1.Advances in transcription activator-like effectors--a review.
Chinese Journal of Biotechnology 2015;31(7):1024-1038
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
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Biotechnology
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DNA
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chemistry
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Endonucleases
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chemistry
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Tandem Repeat Sequences
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Trans-Activators
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chemistry
2.Role of perforin in severe preeclampsia
Jun WEI ; Xueling LI ; Yu SUN ; Lin LIN ; Caixia LIU
Chinese Journal of Obstetrics and Gynecology 2011;46(8):587-590
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.
3.Effect of natural killer cells treated by serum of severe preeclampsia patient on apoptosis and endothelin-1 secretion of endothelial cells
Jun WEI ; Lin LIN ; Yu SUN ; Zhiying YANG ; Tao SHANG
Chinese Journal of Perinatal Medicine 2011;14(5):289-293
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
4.The Reproduction of Nuclear Polyhedrosis Virus in the Midgut Epithelial Cell and the Other Sensitive Cell of Helicoverpa armigera
Yan-jun, ZHANG ; Jun, TAN ; Yu-qin, LIN
Virologica Sinica 2001;16(2):151-154
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.
6.Recent advances in growth factor and hepatocarcinoma
Jun LI ; Xiaofang YU ; Yifei WANG ; Jian LIN
Chinese Journal of Pathophysiology 2001;17(6):589-592
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.
7.Clinical studies of surviving sepsis bundles according to PiCCO on septic shock patients
Nianfang LU ; Ruiqiang ZHENG ; Hua LIN ; Jun SHAO ; Jiangquan YU
Chinese Critical Care Medicine 2014;26(1):23-27
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.
8.The clinical effects of pulse indicator continuous cardiac output in goal-optimized fluid management on patients with acute exacerbation of chronic obstructive pulmonary disease with respiratory failure
Jiangquan YU ; Ruiqiang ZHENG ; Hua LIN ; Jun SHAO ; Daxing WANG
Chinese Journal of Geriatrics 2015;34(9):976-978
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.
10.Significance of MSCT in detecting mediastinal lymph node metastasis in T1 and T2 non-small cell lung cancer
Yinghui ZHANG ; Yu ZHAO ; Lin ZHANG ; Jianjun HAN ; Jun CHEN
Chinese Journal of Clinical Oncology 2014;(15):961-963
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.