral blood of newly diagnosed patients with acute myeloid leukemia (AML) on day 8 of induction chemotherapy and analyze the correlation between day 8 MRD (D8RD) and therapeutic effectiveness. 29 adult patients (13 males and 16 females, aged 16 - 75 years, median 41 years) with AML diagnosed and treated in West China Hospital from September 2009 to June 2010 were analyzed and followed up in the study. The leukemia-associated aberrant immunophenotype (LAIP) of all the patients were detected by multiparameter flow cytometry (FCM) before therapy. The level of MRD in the peripheral blood at day 8 of induction chemotherapy was detected by FCM based on the LAIP. The overall survival curve was drawn by calculation using Kaplan-Meier method using, and the comparison between different groups was carried out by Log-rank test. The results indicated that after first course therapy, the levels of peripheral D8RD in 7 out of 29 AML cases were lower than 0.01% (negative group), and that in another 22 cases were higher than 0.01% (0.08% - 55%, positive group). The sex, age, WBC, LDH, percentage of bone marrow blasts at diagnosis in these groups were not statistically different. 6 cases achieved CR (86%) in D8RD negative group, and also 6 cases achieved CR (27%) in D8RD positive group, CR rate in D8RD negative group was higher than in D8RD positive group (86% vs 27%, P < 0.05). The median follow-up of 29 cases lasted for 15 months; the 1-year overall survival rate of D8RD negative and D8RD positive groups was 100% and 39.4%, respectively (P < 0.01). It is concluded that MRD level in peripheral blood at day 8 of induction chemotherapy is an early index to predict clinical efficacy of induction therapy in AML.
Adolescent ; Adult ; Aged ; Early Diagnosis ; Female ; Flow Cytometry ; Humans ; Leukemia, Myeloid, Acute ; blood ; drug therapy ; mortality ; Male ; Middle Aged ; Neoplasm, Residual ; diagnosis ; drug therapy ; mortality ; Prognosis ; Survival Rate ; Treatment Outcome ; Young Adult

To construct gene vaccine of PPV and to investigate the effects of interleukin 2 (IL-2) as an adjuvant on immune responses in mouse, the recombinant expression plasmid of pCIneo-IL2-VP2 was constructed and transfected into PK-15 cells by lipofectamine, the expressed product was detected by immunofluore assay. To study the immune effects of DNA vaccine in vitro and in vivo, mice were used as the animal model. The recombinant plasmid pCIneo-IL2-VP2, the control plasmid pCI-neo and the PPV live vaccine were immunized by intramuscular injection. Anti-PPV antibodies were measured by ELISA, lymphocyte proliferation activity was detected using MTT method, and the specific killing activities of CTL were assayed too. The results show that the immunized mice produced PPV antibody after one week, and reached to highest after four weeks. Compared with the control group, the pCIneo-IL2-VP2 immunized group produced significant differences in the antibody titers, the lymphocyte proliferation activity and the specific killing activities of CTL. The pCIneo-IL2-VP2 induced humoral and cellular immunity responses similarly to that the live vaccine induced. These results manifested that the PPV DNA vaccine successfully induced humoral and cellular immunity response in mice with the IL-2 gene as an adjuvant.
Adjuvants, Immunologic ; genetics ; Animals ; Antibodies, Viral ; blood ; Antigens, Viral ; genetics ; immunology ; Capsid Proteins ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Immunization ; Interleukin-2 ; genetics ; immunology ; Mice ; Parvovirus, Porcine ; genetics ; immunology ; Random Allocation ; Recombinant Fusion Proteins ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Vaccines, DNA ; immunology ; Viral Vaccines ; immunology