Pyridoxine hcl

HOCAS was established using paired human donor eyes as described previously^{38 (link)}. Briefly, the anterior segments were dissected out after removing vitreous, lens and iris leaving the ciliary body from the donor eyes. They were mounted onto a specially designed Petri dish containing Dulbecco’s Modified Eagle’s Medium (DMEM, containing 4500 mg glucose/L, L-glutamine, NaHCO₃ and pyridoxine HCl, Sigma-Aldrich, MO, USA) supplemented with gentamycin (15 mg/L, Sigma-Aldrich, MO,USA) and antibiotic/antimycotic solution (penicillin G, 100 U/ml; streptomycin sulphate, 100 μg/ml; amphotericin B, 0.25 μg/ml: Sigma-Aldrich, MO, USA)^{27 (link)}. Topical antibiotic solution was applied to the cornea. Stopcocks were closed and the eye segments were transferred to an incubator (37 °C, 5% CO₂) and attached to pressure transducers (APT 300 pressure transducers, Harvard Apparatus, MA, USA). The pump (PHD 2000 Ultra Syringe Infusion only pump, Harvard Apparatus, MA, USA) was started; infusion set at 3 μl/minute and pressure was monitored on a computer connected to the Power Lab system with Lab Chart Pro software (AD Instruments, CO, USA). The eyes that maintained IOP within physiological range (10–21 mm Hg) for ~2 days were subsequently used for drug treatment.

HA-VSMCs and human embryonic lung fibroblasts (HELFs) were obtained from ATCC (Manassas, VA, USA) and cultured with Dulbecco’s modified Eagle’s medium (DMEM) with 10% fetal bovine serum (FBS), 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), l-glutamine, and pyridoxine HCl (Sigma-Aldrich, MO, USA), while HEK293 cells were obtained from the Chinese Academy of Sciences cell bank and cultured with DMEM with 10% FBS. The AD169 strain of human cytomegalovirus (HCMV) and AD169 (ΔUS33) with the US33 gene knocked out were generated using TB40-BAC_KL7-UL32EGFP (gifted by Dr. Katharina Gohring, Tubingen University, Germany) by Dr. Gan from Anhui Medical University, Department of Microbiology [32 (link)]. The virus was propagated in HELF cells and stored in liquid nitrogen.

The vitamin content of G. corticata and G. edulis was determined by the method previously described [20 (link)]; vitamin A or retinol (328 nm), vitamin B1 or thiamine monohydrate (420 nm), vitamin B2 or riboflavin (254 nm), vitamin B6 or pyridoxine HCl (254 nm), vitamin C or ascorbic acid, vitamin E or tocopheryl acetate (520 nm) and folic acid (550 nm) were determined by HPLC methods, and the results were compared with the respective standards retinyl acetate, thiamine monohydrate, riboflavin, pyridoxine HCl, ascorbic acid, tocopheryl, and folic acid (Sigma Aldrich). Vitamin content was determined in triplicate for each seaweed and was expressed as mg/g of DW seaweed.