Objective To determine the optimum water-extracting technology of Mailuoning injection. Methods The orthogonal test was used to select the water extract technology of Mailuoning injection. Results The best technology was that crude drug was added 12 and 8 times water, then soaked 0.5 h, and extracted for l.5 and 1.0 h. Conclusion The study of technology provides the basis for further purification.

Humans ; Medical Staff ; Occupational Exposure ; prevention & control ; Occupational Health

Adult ; Benzene ; toxicity ; Bone Marrow ; drug effects ; pathology ; Bone Marrow Cells ; Humans ; Leukemia, Monocytic, Acute ; chemically induced ; pathology ; Male

Adolescent ; Adult ; Endoscopy ; Female ; Humans ; Male ; Middle Aged ; Orbit ; surgery ; Orbital Fractures ; surgery ; Otorhinolaryngologic Surgical Procedures ; Treatment Outcome ; Young Adult

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

The nonstructural protein 5A (NS5A) of the hepatitis C virus (HCV) has been shown to interact with a variety of cellular proteins and implicated in the regulation of cell growth, interferon resistance, and other cellular signaling pathways. Using the yeast-two hybrid method, we have isolated a clone that encodes a novel NS5A--associated binding protein: NS5ABP37. Reverse transcription polymerase chain reaction (RT-PCR) method was employed to amplify the full fragment,and the plasmid pGADT7-NS5ABP37 with the Saccharomyces cerevisiae vector pGADT7 was constructed. To prove the interaction, yeast cell Y187 transformed with pGADT7-NS5ABP37 was mated with yeast cell AH109 containing pGBKT7-NS5A to verify the interaction between the novel protein coded by the new gene NS5ABP37 and NS5A.

Objective HCV NS3 protein plays an important role in disease caused by HCV. We investigate the gene expression of HCV NS3 in yeast for future study of the function of the protein. Methods PCR was performed to amplify the gene of HCV NS3 from the plasmid pBRTM/HCV containing the whole fragment of HCV and the gene was cloned into pGEM T vector. Thereafter, HCV NS3 gene was cut from pGEM T vector and cloned into yeast expression plasmid pGBKT7, and recombinant pGBKT7∶NS3 was transformed into yeast AH109. The yeast protein was isolated and analyzed with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting. Results HCV NS3 gene was successfully cloned into pGBKT7. The results of SDS PAGE and Western blotting assay showed that the molecular weight of the expressed product was about 22000 Da and HCV NS3 protein was existed within yeast cells.Conclusions HCV NS3 was successfully expressed in yeast expression system.

Objective To evaluate retroperitoneal laparoscopic adrenalectomy. Methods From June of 1999 to January of 2001, retroperitoneal laparoscopic adrenalectomy was performed on 52 patients with adrenal diseases, including 34 cases of aldosterone-preducing adenoma,3 nodular hyperplasia bilateral in 1, 5 Cushing's syndrom,1 Cushing's disease, 3 adrenal pheochromacytoma (bila- teral in 1), 4 nonfunctional adrenal adenoma, 1 myelolipoma and 1 metastasis carcinoma. Results 54 sessions of retroperitoneal adrenalectomy have been carried out for 52 patients (2 on both sides) with success in 51 procedures. The procedure was converted to open operation in 3 occasions because of bleeding or adhesion. The mean operation time was 135 minutes (40～270 min), and the estimated blood loss 45 ml (5～150 ml) with no need of transfusion. The postoperative hospital stay was 6 d (3～14 d) and the mean analgesia consumed 8.5 mg (0～50 mg) of morphine equivalents with no need of any analgesic at all in 17 patients. Conclusions Retroperitoneal laparoscopic adrenalectomy was less traumatic to the patients, with less postoperative discomfort and quicker recovery. The procedure should be considered as the first choice of therapy for benign adrenal diseases.

Objective To evaluate retroperitoneal laparoscopic surgery in the treatment of primary aldosteronism. Methods Retroperitoneal laparoscopic surgery was performed in 90 patients with primary aldosteronism (92 sites on either or both sides).Their clinical indicators were compared with those of 30 cases undergoing open surgery. Results Retroperitoneal laparoscopic procedures were successfully performed in 91 of the 92 sides.In these successful cases,mean operative time was 93 minutes (compared with 114 minutes in open surgery cases),mean volume of blood loss was 36 ml (compare with 87 ml),mean postoperative hospital stay was 6.8 d(compared with 11.0 d),mean time to flatus was 1.6 d (compared with 3.1 d),and mean time to walk around was 2.1 d(compared with 3.3 d), P

0.05).In the 16 cases,17 retroperitoneal laparoscopic excisions of pheochromocytoma were successful,only 1 was converted to open surgery due to intraoperative bleeding. The mean operative time was 85?31 (range,45 to 150) min in laparoscopic surgery group vs 155?39 (90 to 240) in open surgery group (P