Periplaneta Americana (L.) extract activates the ERK/CREB/BDNF pathway to promote post-stroke neuroregeneration and recovery of neurological functions in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Periplaneta americana (L.) (PA) has been used in traditional Chinese medicine for thousands of years for the effect of invigorating blood circulation and removing blood stasis. Modern pharmacological research shown that PA extract exhibits promising effects in promoting wound healing and regeneration, as well as in brain diseases such as Parkinson's disease (PD). However, whether it is effective for neuroregeneration and neurological function recovery after stroke still unknown. AIM OF THE STUDY: This study aims to investigate the potential effect of PA extract to promote brain remodeling through the activation of endogenous neurogenesis and angiogenesis, in addition, preliminary exploration of its regulatory mechanism. METHODS: Firstly, BrdU proliferation assay and immunofluorescence (IF) staining were used to evaluate the effect of PA extract on the neurogenesis and angiogenesis in vitro and in vivo. Subsequently, the effects of PA extract on brain injury in stroke rats were assessed by TTC and HE. While mNSS score, adhesive removal test, rota-rod test, and morris water maze test were used to assess the impact of PA extract on neurological function in post-stroke rats. Finally, the molecular mechanisms of PA extract regulation were explored by RNA-Seq and western blotting. RESULTS: The number of BrdU+ cells in C17.2 cells, NSCs and BMECs dramatically increased, as well as the expression of astrocyte marker protein GFAP and neuronal marker protein Tuj-1 in C17.2 and NSCs. Moreover, PA extract also increased the number of BrdU+DCX+, BrdU+GFAP+, BrdU+CD31+ cells in the SGZ area of transient middle cerebral artery occlusion model (tMCAO) rats. TTC and HE staining revealed that PA extract significantly reduced the infarction volume and ameliorated the pathological damage. Behavioral tests demonstrated that treatment with PA extract reduced the mNSS score and the time required to remove adhesive tape, while increasing the time spent on the rotarod. Additionally, in the morris water maze test, the frequency of crossing platform and the time spent in the platform quadrant increased. Finally, RNA-Seq and Western blot revealed that PA extract increased the expression of p-ERK, p-CREB and BDNF. Importantly, PA extract mediated proliferation and differentiation of C17.2 and NSCs reversed by the ERK inhibitor SCH772984 and the BDNF inhibitor ANA-12, respectively. CONCLUSION: Our study demonstrated that PA extract promoted neurogenesis and angiogenesis by activating the CREB/ERK signaling pathway and upregulating BDNF expression, thereby recovering neurological dysfunction in post-stroke.

Analysis of Clinical Characteristics of 52 Patients with Uveitis before and after Vitrectomy and Factors Affecting Clinical Efficacy.

OBJECTIVE: To investigate the pre- and postsurgical clinical characteristics and clinical efficacy of patients with uveitis. METHODS: The clinical data of patients with uveitis who underwent vitrectomy in our hospital from March 2019 to February 2021 were retrospectively analyzed. There were 52 cases of 64 eyes in total. The data on patient's gender, age, etiology, course of disease, anatomical classification, number of recurrences, changes in vision before and after surgery, changes in eye signs before and after surgery, and occurrence of postoperative complications were collected. The clinical features before and after vitrectomy were compared, and the influencing factors of clinical efficacy were analyzed. RESULTS: The ocular signs of patients with uveitis after vitrectomy were significantly improved compared with before operation, and the difference was statistically significant (P < 0.05). The visual acuity after vitrectomy in patients with uveitis was significantly improved compared with that before operation, and the difference was statistically significant (P < 0.05). There was no significant difference in the surgical treatment of uveitis patients of different gender, age, and etiology (P > 0.05). There are significant differences in the clinical efficacy of vitrectomy in patients with different anatomical classifications. Among them, patients with panuveitis have the best clinical efficacy with vitrectomy and patients with posterior uveitis have the worst clinical efficacy with vitrectomy (P < 0.05). There is a significant difference in the clinical efficacy of vitrectomy in patients with recurrence times. The lower the number of recurrences, the better the clinical efficacy of vitrectomy in patients (P < 0.05). CONCLUSION: There are significant differences in the clinical signs of patients with uveitis before and after vitrectomy. Vitrectomy is effective in the treatment of uveitis. The type of anatomy and the number of recurrences are influencing factors for the clinical efficacy of vitrectomy. For patients with posterior uveitis, the surgical method should be carefully considered or a more reasonable treatment method should be selected, and for patients with uveitis with less recurrence, vitrectomy should be considered for active treatment.

[Role of Wnt/ß-catenin signaling in aging of mesenchymal stem cells of rats].

OBJECTIVE: To investigate the role of Wnt/ß-catenin signaling in aging of mesenchymal stem cells (MSCs) of rats. METHODS: Serum samples were collected from young (8 ≈ 12 w) and aged (64 ≈ 72 w) SD rat. Four experiment groups were assigned: young rat serum (YRS), YRS+Wnt 3a, old rat serum (ORS) and ORS+DKK1 groups. Immunofluorescence and Western blotting were used to detect the expression of intracellular ß-catenin. The senescence-associated changes were examined with SA-ß-galactosidase staining. The proliferation ability was tested by MTT assays. The survived and apoptotic cells were determined by AO/EB staining. The expressions of γ-H2A. X and p53 protein were detected by immunofluorescence and Western blotting. RT-PCR was used to detect the expression of p53 and p21 mRNA. RESULTS: Compared with the YRS group, the intracellular expression of ß-catenin in the ORS group was significantly increased,especially in the nuclei of MSCs. After treatment of DKK1 in ORS, the γ-catenin expression was reduced. The number of SA-ß-galactosidase positive MSCs was significantly higher in the YRS+Wnt 3a group than that in the YRS group (P<0.01), and the proliferative and survival ability of MSCs was significantly decreased in the YRS+Wnt 3a group. The number of SA-ß-galactosidase positive MSCs in the ORS+DKK1 group was significantly decreased compared with that in ORS group (P <0.01), and the proliferative and survival ability of MSCs was significantly increased in the ORS+DKK1 group. The expression of γ-H2A.X, p53 and p21 was markedly increased in the ORS group than that in YRS group, however, after treatment with Wnt/ß-catenin signaling inhibitor DKK1, the expression of γ-H2A.X, p53 and p21 was significantly decreased compared with that in the ORS group. CONCLUSION: Results suggest that the Wnt/ß-catenin signaling is activated in the MSCs cultured with ORS and excessive activation of Wnt/ß-catenin signaling can promote MSCs aging. The DNA damage response and p53/p21 pathway may be main mediators of MSC aging induced by excessive activation of Wnt/ß-catenin signaling.