Exosomes are nanoscale extracellular vesicles that are secreted by a variety of cells in the body. They carry particular miRNA, protein molecules, transcription factors, and other information molecules, and they play a role in the pathophysiological regulation of a number of diseases in the body. Exosomes can persist steadily in biological tissues and bodily fluids. Exosomes have quickly advanced in ophthalmology in recent years due to the extensive studies of exosomes in a variety of fields, such as diabetic retinopathy, age-related macular degeneration, autoimmune uveitis, corneal disease, glaucoma, and other diseases. The number of people who are blind caused by diabetic retinopathy is rising as living standards rise. However, it is still unclear how diabetic retinopathy works. In recent years, many studies have found that exosomes play an important role in diabetic retinopathy. In this paper, the most recent developments in exosome studies as they relate to the pathogenesis and progression of diabetic retinopathy are reviewed.

Though cells are easily damaged and performed metabolic disorders in freezing-thawing process, cryoprotectant can relief the injure, which is widely used in the field of cell frozen biology. As one of osmotic cryoprotectants, in freezing process, proline can penetrate through cell membrane quickly, bind intracellular water hydrogen bonds, inhibit formation of ice crystals at low temperatures, protect structure and function of proteins and cell membranes. It can increase the survival rate in plant cells, animal cells and human somatic cells post freezing-thawing. In germ cell cryopreservation, studies have shown that proline can be used for oocyte cryopreservation, which can improve the survival rate of mouse oocytes after cryopreservation and protect mitochondrial function. However, the reports about the cryopreservation efficiency of proline on sperm are not consistent. By improving the efficiency of cryopreservation in various kinds of cells, proline may be more widely used in the area of cell freezing biology in the future. This paper summarizes the research progress of proline as a natural small molecule cryoprotectant, hoping to help people better understand the cryobiological mechanism of germ cell cryopreservation, and provide a reference for improving fertility cryopreservation technology.

Though cells are easily damaged and performed metabolic disorders in freezing-thawing process, cryoprotectant can relief the injure, which is widely used in the field of cell frozen biology. As one of osmotic cryoprotectants, in freezing process, proline can penetrate through cell membrane quickly, bind intracellular water hydrogen bonds, inhibit formation of ice crystals at low temperatures, protect structure and function of proteins and cell membranes. It can increase the survival rate in plant cells, animal cells and human somatic cells post freezing-thawing. In germ cell cryopreservation, studies have shown that proline can be used for oocyte cryopreservation, which can improve the survival rate of mouse oocytes after cryopreservation and protect mitochondrial function. However, the reports about the cryopreservation efficiency of proline on sperm are not consistent. By improving the efficiency of cryopreservation in various kinds of cells, proline may be more widely used in the area of cell freezing biology in the future. This paper summarizes the research progress of proline as a natural small molecule cryoprotectant, hoping to help people better understand the cryobiological mechanism of germ cell cryopreservation, and provide a reference for improving fertility cryopreservation technology.

Background: The accessory tendon of the extensor hallucis longus (ATEHL) muscle is a common abnormal structure, and its clinical significance remains debatable. In this study, we provide the incidence of the ATEHL and characterize its morphological types in Asian cadavers and investigate its clinical applications. Methods: The tendons from 50 adult cadaveric feet, fixed in 10% formalin, were analyzed. We measured the length and width of both the ATEHL and the extensor hallucis brevis (EHB). Results: All dissected specimens had an ATEHL. The first metatarsophalangeal joint was surrounded by an accessory tendon that inserted onto the joint capsule and the dorsal base of the proximal phalanx. We classified the ATEHL into 3 types based on their directions. Differences in ATEHL type based on sex were not statistically significant. Conclusions We found an ATEHL in all cadaveric specimens in this study. We surmise that the ATEHL acts as an antagonist with the EHB when the toe is extending, which might help prevent the occurrence of hallux valgus deformity.

Oocyte cryopreservation is widely used for clinical and social reasons. Previous studies have demonstrated that conventional slow-freezing cryopreservation procedures, but not storage time, can alter the gene expression profiles of frozen oocytes. Whether vitrification procedures and the related frozen storage durations have any effects on the transcriptomes of human metaphase II oocytes remain unknown. Four women (30-32 years old) who had undergone IVF treatment were recruited for this study. RNA-Seq profiles of 3 fresh oocytes and 13 surviving vitrified-thawed oocytes (3, 3, 4, and 3 oocytes were cryostored for 1,2, 3, and 12 months) were analyzed at a single-cell resolution. A total of 1987 genes were differentially expressed in the 13 vitrified-thawed oocytes. However, no differentially expressed genes were found between any two groups among the 1-, 2-, 3-, and 12-month storage groups. Further analysis revealed that the aberrant genes in the vitrified oocytes were closely related to oogenesis and development. Our findings indicated that the effects of vitrification on the transcriptomes of mature human oocytes are induced by the procedure itself, suggesting that long-term cryostorage of human oocytes is safe.
Adult ; Cryopreservation ; Female ; Humans ; Metaphase ; Oocytes ; RNA-Seq ; Vitrification

Background: The accessory tendon of the extensor hallucis longus (ATEHL) muscle is a common abnormal structure, and its clinical significance remains debatable. In this study, we provide the incidence of the ATEHL and characterize its morphological types in Asian cadavers and investigate its clinical applications. Methods: The tendons from 50 adult cadaveric feet, fixed in 10% formalin, were analyzed. We measured the length and width of both the ATEHL and the extensor hallucis brevis (EHB). Results: All dissected specimens had an ATEHL. The first metatarsophalangeal joint was surrounded by an accessory tendon that inserted onto the joint capsule and the dorsal base of the proximal phalanx. We classified the ATEHL into 3 types based on their directions. Differences in ATEHL type based on sex were not statistically significant. Conclusions We found an ATEHL in all cadaveric specimens in this study. We surmise that the ATEHL acts as an antagonist with the EHB when the toe is extending, which might help prevent the occurrence of hallux valgus deformity.

ObjectiveTo assess the diagnostic performance of deep learning elastography （DLE） for liver fibrosis. MethodsTotally 545 chronic liver disease patients with liver biopsy were retrospectively enrolled. The results of DLE， two dimensional shear wave elastography （2D-SWE）， serological markers and transient elastography （TE） were collected. Area under receiver operating curve （AUC） was calculated and inter-compared while evaluating liver fibrosis. Then we tested the diagnostic performance of the trained DLE model in different validation groups when evaluating the same liver fibrosis stage， respectively， to assess the stability of DLE. ResultsDLE showed statistically significantly （P＜0.05） better results than any other methods. When evaluating F=4， F≥3 and F≥2， the AUC of DLE was 0.99， 0.98 and 0.92， respectively. 2D-SWE showed a second high diagnostic performance， while the AUCs were 0.89， 0.86 and 0.86， respectively. Little difference in diagnostic performance was showed among other methods， while the highest AUC was no more than 0.81. Besides， no statistical difference was showed among the three validation groups， while accessing the same liver fibrosis stage. ConclusionsDLE can be used to accurately assess liver fibrosis， whose diagnostic performance is higher than that of 2D-SWE， serological markers and TE. Moreover， with good stability， DLE is expected to become a new method for noninvasive assessment of liver fibrosis.

OBJECTIVETo compare the expression of pinopodes, the marker of endometrial receptivity, during the implantation window in Kunming mice stimulated with two different doses of raloxifene (RAL).

METHODSForty-eight 8-week-old female Kunming mice were randomly divided into 4 groups (n=12), namely saline group, clomiphene citrate (CC, 18 mg/kg) group, RAL (33 mg/kg) group and RAL (44 mg/kg group). In each group, the mice received intragastric administration of 1 mL of normal saline containing CC or RAL at the specified doses or saline only as indicated for ovulation induction, once daily for 2 days. The mice received then injection with 5 IU human chorionic gonadotropin (HCG) and mated and on day 4.5 of gestation, the pregnant mice were sacrificed for examination of the uterus with scanning electron microscopy.

RESULTSAbundant and well developed pinopodes were observed in the endometrium of the mice in the 2 RAL groups and in the saline control group. The mice in CC group showed obviously reduced endometrial pinopodes with poor development.

CONCLUSIONSRAL at two different doses does not obviously affect the expression of pinopodes in the uterine epithelium of mice, suggesting the safety of RAL at these two doses for ovulation induction without causing adverse effects on endometrial receptivity.

PURPOSE: In this study, we aimed to evaluate lymphocyte-activation gene-3 (LAG-3) expression and its prognostic value in neoadjuvant-treated triple-negative breast cancer (TNBC). METHODS: LAG-3, programmed death-1 (PD-1), programmed death ligand-1 (PD-L1), and CD8⁺ tumor-infiltrating lymphocyte (TILs) levels were examined using immunohistochemistry in 148 preand 114 post-neoadjuvant chemotherapy (NACT) specimens of human TNBC tissue. Correlations between expression levels and clinicopathological features were analyzed. Prognostic values for combined detection in TNBC following NACT were evaluated. RESULTS: In pre-NACT specimens, LAG-3 expression showed a significant association with pathological complete response (pCR, p=0.038) and was correlated with PD-1 (p<0.001) and PD-L1 (p=0.008). In post-NACT specimens, high expression of LAG-3 showed significant effects on nodal status (p=0.023) and PD-1 (p<0.001). The expression of immune markers on TILs significantly increased following NACT. Multivariate analysis indicated that only nodal status (odds ratio [OR], 0.226; 95% confidence interval [CI], 0.079–0.644; p=0.005) and high quantities of CD8⁺TILs (OR, 3.186; 95% CI, 1.314–7.721; p=0.010) are independent predictors of pCR. Nodal status (hazard ratio [HR], 2.666; 95% CI, 1.271–5.594; p=0.010), CD8⁺TILs (HR, 0.313; 95% CI, 0.139–0.705; p=0.005), and the LAG-3-high/PD-L1-high group (HR, 2.829; 95% CI, 1.050–7.623; p=0.040) provided prognostic values for patients with TNBC following NACT. CONCLUSION: CD8+TILs were sensitive predictive markers in response to NACT. High expression of LAG-3 in residual tissues, especially in combination with PD-L1, was associated with poor prognosis.
Biomarkers ; Drug Therapy ; Humans ; Immunohistochemistry ; Lymphocytes, Tumor-Infiltrating ; Multivariate Analysis ; Neoadjuvant Therapy ; Polymerase Chain Reaction ; Prognosis ; Triple Negative Breast Neoplasms*

OBJECTIVETo compare the expression of DKKL1 in ejaculated spermatozoa of normal fertile men and men with asthenospermia and investigate the role of DKKL1 in the pathogenesis of asthenospermia.

METHODSThe characteristics of semen samples collected from normal fertile men and men with asthenospermia were analyzed using computer-assisted sperm analysis according to WHO criteria. The ejaculated sperms were isolated by Percoll discontinuous density gradients to detect the expression of DKKL1 mRNA and protein using real-time PCR and Western blotting.

RESULTSThe expression of DKKL1 mRNA was significantly down-regulated by 11.1 times in asthenospermic men as compared with that in normal fertile men (P<0.01). Western blotting showed that the expression of DKKL1 protein was down-regulated by 2.4 times in asthenospermic men compared to normal fertile men.

CONCLUSIONThe expression of DKKL1, which may play an important role in sperm motility,is significantly decreased in ejaculated spermatozoa of men with asthenospermia.