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

Metabolic syndrome (MetS) is a serious threat to public health worldwide with an increased risk of developing type 2 diabetes, cardiovascular diseases and all-cause morbidity and mortality. In this study, a urinary metabolomic approach was performed on high performance liquid chromatography quadrupole time-of-flight mass spectrometry to discriminate 36 male MetS patients and 36 sex and age matched healthy controls. Pattern recognition analyses (principal component analysis and orthogonal projections to latent structures discriminate analysis) commonly demonstrated the difference between MetS patients and no-MetS subjects. This study found 8 metabolites that showed significant changes in patients with MetS, including branch-chain and aromatic amino acids (leucine, tyrosine, phenylalanine and tryptophan), short-chain acylcanitine (tiglylcarnitine), tricarboxylic acid (TCA) cycle intermediate (cis-aconitic acid) and glucuronidated products (cortolone-3-glucuronide and tetrahydroaldosterone-3-glucuronide). The candidate biomarkers revealed in this study could be useful in providing clues for further research focusing on the in-depth investigation of the cause of and cure for MetS.

Excessive and fast rise of medical expenses in China recent years has worsened the financial burden of both the government and individuals alike,making it a hotspot issue countrywide.To alleviate the difficulties of "accessibility to medical services" and "poverty or repoverty resulting from sickness" faced by the Chinese people at large,we built a multi-factor analysis model to identify key factors contributing to such a fast rise of medical expenses based on an analysis of the medical expenses in Shandong Province.In addition,we discussed the causes for GDP and headcount of medical staff in relation to the rise of medical expenses,and the strategies to curb expenses.

Objective To investigate the roles of a variety of cytokines including transforming growth factor beta (TGF-β),interleukin-6 (IL-6) and interleukin-2 (IL-2) in the differentiation of CD+4 Tlymphocyte cells.Methods T lymphocyte cells either in human peripheral blood or routine spleen were cultured in vitro under different stimulation conditions.Flow cytometry was used to analyze the percentages of CD+4IL-17+ T-helper 17(Th17) cells,CD+8 IL-17+ T cells,CD+4 CD+25 FOXP+3 T regulatory (Treg) cells among activated T cells.Results Differentiation of Treg cells,Th17 cells and CD+8 IL-17+ T lymphocyte cells was enhanced when murine splenic T cells were cultured with TGF-β.The levels of expression were (7.8±2.2)%,(12.6±3.1)%,(10.1±2.6)% ,respectively.Experimental control group was severally same type of T cells without cytokine treatment.The levels of expression were (4.8±0.6) %,(1.7±0.5) %,(1.0±0.4) %,respectively.There were statistically significant differences among them (q=4.09,8.80,9.61.P<0.05 or P<0.01).Under combination treatment with IL-6 and TGF-β,(17.8±5.3) % Th17 cells and (15.0±4.2)% CDCD+8 IL-17CD+ T cells were induced,whereas the levels of Treg cells whose differentiation were restrained were (4.1±1.2) %.The differences were statistically significant compared with the level of same type of T cells in TGF-β group (q=5.03,5.17,5.04,P<0.01).Moreover,combination treatment with IL-2 and TGF-β decreased percentages of Th17 and CDCD+8 IL-17CD+ T cells and increased percentages of Treg cells in T cell population.There was an opposite effect when anti-IL-2 was apphed.The percentages of Th17 and CD+8 IL-17+ T cells were increased and the percentages of Treg cells were reduced The regulation trend of T lymphocyte cells in human peripheral blood was similar with those in routine spleen.Conclusion Various cytokines are of great importance in the regulation of the balance between Th17 and Treg cell.

Objective To evaluate anesthesia efficacy of sevoflurane combined midazolam and remifentanil and propofol in patients undergoing short surgery. Methods Fifty ASA Ⅰ or Ⅱ patients who scheduled for elective surgery under general anesthesia were enrolled in study. The anesthesia was induced with intravenous injection of midazolam 0.03 mg/kg and sevoflurane inhalation. Induction of anesthesia was started with 6% sevoflurane and fresh gas flow rate of 6 L/min. The inspired concentration of sevoflurane was reduced to 2% as soon as eyelash reflexes of these patients were abolished. At the same time, remifentanil 2 μg/kg was injected slowly over 1 min,and 30 s later,patients were mechanically ventilated after tracheal intubation. 2% sevoflurane was kept inhaling, and remifentanil and propofol were kept pumping for anesthesia maintenance. The mean arterial pressure (MAP), heart rate, pulse oxygen saturation (SpO_2) and bispectral index (BIS)were recorded before administration,at the time of the disappearance of eyelash reflex, immediately before tracheal intubation,and immediately after tracheal intubation. The time period from sevoflurane inhalation to the disappearance of eyelash reflex was also recorded. And conscious recovery, spontaneous breathing recovery,the endotracheal extubafion time and OAAS score were recorded. Results All patients were successfully intubated at the first attempt. The satisfactory intubation conditions were found in 82% (41/50) of cases. The time period from sevoflurane inhalation to the disappearance of eyelash reflex was (73 ± 12) s. MAP,heart rate and SpO_2 remained stable and BIS was maintained at 45-55 before and after tracheal intubation, and all patients were quick recovery. Conclusion The anesthesia efficacy of sevoflurane combined midazolam and remifentanil and propofol without muscle relaxant in patients undergoing short surgery is smooth, safe and reliable.

Objective To discuss the clinical value of application of three-dimensional reconstruction technique of MSCT in examination of colonic tumours.Methods Thirty patients underwent scanning using helical CT and post-processing on the workstation.All CTC images were reconstructed using shaded surface display (SSD) on workstation.Perspestive images such as the ones from DCBE were generated via raysum.The images could clearly demonstrate the extent and detail of the disorder by adjusting the CT value threshold.These results and that of barium enema and colon endoscopy were compared.Results Thirty cases had completely clinical and pathological data.3D images could correctly reveal the macroscopic morphology of colon tract and location of lesions.The correctly diagnostic rate was 100％ by MSCT,while the diagnostic rate by CTVE and colon endoscopy was basicly identical.Conclusion With the utilization of multiple methods,3D reconstruction can effectively demonstrate colonic tumours,and can be applied routinely in examination of colonic tumours.

SUN Shu-chun,the chief researcher,is one of the famous expert of osteo-traumatology of TCM.He makes systematic sort and study on chirismus on basis of succeeding experience predecessors and his own years of clinic experience.He is good at treating cervical syndrome with TCM chirismus,especially combine it with drugs.In osteo-traumatology academic circles,he is called "the best doctor at home".Professor SUN advocates that the rule of applying various therapies according to different conditions,combination of rigid-flexible,light,skill,gentle,harmony should be obey when we use chirismus.His diagnostic and therapeutic methods and thoughts are original and ingenious.In this article,the author analyzes the academic experience of tendon trauma of professor SUN.

Objective To study the effect of low molecular weight heparin on the growth and metastasis of hepatocarcinoma in vivo Methods Metastatic model simulating human hepatocarcinoma was established by orthotopic implantation of the tumor tissue into nude mice Animals were randomly divided and submitted to different treatment groups: control group treated with nornal saline, chemotherapy group with 5 FU and CDDP i p , Fragmin group (low molecular heparin), combination group After 5 weeks of treatment, tumor volume、 microvessel density (MVD)、 VEGF smoothmuscle actin (SMA) and metastasis in Fragmin group significantly decreased and tumor inhibitive rate increased ( P

AIM To testify the special cytotoxicity of TGF? SAP on proliferating vascular smooth muscle cells and endothelial cells. METHODS Conjugation of saporin to TGF? was accomplished after derivatization of saporin and TGF? with N succinimidyl 3 (2 pyridyldithio) proprionate. Cytotoxicity assays were measured by cell count. The studies of influence of TGF? SAP on values of thymidine and leucine incorporation into SMCs and ECs were measured by 3H thymidine uptake and 3H leucine uptake, respectively. RESULTS Cytotoxicity assays testified TGF? SAP conjugate could inhibit remarkably proliferation of SMCs in culture. The values of thymidine of TGF? SAP group (1?10 -9 mol?L -1 and 1?10 -7 mol?L -1 ) in comparison significantly decreased to 60 9% and 56 0% of the control group respectively, suggesting that cellular DNA synthesis obviously decreased as TGF? SAP was added. But saporin did not affect cellular DNA synthesis at higher level. The rate of 3H leucine incorporation of TGF? SAP group significantly decreased to 47 3% of the control group, suggesting that SMCs protein synthesis obviously decreased as TGF? SAP was added. But TGF? SAP at the same level did not affect DNA synthesis and protein synthesis of ECs compared with the control group. CONCLUSION TGF? SAP possesses the more effective cytotoxicity than saporin and the more specific citotoxicity on proliferating vascular smooth muscle cells than on proliferating endothelial cells.

Objective To obtain highly purified oligodendrocyte precursor lineage cells in vitro and make identification.Methods The oligodendrocyte precursors were separated from astrocyte by orbital shaker and further purified by differential adhesion,and finally cultured in chemically defined serum-free medium,with appended neurotrophin 2(N2),platelet-derived growth factor(PDGF),basic fibroblast growth factor(bFGF).Immunofluorescence assay was applied to identify the separated cells with A2B5,O4,O1 and glial fibrillary acidic protein(GFAP) antibodies.Results Over 95% of cultured oligodendrocyte precursor cells were obtained.The oligodendrocyte progenitors were A2B5 and O4 positive,immature oligodendrocytes were O4 and O1 positive while GFAP were negative.Conclusions Separation and purification by shaking and differential adhesion and chemically defined medium are suitable and effective to obtain highly purified oligodendrocyte precursor cells.Cell output will increase notabily and rest in immature phase by appending both N2,PDGF and bFGF.