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.

BACKGROUND: Compound antitumor coral hydroxyapatite (CCHA) has a good delayed-release and anti-tumor effect. However, whether the high-dose drug contained in the CCHA influences normal induction, conduction and growth of bone tissues at the implant site is unclear.OBJECTIVE: To establish an osteogenesis model of CCHA and to investigate the osteogensis effect and rule of self-made CCHA in vivo. METHODS: Implants of CCHA (20%CDDP-CHA w/w) and CHA(control, 0% CDDP w/w) were implanted into the metaphyseal holes of rabbit femur. X-rays and decalcified histological section of rabbit femoral bone with hematoxylin and eosin staining were used regularly to investigate the degradation of CCHA and CHA, and how bone tissues grow at the implant site. RESULTS AND CONCLUSION: After implantation, CHA crystals were faster than CCHA in connecting with surrounding bone tissues and forming bone bridges. The borderlines of implanted CHA became obscure in 4 weeks. Loose connective tissues were found in pores of the CHA and osteoblasts were growing on the surface. Bone tissues of the surrounding gradually grew into the CHA, finally repaired the bone defects. At the beginning of implantation, CCHA mainly inhibited the growth of surrounding tissues until 6-12 weeks later, normal bone tissues gradually grew into pores of CCHAs, and healed bone defects at 26 weeks. CCHA can inhibit the osteogenesis effects at early stage; however, it can achieve bone healing with surrounding bone defect ultimately.

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 clinical effects of the pulse indicator continuous cardiac output (PiCCO) in goal-optimized fluid management on patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) with respiratory failure.Methods A total of 71 AECOPD patients with respiratory failure who were hospitalized in department of Critical Care Medicine in our hospital were selected from October 2010 to September 2013.The PiCCOtechnology was applied to monitor intrathoracic blood volume index (ITBI) and extravascular lung water index (ELWI).We compared the relationship of ELWI with ITBI and oxygenation index,and a simple correlation analysis was used for statistical analysis.Results A significantly negative correlation was found between ELWl and oxygenation index (r=-0.743,P＜0.01).ELWI =14 ml/kg was defined as the cutoff value for the subgroup analysis.A negative correlation still was found between ELWI and oxygenation index in the subgroup with ELWI ＜ 14 ml/kg,but it had no significant difference (r=-0.533,P=0.080),while a significantly negative correlation was found in the subgroup with ELWI≥14 ml/kg (r=-0.961,P＜0.01).There was no significant correlation between ELWI and ITBI (r=0.477,P=0.072).ITBI=1 000 ml/m2 was defined as the cutoff value for the subgroup analysis.There is no significant correlation between ELWI and ITBI in the subgroup with ITBI＜1 000 ml/m2(r=0.338,P=0.116),but in the subgroup with ITBI≥1 000 ml/m2,a significantly positive correlation between ELWI and ITBI was found (r=0.677,P＜ 0.01).Conclusions The results suggest that when ELWI is higher than 14 ml/kg,ELWI becomes an important influencing factor for oxygenation,which can be decreased to improve oxygenation in patients with AECOPD,and when ITBI≥ 1 000 ml/m2,ITBI can be reduced by decreasing ELWI.PiCCO has an important clinical significance on a goal-optimized fluid management in AECOPD patients with respiratory failure.

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.

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

Chronic Disease ; Graft Rejection ; therapy ; Humans ; Integrative Medicine ; Kidney Diseases ; etiology ; therapy ; Kidney Transplantation ; adverse effects