Selectable marker genes that usually encode antibiotic or herbicide resistances are widely used for the selection of the transgenic plants, but they become unnecessary and undesirable after transformation selection. An important strategy to improve the transgenic plants' biosafety is to eliminate the marker genes after successful selection. In the FLP/frt site-specific system of the 2 microm plasmid of Saccharomyces cerevisiae, the FLP enzyme efficiently catalyzes recombination between two directly repeated FLP recombination target (frt) sites, eliminating the sequence between them. By controlled expression of the FLP recombinase and specific allocation of the frt sites within transgenic constructs, the system can be applied to eliminate the marker genes after selection. Through a series of procedures, the plant FLP/frt site-specific recombination system was constructed, which included the frt containing vector pCAMBIA1300-betA-frt-als-frt and the FLP expression vector pCAMBIA1300-hsp-FLP-hpt. The FLP recombinase gene was introduced into transgenic (betA-frt-als-frt) tobacco plants by re-transformation. In re-transgenic plants, after heat shock treatment, the marker gene als flanked by two identical orientation frt sites could be excised by the inducible expression of FLP recombinase under the control of hsp promoter. Excision of the als gene was found in 41% re-transgenic tobacco plants, which indicated that this systerm could make a great contribution to obtain the marker free transgenic plants.
Base Sequence ; DNA Nucleotidyltransferases ; metabolism ; Molecular Sequence Data ; Plants, Genetically Modified ; genetics ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; Recombination, Genetic ; Tobacco ; genetics

AIM: To evaluate the safety, efficacy and stability of posterior chamber phakic intraocular lens ( ICL ) implanation and clear lens extraction for the correction of high myopia.
METHODS: The study enrolled 56 cases ( 100 eyes ) of high myopia. Group I comprised 32 cases ( 58 eyes ) receiving ICL implantation and Group II comprised 24 cases (42 eyes) undergoing clear lens extraction. In this study, we evaluated the two groups of subject's the visual and refractive results, intraocular pressure ( IOP ) , endothelial cell density ( ECD ) , anterior chamber depth ( ACD) , lens transparency, the surgical complications as well as visual adverse symptoms before and after surgery.
RESULTS: The postoperative subjects in group I and group II were followed, uncorrected vision acuity ( UCVA)>0. 5 were 69. 0% in group I and 71. 4% in group II after 3mo. UCVA>0. 5 were 72. 4% in group I and 73. 8% in group II after 1a. Predictability of the manifest spherical equivalent refraction within±1. 00D was achieved in 62. 1%of eyes in group I and 57. 1% in group II after 1a. The central vault of the ICL ( distance from posterior surface of ICL to the crystalline lens ) measured with anterior segment optical coherence tomography ( AS-OCT ) was 0. 35-0. 54 (0. 40±0. 16) mm. Twelve point one percent of eyes in group I and 7. 1% of eyes in group II had transient mild increase in IOP. Here were statistically significant differences between preoperative and postoperative ECD (P<0. 001 ). Complications of surgery: 1 eye had ICL spontaneous rotation, 2 eyes had anterior subcapsular cataract, 4 eyes noticed halos around lights at night in group I. Three eyes had posterior capsule mild opacification, 3 eyes noticed halos around lights at night, 12 eyes had difficulty in near vision in group II.
CONCLUSION: ICL implantation and clear lens extraction are effective, safe and predictable surgical option for the management of high myopia. No severe complications occurred, but its long time effect and safety still need more time to prove.

Aged ; Drug-Related Side Effects and Adverse Reactions ; blood ; therapy ; Emergency Treatment ; Female ; Hemoperfusion ; Humans ; Hypnotics and Sedatives ; poisoning ; Male ; Middle Aged ; Serum ; chemistry

Animals ; Apoptosis ; drug effects ; Cell Division ; drug effects ; Cell Separation ; Cells, Cultured ; Interferon-alpha ; pharmacology ; Liver ; cytology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Rats ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics ; bcl-2-Associated X Protein ; fas Receptor ; biosynthesis ; genetics

OBJECTIVETo observe the effect of electroacupuncture at Zusanli (ST 36) on phosphorylated extracellular signal-regulated kinase 1/2 (pERK1/2) in the dorsal horn of spinal cord induced by plantar inflammation in the rat.

METHODSAll the rats were randomly divided into 5 groups: normal control group, simple electroacupuncture group, formalin group, formalin plus ipsilateral electroacupuncture group and formalin plus contralateral electroacupuncture group. The acute inflammation animal model was made by injection of 100 microL of 4% formalin into the right posterior foot pad. Electroacupuncture was given at "Zusanli" (ST 36) for 30 min, with sparse-dense waves, frequency 2-15 Hz, and intensity 2-3 mA. One and a half hours latter, the rats were killed under anesthesia, and pERK1/2 expression in the lumbar dorsal horn were investigated with immunohistochemical method.

RESULTSThe positive cells were rarely seen (6.45 +/- 1.05) in the superficial spinal cord in the control group; a few cells (14.07 +/- 3.19) in ipsilateral superficial spinal cord were found in the electroacupuncture group. The number of pERK1/2-positive neurons (26.57 +/- 4.93) in lamina I - II0 of the ipsilateral dorsal horn in the formalin group increased significantly. After electroacupuncture at ipsilateral Zusanli (ST 36), the number of positive cells (20.79 +/- 5.21) had a tendency to decrease, but with no statistically significant difference. However, after electroacupuncture at contralateral Zusanli (ST 36), the number of positive cells (14.75 +/- 3.03) significantly decreased as compared with the non-acupuncture group (P < 0.05).

CONCLUSIONThe inhibition of ERK1/2 phosphorylation in the spinal cord dorsal horn by electroacupuncture is possibly involved in acupuncture analgesic effect.

Acupuncture Analgesia ; Acupuncture Points ; Animals ; Electroacupuncture ; Male ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Posterior Horn Cells ; enzymology ; Rats ; Rats, Sprague-Dawley

Objective To analyze the causes of severe adverse reaction after painless abortion and to offer some advices for prevention and cure.Methods Clinical files and results of 3 patients with severe adverse re-action after painless abortion were analyze.Results Total of 130-140 mg· kg -1 propofol intravenous injection and oxytocin 10 u intramuscular injection was used in three patients underwent painless abortion.Three patients have the symptoms of chills , fever , nausea , vomiting , low blood pressure , damages of multiple organs and elevation of peripheral white blood cell and neutrophilic granulocyte.The shock occurred within 0.5-4 h after operation.Conclusion The symptoms of severe adverse reac-tion after transfusion are very serious.Early diagnosis and rational man-agement are the key to the successful salvage.

Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Female ; Hepatitis B e Antigens ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Interferon-alpha ; administration & dosage ; therapeutic use ; Male ; Nucleotides ; administration & dosage ; therapeutic use ; Polyethylene Glycols ; administration & dosage ; therapeutic use ; Recombinant Proteins ; administration & dosage ; therapeutic use ; Treatment Outcome ; Young Adult

OBJECTIVETo summarize methods for repair of claw hand deformity after burn.

METHODSNinety-seven patients with 136 claw hands after burn hospitalized from May 1992 to May 2007 were repaired with skin grafting (104 hands) and transposition of skin flap (32 hands), among which 21 hands were minor-grade, 92 hands moderate, 23 hands severe. The metacarpophalangeal joint was repaired after scar release in dorsum of hand with manual extraction reduction, release of collateral ligament and joint capsula, separation of adhesion in joint, tendon lengthening for obvious contracture. Restitution of finger flexion deformity, lysis of adhesion and grafting among first web and finger webs, repair of central slip extensor tendon or phalangeal arthrodesis were performed according to the abnormal condition after lysis of dorsal scar of hand. The metacarpophalangeal joint from 31 patients were not repaired with above methods for severe finger flexion deformity, their palmar scar were loosened and transplanted firstly, then scar in dorsum of hand were loosened, metacarpophalangeal joint were repaired, flap or skin were transferred or transplanted. General rehabilitation were performed routinely after operation.

RESULTSThe ending of flaps (4 hands) due to the scar were necrosis after transposition and healed through dressing change, other skins or flaps all survived. Most articular deformities were corrected completely or basically. Functions including palmar opposition, grasp were also recovered with satisfactory results.

CONCLUSIONSkin transplantation and transferring of skin flap with overall planning and individual isatin are the key points for repair of claw hand after burn.

Adolescent ; Adult ; Burns ; complications ; Child ; Child, Preschool ; Cicatrix ; etiology ; surgery ; Female ; Hand Deformities, Acquired ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; Surgical Flaps ; Young Adult

OBJECTIVETo study transcription, expression, and bioactivity of expressive protein of vascular endothelial growth factor 165 (VEGF(165)) gene, after bone marrow stromal cells (BMSCs) were transferred by VEGF(165) gene mediated by adenovirus vector ex vivo.

METHODSBMSCs were Isolated and cultured by gradient centrifugation method. The cells cultured are transferred by recombinant adenovirus vector that carry VEGF(165) gene. Transcription and expression of VEGF(165) gene in BMSCs and secretion of VEGF protein in culture medium were measured by reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot and enzyme linked immunosorbent assay (ELISA) methods. The activity of VEGF protein in culture media was detected by proliferation effects on vascular endothelial cells.

RESULTSWhen Ad. VEGF(165) transferred into BMSCs, there was an effective transcription and expression. The expressed product in the media of transferred cells had highly biological activity on proliferation of rat aortic vascular endothelial cells (P < 0.01).

CONCLUSIONSAdenovirus vector can be transferred into BMSCs efficiently and safely. Adenovirus mediated VEGF(165) gene transferred into BMSCs could express VEGF protein with highly biological activity, which provided foundation on BMSCs based gene therapy for ischemic disease.

Adenoviridae ; genetics ; Animals ; Bone Marrow Cells ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Genetic Vectors ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; biosynthesis ; pharmacology ; Stromal Cells ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics ; pharmacology

OBJECTIVETo explore repair methods of skin and soft tissue defects in lower extremities with free latissimus dorsi flaps.

METHODSForty-two patients with wounds and soft tissue defects in lower extremities, including 4 cases on knee, 22 cases on leg, 15 cases on ankle and foot, 1 case with extensive avulsion from knee to dorsum of foot, were hospitalized in our unit from February 1996 to February 2008. Wounds or soft tissue defects were respectively repaired with latissimus dorsi musculocutaneous flaps, latissimus dorsi muscle flaps, latissimus dorsi perforator flaps with preserved vascular sleeves, 2 double-leaf segmental latissimus dorsi compound flaps after debridement. The flaps ranged from 18 cm x 8 cm to 40 cm x 18 cm in size. The donor sites were covered by skin grafting in 19 cases.

RESULTSAll wounds were healed primarily except vascular crisis occurred in 3 cases, partial necrosis of skin at donor site in 2 cases, and graft site (1 case). Follow-up for 3 to 24 months of 31 patients showed: six cases received two-stage plastic operation on account of bulkiness with trouble in wearing shoes, and mild contraction of muscular flap in 3 cases.

CONCLUSIONSLatissimus dorsi flap in various forms can be satisfactory for repair of large skin and soft tissue defects in lower extremities.

Adolescent ; Adult ; Child ; Female ; Humans ; Lower Extremity ; injuries ; surgery ; Male ; Middle Aged ; Muscle, Skeletal ; transplantation ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; Young Adult