Objective To investigate the antibacterial activity of the alkaloids extract from Oxytropis falcata Bunge against 9 common pathogens in vitro. Methods Alkaloids was extracted from Oxytropis falcata Bunge. The agar plate dilution method was adopted to evaluate the activity against bacteria of 4 Gram-negative test strains and 5 Gram-positive test strains, while the MIC and MBC were determined by tube double dilution method. Results The alkaloids extract from Oxytropis falcata Bunge had the antibacterial activity against 9 pathogens and there was a good dose-response relationship between sensitivity and concentration. Antibacterial activities against bacillus subtilis and stentrophmomas maltophilia showed stronger than other tested bacteria, the MIC and MBC were 0.95 mg/mL and 1.91 mg/mL, respectively. In other tested bacteria, MIC and MBC were ranged in 1.91-7.63 mg/mL. Conclusion The alkaloids extract showed a broad spectrum of a stronger antibacterial activity against all tested pathogens. Gram-positive strains were more sensitive than Gram-negative strains to the inhibitory effects of the alkaloids extract.

Objective To establish a method of HPLC for determination of rhamnocitrin in Oxytropis falcate Bunge.Methods The HPLC method was used.Chromatographic column:Lichrospher C18(4.6 mm? 250 mm,5 ?m) reversed-phase column;mobile phase:0.4% phosphoric acid in water-methanol(20∶80);flow rate:1.0 mL/min;detective wavelength:350 nm.Results Rhamnocitrin was liner at the range of 4～ 30 ?g(r =0.999 8),and its average recovery rate was 99.7%,RSD=1.4%.Conclusion The method was simple and accurate,so it can be used for the quality control of Oxytropis falcate Bunge.

Proteins extracted from two varieties of Chinese roses leaves were separated by two- dimensional polyacrylamide gel electrophoresis (2-DE) with immobilized pH gradient (IPG). Many difference proteins were isolated with molecular weights ranging 10-30 kDa and pI5-6. Three proteins of high levels observed in a gel were excised and identified using peptide mass fingerprinting and MS-MS. A summary of the identified proteins and their putative functions are presented. They are identified as eIF-5A、LEA protein and Hsp17. 5. Functions of these proteins in plant tolerance to high temperature were discussed.

BACKGROUND: Transfection is the most important beginning component of research about gene function. It is a problem to find a transfection agent with high efficiency and safety. Nanosized materials have high surface activity, are easy to be modified and easier to pass the biomembrane. Researchers are studying how to use nanosized materials as a transfection agent.OBJECTIVE: To study the transfection efficiency of different basic polymers with different molecular mass and degree of substitute, and to find the optimized transfection agent.DESIGN: Controlled study.SETTING: Institute of Genetic Engineering, Southern Medical University.METHODS: This study was performed in the Institute of Genetic Engineering, Southern Medical University from March 2006 to June 2007. Using the lipofectamine reagent as the positive control, we transfected siRNA (0.2 nmol/L), FITC-labeling targeting bcl-2, by nine nanometers (polylactic acid-polyglycolic acid, chitosan-poly-caprolactone, polyethyleneimin-macrogol) into leukemic cell K562 cultured without serum. Six hours post-transfection, 20% FBS serum was added. The cell proliferation was measured at 24, 48 and 72 hours by using the MTT method. After transfecting for 48 hours, the cells were collected to detect transfection ratio by using fluorescent microscopy, apoptosis ratio and expression of K562 and bcl-2 protein by flow cytometer (FCM). RESULTS: ① Fluorescence microscope detection showed there were significant differences of transfection efficiency between different materials (P < 0.05). Moreover there were statistical differences between different degrees of substitute, although they were the same material (P < 0.05). ② MTT method indicated the cell proliferation ratio was positively related to transfection ratio. ③ Flow cytometry results showed the suppression of expression of targeting gene and apoptosis ratio were positive correlated with transfection ratio. CONCLUSION:The nanometer poly-ethylene glycol combined with poly-ethylene imine (PEG-PEI), whose molecular mass is 1 800/2 000 and degree of substitute is 29%, has a high efficiency and low toxicity.

AIM: To investigate the efficacy, safety and localization of multilayer amniotic membrane transplantation (AMT) for the management of corneal perforation associated with ulceration.METHODS: Six eyes (6 patients) were treated with multilayer AMT for corneal perforation associated with ulceration. The perforation size was 0.5-2mm in diameter. The amniotic membrane (AM) was applied to tamp the perforation, fill the ulcer and cover the surface of ulcer. The follow-up ranged from 5-19 months.RESULTS: The anterior chamber reformed at the first postoperative day in all patients, and kept in normal depth in the follow- up time. The epithelialization of the AM grafts was observed in about 3 weeks after surgery. During the first postoperative month, the corneal thickness at the perforation site gradually increased, and the stromal inflammatory cell invasions surrounding the corneal ulcer were gradually subsided.Two months postoperatively, the ulcer healed with scar with part neovascutarization; the corneal thickness of the ulcer area was stable and recovered almost normal. The vision was not improved in all patients after operation. During-follow up,no severe complications or recurrence of ulceration was noted.CONCLUSION: Multilayer AMT is an effective and safe method for the management of small corneal perforation associated with ulceration, but the ulcer healed with scar.

AIM: To evaluate the efficacy and safety of annular bubble conjunctival excision combining with amniotic membrane transplantation(AMT) for vernal keratoconjunctivitis(VKC) with gelatinous swellings(papillae) at the limbus.METHODS: Twenty eyes(10 patients) with VKC, were characterized by gelatinous swellings(papillae) at limbus and were underwent annular bubble conjunctival excision associated with AMT. The follow up ranged 5-21months.RESULTS: After operation, the symptoms were subsided remarkably: itching,discharge and foreign body sensation were almost disappear; photophobia and tearing were obvious in the first week after surgery, but were gradually alleviated, and almost disappear after one month of postoperative.Yellow-brown or filthy red gelatinous protuberances at limbus were cleaned away by operation and did not recur during the follow up. Corneal superficial punctuate keratitis(11 eyes) and corneal ulcers(3 eyes) were healed during the first week after surgery and did not recur. Conjunctival congestion was gradually lessened after surgery, which could not be noted 1months after surgery in 19 eyes, and the conjunctiva around the limbus, where AMT was performed, was whiter than the normal. But conjunctival congestion in 1 eye lasted for about 5mon accompanying with a little of itch, and this eye was treated with eyedrops with improvement but recurrence. No severe complications had been seen during the follow up.CONCLUSION: Annular bubble conjunctival excision combining with amniotic membrane transplantation is an effective and safe method for the treatment of vernal keratoconjunctivitis with gelatinous swellings (papillae) at limbus. But the candidates must be chosen with discretion.

AIM: To investigate the efficacy and safety of annular conjunctival excision combined with amniotic membrane transplantation for the management of earlier Mooren's ulcer.METHODS: Thirty eyes (24 patients) with earlier Mooren's ulcer were treated with this method, followed-up for 5-30mo. The characteristic of these patients was localized periphery corneal ulcer: the ulcer involved 30-120° corneal limbus with depth of 1/3-1/2 cornealthickness and width of 2-5mm.RESULTS: After surgery, symptoms subsided remarkably and immediately in all patients,and almost disappeared after stitches were removed. Corneal melts were controlled. At postoperative 2-3mo, the corneal thickness of the ulcer area became almost stable, which in some patients recovered normal and in others were still thinner than normal, and the average increase was about 1/3 of the corneal thickness. Corneal transparence and vision were improved. During follow-up, no severe complications or recurrence were noted.CONCLUSION: Conjunctival peritomy combined with amniotic membrane transplantation is an effective and safe method for the treatment of earlier Mooren's ulcer.

Objective To investigate the potential mechanism of inhibition of ursolic acid(UA) on growth of human gastric cancer cell lines SGC7901 in vitro.Methods SGC7901 cells were cultured in vitro,MTT assay was used to observe the effect of UA on growth of SGC7901 cells in various concentrations for different times.After SGC7901 cells were treated by 0—40 ?mol/L UA for 24 h,morphological changes were observed by inverted microscope.Apoptotic changes were detected by fluorescence microscopy and flow cytometry (FCM).Protein expressions of Bcl-2 and Bax were determined by Western blotting.Results UA(20—40 ?mol/L) could significantly inhibit the growth of SGC7901 cells in a(dose-and) time-dependent manner,the IC_(50) value of UA for SGC7901 cells for 12,24,36,and 48 h were((57.50?)1.18),(34.28?2.05),(27.54?1.11),and(24.83?1.02) ?mol/L,respectively.After UA((20—)40 ?mol/L) treatment for 24 h,SGC7901 cells turned round and floated at different levels;SGC7901 cells were arrested at G_0/G_1 phase and apoptosis was induced,and the apoptotic rate was increased along with the increase of UA concentration.Meanwhile Bcl-2 protein expression decreased,whereas Bax protein expression unchanged.Conclusion UA has a stronger antitumor effect on SGC7901 cells.Cytotoxic effect,proliferation inhibition,and apoptosis may be involoved in the mechanism of UA,and the apoptosis caused by UA may be enhanced by decrease of Bcl-2 protein expression.

0.05). Conclusion Enough high-purified LECs can be isolated by collagenase digestion procedure followed by immunomagnetic beads sorting. Post-thawed endothelial cells are proved to have high vitality and growth potential in vitro without significant morphological changes. Cryopreserved LECs may serve as a cell choice for research of lymphangiogenesis and lymphatic patterning.

Objective To investigate the value of intraoperative neuromonitoring(IONM) for preventing recurrent laryngeal nerve(RLN) injury in thyroid surgery.Methods 1087 consecutive patients undergoing thyroid surgery at Wuhan Central Hospital from Oct 2010 to Dec 2012 were evaluated.277 cases(the study group) underwent thyroidectomy with RLN identification by naked eye plus IONM.810 cases(the control group)underwent thyroidectomy with RLN identification by naked eye only.Results In the control group,733 RLNs (90.49％,733/810)were successfully identified and 77 RLNs were failed to be identified.In the study group,all the 277 RLNs(100％,277/277)were successfully identified.28 cases had postoperative temporal RLN injury,among whom 26 cases were in the control group and 2 cases were in the study group.18 cases in the control group and 2 cases in the study group recovered in 2 weeks after surgery.The rest recovered in 2 months after surgery.RLN injury rate was significantly lower in the study group than in the control group.There was no significant difference in RLN injury for low-risk surgical patients between the 2 groups; however,the study group had advantages in RLN injury than the control group for high-risk patients.Conclusions IONM in thyroid surgery can improve the recognition rate of RLN during thyroidectomy.IONM can significantly reduce the incidence of RLN injury,especially in high-risk surgery.