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

BACKGROUND: The upward attack of drug-heat is a special type of syndromes of coexistence of cold and heat, it influence the treatment and conversion of the primary disease.OBJECTIVE: To investigate the pathophysiological basis of upward attack of drug-heat.DESIGN: A randomized and controlled observation. SETTING: Department of Traditional Chinese Medicine (TCM) of the People' s Central Hospital of Huizhou City.PARTICIPANTS: Totally 427 patients with cold syndrome, visiting on Department of TCM of the People's Central Hospital of Huizhou City from January 1996 to December 2001, were given warm-heat remedy. Among them 66 cases, 30 males and 36 females, got upward attack of drug-heat during the process of treatment were enrolled. All these subjects, according to their visiting order, at a ratio of 2:1, were chosen as treatment group (44 cases) and control group (22 cases).METHODS: For the patients in treatment group: Rhubarb 6 g was added boiling water 100 mL and soaked for 8 minutes, then the soaking solution was given to the patients, once a day, for 3 days successively. For patients in control group: Table salt 6 g was added boiling water 100 mL for soaking for 8 minutes, after the salt was completely dissolved, the solution was given to the patients, once a day for 3 days successively. The patients of the two groups were at the same time given remedy to treat cold syndrome. The changes of free triiodothyronine (FT3), free thyronine (FT4), high sensitizing thyroid-stimulating hormone (HS-TSH) and cortisol of the patients were detected before and after administration of warm-heat remedy, before and after medication with rhubarb. MAIN OUTCOME MEASURES: ① The changes of the relevant hormones of the patients with upward attack of drug-heat before and after ad ministration of warm-heat remedy. ② The changes of the relevant hormones of the patients with upward attack of drug-heat before and after ad ministration of rhubarb. ③ Comparison of the therapeutic effects in the two groups.RESULTS: All 66 patients involved entered the final result analysis. ①The changes of the relevant hormones of the patients with upward attack of drug-heat before and after administration of warm-heat remedy: FT3 and FT4 were decreased as compared with those before treatment [(2.51±1.20),(6.50±2.30); (10.01±3.21), (15.50±6.31) pmol/L], but HS-TSH and cortisol were increased as compared with those before treatment [(8.25 ±3.75),(4.11±1.75) mU/L; (0.56±0.17), (0.43±0.10) μmol/L]. ② The changes of the relevant hormones of the patients with upward attack of drug-heat before and after administration of rhubarb: FT3 and FT4 ywere increased as compared with those before treatment[(4.71 ±1.56), (2.45±1.25); (14.21 ±4.61),(10.21±3.52) pmol/L], but HS-TSH and cortisol were decreased as compared with those before treatment [(6.24±2.25), (8.35±3.51) mU/L; (0.48±0.10),(0.60±0.17) μmol/L]. ③ Comparison of the therapeutic effects in the two groups: The rate of excellenly effect in treatment group was obviously higher than that in control group (33%, 5%).CONCLUSION: The pathophysiological basis of upward attack of drug heat is stress state of the body, the secretions of the pituitary-thyroid axis play a role of physiological protection, the adrenocortical function is hyperactive, and rhubarb can control this stress state through inhibiting the metabolism of the body.

OBJECTIVE: To observe the effects of ginkgolide (GL) on the cerebral blood flow in dogs. METHODS: Dogs anesthetized with sodium pentobarbital were randomly divided into 5 groups, with 4 dogs in each group. Ginkgolide of 4.86, 14.6 and 43.7 mg/kg and Xingling Granule of 0.22 g/kg were administered by gavage to the dogs in each of 4 groups. The dogs in the other group were administered with edible oil (1 ml/kg) as control group. The cerebral blood flow, systolic blood pressure, diastolic blood pressure, mean arterial blood pressure and electro-cardiogram of the dogs were measured at different times after the administration. RESULTS: Ginkgolide of 4.86, 14.6 and 43.7 mg/kg had no obvious effects on the blood pressure and the heart rate. Ginkgolide of 14.6 and 43.7 mg/kg increased the cerebral blood flow 90 minutes after administration, and ginkgolide of 43.7 mg/kg obviously decreased the cerebral vascular resistance 150 minutes after administration. CONCLUSION: Ginkgolide can increase the cerebral blood flow and decrease the cerebral vascular resistance, and it has no obvious effects on blood pressure and heart rate in dogs.

AIM: To study the therapeutical effects of salvianolic acid B (Sal B) on myocardial infarction(MI) in dogs. METHODS : The models of myocardial infarction were produced by ligation of the left anterior descending (LAD) coronary artery in the anaesthetized dogs. The extent of myocardial ischemia was detected by epicardial-electrocardiogram and the extent of infarction was determhled by N-BT staining method. The parameter of lactate dehydrogenase was determined in the serums of dogs. RESULTS : It was indicated that the adminstration of Sal B(1、3 and 10mg/kg) by the way of intravenous injection reduced the extent of myocardial infarction (N-ST) ( P

Objective To investigate the effect of exercise therapy on I° adolescent idiopathic scoliosis (AIS). Methods 70 cases with I°AIS were included. 32 patients correcting posture themselves for 0.5 year or above were as control group, and the other 38 cases who accepted exercise therapy for 0.5 year were as treatment group. Their X-ray of spine before and after treatment were compared. Results The Cobb's angle, rotation of the spine, and compensatory improved after treatment in the treatment group (P<0.01), but no significant improvement was obsereved in the control group after 0.5 year. Conclusion Exercise therapy is safe and effective on I° AIS.

Objective To explore the percentage changes of peripheral T , B cell subsets and NK cells in chronic HBV infectors under different immune states and hepatitis B cirrhosis . Methods Seventy-five chronic HBV infectors, including 20 cases with immune clearance, 20 cases with immunodeficiency (inactive) and 35 cases with cirrhosis, and 20 healthy control were enrolled. The percentages of peripheral T and B lymphocyte subsets and NK cells were detected by Flow Cytometry. The differences of the groups were analyzed. Results Comparing with the control group, CD4+T cells were decreased in the other four groups (P<0.05). The sequence of CD4+T cells, from high to low, was the control group, the immunodeficiency group, the immune clearance group, the compensated cirrhosis group and the de-compensated cirrhosis group. CD4+/CD8+T cell and NK cell were lower , but CD8+T cell and B cell were higher in immune clearance group than that of the control group (P < 0.05). Patients in immunodeficiency group had lower ratio of CD4+/CD8+T cell and higher CD8+T cell than those in the control group (P < 0.05). In all the groups, patients with de-compensated cirrhosis showed highest ratio of CD4+ to CD8+ T cells and B cells, but lowest CD3+T, CD8+ T and NK cells (P < 0.05). Conclusions Results suggests immune dysfunction exists in patients with chronic HBV infection. It has potential clinical value in understanding patients′ immune states and progression of disease by detecting peripheral blood lymphocyte subsets and NK cells.

Aim To obtain two-dimensional gel electrophoresis maps of the prefrontal cortex(PFC)proteins of normal and morphine-addicted rats for identifying the diferentially expressed proteins in the addicted rats.Methods Rat models of morphine addiction were established.The proteins in the PFC underwent two-dimensiona1 gel electrophoresis with immobiline pH gradient isoelectric focusing as the first and vertica1 SDS-PAGE as the second dimension.Analysis of 2-DE maps was used to determine differential expressions of proteins between the two groups by ImageMaster 2D Platinum v5.0,and four protein spots expressed differently were picked up for further identification by MALDI-TOF-MS.Results Matched and compared with those of control group,87 protein spots had been determined with differently expressive levels in morphine addiction group.By MALDI-TOF-MS,two protein spots of them had been identified as Snap25 Isoform ?-Snap 25 of Synaptosomal-associated protein 25 and Actb Actin,cytoplasmic 1.Conclusions There is obvious difference in expressive proteomes in PFC between normal and morphine-addicted rats. The functions of those identified proteins are involved in cell growth,apoptosis,differentiation,signal transduction,axon growth and cellular energy metabolism,so proteomics can serve as a new approach in the study of morphine dependence to discover new therapeutic targets.

Objective To analyze re-treatments of recurrence after the pelvic floor repair surgery.Methods The protocol and the effect of re-treatments were investigated by reviewing and analyzing the clinical data of 81 recurrent patients (grade Ⅱ and above),who had received the pelvic floor repair surgery from January 2011 to January 2016.Pelvic organ prolapse quantitation system (POP-Q) and two questionnaires about quality of life [pelvic floor distress inventory-short form 20 (PFDI-20) and pelvic floor impact questionnaire short form (PFIQ-7)] were used to evaluate objective and subjective efficacy,respectively.Results Among 81 recurrent patients who were followed up for a median of 35 months (10-69 months),78 cases (with prolapse up to grade Ⅲ or Ⅳ) were treated by surgical operation with both objective cure rate and subjective satisfaction being 100％ (78/78);3 cases (with grade Ⅱ prolapse) were treated by pelvic floor electrical stimulation biofeedback,and 1 case among the three cases had the vaginal foreign body sensation,the subjective satisfaction was 2/3.The methods of surgical operation for the 78 recurrent patients included:total pelvic floor reconstructive surgery (55 cases;3 of which involve trachelectomy),anterior pelvic reconstructive surgery (2 cases),posterior pelvic reconstructive surgery (3 cases),Y-mesh sacral colpopexy (2 cases),colpocleisis (11 cases),vaginal hysterectomy combined posterior fornix forming (3 cases),and vaginal hysterectomy combined posterior pelvic reconstructive surgery (2 cases).Conclusion The extent of recurrence,the recurrent site and complications must be carefully considered and evaluated for re-treatments of recurrence after pelvic floor repair surgery,and then an appropriately individualized re-treatment protocol could be designed for each of the patients.

Objective To investigate the different immune status of TCM classification and the levels of T and B cells and the re-lationship between the NK cells .Methods Three different immune state of patients with chronic HBV were divided into asthenia syndrome(AS) group and sthenia syndrome(SS) group ,and then we detected the peripheral blood lymphocyte subsets and NK cells ,and compared the above each index analysis .Results In three different immune status ,all patients with AS and SS groups CD4+ T cell percentages are lower than normal control group (P<0 .05) ,and in a state of immune tolerance ,CD4+ T cells of SS group were lower than the AS group (P<0 .05) ,in a state of immune clearance ,CD8+ T cell percentage were SS group> control group> the AS group (P<0 .05) .Three different immune status ,the ratio of CD4+ /CD8+ SS group were significantly lower than the AS group or control group ,in a state of immune clearance the SS and the AS group B lymphocyte percentage were higher than the control group ,the percentage of NK cells was lower than the control group (P<0 .05) .Conclusion Chronic HBV infection have immune dysfunction ;HBV infected different immune status of TCM classification have relationship with T and B cell subsets and NK cells ,and the immune function test has certain clinical application value for the judgment of TCM syndrome type .

The purpose of this study was to investigate the natural infection of Angiostrongylus cantonensis (A. cantonensis) in rodents in Fujian Province and provide more scientific evidences for control and prevention.?Rats captured with cages were numbered, and the species, gender and maturity were identified in each village every day. Rats' hearts and lungs were taken out and soaked into physiological saline solution to tear to pieces. The detected A. cantonensis were put into physiological saline solution in other glassware then the number and gender were identified. This investigation was conducted in 67 villages in 35 towns, covering 23 counties in 9 cities and regions. Except Shanghang County in Longyan city and Pingtan County in Fuzhou city, rodents infected with A. cantonensis was found in 21 counties. Of 4 890 rats were examined and 434 infected rats were identified with an infection rate of 8.90%. Rattus norvegicus had the highest infection rate of 17.75% (315/1 775), following by Rattus flavipectus with an infection rate of 6.21% (49/789). Additionally, the infection rates of the Musk shrew and Mus musculus were 2.81% (29/1 032) and 2.52%(3/119), respectively (χ2＝185.06, P＜0.001). Moreover, the infection rate of adult rats (21.37%) was much higher than that of immature rats (11.11%) (χ2＝14.50, P＜0.05). These results indicated that the A. cantonensis infection of rodents has covered most area in Fujian Province and the geographical distribution of infected rats was similar with that of Ampullaria gigas.