Rosmarinic acid activates the Ras/Raf/MEK/ERK signaling pathway to regulate CD8+ T cells and autophagy to clear Chlamydia trachomatis in reproductive tract-infected mice.

Chlamydia trachomatis (CT) is the leading cause of bacterial sexually transmitted diseases worldwide, which can cause diseases such as pelvic inflammatory disease, and cervical and fallopian tube inflammation, and poses a threat to human health. Rosmarinic acid (RosA) is an active ingredient of natural products with anti-inflammatory and immunomodulatory effects. This study aimed to investigate the role of RosA in inhibiting autophagy-regulated immune cells-CD8+ T cells via the Ras/Raf/MEK/ERK signaling pathway in a CT-infected mouse model. Mice were inoculated with CT infection solution vaginally, and the mechanistic basis of RosA treatment was established using H&E staining, flow cytometry, immunofluorescence, transmission electron microscopy, and western blot. The key factors involved in RosA treatment were further validated using the MEK inhibitor cobimetinib. Experimental results showed that both RosA and the reference drug azithromycin could attenuate the pathological damage to the endometrium caused by CT infection; flow cytometry showed that peripheral blood CD8+ T cells increased after CT infection and decreased after treatment with RosA and the positive drug azithromycin (positive control); immunofluorescence showed that endometrial CD8 and LC3 increased after CT infection and decreased after RosA and positive drug treatment; the results of transmission electron microscopy showed that RosA and the positive drug azithromycin inhibited the accumulation of autophagosomes; western bolt experiments confirmed the activation of autophagy proteins LC3Ⅱ/Ⅰ, ATG5, Beclin-1, and p62 after CT infection, as well as the inhibition of Ras/Raf/MEK/ERK signaling. RosA and azithromycin inhibition of autophagy proteins activates Ras/Raf/MEK/ERK signaling. In addition, the MEK inhibitor cobimetinib attenuated RosA's protective effect on endometrium by further activating CD8+ T cells on a CT-induced basis, while transmission electron microscopy, immunofluorescence, and western blots showed that cobimetinib blocked ERK signals activation and further induced phagocytosis on a CT-induced basis. These data indicated that RosA can activate the Ras/Raf/MEK/ERK signaling pathway to inhibit autophagy, and RosA could also regulate the activation of immune cells-CD8+T cells to protect the reproductive tract of CT-infected mice.

Clinical characteristics of glaucoma associated with primary retinitis pigmentosa / 中华眼底病杂志

Objective:To observe the clinical characteristics of primary retinitis pigmentosa (RP) complicated with glaucoma.Methods:A retrospective clinical study. From June 2008 to March 2020, the diagnosis of primary RP were included in the diagnosis confirmed by the eye examination of West China Hospital of Sichuan University included 4794 eyes of 2432 patients. Among them, 4679 eyes (97.2%, 2364/2432) were in 2364 cases with RP alone, and 115 eyes were in 68 cases with RP combined with glaucoma (2.80%, 68/2432). All affected eyes underwent best corrected visual acuity (BCVA) and intraocular pressure examination. The BCVA examination was carried out using the international standard visual acuity chart, which was converted into the logarithmic minimum angle of resolution (logMAR) visual acuity during statistics. The 67 eyes of 40 patients with RP and glaucoma with complete follow-up data were analyzed to observe the proportion of different glaucoma types, logMAR BCVA, intraocular pressure and other clinical characteristics, as well as treatment methods and post-treatment intraocular pressure control. After treatment, the intraocular pressure ≤21 mm Hg (1 mm Hg=0.133 kPa) was regarded as intraocular pressure (IOP) control; >21 mm Hg was regarded as uncontrolled IOP.Results:Among the 67 eyes of 40 cases with complete follow-up data, 5 cases (7 eyes) with primary open-angle glaucoma (10.45%, 7/67), 56 cases (58 eyes) with angle-closure glaucoma (ACG) (86.57%, 58/67), 4 cases (4 eyes) with neovascular glaucoma (5.97%, 4/67), 2 of them had both ACG and neovascular glaucoma. Among 58 ACG eyes, 17 eyes were acute ACG (25.37%, 17/67), 21 eyes were chronic ACG (31.34%, 21/67), and 2 eyes were suspicious angle closure (2.99%, 2/67), lens dislocation secondary to angle-closure glaucoma in 8 eyes (11.94%, 8/67), chronic angle-closure glaucoma after anti-glaucoma surgery, intraocular lens shift in 5 eyes (7.46%, 5/67), 5 eyes (7.46%, 5/67) secondary to glaucoma with true small eyeballs. The logMAR BCVA 3.50 of the affected eye,<3.50->2.00, ≤2.00-≥1.30,<1.30->1.00, ≤1.00- 0.52,<0.52 were 9 (13.43%, 9/67), 30 (44.78%, 30/ 67), 7 (10.45%, 7/67), 4 (5.97%, 4/67), 11 (16.42%, 11/67), 6 (8.96%, 6/67) eyes, which correspond to mean intraocular pressure were 32.31±11.67, 30.15±14.85, 28.17±13.19, 31.50±17.25, 18.71±8.85, 14.12±4.25 mm Hg. Among 67 eyes, 37eyes (55.22%, 37/67), 18eyes (26.86%, 18/67), and 6 (8.96%, 6/67) eyes underwent surgery, medication alone, and peripheral iris laser perforation treatment, respectively. The treatment of 6 eyes was abandoned (8.96%, 6/67). Malignant glaucoma occurred in 3 eyes (8.11 %, 3/37) after the operation, all of which were after trabeculectomy of the ACG eye. After treatment, intraocular pressure was controlled in 37 eyes (55.22%, 37/67), 19 eyes were not controlled (28.36%, 19/67), and 11 eyes were lost to follow-up (16.42%, 11/67).Conclusions:The incidence of glaucoma in patients with primary RP is 2.80%. ACG is more common, and the combined lens dislocation or intraocular lens shift is more common.

Research progress of programmed cell death 1 and its receptor programmed cell death 1 ligand 1 in the pathogenesis of eye diseases / 中华实验眼科杂志

Programmed cell death 1 (PD-1) is an important negative costimulatory molecule discovered in recent years, which is expressed on the surface of T and B cells and plays an important role in the regulation of cellular immune response and immune tolerance.After the combination of PD-1 and its receptor programmed cell death 1 ligand 1 (PD-L1), negative regulatory signals are transmitted to suppress the immune response.Under the activation of stimulating factors, PD-1 and PD-L1 are combined to weaken the conduction of downstream signaling pathways such as PI3K/Akt and ERK through recruitment of phosphorylated SHP2, thereby inhibiting the proliferation of T cells and the production of cytokines, inhibiting the immune response and participating in the occurrence and development of a large number of inflammatory diseases.PD-1 in the field of ophthalmology research is still in its infancy.As far as we know, PD-1 participates in ocular inflammatory diseases such as uveitis, sympathetic ophthalmia, allergic conjunctivitis, also is involved in corneal transplant rejection, optic nerve crush injury and optic myelitis, diabetic retinopathy, thyroid related ophthalmopathy, melanoma and other diseases, thus preventing the interactions between PD-1 and its receptor PD-L1 may become a new potential target for the treatment of ocular tumor, inflammation, autoimmune and neurodegeneration disease.In this article, the latest research progress of PD-1 and its receptor PD-L1 in the pathogenesis of ocular diseases were reviewed.

The oncosis and apoptosis-inducing effect of artesunate on the K562 human leukemia cell line / 重庆医科大学学报

Objective:To investigate the effect of artesunate(ART) on human leukemia cell line K562. Methods:Inhibition of proliferation was measured by a colorimetric MTT assay. The morphological changes of K562 were observed under a light microscope and an electron microscope. Flow cytometry assay detected the ratio of apoptosis and oncosis of K562 cell lines which were induced at three different times by three different concentrations of ART. Results:ART extract clearly inhibited the proliferation of K562. The 50% effective dose evaluated on 24h,48h,and 72h of ordinal exposure to the extract,were 65.17?g/ml,31.63?g/ml,and 10.51?g/ml,respectively,ART extract-treated cells exhibited morphological changes typical of oncosis and apoptosis. Futheremore,the ratio of apoptosis was not different from to that of oncosis. Conclusion:Artesunate can control proliferation of the K562 human leukemia cell line in two forms:oncosis and apoptosis. The ratio of oncosis and apoptosis present typical dose-dependence and time-dependence.