Liuwei Dihuang pills attenuate ovariectomy-induced bone loss by alleviating bone marrow mesenchymal stem cell (BMSC) senescence via the Yes-associated protein (YAP)-autophagy axis.

CONTEXT: Liuwei Dihuang pill (LWDH) has been used to treat postmenopausal osteoporosis (PMOP). OBJECTIVE: To explore the effects and mechanisms of action of LWDH in PMOP. MATERIALS AND METHODS: Forty-eight female Sprague-Dawley rats were divided into four groups: sham-operated (SHAM), ovariectomized (OVX), LWDH high dose (LWDH-H, 1.6 g/kg/d) and LWDH low dose (LWDH-L, 0.8 g/kg/d); the doses were administered after ovariectomy via gavage for eight weeks. After eight weeks, the bone microarchitecture was evaluated. The effect of LWDH on the differentiation of bone marrow mesenchymal stem cells (BMSCs) was assessed via osteogenesis- and lipogenesis-induced BMSC differentiation. The senescence-related biological indices were also detected using senescence staining, cell cycle analysis, quantitative real-time polymerase chain reaction and western blotting. Finally, the expression levels of autophagy-related proteins and Yes-associated protein (YAP) were evaluated. RESULTS: LWDH-L and LWDH-H significantly modified OVX-induced bone loss. LWDH promoted osteogenesis and inhibited adipogenesis in OVX-BMSCs. Additionally, LWDH decreased the positive ratio of senescence OVX-BMSCs and improved cell viability, cell cycle, and the mRNA and protein levels of p53 and p21. LWDH upregulated the expression of autophagy-related proteins, LC3, Beclin1 and YAP, in OVX-BMSCs and downregulated the expression of p62. DISCUSSION AND CONCLUSIONS: LWDH improves osteoporosis by delaying the BMSC senescence through the YAP-autophagy axis.

Efficacy of MP-3 microperimeter biofeedback fixation training for low vision rehabilitation in patients with maculopathy.

BACKGROUND: To evaluate the efficacy of MP-3 microperimeter biofeedback fixation training (MBFT) in vision rehabilitation of low-vision patients affected by macular disease with central vision loss. METHODS: Seventeen eyes (7 age-related macular degeneration, 10 myopic maculopathy) of 17 patients were included in this prospective, interventional study. The preferred retinal locus was determined by comprehensive ophthalmoscopic fundus evaluation including fundus photography, autofluorescence, optical coherence tomography, and microperimetry. The rehabilitation consisted of three 10-min sessions per eye to be performed twice per week for 20 consecutive weeks using the MP-3 microperimeter. Best corrected visual acuity (BCVA), reading speed, mean central sensitivity, the percentages of fixation points within specified regions, bivariate contour ellipse area (BCEA) and the 25-item National Eye Institute visual function questionnaire (NEI-VFQ-25) were recorded pre- and post-training. RESULTS: The final BCVA, reading speed and mean central sensitivity all showed significant improvements after rehabilitation (P <  0.0001, P = 0.0013, and P = 0.0002, respectively). The percentages of fixation points located within 2° and 4° diameter circles both significantly increased after training (P = 0.0008 and P = 0.0007, respectively). The BCEA encompassing 68.2, 95.4, 99.6% of fixation points were all significantly decreased after training (P = 0.0038, P = 0.0022, and P = 0.0021, respectively). The NEI-VFQ-25 scores were significantly increased at the end of the rehabilitation training (P <  0.0001). CONCLUSION: Rehabilitation with MP-3 MBFT is a user-friendly therapeutic option for improving visual function, fixation stability, and quality of life in advanced macular disease. TRIAL REGISTRATION: The prospective study was registered with the Chinese Clinical Trial Registry ( http://www.chictr.org.cn/ ). TRIAL REGISTRATION NUMBER: ChiCTR2000029586 . Date of registration: 05/02/2020.

Quercetin protects retina external barrier from oxidative stress injury by promoting autophagy.

Purpose: This study aimed to investigate the protective effects of quercetin on the tight junction proteins of human retinal pigment epithelial cells (ARPE-19 cells) suffering from oxidative stress injury and explore the possible mechanism.Methods: H2O2 (300 µM) was used to establish an oxidative stress model of ARPE-19 cells. ARPE-19 cells were pretreated with different concentrations (0-80 µM) of quercetin before H2O2 exposure. The expression and distribution of tight junction proteins and autophagy-related proteins were detected by Western blot and immunostaining. ARPE-19 cells were pretreated with 5 mM 3-methyladenine (3-MA).Results: The cell viability weakened in the H2O2 group compared with the control group. However, it was preserved after pretreatment with quercetin. It was observed that the expression levels of occludin, claudin-1 were decreased in the H2O2 group. Quercetin treatment significantly enhanced the expression levels of them as compared to the H2O2 group. H2O2 alone strongly decreased the Zonula occludens protein 1 (ZO-1) expression in the cytomembrane. Quercetin supplementation enhanced the accumulation of ZO-1 in ARPE-19 cells. The expression levels of Beclin-1 and Microtubule associated protein light chain 3 II (LC-3II) increased, and that of P62 decreased in the quercetin protection group. The appearance of LC-3II, which examined by immunofluorescence experiments, enhanced in the quercetin protection group as compared with the control group. The expression levels of beclin-1 and LC-3II increased, and that of P62 increased in the autophagy-inhibited group compared with the quercetin protection group. The levels of occludin and claudin-1 also decreased.Conclusion: Quercetin prevents the loss of tight junction proteins by upregulating autophagy after oxidative stress in ARPE-19 cells.

Computed tomography (CT) imaging evaluation of integrated traditional Chinese medicine cooperative therapy in treating acute cerebral infarction: A randomized controlled trial.

INTRODUCTION: Acute cerebral infarction is a clinically common cerebrovascular disease. Acute cerebral infarction is characterized by sudden onset, dangerous illness, high risk of death, and disability. Computed tomography perfusion imaging can detect abnormal brain tissue perfusion 30 minutes after the onset of cerebral ischemia, providing the earliest and most valuable information for clinical diagnosis and treatment. In recent years, the effect of traditional Chinese medicine on acute cerebral infarction has been remarkable. METHODS/DESIGN: This study plan randomly divided eligible acute cerebral infarction patients into two groups. Patients in the control group will be treated with conventional Western medicine; patients in the intervention group will be treated with traditional Chinese medicine cooperative therapy on the basis of conventional Western medicine. The curative effects will be selected before treatment, 2 weeks after treatment, and 3 months follow-up. The changes in CT imaging evaluation, NIHSS score, and BI index of the two groups of patients will be observed. DISCUSSION: We aim to provide higher evidence-based medical evidence for traditional Chinese medicine treatment of acute cerebral infarction. And clarify the application value of computed tomography perfusion imaging in the diagnosis and efficacy evaluation of acute cerebral infarction. TRIAL REGISTRATION: ClinicalTrials.gov, ChiCTR2000030230, Registered on 03 March 2020.

Effect of Moxibustion on HIF-1α and VEGF Levels in Patients with Rheumatoid Arthritis.

Background: Moxibustion has a therapeutic effect of reducing swelling and relieving pain in patients with rheumatoid arthritis (RA) but its mechanism is uncertain. Objective: To evaluate the effect of moxibustion on serum levels of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in patients with RA and to explore the possible mechanism of moxibustion. Methods: This study involved 46 RA patients who had fulfilled the inclusion criteria and were randomly assigned to a treatment group and a control group in an equal ratio. The control group was treated with methotrexate or leflunomide, while the treatment group received methotrexate or leflunomide and moxibustion at ST 36 (Zusanli), BL 23 (Shenshu), and Ashi points. Patients' clinical symptoms, RA-associated serum markers, and serum levels of TNF-α, IL-1ß, HIF-1α, and VEGF were compared in the two groups before and after intervention. Statistical analysis was performed using SPSS 21.0 statistical software. Results: 37 of 46 RA patients eventually completed the whole treatment course. Compared with the control group, the treatment group significantly improved the clinical symptoms (P < 0.05) but with no significant differences in RA-associated serum markers (P > 0.05). There were significant differences in TNF-α and IL-1ß among the groups after 8 weeks of treatment (P < 0.05). HIF-1α and VEGF were decreased in the treatment group after therapy (P < 0.05). VEGF was reduced in the control group (P < 0.05), while HIF-1α was not significantly improved (P > 0.05). The reductions of HIF-1α and VEGF in the treatment group were superior to the control group (P < 0.05). Conclusions: Moxibustion enhanced the anti-inflammatory and analgesic effects of conventional medicine and can enhance the effect of conventional medicine, downregulating HIF-1α/VEGF contents to inhibit angiogenesis.

Effect of charred Radix et Rhizoma Rhei in a laser-induced choroidal neovascularization murine model.

A pharmaceutical composition (patent no. WO2012079419) exhibited favorable outcomes in a clinical trial of wet age­related macular degeneration. The aims of the present study were to explore the effects of one composition component, charred Radix et Rhizoma Rhei (CRRR), in a laser­induced choroidal neovascularization (CNV) murine model. A total of 30 eight­week­old C57BL/6 mice were subjected to diode laser treatment, and CNV was induced by rupturing the Bruch's membrane. The mice were then randomly divided into two groups: the CRRR­treated group that was administered CRRR water extract (concentration, 0.6 g/100 ml; dose, 1 ml/0.1 kg twice a day for 21 days); and the control group that was treated with saline (dose, 1 ml/0.1 kg twice a day for 21 days). The retinal tissue was subjected to quantitative polymerase chain reaction (qPCR) and western blot analysis to determine the expression levels of interleukin­10 (IL­10) and vascular epithelial growth factor (VEGF) at day seven following laser treatment. At weeks 2 and 3 after laser treatment, fundus fluorescein angiography was performed and graded to assess the severity of lesion leakage. Retinal flat mounts were prepared for three­dimensional confocal microscopy at day 22 after laser treatment. At days 14 and 21 after laser treatment, no statistically significant differences were observed between the clinically relevant lesions of the CRRR­treated and control mice. CNV volumes were not found to be significantly different between the CRRR­treated and control mice. The expression levels of IL­10 were significantly increased in the CRRR­treated mice (P<0.05). However, no statistically significant differences were observed between the VEGF expression levels of the CRRR­treated and control mice. In conclusion, CRRR did not appear to significantly inhibit CNV in this murine model. The function of CRRR in the pharmaceutical composition may be due to the effects of IL­10 and a synergistic effect with other components of the composition. However, further investigation is required.

[Chinese herbal medicine Lingqi Huangban Granule protects retinal pigment epithelial cells against oxidative stress-induced injury in vitro].

OBJECTIVE: To observe the protective effects of drug-contained serum of Lingqi Huangban Granule (LQHBG), a compound traditional Chinese herbal medicine, on oxidative stress-induced injury in rabbit retinal pigment epithelial (RPE) cells in vitro. METHODS: The oxidative stress of rabbit RPE cells in vitro was induced with hydrogen peroxide (500µmol/L) and different concentrations of LQHBG were administered to rats to prepare medicated serum. RPE cells were randomized into normal control group (no hydrogen peroxide), model group (hydrogen peroxide), model plus serum group (hydrogen peroxide and 10% control serum), model plus low-dose LQHBG group (hydrogen peroxide and low-dose LQHBG-medicated serum) and model plus high-dose LQHBG group (hydrogen peroxide and high-dose LQHBG-medicated serum). Teminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assay and flow cytometry (FCM) were used to measure apoptosis of cultured rabbit RPE cells. Protein expressions of caspase-3 and Bcl-X(L) were observed by Western blot method. RESULTS: FCM results showed that the apoptotic rates of the normal control group, model group, control serum group and serum containing low- and high-dose LQHBG groups were (4.85±0.26)%, (20.02±1.37)%, (21.84±0.94)%, (13.56±0.55)%, and (8.58±0.39)%, respectively; compared with the model group, the apoptotic rates of RPE cells in the low- and high-dose LQHBG groups were obviously reduced in a dose-related manner (P<0.05). TUNEL results showed that nuclei of apoptotic cells were stained brown; the number of apoptotic cells in the low- and high-dose LQHBG groups was obviously less than that in the model group. The protein expression of caspase-3 was up-regulated in the model and control serum groups, which was higher than that in the high-dose LQHBG group (P<0.05). The protein expression of Bcl-X(L) was down-regulated in the model and control serum groups, which was lower than that in the low- and high-dose LQHBG groups (P<0.01). CONCLUSION: Drug-contained serum of LQHBG obviously reduces apoptosis and partly protects rabbit RPE cells from oxidative stress-induced injury. The protective function is due to an improvement in antioxidant abilities, down-regulation of the expression of caspase-3 and up-regulation of the expression of Bcl-X(L).

[Protective effect of Huangban Granule against light-induced retinal damage in rats].

OBJECTIVE: To observe the protective effect of Huangban Granule, a compound of traditional Chinese herbal medicine, on rats with retinal damage induced by light. METHODS: A total of 24 male Sprague-Dawley (SD) rats were randomly divided into normal control group, untreated group and Huangban Granule group. Retinal light damage was induced by exposure to constant white fluorescent light for 5 hours at an illumination of 2,800 Lux. The Huangban Granule was given 10 days before light exposure until the animals were sacrificed in Huangban Granule group, and an equal volume of distilled water for the rats in untreated group. Electroretinogram (ERG) was recorded in all animals 2 weeks after light exposure and the animals were sacrificed for histopathological examination of retina. The outer nuclear layers (ONLs) on the superior and inferior retina were counted. RESULTS: Fourteen days after light exposure, the ONLs on the superior retina were 3 to 6 in the untreated group and 7 to 9 in treatment group. There were 9 to 11 layers in normal group. The mean number of ONLs in the untreated group (4.68+/-1.64) was less than that in the treatment group (8.23+/-1.35) (P<0.01). B-wave amplitudes were (319.38+/-71.96) muV and (135.16+/-42.30) muV in Huangban Granule group and the untreated group respectively (P<0.01). A-wave amplitudes were (184.63+/-47.23) muV and (83.35+/-27.75) muV (P<0.01), and oscillatory potential amplitudes were (239.38+/-20.19) muV and (125.44+/-26.23) muV (P<0.01) respectively in the two group. There was no significant difference in implicit times of a-wave and b-wave among the three groups. CONCLUSION: Huangban Granule obviously protects both function and morphology of the retina from light-induced retinal damage in rats.

Intraperitoneal injection of Ginkgo biloba extract enhances antioxidation ability of retina and protects photoreceptors after light-induced retinal damage in rats.

PURPOSE: To investigate the effect of Ginkgo biloba extract EGb 761, a free-radical scavenger, on the antioxidation capability of retina after light-induced retinal damage in rats in an attempt to understand the mechanism by which EGb 761 protects the photoreceptors after light-induced retinal damage. METHODS: Seventy-two female Sprague-Dawley (SD) rats were evenly randomized into normal control group (NC group), light-induced retinal damage model group (M group), model + normal saline group (MN group), and model + EGb 761 group (ME group). Light-induced retinal damage model was induced via exposure to white light at 2740 +/- 120 lux for 6 hr. Rats in MN group and ME group were intraperitoneally injected daily with normal saline and 0.35% EGb 761 (100 mg/kg), respectively, 1 week before and 2 weeks after light exposure. The levels of malondialdehyde (MDA), total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) in the retinal tissues were determined 24 hrs after light exposure; photoreceptor apoptosis was detected 4 days after light exposure. One and 2 weeks after light exposure, histopathologic examination was carried out, and the outer nuclear layer (ONL) thickness (number of nuclei) in the superior and inferior retina was counted. RESULTS: Twenty-four hours after exposure, the MDA levels in the other three groups were significantly higher than that in the NC group (p < 0.05); those in the M and MN groups were similar to each other (p > 0.05); and that of the ME group was significantly lower than those in the M and MN group (p < 0.05). The activities of T-SOD, GSH-Px, and CAT were similar in the M and MN groups (p > 0.05); the activities in the M and MN groups were significantly lower than those in the NC and ME groups (p < 0.05); and the activities in the ME group were significantly higher than those in the M and MN groups (p < 0.05). Four days after exposure, the apoptotic photoreceptors within the ONL in the ME group were obviously fewer than those in the M and MN groups. One week and 2 weeks after exposure, the ONL thickness (number of nuclei) in the ME group was more than that in the M and MN groups but less than that in the NC group. CONCLUSIONS: Intraperitoneal injection of EGb 761 can enhance the antioxidation ability of retina and partially inhibit the apoptosis of photoreceptors, thus exert a protective effect on photoreceptors.