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

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

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.

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.

There are multimechanism and multipathogens in the course of the progress and the metastasis of hepatocarcinoma. Growth factors play an important role in the process. In this review, the relationship between growth factors and hepatocarcinoma are summarized.

Objective To explore the effect of early goal-directed therapy (EGDT) according to pulse indicated continuous cardiac output (PiCCO) on septic shock patients.Methods Eighty-two septic shock patients in Subei People's Hospital of Jiangsu Province from January 2009 to December 2012 were enrolled and randomly divided into two groups using a random number table,standard surviving sepsis bundle group (n=40) and modified surviving sepsis bundles group (n =42).The patients received the standard EGDT bundles in standard surviving sepsis bundle group.PiCCO catheter was placed in modified surviving sepsis bundles group.Fluid resuscitation was guided by intrathoracic blood volume index (ITBVI) with the aim of 850-1 000 mL/m2.Dobutamine was used to improve the heart function according to left ventricular contractile index (dPmax) and stroke volume index (SVI).The mean arterial blood pressure (MAP) was maintained 65 mmHg (1 mmHg=0.133 kPa) or above with norepinephrine.Extra-vascular lung water was monitored for the titration of liquid and diuretics.The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score,sequential organ failure assessment (SOFA) score,the number of patients needed vasopressor,serum procalcitonin (PCT),lactic acid and lactate extraction ratio,the amount of fluid resuscitation,duration of mechanical ventilation,duration of intensive care unit (ICU) stay,hospital mortality were recorded in both groups.Results After treatment,the APACHE Ⅱ score,SOFA score and the number of patients needed vasopressor were gradually reduced in both groups,and those in modified surviving sepsis bundle group were significantly lower than those of standard sepsis bundle group at 72 hours (APACHE Ⅱ score:13.1 ± 6.5 vs.20.9 ± 7.5,SOFA score:8.8 ± 4.3 vs.14.6 ± 4.9,the number of patients needed vasopressor:8 vs.17,all P＜0.05).Arterial blood lactate clearance rate was gradually increased after treatment in both groups.Lactate clearance rate in modified surviving sepsis bundle group was significantly higher than that of standard surviving sepsis bundle group [6 hours:(18.2 ± 8.3)％ vs.(10.8 ± 7.5)％,t=-6.036,P=0.001 ; 12 hours:(22.6 ± 7.3)％ vs.(12.4 ± 8.1)％,t=-4.536,P=0.001 ; 24 hours:(27.8 ± 5.6)％ vs.(16.4 ± 9.5)％,t=-5.882,P=0.000].The amount of fluid resuscitation within 6 hours in modified surviving sepsis bundle group increased significantly compared with standard surviving sepsis bundle group (mL:3 608 ± 715 vs.2 809 ± 795,t=-3.865,P=0.033).The amount of fluid resuscitation within 24,48 and 72 hours in modified surviving sepsis bundle group was significantly less than that of standard modified surviving sepsis bundle group with the nadir at 72 hours (mL:918 ± 351 vs.1 805 ± 420,t=5.907,P=0.037).Duration of mechanical ventilation (hours:98.4 ± 20.3 vs.143.3 ± 29.6,t=9.766,P=0.001) and ICU stay (days:7.1 ± 3.1 vs.9.5 ± 2.5,t=2.993,P=0.004) were significantly reduced in modified surviving sepsis bundle group compared with standard surviving sepsis bundle group.The hospital mortality in modified surviving sepsis bundle group was slightly lower than that in standard surviving sepsis bundle group [16.7％(7/42)比 17.5％(7/40),x2=0.010,P=0.920].Conclusions Modified surviving sepsis bundle treatment according PiCCO can reduce the severity of disease in patients with septic shock,can make more accurately guide fluid resuscitation,and can reduce lung water and duration of mechanical ventilation and ICU stay.It has great clinical significance.

There are multimechanism and multipathogens in the course of the progress and the metastasis of hepatocarcinoma Growth factors play an important role in the process. In this review, the relationship between growth factors and hepatocarcinoma are summarized.[

Objective To investigate the effects of ketamine on anoxia-reoxygenation(A/R)induced glutamate release from cerebral cortex neurons.Methods Primary cultured neurons obtained from cerebral cortex of fetal Wistar rats(16-18 d)were randomly divided into 3 groups:Ⅰcontrol group;ⅡA/R group andⅢketamine pretreatment+I/R group.The control group was not subjected to A/R while A/R group was exposed to anoxic air(95% N_2+5% CO_2)for 5 h followed by 24 h reoxygenation.In groupⅢdifferent doses of ketamine were added to the culture media before anoxia and the final ketamine concentrations were 1,20 and 100?mol?L~(-1) respectively.The extracellular glutamate concentration was detected at the end of 24 h reoxygenation.Results The extracellular glutamate concentration was significantly higher after 24 h reoxygenation in A/R group than in control group.Ketamine 20 and 100?mol?L~(-1) significantly inhibited glutamate release from the neurons induced by A/R in a dose-dependent manner.Conclusion Ketamine can inhibit glutamate release from neurons induced by A/R in a dose-dependent manner.

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.