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

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.

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

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.

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.