Journal: RNA

Article Title: Serum-stable RNA aptamers to urokinase-type plasminogen activator blocking receptor binding

doi: 10.1261/rna.2338210

Figure Lengend Snippet: The binding of upanap-12 to uPA requires the GFD. The relative association of upanap-12 to different variants of uPA is indicated: The inactive zymogen form of uPA (pro-uPA) active uPA (uPA), uPA without the growth factor domain (ΔGFD), and uPA without the entire ATF (also called low molecular weight uPA; LMW-uPA). The domain structure of the different variants is shown at each bar, while the inset illustrates pro-uPA with domain designations: (GFD) growth factor domain, (KD) kringle domain, and (SPD) serine protease domain. uPA variants were captured on an SPR sensor surface containing immobilized polyclonal anti-uPA antibody to the following levels: pro-uPA (215–280 RU), uPA (210–260 RU), ΔGFD-uPA (210–230 RU), and LMW-uPA (140–160 RU). The binding level of 50 nM upanap-12 to each captured variant was subsequently recorded. Using the molecular weight of the different variants, the binding level of aptamers per mole of captured variant was calculated and presented relative to the pro-uPA result. Open bars represent mean values and standard deviations derived from three independent experiments.

Article Snippet: Human two-chain uPA was from Wakamoto Pharmaceutical Company, while human pro-uPA and mouse uPA were from Molecular Innovations.

Techniques: Binding Assay, Molecular Weight, Variant Assay, Derivative Assay

Journal: American Journal of Translational Research

Article Title: SQYZ granules, a traditional Chinese herbal, attenuate cognitive deficits in AD transgenic mice by modulating on multiple pathogenesis processes

doi:

Figure Lengend Snippet: SQYZ treatment improved AD behavioral performances of 5XFAD mice. In the Morris water maze, during the hidden platform task, the escape strategies on the fifth day of training (A) of the WT+NS, WT+SQYZ, Tg+NS, Tg+SQYZ, and Tg+HupA groups were detected by using a camera. (B) The escape latency required for the mice to find the hidden platform on each training day was recorded. During the probe trial, one day after finishing the acquisition task, the time required to cross over to the target quadrant (C) was originally used to determine the memory retention of the mice. In the passive avoidance test, one day after fear training, the error times of mice entering the dark field (D), the step-through latency of mice from light side to dark side (E) and the time spent in light compartment (F) were recorded to test the fear memory abilities of the mice. During the nest construction test, the nesting scores of mice (G) were evaluated to reflect the abilities of daily living. All values were represented as the mean ± SEM, n = 13-15/group. *, P < 0.05, **, P < 0.01 vs. Tg+NS, one-way ANOVA with Tukey’s post hoc test.

Article Snippet: Huperzine-A (HupA), as a positive control drug, was purchased from Beijing Century Aoke Biotechnology Co. Ltd. (lot number: MUST-13102808).

Techniques:

Journal: American Journal of Translational Research

Article Title: SQYZ granules, a traditional Chinese herbal, attenuate cognitive deficits in AD transgenic mice by modulating on multiple pathogenesis processes

doi:

Figure Lengend Snippet: SQYZ reduced the levels of Aβ42 in the cortex of the AD transgenic mice. The soluble (A) and the insoluble Aβ42 levels (B) in the cortex of Tg+NS, Tg+SQYZ, and Tg+HupA mouse groups were tested by the ELISA assay. The Aβ concentration was normalized to total protein level in cortex. All values were represented as the mean ± SEM, n = 6/group. *, P < 0.05 vs. Tg+NS, one-way ANOVA with Tukey’s post hoc test.

Article Snippet: Huperzine-A (HupA), as a positive control drug, was purchased from Beijing Century Aoke Biotechnology Co. Ltd. (lot number: MUST-13102808).

Techniques: Transgenic Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Applied Microbiology and Biotechnology

Article Title: Microbial acetylcholinesterase inhibitors for Alzheimer’s therapy: recent trends on extraction, detection, irradiation-assisted production improvement and nano-structured drug delivery

doi: 10.1007/s00253-020-10560-9

Figure Lengend Snippet: Comparative and survey studies regarding the incorporation of HupA with different nanostructure compounds

Article Snippet: Recently, a complex of HupA-loaded muco-adhesive and polylactide-co-glycoside (PLGA-NPs) with external adjustment by lactoferrin-conjugated N-trimethylated chitosan (Lf-TMC NPs) was generated for effective intranasal distribution of HupA to the brain for potential Alzheimer’s disorder therapy (Sheng et al. ; Meng et al. ).

Techniques: Homogenization, In Vitro, Emulsion, Evaporation

Journal: Annals of Translational Medicine

Article Title: Huperzine A lowers intraocular pressure via the M3 mAChR and provides retinal neuroprotection via the M1 mAChR: a promising agent for the treatment of glaucoma

doi: 10.21037/atm-20-8093

Figure Lengend Snippet: The effects of huperzine A (HupA) on the rabbit pupil diameter. After treatment with different concentrations of HupA, the diameter of the rabbit pupil was significantly contracted over time in a dose-dependent manner which was reversed by the M3 mAChR antagonist atropine (Atr) and 4-DAMP. The positive drug control groups were 2% pilocarpine (Pilo) and 0.25% physostigmine (Phy). (A) The effects of HupA on the rabbit pupil diameter; (B) the effects of the nonselective mAChR antagonist atropine (Atr), the selective M1 mAChR antagonist pirenzepine (PZ), the M2 mAChR antagonist gallamine (GM), and the M3 mAChR antagonist 4-DAMP on pupil diameter after HupA treatment. Data are expressed as mean ± SEM, n≥5. *P<0.05, compared with the corresponding control group. SEM, standard error of the mean.

Article Snippet: HupA (WuXi AppTec, China), pilocarpine, physostigmine, atropine, and pirenzepine were added to the cells 30 minutes prior to OGD treatment.

Techniques: Control

Journal: Annals of Translational Medicine

Article Title: Huperzine A lowers intraocular pressure via the M3 mAChR and provides retinal neuroprotection via the M1 mAChR: a promising agent for the treatment of glaucoma

doi: 10.21037/atm-20-8093

Figure Lengend Snippet: HupA did not cause the isolated iris muscle to contract, nor did it affect intracellular calcium flow. (A) Iris muscle contraction after treatment with HupA, carbachol, Phy, and Pilo; (B) detection of the intracellular calcium flow in CHO-M3 after HupA, Phy, and Pilo treatment. Data are expressed as mean ± SEM, n≥5 rabbits or 3 independent experiments. HupA, huperzine A; CCh, carbachol; Phy, physostigmine; Pilo, pilocarpine; RFU, relative fluorescence units; SEM, standard error of the mean.

Article Snippet: HupA (WuXi AppTec, China), pilocarpine, physostigmine, atropine, and pirenzepine were added to the cells 30 minutes prior to OGD treatment.

Techniques: Isolation, Fluorescence

Journal: Annals of Translational Medicine

Article Title: Huperzine A lowers intraocular pressure via the M3 mAChR and provides retinal neuroprotection via the M1 mAChR: a promising agent for the treatment of glaucoma

doi: 10.21037/atm-20-8093

Figure Lengend Snippet: HupA significantly protected the retinal neuronal injury induced by I/R and OGD. (A) H&E staining of rat retinal slices from the different groups; (B,C,D) thickness of the whole retinas, inner platform layer (IPL) and inner nuclear player (INL) of each different group; (E) cell count of RGCs in the ganglion cell layer (GCL); (F,I) the mRNA expression levels of retinal selected markers including Sncg, Fem1c, Thy1, Nrn1, and Nfl as detected by PCR in I/R-induced retinal tissue or OGD-induced PRNs; (G) cell viability of OGD-induced PRNs detected by CCK-8 kit; (H) cytotoxicity of PRNs induced by OGD detected using the LDH kit. Data are expressed as mean ± SEM, n≥5 rats or 3 independent experiments. *P<0.05, **P<0.01, ***P<0.001, compared with the control group; #P<0.05, ##P<0.01, compared with the OGD or I/R group. HupA, huperzine A; I/R, ischemia/reperfusion; OGD, oxygen glucose deprivation; H&E, hematoxylin and eosin; PRN, primary retinal neuron; CCK-8, cell count kit 8; LDH, lactate dehydrogenase; Pilo, pilocarpine; Phy, physostigmine; Latan, latanoprost; SEM, standard error of the mean.

Article Snippet: HupA (WuXi AppTec, China), pilocarpine, physostigmine, atropine, and pirenzepine were added to the cells 30 minutes prior to OGD treatment.

Techniques: Staining, Cell Counting, Expressing, CCK-8 Assay, Control

Journal: Annals of Translational Medicine

Article Title: Huperzine A lowers intraocular pressure via the M3 mAChR and provides retinal neuroprotection via the M1 mAChR: a promising agent for the treatment of glaucoma

doi: 10.21037/atm-20-8093

Figure Lengend Snippet: HupA regulated oxidative stress induced by I/R and OGD in retinal neuronal cells. (A,B) GSH levels in retinal tissue subjected to I/R and in PRNs subjected to OGD; (C,D) SOD activity in retinal tissue and neurons; (E,F) MDA level in retinal tissue and neurons. Data are expressed as mean ± SEM, n≥5 rats or 3 independent experiments. *P<0.05, **P<0.01, ***P<0.001, compared with the control group; #P<0.05, ##P<0.01, compared with the I/R or OGD group. HupA, huperzine A; I/R, ischemia/reperfusion; OGD, oxygen glucose deprivation; PRN, primary retinal neuron; Pilo, pilocarpine; Phy, physostigmine; Latan, latanoprost; GSH, glutathione; SOD, superoxide dismutase; MDA, malondialdehyde; SEM, standard error of the mean.

Article Snippet: HupA (WuXi AppTec, China), pilocarpine, physostigmine, atropine, and pirenzepine were added to the cells 30 minutes prior to OGD treatment.

Techniques: Activity Assay, Control

Journal: Annals of Translational Medicine

Article Title: Huperzine A lowers intraocular pressure via the M3 mAChR and provides retinal neuroprotection via the M1 mAChR: a promising agent for the treatment of glaucoma

doi: 10.21037/atm-20-8093

Figure Lengend Snippet: HupA protected against retinal neurons apoptosis induced by I/R and OGD. (A,B) The expression and analysis of apoptosis-related proteins including caspase-3, Bcl-2, and Bax in I/R-induced rat retinal tissue detected by western blot analysis; (C,D) caspase-3 expression and analysis as detected by immunofluorescent staining of rat retinal tissue subjected to I/R treatment; (E,F) the expression and analysis of caspase-3, Bcl-2, and Bax in OGD treated PRNs detected by western blotting. Data are expressed as mean ± SEM, n≥5 rats or 3 independent experiments. **P<0.01, *P<0.05, compared with the control group; #P<0.05, ##P<0.01 compared with the OGD group. HupA, huperzine A; I/R, ischemia/reperfusion; OGD, oxygen glucose deprivation; PRN, primary retinal neuron; Pilo, pilocarpine; Phy, physostigmine; Latan, latanoprost; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; IHC, immunofluorescence; OPL, outer platform layer; IPL, inner platform layer; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer; SEM, standard error of the mean.

Article Snippet: HupA (WuXi AppTec, China), pilocarpine, physostigmine, atropine, and pirenzepine were added to the cells 30 minutes prior to OGD treatment.

Techniques: Expressing, Western Blot, Staining, Control, Immunofluorescence

Journal: Annals of Translational Medicine

Article Title: Huperzine A lowers intraocular pressure via the M3 mAChR and provides retinal neuroprotection via the M1 mAChR: a promising agent for the treatment of glaucoma

doi: 10.21037/atm-20-8093

Figure Lengend Snippet: The disorder of the cholinergic system in retinal tissue and neurons induced by I/R and OGD could be reversed by HupA. This protective effect of HupA in OGD treated PRNs was reversed by the nonselective mAChR antagonist atropine and the M1 mAChR antagonist perizipine. (A,C) The AChE activity in rat retinal tissue and in PRNs subjected to OGD; (B,D) the levels of ACh detected by HPLC-MS/MS in rat retinal tissue and in PRNs subjected to OGD; (E) the photographs of PRNs in the different groups; (F) the retinal neuronal cell viability was detected by CCK-8 assay; (G) the cytotoxicity of the PRNs was detected by the LDH kit; (H) GSH levels in the OGD-treated PRNs; (I) SOD activity in retinal neurons; (J) MDA levels in retinal neurons; (K, L) the expression and analysis of caspase-3, Bcl-2, and Bax in PRNs subjected to OGD detected by western blotting after treatment with the M1 mAChR antagonists. (M) detection of the intracellular calcium flow in CHO-M1 after HupA, Phy, and Pilo treatment. Data are expressed as mean ± SEM, n≥5 rats or 3 independent experiments. *P<0.05, **P<0.01, ***P<0.001, compared with the control group; #P<0.05, ##P<0.01, compared with the I/R or OGD groups; $P<0.05, $$P<0.01, compared with the HupA group. HupA, huperzine A; I/R, ischemia/reperfusion; OGD, oxygen glucose deprivation; PRN, primary retinal neuron; Atr, atropine; PZ, perizipine; mAChR, muscarinic acetylcholine receptor; ACh, acetylcholine; HPLC-MS/MS, liquid chromatography-tandem mass spectrometry; CCK-8, cell counting kit 8; LDH, lactate dehydrogenase; GSH, glutathione; SOD, superoxide dismutase; MDA, malondialdehyde; Pilo, pilocarpine; Phy, physostigmine; Latan, latanoprost; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; SEM, standard error of the mean.

Article Snippet: HupA (WuXi AppTec, China), pilocarpine, physostigmine, atropine, and pirenzepine were added to the cells 30 minutes prior to OGD treatment.

Techniques: Activity Assay, Tandem Mass Spectroscopy, CCK-8 Assay, Expressing, Western Blot, Control, Liquid Chromatography, Mass Spectrometry, Cell Counting

Journal: Annals of Translational Medicine

Article Title: Huperzine A lowers intraocular pressure via the M3 mAChR and provides retinal neuroprotection via the M1 mAChR: a promising agent for the treatment of glaucoma

doi: 10.21037/atm-20-8093

Figure Lengend Snippet: Involvement of AKT/MAPK signal pathway in the retinal neuroprotection of HupA. (A,C) Western blot analyses of the levels of phosphorylation of AKT, ERK, P38, and JNK in OGD- treated PRNs at different time points; (B,D) analyses of the level of phosphorylation of AKT, ERK, P38, and JNK treated by HupA and M1 mAChR antagonists; (E) effects of the AKT/MAPK pathway inhibitors on retinal neuronal cell viability. Data are expressed as mean ± SEM, n≥3 independent experiments. *P<0.05, **P<0.01, compared with the control group; #P<0.05, ##P<0.01, compared with the OGD group; $P<0.05, compared with the HupA group. HupA, huperzine A; OGD, oxygen glucose deprivation; PRN, primary retinal neuron; Atr, atropine; PZ, perizipine; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; P-AKT, phosphorylated AKT; P-P38, phosphorylated P38; ERK, extracellular-signal-regulated kinase ½; P-ERK, phosphorylated ERK; JNK, c-Jun N-terminal kinases; P-JNK, phosphorylated JNK; LY204002, AKT inhibitor; SB203580, P38 inhibitor; SP600125, JNK inhibitor; U0126, ERK inhibitor; SEM, standard error of the mean.

Article Snippet: HupA (WuXi AppTec, China), pilocarpine, physostigmine, atropine, and pirenzepine were added to the cells 30 minutes prior to OGD treatment.

Techniques: Western Blot, Phospho-proteomics, Control