As a protein originally found in plant pathogenic bacteria, transcription activator-like effectors (TALEs) can be fused with the cleaving domain of restriction endonuclease (For example Fok I) to form artificial nucleases named TALENs. These proteins are dependent on variable numbers of tandem Repeats of TALEs to recognize and bind DNA sequences. Each of these repeats consists of a set of approximately 34 amino acids, composed of about 32 conserved amino acids and 2 highly variable amino acids called repeat variant di-residues (RVDs). RVDs distinguish one TALE from another and can make TALEs have a simple cipher for the one-to-one recognition for proteins and DNA bases. Based on this, in theory, artificially constructed TALENs could recognize and break DNA sites specifically and arbitrarily to perform gene knockout, insertion or modification. We reviewed the development of this technology in multi-level and multi species, and its advantages and disadvantages compared with ZFNs and CRISPR/Cas technology. We also address its special advantages in industrial microbe breeding, vector construction, targeting precision, high efficiency of editing and biological safety.
Amino Acid Motifs ; Biotechnology ; DNA ; chemistry ; Endonucleases ; chemistry ; Tandem Repeat Sequences ; Trans-Activators ; chemistry

Objectives To investigate the possible role of perforin (PFN) in the pathogenesis of severe preeclampsia.Methods Thirty-two cases of severe preeclampsia were included in the study.Thirtytwo cases of normal pregnancy were selected as control group in random.The expression of PFN mRNA in the peripheral blood mononuclear cells (PBMC) was detected by reverse transcription (RT)-PCR, and its correlation with mean arterial pressure was analyzed in severe preeclamptic patients.The expression of PFN protein in the decidua was detected by immunohistochemistry.Results ( 1 ) The expression of PFN mRNA in PBMC:the PFN mRNA level in severe preeclamptic group was 1.19 ± 0.31, and that in normal pregnancy group is 0.82 ± 0.28.The PFN mRNA level in severe preeclamptic group was significantly higher than that of control group (P ＜ 0.0l ).(2)Correlation analysis:the mean blood pressure in severe preeclampsia group was (133 ±5) mm Hg( 1 mm Hg =0.133 kPa).There was significant positive correlation between level of PFN mRNA in PBMC and mean blood pressure in severe preeclamptic patients ( r = 0.701, P = 0.000).(3)Decidual PFN protein expression:PFN protein was mainly expressed in lymphocytes and the cytoplasm of decidual stromal cells.The positive ratio of PFN in the decidua of severe preeclamptic patients was 84% ( 27/32), significantly higher than that of control group (53%, 17/32, P ＜ 0.01 ).Conclusions Expression of PFN was significantly increased in severe preeclampsia, and it was of significant positive correlation with mean blood pressure.PFN may participate in the pathogenesis of severe preeclampsia.

Objective To investigate the effect of natural killer (NK) cells treated by serum of severe preeclampsia patient on the function of endothelial cells.Methods Fifteen patients with severe preeclampsia and 15 normal pregnant women from the Obstetrics department,Shengjing Hospital,China Medical University were admitted into this case-control study from January 1,2006 to December 31,2008.NK cells from healthy non-pregnant woman were incubated with 20% serum from severe preeclampsia patients or normal pregnant women for 20 hours.Then,human umbilical vein endothelial cells ( HUVEC) and serum-treated NK cells were co-incubated for 24 hours.Apoptosis of HUVEC was checked by flow cytometry and electronic microscope.Endothelin-1 (ET-1) levels in the supernatants of HUVEC and NK cells were examined by radioimmunologic method.Results In severe preeclampsia group,the percentage of early apoptosis cell (Annexin V-FITC+ /PI+ ) was (23.81±4.79)%,that of late apoptosis cell (AnnexinV-FITC+/PI+ ) was (3.29±1.04) %,while those were (16.59±5.13)% and (2.24±0.72)% respectively in normal pregnant group (P＜0.01).There was no significant difference in dead cells (Annexin V-FITC- /PI+ ).Under electronic microscope,typical morphologic changes of apoptosis were shown in severe preeclampsia group.Level of ET-1 in

The reproduction of Helicoverpa armigera nucleopolyhedrovirus in the midgut epithelia cells and the other sensitive tissues was observed by electron microscopy. The reproducing viruses in the midgut epithelia cells were mostly without envelopes, and thte polyhedrons were seldom formed. The reproduciing viruses in the other sensitive cells were with envelopes, and packed in polyhedrons.

Angiogenesis Inhibitors ; therapeutic use ; Endostatins ; therapeutic use ; Histiocytic Sarcoma ; drug therapy ; Humans ; Male ; Middle Aged ; Recombinant Proteins ; therapeutic use

To evaluate multi-slice computer tomography (MSCT) in mediastinal lymph node metastasis of T1 and T2 non-small cell lung cancer (NSCLC). Methods:A total of 32 patients with T1 and T2 NSCLC from February 2004 to October 2012 were selected. Preoperative MSCT assessment of mediastinal lymph nodes was performed on basis of the pathological results. Results:Lymph nodes with diameters of≥10 mm were evaluated, and the sensitivity and specificity of the MSCT mediastinal lymph node me-tastases were 82.4%and 92.4%, respectively. Lymph node size, primary tumor location, and visceral pleural invasion showed statistical significance in forecasting mediastinal lymph node metastases (P<0.05). Conclusion:MSCT can be used for the effective evaluation of mediastinal lymph node metastasis, lymph node size, and position of primary tumor. and visceral pleural invasion of the tumor had a higher risk of mediastinal lymph node metastasis.

As one of the risk factors for atherosclerosis,human cytomegalovirus infection has received increasing attention,while cytomegalovirus infection of vascular endothelial cells is considered as the initial stage of causing atherosclerosis.Therefore,elucidating the mechanism of human cytomegalovirus infection of vascular endothelial cells will help further confirm the viral and etiological theories of atherosclerosis,and may block its occurrence and development from the initial stage of infection.

Objective To explore the risk factors for asthma in children.Methods A 1:1 matched and hospital-based case-control study was conducted to analyses risk factors for asthma in 300 pairs of children by logistic regression analysis. Results The result of univariate Logistic regression analysis showed that there were 17 related factors for children asthma, including disease history of parents in respiratory system,family income,atopie character,history of acute respiratory infections, eating habit,the amount of sea foods intakes,foam plastics,family decoration,the way of exhaust fume in kitchen,the exhaust effectiveness,raising pet in house,family history of asthma,family history of allergic rhinitis,family history of food allergy,dust allergy of parents,systemic therapy after the first attack.With multivariate Logistic regression analysis,7 factors were entered the model,6 risk factors including father's history of respiratory diseases(OR 3.771,95%CI 1.533～9.278),low family income(OR 1.503, 95%CI 1.258～1.795),atopy(OR 3.788,95%CI 2.368～6.058),meat-eating habit(OR 2.042,95%CI 1.481～2.815),asthma history of family members(OR 1.710,95%CI 0.988～2.958),the family history of allergic rhinitis(OR 1.991,95%CI 1.234～3.211), and 1 protective factor of raising pet in house(OR 0.443,95%CI 0.265～0.739).The coefficients of these factors in multivariate logistic regression model were 1.327、0.407、1.322、0.714、0.536、0.689、and-0.814 respectively.Conclusion Children asthma was a multi-factorial complex disease,and the interaction of environmental and genetic risk factors played an important role in the onset of this disease.

A solid-phase cell enzyme-linked immunoassay is described for screeningand analyzing monoclonal antibodies against the cell surface antigensof human colorectal adenocarcinoma.Horse-radish peroxidase conjugatedProtein A was used to detect the binding of mouse monoclonal antibodiesto human colorectal adenocarcinoma.HR_(8348) cells which had been cultured inand then fixed to the wells of microtiter plates by using glutaraldehyde.Thismethod was found to be specific,reproducible and practical,and especiallyto be advantageous for the large scale screening and analyzing of monoclo-nal antibodies with a panel of cell types.

AIM To testify the special cytotoxicity of TGF? SAP on proliferating vascular smooth muscle cells and endothelial cells. METHODS Conjugation of saporin to TGF? was accomplished after derivatization of saporin and TGF? with N succinimidyl 3 (2 pyridyldithio) proprionate. Cytotoxicity assays were measured by cell count. The studies of influence of TGF? SAP on values of thymidine and leucine incorporation into SMCs and ECs were measured by 3H thymidine uptake and 3H leucine uptake, respectively. RESULTS Cytotoxicity assays testified TGF? SAP conjugate could inhibit remarkably proliferation of SMCs in culture. The values of thymidine of TGF? SAP group (1?10 -9 mol?L -1 and 1?10 -7 mol?L -1 ) in comparison significantly decreased to 60 9% and 56 0% of the control group respectively, suggesting that cellular DNA synthesis obviously decreased as TGF? SAP was added. But saporin did not affect cellular DNA synthesis at higher level. The rate of 3H leucine incorporation of TGF? SAP group significantly decreased to 47 3% of the control group, suggesting that SMCs protein synthesis obviously decreased as TGF? SAP was added. But TGF? SAP at the same level did not affect DNA synthesis and protein synthesis of ECs compared with the control group. CONCLUSION TGF? SAP possesses the more effective cytotoxicity than saporin and the more specific citotoxicity on proliferating vascular smooth muscle cells than on proliferating endothelial cells.