ABSTRACT
AIM: To construct the prokaryotic expression system containing protein transduction domain (PTD) with heat shock protein 27 (HSP27) in order to prepare and purify the recombinant protein , and to verify whether the recombinant protein PTD-HSP27 has the ability to penetrate the human lens epithelial cell ( HLEC) membrane and the rabbit cornea.METHODS:The plasmid pKYB-PTD-HSPB1-6His was constructed by the technique of overlap extension PCR.The plasmid was transformed and PTD-HSP27 was purified through nickel affinity chromatography column and identi-fied by Western blotting.PTD-HSP27-6His was labeled with the fluorescein isothiocyanate (FITC).The penetrating ability of PTD-HSP27 into HLECs and rabbit cornea was tested .RESULTS:The recombinant PTD-HSP27 plasmid was success-fully cloned and effectively expressed .The correctness of the recombinant protein PTD-HSP27 was demonstrated .Fluores-cence microscopic examination showed that PTD-HSP27-FITC was internalized by HLECs .Fluorescent labeled PTD-HSP27 was then observed in the rabbit aqueous humor .CONCLUSION:The recombined gene PTD-HSPB1 was constructed by o-verlap extension PCR technique and the PTD-HSP27 fusion protein was prepared and purified by nickel affinity chromatog-raphy column.Using the technique of PTD-fusion protein, HSP27 was transduced into HLECs and passed through the cor-nea .
ABSTRACT
Progressive loss of retinal ganglion cells (RGCs) and their axons is the main pathogenesis of glaucoma. The cause of glaucoma is not fully understood, but the neurodegeneration of glaucoma involves many mechanisms such as oxidative stress, glutamate toxicity and ischemia/reperfusion insult. In order to target these mechanisms, multiple neuroprotective interventions have been investigated to prevent the death of RGCs. Of note are some tonic herbs from the traditional Chinese medicine (TCM) pharmacopeia that have shown neuroprotective effects in glaucoma. TCM differs from Western medicine in that TCM exhibits complicated bioactive components, triggering many signaling pathways and extensive actions on vital organs. Modern scientific approaches have demonstrated some of their underlying mechanisms. In this review, we used Lycium barbarum and Ginkgo biloba as examples to elaborate the characteristics of TCM and their potential applications in neuroprotection in glaucoma.
ABSTRACT
[Objective] To observe the effect of Shuhou Kangyan Decoction (SKD), a liver-kidney nourishing, Qi-strengthening and blood-activating, wind-dispelling and heat-clearing prescription mainly composed of Radix Astragali, Radix Angelicae Sinensis, Fructus Lycii, Radix Rehmanniae, Radix Saposhnikoviae, Rhizoma Atractylodis Macrocephalae, etc. , on interleukin-2 (IL-2) and soluble IL-2 receptor (SIL-2R) in serum and tears of patients with cataract after operation. [Methods] Forty-seven cases of cataract after ultrasonic emulsification plus artificial lens implantation were randomized into two groups. Group A (n = 26) was treated with routine operation and eye drops and group B ( n = 21) with SKD additionally and other 15 healthy aged people ( n = 15) receiving physical examination in the out-patient department served as normal controls. [Results] Post-operative serum SIL-2R level was higher in groups A and B than that before operation ( P 0.05). Post-operative IL-2 and SIL-2R levels in the serum and tears were higher in group A than that in groups B and C (P
ABSTRACT
AIM: To observe the immunoregulation of all ogeneic cornea on the human peripheral blood T lymphocytes in vitro. METHODS: After Co-culture of human peripheral blood lymphocytes and allogeneic cornea in vitro, T lymphocytes were labeled by monoclonal ant ibody, and analyzed by fluorescent activated cell sorter (FACS). RESULTS: CD25 expression on T lymphocytes in control was 25.2%, after stimulated by the allogeneic cornea or PDB, CD25 expression on T lymphocy tes was 56.8% and 80.9%, respectively. After stimulated by the allogeneic corn ea, CD25 expression on CD 4+ or CD 8+ T lymphocytes were 67.3% and 52.3% , respectively. CONCLUSION: Allageneic cornea stimulates CD25 expression on huma n peripheral blood T lymphocytes, and the CD25 expression on CD 4+ T lymphocy tes is more prominent than CD 8+ T lymphocytes.