Lithium promotes proliferation and suppresses migration of Schwann cells

Schwann cell proliferation,migration and remyelination of regenerating axons contribute to regeneration after peripheral nervous system injury.Lithium promotes remyelination by Schwann cells and improves peripheral nerve regeneration.However,whether lithium modulates other phenotypes of Schwann cells,especially their proliferation and migration remains elusive.In the current study,primary Schwann cells from rat sciatic nerve stumps were cultured and exposed to 0,5,10,15,or 30 mM lithium chloride(LiCl)for 24 hours.The effects of LiCl on Schwann cell proliferation and migration were examined using the Cell Counting Kit-8,5-ethynyl-2′-deoxyuridine,Transwell and wound healing assays.Cell Counting Kit-8 and 5-ethynyl-2′-deoxyuridine assays showed that 5,10,15,and 30 mM LiCl significantly increased the viability and proliferation rate of Schwann cells.Transwell-based migration assays and wound healing assays showed that 10,15,and 30 mM LiCl suppressed the migratory ability of Schwann cells.Furthermore,the effects of LiCl on the proliferation and migration phenotypes of Schwann cells were mostly dose-dependent.These data indicate that lithium treatment significantly promotes the proliferation and inhibits the migratory ability of Schwann cells.This conclusion will inform strategies to promote the repair and regeneration of peripheral nerves.All of the animal experiments in this study were ethically approved by the Administration Committee of Experimental Animal Center of Nantong University,China(approval No.20170320-017)on March 2,2017.

Aquaporin-1 down regulation associated with inhibiting cell viability and inducing apoptosis of human lens epithelial cells

AIM：To investigate the role of Aquaporin-1（AQP-1）in lens epithelial cells（LECs）and its potential target genes.AQP-1 is specifically expressed in LECs of eyes and is significant for lens homeostasis and transparency maintenance.Herein,AQP-1 expression in LECs was investigated to evaluate its influence on cell survival in association with its potential role in cataract formation.·M ETHODS：LECs were transfected with lentivirus carrying AQP-1 small interfering RNA（si RNA）.Real-time polymerase chain reaction（PCR）and Western blotting were conducted to detect AQP-1 expression in LECs from different groups.Meanwhile,cell counting kit-8（CCK-8）assay and flow cytometry were performed to measure LEC proliferation and apoptosis,respectively.·RESULTS：AQP-1 expression was significantly reduced in LECs,both at m RNA and protein levels（〈0.05）,after si RNA treatment.Decreased cell viability was detected by CCK-8 assay in LECs with si RNA interference,compared to control cells（〈0.05）.The apoptosis rate significantly increased in cells after si RNA interference（〈0.05）.·CONCLUSION：The decreased cell viability following AQP-1 down regulation is largely due to its induction of apoptosis of LECs.AQP-1 reduction might lead to changes of physiological functions in LECs,which might be associated with the occurrence and development of cataracts.

Effects of curcumin nanoparticles on proliferation and VEGF expression of human retinal pigment epithelial cells

AIM:To investigate the effects of curcumin(Cur)nanoparticles loaded with chitosan derivatives grafted by deoxycholic acid(Chit-DC)on human retinal pigment epithelial(h RPE)cell proliferation and vascular endothelial growth factor(VEGF)m RNA expression.METHODS:Cur nanoparticles were synthesized with Chit-DC as the carrier and Cur as the supported drug.Cell counting kit-8(CCK-8)method was used to detect the effects of different concentrations of Cur/Chit-DC,Chit-DC,and Cur on the proliferation of h RPE cells for different times.The changes of Cur/Chit-DC and Cur on h RPE cell cycle were determined by flow cytometry.Semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the m RNA expression levels of VEGF in h RPE cells treated with Cur,Chit-DC and Cur/Chit-DC at 10μg/m L for 24 h.RESULTS:Different concentrations of Chit-DC nanoparticle treated h RPE cells had no significant difference in terms of optical density(OD)values compared with the control group at 24 h and 48 h.Moreover,there was no change in the cell morphology under a light microscope.After 24 h treatment with Cur/Chit-DC and Cur,the percentage of G0-G1 phase cells increased and the percentage of S phase cells decreased in all concentration groups.Cur/Chit-DC and Cur in all concentration groups inhibited the proliferation of h RPE cells in a time and dose dependent manner,and reduced the expression level of VEGF m RNA.CONCLUSION:The Cur/Chit-DC nanoparticles can release Cur continuously and have sustained release function.Both Cur/Chit-DC nanoparticles and Cur could inhibit h RPE cells cultured in vitro,and could reduce the expression level of VEGF m RNA in h RPE cells.

新型根管充填材料HiFlow的体外细胞毒性研究

目的:评价EndoSequence BC Sealer TM HiFlow(HiFlow)的细胞毒性。方法:体外培养L929细胞,制备HiFlow(A组)、AH Plus(B组)、iRoot SP(C组)、MTA(D组)4种根充材料浸提液,细胞培养液作为阴性对照(E组)。分别与L929接触12、24、48 h,CCK-8法对比HiFlow、AH Plus、iRoot SP、MTA 4种根充材料对L929细胞的毒性作用。结果:CCK-8实验显示不同根充材料12、24、48 h A值差异有统计学意义(P<0.05);其中B组A值低于A、C、D组(P<0.05);A组、C组和D组差异无统计学意义(P>0.05)。结论:HiFlow对L929细胞的细胞毒性作用与MTA、iRoot SP类似,均小于AH Plus,认为可临床用于根管充填。

The Characterization of Steam Distillation as an Extraction Method to Extract Volatile Compounds from <i>Prunella vulgaris</i>and the Investigation of Their Anti-Tumorous Effect

Prunella vulgaris (PV) is a perennial plant which is widely grown around the world. It has been widely used as a medicinal treatment for generations. Previous studies showed extracts from this plant had a wide range of therapeutic efficacy, including anti-tumorous effect. However, the volatile organic compounds (VOCs) extracted from it were rarely explored. This paper reports on the characterization of a steam distillation process to extract VOCs in PV and also the anti-tumorous effects of the PV distillate using the tetrazolium-based Cell Counting Kit-8 (CCK-8) as the test agent, when the VOCs were used to treat oral squamous cancer cells, SSC154. It was found that most abundant VOCs came out steadily and continuously for as long as the duration of the steam extraction could extend. However, some compounds such as benzaldehyde did show depletion as the distillation process progressed, while some compounds such as caryophyllene oxide was only sparsely found at the beginning of distillation. The PV distillate was mildly effective in its cytotoxicity to cancer cells SCC154, in a dosage dependent manner.

Cytotoxicity of six copper-bearing intrauterine devices on Chinese hamster ovary cells: the influence of frame, shape and copper surface area

Objective To evaluate the cytotoxicity of six commonly used copper-bearing intrauterine devices （Cu-IUDs） on Chinese hamster ovary （CHO-K1） cells and to investigate the influence of frame, shape and copper surface area of Cu-IUDs on cell toxicity.Methods Cu-IUDs were incubated in 10% FBS-DMEM/F12 culture medium at 37 ℃ for 24 h. The extracts were analyzed by flame atomic absorption spectrometer and were then diluted into different concentrations with culture medium. Finally, cytotoxicity of these original and diluted extracts on CHO-K1 cells was detected by cell counting kit-8 （CCK-8） assay.Results The viabilities of cells treated with the original extracts of six Cu-IUDs （TCu220C bulb, TCu220C, GCu220, GCu300, Yuangong Cu270 and Yuangong Ⅱ- 300） were all below 10% and the cupric ion concentrations in these extracts were 28.22 mg/L, 31.80 mg/L, 92.80 mg/L, 99.74 mg/L, 114.90 mg/L and 119.20 mg/L, respectively. After these original extracts were diluted, significant differences in cytotoxicity were exhibited. IUDs with larger copper surface areas （GCu300 and Yuangong Ⅱ-300） showed more cytotoxicity than those with smaller areas （GCu220 and Yuangong Cu270） respectively; When different shapes of Cu-IUDs were compared, TCu220C bulb showed lower cytotoxicity than TCu220C, and GCu300 exhibited higher toxicity than Yuangong Ⅱ-300; TCu220C displayed significantly lower cytotoxicity than GCu220 due to their differences in frames.Conclusion We presented evidence on the cytotoxic effects of copper ions released from Cu-IUDs on CHO-K1 cells and found that shape, frame together with copper surface area of Cu-IUDs had obvious influence on the cytotoxicity.

Studies on New Steroidal Saponins from Allii macrostemonis Bulbus and Their Antitumor Activities

Two new steroidal saponms（2, 3） and two known compounds（1, 4） were isolated from 75% ethanol extract ofAllii macrostemonis bulbus by various spectral methods. The two new steroidal saponins were respectively elucidated as （25S）-26-O-β-D-glucopyranosyl-5α-furostanol-2α,3β,22,26-tetrol-3-O-β-D-glucopyranosyl（1→2）-[β-D- glucopyranosyl（1→3）]-β-D-glucopyranosyl（1→4 ）-β-D-galactopyranoside（2 ） and 26-O-β-D-glucopyranosyl-5α- furostanol-25（ 27）-ene-3β,22,26-tetrol-3-O-β-D-glucopyranosyl（ 1→2 ）-[β-D-glucopyranosyl（ 1→3 ）] -β-D-glucopyranosyl- （1→4）-β-D-galactopyranoside（3）. Compound 4 was obtained from this medical plant for the first time. The activity experiments of proliferation inhibition on tumor cells were performed by cell counting kit-8（CCK-8） method for compound 2（25S-configuration） and its isomer（compound 1, 25R-configttration）. The results show that compounds 1 and 2 have weak inhibition on lung cancer A549 cells, melanoma A375 cells and breast cancer MCF-7 cells as well as certain inhibitory effect on liver cancer HepG2 cells, and the inhibitory effect of compound 1 is stronger than that of compound 2.