Single-cell profiling reveals Müller glia coordinate retinal intercellular communication during light/dark adaptation via thyroid hormone signaling

Light adaptation enables the vertebrate visual system to operate over a wide range of ambient illumination. Regulation of phototransduction in photoreceptors is considered a major mechanism underlying light adaptation. However, various types of neurons and glial cells exist in the retina, and whether and how all retinal cells interact to adapt to light/dark conditions at the cellular and molecular levels requires systematic investigation. Therefore, we utilized single-cell RNA sequencing to dissect retinal cell-type-specific transcriptomes during light/dark adaptation in mice. The results demonstrated that, in addition to photoreceptors, other retinal cell types also showed dynamic molecular changes and specifically enriched signaling pathways under light/dark adaptation. Importantly, Müller glial cells (MGs) were identified as hub cells for intercellular interactions, displaying complex cell‒cell communication with other retinal cells. Furthermore, light increased the transcription of the deiodinase Dio2 in MGs, which converted thyroxine (T4) to active triiodothyronine (T3). Subsequently, light increased T3 levels and regulated mitochondrial respiration in retinal cells in response to light conditions. As cones specifically express the thyroid hormone receptor Thrb, they responded to the increase in T3 by adjusting light responsiveness. Loss of the expression of Dio2 specifically in MGs decreased the light responsive ability of cones. These results suggest that retinal cells display global transcriptional changes under light/dark adaptation and that MGs coordinate intercellular communication during light/dark adaptation via thyroid hormone signaling.

Pathogenesis of glucocorticoid-induced osteoporosis based on label-free mass proteomics / 中国骨伤

OBJECTIVE@#To explore pathogenesis of glucocortocoid-induced osteoporosis(GIOP) based on label-free mass proteomics.@*METHODS@#Twevle female Sprague-Dawley(SD) rats were randomly divided into two groups, named as sham group and GIOP group. After one-week adaptive feeding, the rats of GIOP group were administered with dexamethasone via intramuscular injection according to 2.5 mg/kg weighting, while the rats of sham group were administered with the same amount of saline, twice a week. The tibias of each group were collected after 8-week modeling and made pathological sections to confirm the success of modeling. Three samples of each group were picked up to perform label-free mass proteomics. After quality control, differentially expressed proteins were identified according to qualitative and quantitative analyses. Then gene ontology(GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis, cluster analysis as well as protein-protein interaction analysis were performed using bioinformatics analysis.@*RESULTS@#Compared with sham group, the structure of bone trabecular in GIOP group showed abnormal arrangement, uneven distribution and obvious fragmentation, which could demonstrate successful modeling. A total of 47 differentially expressed proteins (DEPs) were identified including 20 up-regulated and 27 down-regulated proteins. The expression of protein nucleophosmin 1(NPM1), adipocyte plasma membrane associated protein (APMAP), cytochromec oxidase subunit 6A1 (COX6A1) and tartrate-resistant acid phosphatase (ACP5) showed a significant difference between two groups. KEGG results showed DEPs were enriched on metabolism-related pathways, immune-related pathways and AMP-activated kinase (AMPK) signaling pathway.@*CONCLUSION@#Protein NPM1, APMAP, COX6A1 and ACP5 showed a close relationship with pathogenesis of GIOP, which could serve as potential biomarkers of GIOP. AMPK signaling pathway played an important role in the occurrence and development of GIOP, which could be regarded as potential signaling pathway to treatment GIOP.

Chemical constituents from Callicarpa kwangtungensis and their hemostatic activities / 中国中药杂志

This study aims to identify the chemical constituents from Callicarpa kwangtungensis and determine their activities. MCI, ODS, and Sephadex LH-20 chromatography and semi-preparative HPLC were employed to separate the chemical constituents. A total of 15 compounds were separated, and their structures were identified on the basis of spectroscopic analysis and comparison with the data in relevant literature. Specifically, the 15 compounds were 3-O-α-L-rhamnopyranosyl-6-O-β-D-apiofuranosyl-4-O-E-caffeoyl-D-glucopyranoside(1), 3,6-O-α-L-dirhamnopyranosyl-4-O-E-caffeoyl-D-glucopyranoside(2), β-OH-forsythoside B(3), β-OH-poliumoside(4),(+)-lyoniresinol-3α-O-β-D-apiofuranosyl-(1→2)-β-D-glucopyranoside(5),(+)-lyoniresinol-3α-O-β-D-glucopyranoside(6),(-)-lyoniresinol-3α-O-β-D-glucopyranoside(7), kelampayoside A(8), descaffeoylpoliumoside(9), acteoside(10), alyssonoside(11), poliumoside(12), isacteoside(13), acetyl forsythoside B(14), and forsythoside B(15). Compounds 1 and 2 were novel, and the NMR data of compounds 3 and 4 were reported here for the first time. Furthermore, the hemostatic activities of the extract and abundant ingredients(compounds 12 and 15) of C. kwangtungensis were determined with Yunnan Baiyao as the positive control and normal saline as the negative control. The extract and compounds 12 and 15 significantly shortened the tail tip bleeding time in mice.

Fluid?attenuated inversion recovery diffusion?weighted imaging (DWI) for evaluating chemotherapy response in patients with acute leukemia: Comparison with conventional DWI

Background: At present, the diagnosis and efficacy evaluation of acute leukemia (AL) are assessed by bone marrow aspiration, which is invasive and subject to sampling errors. Therefore, there is a pressing need to develop a noninvasive and accurate imaging method to evaluate bone marrow changes in patients with AL. This study aimed to compare the apparent diffusion coefficient (ADC) values obtained from fluid?attenuated inversion recovery diffusion?weighted imaging (FLAIR?DWI) and conventional DWI in the lumbar bone marrow of patients with AL and to investigate their performance for evaluating response to induction chemotherapy. Methods: A total of 28 patients with newly diagnosed AL and 25 patients with AL after induction chemotherapy underwent MRI scans at 1.5 Tesla using a conventional DWI and a FLAIR?DWI sequence on sagittal planes covering the lumbar bone marrow. Further, the ADC values from these two sequences, denoted as ADCCON and ADCFLAIR, were measured on multiple vertebrae. The percentage of leukemia cells in bone marrow was recorded, and bone marrow aspiration was performed on treated patients to determine complete remission (CR) and nonremission (NR). Results: ADCFLAIR [(0.453 ± 0.103) × 10?3 mm2/s] was significantly lower than ADCCON [(0.486 ± 0.096) × 10?3 mm2/s] in the 28 untreated patients (t = 3.051, P = 0.005). In the 25 treated patients, ADCFLAIR and ADCCON values [(0.566 ± 0.239) × 10?3 mm2/s] and [(0.716 ± 0.235) × 10?3 mm2/s], respectively, were higher compared with the untreated patients. The ADCCON values showed a nonsignificant difference between the CR (n = 18) and NR (n = 7) groups (t = 1.409, P = 0.305). However, the ADCFLAIR values exhibited statistically significant difference (t = 2.542, P = 0.018) between the two groups. In a receiver operator characteristic (ROC) analysis, the area under the curve (AUC) using ADCFLAIR (0.770) was larger than that of ADCCON (0.611) in distinguishing the CR and NR patients following the chemotherapy. Conclusion: Although both ADCCON and ADCFLAIR are sensitive to tissue changes induced by chemotherapy, FLAIR?DWI outperformed conventional DWI in separating AL patients with CR from NR after chemotherapy. A possible mechanism is that FLAIR?DWI suppresses signals from free water, making the ADC measurement more sensitive to structural changes in the bone marrow

Transplantation of retinal organoids in vivo and long-term survival of photoreceptor cells / 中华实验眼科杂志

Objective:To investigate the differentiation process of human-induced pluripotent stem cells (hiPSCs) into retinal organoids (ROs) in vitro and its potential application in cell transplantation, and to provide a fundamental study for stem-cell therapy. Methods:BC1-eGFP hiPSCs were differentiated into neurospheres in directional differentiation medium via suspension culture.On day 7, hiPSCs-induced-neurospheres were seeded onto Geltrex-coated dishes to induce neural retinal (NR) domains.On day 28, the NR domains were manually detached and collected.These NRs were cultured until the maturation of ROs.The quantitative real-time PCR (at week 0, week 7, week 15, week 21 and week 30 group) and immunohistochemistry (at day 8, day 15, week 15, week 21 and week 30 group) were used to characterize the differentiation process of hiPSCs.For ROs transplantation, the ROs were digested, and the cell suspension was injected into the subretinal cavity of Gnat1-/- mice with the outer limiting membrane damaged in retina.Immunohistochemistry was also used to detect the survival and integration ability of the implanted cells 5 months after transplantation. Results:The morphology and immunofluorescence staining illustrated that the cells induced from hiPSCs highly expressed the neural-retinal-epithelial specific markers PAX6 and SOX1 in the early stage, then the cells expressed the retinal-progenitor-cell specific marker LHX2 and a transparent and horseshoe-shaped NR domain was formed at the outer region of the colony.ROs was obtained by manual isolation and suspension culture.The diameter of ROs was about 1 millimeter.The retinal-like tissue gradually became thicker, even formed retinal pigment epithelial cells.Quantitative real-time PCR results showed that the expression of retinal-progenitor marker VSX2 peaked at week 7 and maintained its high expression thereafter ( F=168.30, P<0.01); the expression of retinal-precursor marker RCVRN also appeared at week 7 and increased thereafter ( F=271.60, P<0.01); the expressions of RHO was detected at week 15 ( F=95.02, P<0.01), and the expression of OPN1LW/MW was detected at week 21 ( F=40.57, P<0.01). Moreover, the expression of photoreceptor protein RHO maintained in a relatively high expression state at week 30.Three weeks after the transplantation of RO cells, cells with green fluorescence were successfully moved into the outer nuclear layer of the host retina.Four to six months after transplantation, the implanted cells expressed the functional light signal transduction protein GNAT1. Conclusions:Transplantation of retinal organoids in vivo can recapitulate the development of human retina.Transplanted RO cells can effectively move into the outer nuclear layer, differentiate into photoreceptors and survive in the recipient mice's retina over several months.

Principle and applications of fluorescent probes for intracellular redox detection / 上海交通大学学报（医学版）

Redox state indicated by reactive oxygen species (ROS) level is a cellular physiological feature.Redox status is exactly controlled by conserved system.Its alterations consequently modulate various biological activities and play important roles in cells and organism under physiological and pathological conditions.Hence detection of redox state is deemed as an important technique both in basic and clinical researches.The present article reviews the principle and current applications of fluorescent probes or sensors for redox detection.With comparison of properties of various probes,especially genetic coding fluorescence probes,much understanding will be gained.And the review helps us to understand how to apply the redox probes on the biological and clinical researches and eventually look forward to new applications in the cell and whole animal level.

Optimization of extraction process of total triterpenoid from root of Rose odorata var.gigantean by orthogonal test and its inhibitory effect on contraction of isolated rat intestinal smooth muscle / 药物评价研究

Objective To optimize the extraction technology of total triterpenoid from root of Rose odorata var.gigantean (TTROG) by orthogonal test combined with the contraction effect of TTROG on the isolated intestinal smooth muscle of rats in vitro.Methods UV spectrophotometric method was used to determine the contents of total triterpenoids in the TTROG extractive at the wavelength of 550 nm by taking ursolic acid as standard substance,and vanillin acetic acid as chromogenic reagent.The extraction rate of total triterpenoids was used as index to evaluate the technology based on single factor test,in which three factors were considered as follows:the concentration of extraction solvent,ratio of material to liquid,extraction time,and their interaction on extraction were studied by orthogonal experimental design.The inhibition effect of different extracts obtained from the optimized extraction process on the contraction of intestinal smooth muscle were recorded by tension transducer to the BL-420 biological experimental multi-channel physiological signal acquisition and processing system.The extraction process of TTROG was evaluated by the combination of biological activity and extraction rate with weighting method.Results The optimal extraction conditions of TTROG were as follows:extraction solvent 80％ ethanol,solid-liquid ratio 1∶10,extraction time for 2 h,three times and extraction temperature of 80 ℃.The optimized extraction rate could reach 42.12 mg/g.TTROG obtained using the optimized method showed significantly contraction effect on rat intestinal smooth muscle with dose effect dependence,and the effect on jejunum was the strongest,and the inhibition rate was 41.96％.Conclusion The optimized extraction technology is stable and effective with high extraction rate.TTROG showed the significant inhibitory function on contraction of isolated rat intestinal smooth muscle.

Effect of proline rich domain of an RNA-binding protein Sam68 in cell growth process, death and B cell signal transduction / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Sam68 plays an important role as a multiple functional RNA binding nuclear protein in cell cycle progress, RNA usage, signal transduction, and tyrosine phosphorylation by Src during mitosis. However, its precise impact on these essential cellular functions remains unclear. The purpose of this study is to further elucidate Sam68 functions in RNA metabolism, signal transduction regulation of cell growth and cell proliferation in DT40 cell line.</p><p><b>METHODS</b>By using gene targeting method, we isolated a mutation form of Sam68 in DT40 cells and described its effect on cell growth process and signal transduction. Southern, Northern, and Western blot, phosphorylation and flow-cytometric analyses were performed to investigate the Sam68 functions.</p><p><b>RESULTS</b>A slower growth rate (2.1 hours growth elongation) and longer S phase (1.7 hours elongation) was observed in the Sam68 mutant cells. Serum depletion resulted in increased amounts of dead cells, and expansion of S phase in mutant cells. Upon B cell cross-linking, the maximal level of tyrosine phosphorylation on BLNK was observed to be significantly lower in mutant cells.</p><p><b>CONCLUSIONS</b>The proline rich domain of Sam68 is involved in cell growth control by modulating the function of mRNAs in S phase or earlier and the functions as an adaptor molecule in B cell signal transduction pathways.</p>

Interventional therapy for the chronic seminal vesiculitis / 中华男科学杂志

<p><b>OBJECTIVES</b>To investigate the efficacy of transperineal seminal vesicle puncture under ultrasound guidance and continuous transcatheter antibiotic drugs instillation for the treatment of chronic seminal vesiculitis.</p><p><b>METHODS</b>Forty-two patients with hemospermia were treated from April 1988 to January 2001. Of them 35 patients with urogenital inflammation were treated by transperineal seminal vesicle puncture under ultrasound guidance and continuous transcatheter antibiotic drugs instillation.</p><p><b>RESULTS</b>Transperineal seminal vesicle puncture and continuous transcatheter antibiotic drugs instillation therapy was adopted for 35 patients with urogenital inflammation and the cure rate was 91.43%.</p><p><b>CONCLUSIONS</b>Transperineal seminal vesicle puncture under ultrasound guidance and continuous transcatheter antibiotic drugs instillation was an effective method for diagnosis and treatment of chronic seminal vesiculitis.</p>