Progress of neoadjuvant therapy for high-risk prostate cancer in the era of novel hormonal therapy / 现代泌尿外科杂志

In the past, the use of neoadjuvant androgen deprivation therapy (ADT) for prostate cancer did not exhibit survival benefits and was not recommended by the practicing guidelines. In recent years, with the emergence of novel hormonal therapeutics such as Abiraterone, Enzalutamide, Apalutamide and Darolutamide, the interest for neoadjuvant therapy has been reignited. Here, we summarize the four categories of neoadjuvant therapy with new hormonal agents, and discuss how to evaluate the efficacy and explore the molecular mechanism after neoadjuvant therapy.

Therapeutic effect of small extracellular vesicles derived from mesenchymal stem cells on retinal light injury and its mechanism / 中华实验眼科杂志

Objective:To investigate the effect of small extracellular vesicles (sEVs) derived from mesenchymal stem cells (MSCs) in mouse model of retinal light injury and the possible mechanism.Methods:Human umbilical cord derived MSCs were identified by flow cytometry.Supernatants of passage 3-5 MSCs were collected.sEVs were harvested by ultracentrifugation and were identified by transmission electron microscopy.Sixty-five healthy female SPF-grade BALB/c mice aged 8-10 weeks were randomly divided into normal group (17 mice), phosphate buffered saline (PBS) group (24 mice) and sEVs group (24 mice). Mice in PBS and sEVs groups were intravitreally injected with 2 μl of PBS and sEVs, respectively, and were exposed to 930 lx blue light for 6 hours.No intervention was administered to the normal group.Three days after lighting, mice retinal structure was observed by hematoxylin-eosin staining.Apoptotic retinal cells were detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Retinal function was tested by electroretinogram.Differentially expressed mRNAs between PBS group and sEVs group were assayed by mRNA transcriptome sequencing and were analyzed through KEGG cluster analysis.The differential mRNAs were verified via real-time quantitative PCR.The study protocol was approved by the Animal Ethics Committee of Tianjin Medical University Eye Hospital (No.TJYY20201221035).Results:MSCs were positive for CD90 and CD105, negative for CD34 and CD45.The extracted MSC-sEVs showed a bilayer membrane vesicle with a diameter of 80-140 nm.Hematoxylin-eosin staining showed the arrangement of photoreceptor nuclei was disordered in outer nuclear layer in PBS group.The disorder of photoreceptor nuclei arrangement of sEVs group was slighter than that of PBS group.The apoptotic cell number of sEVs group was (14.60±4.04)/visual field, which was lower than (24.00±8.52)/visual field of PBS group, with a statistically significant difference ( t=2.37, P<0.05). The a-wave amplitude of sEVs group was (64.38±16.70)μV, which was higher than (16.78±6.37) μV of PBS group, showing a statistically significant difference ( P<0.05). The b-wave amplitudes of PBS and sEVs groups were (132.40±39.41) μV and (154.86±34.08) μV, respectively, which were lower than (338.38±27.41) μV of normal group, and the differences were statistically significant (both at P<0.05). A total of 110 differentially expressed mRNAs were detected.There were 109 downregulated mRNAs in sEVs group.Differentially expressed mRNAs were mainly inflammation- and immune-related pathways.PCR showed that the expression level of C-C motif chemokine ligand 2, C-C motif chemokine receptor 2, leukotriene B4, leukocyte Ig-like receptor A6 and interleukin-1β in sEVs group were significantly decreased in comparison with PBS group (all at P<0.05). Conclusions:MSC-sEVs can ameliorate blue light-induced retinal structural and functional damage.The protective effect may be achieved through inhibiting inflammatory response.

Ideal cardiovascular health and mortality: pooled results of three prospective cohorts in Chinese adults / 中华医学杂志(英文版)

BACKGROUND@#Evidence on the relations of the American Heart Association's ideal cardiovascular health (ICH) with mortality in Asians is sparse, and the interaction between behavioral and medical metrics remained unclear. We aimed to fill the gaps.@*METHODS@#A total of 198,164 participants without cancer and cardiovascular disease (CVD) were included from the China Kadoorie Biobank study (2004-2018), Dongfeng-Tongji cohort (2008-2018), and Kailuan study (2006-2019). Four behaviors (i.e., smoking, physical activity, diet, body mass index) and three medical factors (i.e., blood pressure, blood glucose, and blood lipid) were classified into poor, intermediate, and ideal levels (0, 1, and 2 points), which constituted 8-point behavioral, 6-point medical, and 14-point ICH scores. Results of Cox regression from three cohorts were pooled using random-effects models of meta-analysis.@*RESULTS@#During about 2 million person-years, 20,176 deaths were recorded. After controlling for demographic characteristics and alcohol drinking, hazard ratios (95% confidence intervals) comparing ICH scores of 10-14 vs. 0-6 were 0.52 (0.41-0.67), 0.44 (0.37-0.53), 0.54 (0.45-0.66), and 0.86 (0.64-1.14) for all-cause, CVD, respiratory, and cancer mortality. A higher behavioral or medical score was independently associated with lower all-cause and CVD mortality among the total population and populations with different levels of behavioral or medical health equally, and no interaction was observed.@*CONCLUSIONS@#ICH was associated with lower all-cause, CVD, and respiratory mortality among Chinese adults. Both behavioral and medical health should be improved to prevent premature deaths.

Expression and determination of biomarkers in diabetic retinopathy / 中华实验眼科杂志

Diabetic retinopathy (DR) is a kind of common vascular complications in diabetes and one of the leading causes of irreversible blindness.It is characterized by leaky retinal vasculature, neovascularization and fiber membrane proliferation.Although there are various DR treatments, most of them aim at middle and late stage of the disease and have limited efficacy.Therefore, early diagnosis and precise treatment of DR are important.In recent years, the research of biomarkers related to DR has developed rapidly, which plays an important role in the risk assessment and early intervention of the disease.Nowadays, classic biomarkers such as HbA1c have been widely used in clinical practice.With the further study on DR, inflammatory biomarkers and angiogenesis-related biomarkers have been found to be closely related to the occurrence and development of the disease.At the same time, with the progress of modern technology, advanced equipment have built a broad platform for the exploration of DR biomarkers.Omics biomarkers, imaging biomarkers and artificial intelligence have become the focus of research and made important contributions to the early treatment of DR.This article summarized the key progress related to the DR biomarkers research to provide new clinical strategies in the efficient prevention, control and treatment of DR.

Regulatory role of non-coding RNA in the pathogenesis of uveitis and future perspectives / 中华实验眼科杂志

Non-coding RNA (ncRNA) is a type of RNA that has multiple biological functions but is not translated into proteins.Uveitis, a common blindness-causing disease, is susceptible to relapse and difficult to treat, but its pathogenesis is not completely elucidated.Recent studies have shown that ncRNA plays an important role in the pathogenesis of human uveitis and rabbit experimental autoimmune uveitis.ncRNAs participate in the pathogenesis of uveitis by regulating important signaling pathways related to immunity, the immune response of T lymphocytes or antigen-presenting cells, and the secretion of inflammatory factors, so targeting some ncRNAs is of certain value for the treatment of uveitis.The single nucleotide polymorphisms and copy number variation of ncRNA are highly correlated with the genetic susceptibility of uveitis.Therefore, ncRNA may become a biomarker for the diagnosis and prognosis of uveitis and targeting ncRNA may become a new treatment strategy in uveitis.The regulatory roles of microRNA and long non-coding RNA in the pathogenesis of uveitis were reviewed in this article.

Therapeutic effect of anti-IL-12/IL-23 p40 on experimental autoimmune uveitis and its mechanism / 中华实验眼科杂志

Objective:To explore the therapeutic effect of anti-interleukin (IL)-12/IL-23 p40 antibody on experimental autoimmune uveitis (EAU) and its mechanism.Methods:Sixty-six SPF female C57BL/6N mice aged 6-8 weeks were selected.EAU model was established in 24 mice through immunization with the interphotoreceptor retinoid-binding protein (IRBP) 651-670.The 24 mice were sacrificed before immunization, and on the 3rd, 12th, and 18th day after immunization, with 6 at each time point.Flow cytometry was used to detect the proportion of IL-17A + interferon-γ (IFN-γ) + CD4 + T cells in the spleen, lymph nodes and eyeballs.Another 6 mice were selected to establish EAU model, and fundus images of the mice were taken with a small animal imaging instrument and optical coherence tomography (OCT) 18 days after immunization.The 6 mice were sacrificed after OCT examination and the eyeballs were collected.Hematoxylin-eosin staining was used to observe the retinal inflammation and morphological changes in tissue structure.Flow cytometry was employed to detect the proportion of IL-17A + IFN-γ + CD4 + T cells in lymph nodes.The 6 mice were divided into IL-17A + IFN-γ + highly expressed group and IL-17A + IFN-γ + lowly expressed group according to flow cytometry results, and the retinal injury was compared between the two groups.EAU model was established in another 36 mice, which were divided into anti-IL-12/IL-23 p40 group and IgG group by random number table method, with 18 mice in each group.Anti-IL-12/IL-23 p40 or IgG was injected by tail vein at a 3-day inteval according to grouping.On the 12th and 18th day after immunization, 6 mice were selected from each group to collect lymph nodes and eyeballs, and the proportion of T cell subsets was detected by flow cytometry.Eyeballs of 6 mice in each group were extracted on the 24th day after immunization and retinal damage was observed by hematoxylin-eosin staining.The induced differentiation of CD4 + T cells in vitro was assayed by flow cytometry.The expressions of IL-17 and IFN-γ were detected by enzyme-linked immunosorbent assay (ELISA) after induced differentiation of IL-17A + IFN-γ + CD4 + T cells.The relative expression levels of Th1 transcription factor T-bet and Th17 transcription factor retinoid acid-related orphan nuclear receptor γt (ROR-γt) after induced differentiation of IL-17A + IFN-γ + CD4 + T cells were detected by real-time quantitative PCR.The use and care of animals followed the ARVO statement and this study protocol was approved by an Ethics Committee of Experimental Animals of Tianjin Medical University Eye Hospital (No.TJYY2019111019). Results:There were significant differences in the proportion of IL-17A + IFN-γ + CD4 + T cells in lymph nodes, spleen and eyeballs between wild-type mice and EAU mice at the 3rd, 12th and 18th day after immunization ( H=9.642, 16.531, 10.385; all at P<0.05). Compared with before immunization, the proportion of IL-17A + IFN-γ + CD4 + T cells was significantly increased in lymph nodes of EAU mice on the 12th day following immunization and was significantly increased in spleen and lymph nodes on day 18 after immunization (all at P<0.05). Severe retinal exudation, retinal detachment, severe inflammatory cell infiltration and extensive retinal folds were detected in IL-17A + IFN-γ + highly expressed mice.Mild retinal edema, focal inflammatory cell infiltration and mild retinal folds were found in IL-17A + IFN-γ + lowly expressed mice.The proportion of CD3 and IL-17A + IFN-γ + CD4 + T cells in the eyeballs of anti-IL-12/IL-23 p40 group was lower than that in IgG group at the 18th day after immunization, and the differences were statistically significant ( t=15.304, 8.080; both at P<0.05). On day 12 after immunization, the percentage of IL-17A + IFN-γ + CD4 + T cells in anti-IL-12/IL-23 p40 group was (0.33±0.18)%, which was significantly lower than (4.83±0.45)% in IgG group ( t=15.974, P<0.001). Compared with IgG group, the percentage of Th1, Th17, IL-17A + IFN-γ + CD4 + T cells and the expression levels of IL-17, IFN-γ, T-bet, ROR-γt in anti-IL-12/IL-23 p40 group were significantly decreased, with statistical significances (all at P<0.05). Conclusions:Anti-IL-12/IL-23 p40 has a therapeutic effect on EAU by inhibiting IL-17A + IFN-γ + CD4 + T cells.

Value of the biomarkers derived from cerebrospinal fluid in the diagnosis of Alzheimer disease / 中华检验医学杂志

Alzheimer disease (AD) is a neurodegenerative disease in the elderly. Early intervention is the only reliable means to delay the progress of the disease. Referring to the diagnostic criteria of AD, this paper summarizes and analyzes the representative literatures of various AD biomarkers derived from cerebrospinal fluid in recent years, and reviews the efficacy of various cerebrospinal fluid biomarkers in the diagnosis and differential diagnosis of AD. Results show that CSF Aβ42, Aβ42/Aβ40, T-tau, p-tau, Aβ42/p-tau, growth-associated protein 43, synaptosomal-associated protein 25, neurogranin and visinin-like protein-1 are of great value in the diagnosis and differential diagnosis of AD. The above CSF biomarkers can be used in the diagnosis and differential diagnosis of AD. The laboratory should select appropriate AD CSF biomarkers according to its own conditions in daily laboratory works.

Analysis of clinical characteristics and genetic susceptibility of steroid-induced ocular hypertension in patients with uveitis / 中华眼底病杂志

Objective:To observe the clinical characteristics of steroid-induced ocular hypertension (SIOH) in patients with uveitis, and explore the relationship between its clinical phenotype and gene polymorphism.Methods:A retrospective case-control study. From July 2019 to December 2020, 576 patients with uveitis who were treated with glucocorticoid eye drops in Tianjin Medical University Eye Hospital were included in the study. Among them, there were 175 confirmed glucocorticoid responders (SRs) and 401 glucocorticoid non-responders (NRs). Seventy cases of SRs (age ≥18 years) using 1 % prednisone acetate eye drops were selected as the experiment group and 64 cases of NRs were selected as the control group. The polymorphism of rs2523864 and rs3873352 of human leukocyte antigen complex group ( HCG) 22 gene were detected by Sanger sequencing. To observe the clinical characteristics of SIOH after the use of glucocorticoid eye drops, and the correlation between rs2523864 and rs3873352 and the occurrence of SIOH. Differences among groups were compared with the Chi-square test or Fisher's exact test. The correlation between the occurrence of SIOH and the range of intraocular pressure increases after glucocorticoid use and the rs2523864 and rs3873352 loci were compared using the odds ratio ( OR) and its 95% confidence interval ( CI). Results:SIOH occurred in 175 (30.4%, 175/576) of 576 patients. Among them, there were 96 males (54.9%, 96/175) and 79 females (45.1%, 79/175); the average age was 33.64±17.40 years. Steroid high responders (HRs) and steroid moderate responders (MRs) were 58 (33.1%, 58/175) and 117 (66.9%, 117/175) cases. The medication time for the increase in intraocular pressure in MRs that was 33 (19, 56) days, and in HRs that was 28 (14, 36) days, the difference of which was significant ( Z=-1.999, P=0.046). No differences were found in daily doses of ocular hypertension induced by 1% prednisone acetate eye drops between MRs which was 4.24 (3.46, 4.66) drops/day and HRs that was 4.32 (3.84, 5.36) drops/day ( Z=-1.676, P=0.094). The genotype and allele frequency distribution of the rs3873352 locus in the case group and HRs group were significantly different from those in the control group ( P<0.05). The intraocular pressure with rs3873352 GG genotype after the medication was higher than that with GC and CC genotype ( Z=2.855, 2.628; P=0.013, 0.026), whereas there was no significant difference between different genotypes of rs2523864 ( Z=3.580, P>0.05). Genetic model analysis revealed the risk of SIOH in rs3873352 G allele carriers (GG+GC) was 2.048 times that of non-G allele carriers ( OR=2.048, 95% CI: 1.027-4.081, P=0.041). The genotype and allele frequency of rs2523864 locus showed no significant difference between different group ( P>0.05). Conclusions:After the use of glucocorticoid eye drops, HRs have an earlier increase in intraocular pressure than MRs. HCG22-rs3873352 gene polymorphism is related to the occurrence of SIOH, GG genotype increases the risk of SIOH, and G allele is a risk gene for SIOH.

Study on the application of continuous nursing led on the theory of interactive standard theory after heart transplantation / 中国实用护理杂志

Objective:To explore the application research of continuity nursing based on the interactive reaching standard theory after heart transplantation.Methods:The clinical data of 104 patients after heart transplantation in Nanjing Hospital Affiliated to Nanjing Medical University (Nanjing First Hospital) from June 2012 to June 2020 were collected for retrospective study. According to the file order, they were divided into the control group and the experimental group with 52 cases in each group. The control group took regular nursing care, and the experimental group took continuation nursing based on the interactive standard theory on the basis of the control group. The intervention continued for 24 weeks. The compliance behavior, psychological state and other indexes were compared between two groups.Results:The scove of Positive and Negative Affect Scale: after 12 weeks and 24 weeks of nursing, the experimental group′s positive emotion scores (27.71 ± 4.42, 35.11 ± 4.19) were higher than those of the control group (24.45 ± 4.03, 28.87 ± 4.36). The negative emotional scores (24.52 ± 3.71, 16.66 ± 2.28) were lower than those of the control group (28.84 ± 4.14, 23.31 ± 3.46), the differences were statistically significant ( t=5.60, 11.57, P<0.05). After 12-week and 24-week nursing, the experimental group′s health knowledge level score (44.12 ± 4.43, 53.64 ± 4.55) , self-care skills score(35.35 ± 4.01, 44.78 ± 4.22) , self-responsibility score (21.15 ± 2.38, 26.11 ± 1.44) , and self-concept score (18.56 ± 6.25, 23.58 ± 2.58) were higher than the control group (37.78 ± 4.52 and 45.56 ± 5.13, 31.11 ± 3.64 and 36.65 ± 3.91, 18.82 ± 2.46 and 22.35 ± 1.29, 15.96 ± 4.10 and 18.12 ± 3.10), the differences were statistically significant ( t values were 2.51-14.03, P<0.05). Conclusions:Continuing care led by the interactive standard theory can improve the health literacy of patients after heart transplantation, relieve negative emotions, promote post-traumatic growth, enhance self-management capabilities, establish compliance behaviors, and improve heart function.

Practice and thinking of the multi-district hospital management under the new pattern of national regional medical center / 中华医院管理杂志

The authors took the management practice of " one hospital with multiple districts" in Children′s Hospital Affiliated of Zhengzhou University as the research object, analyzed the main problems and challenges faced by the multi-district hospital management under the new pattern of national regional medical center. Through coordinating the hospital′s strategic planning and the development of discipline layout, building an integrated management system, improving the level of homogeneous service and other key countermeasures, the hospital has significantly improved its management efficiency and operation efficiency, and the medical service capacity of each district has developed in a balanced way. It could give full play to the pilot value for the construction of national regional medical center, hoping to provide reference for hospital administrators.

Single-molecule imaging reveals stoichiometry of vascular endothelial growth factor receptor 2 in retinal vascular endothelial cells / 中华眼底病杂志

Objective:To observe the stoichiometry of vascular endothelial growth factor receptor 2 (VEGFR2) on the retinal vascular endothelial cell membrane by single-molecule fluorescence imaging.Methods:Rhesus monkey retinal vascular endothelial cells (RF/6A) were divided into blank control group (normal culture) and plasmid transfection group [transfected with VEGFR2-green fluorescent protein (GFP) recombinant plasmid]. The expression of GFP in the plasmid transfected group was observed by confocal microscope, and the expression of VEGFR2 in the cells was detected by real-time fluorescent quantitative polymerase chain reaction (qPCR) and Western blot. The fluorescence intensity distribution and bleaching steps of single VEGFR2-GFP molecule on the cell membrane were recorded by single-molecule imaging. The distribution of fluorescence intensity and the number of fluorescence bleaching steps of GFP were recorded.Results:GFP green fluorescence was observed in the transfected cells 12 hours after transfection. qPCR results showed that the expression of VEGFR2 and GFP mRNA in the plasmid transfected group was significantly higher than that in the blank control group ( t=11.240, 12.330; P<0.001, 0.001). Western blot results showed that the expression of VEGFR2 protein in the plasmid transfected group was significantly higher than that in the blank control group ( t=8.346, P<0.01). The results of single-molecule imaging showed that the fluorescence intensity distribution of VEGFR2-GFP on the surface of RF/6A cell membrane without ligand stimulation was bimodal, in which monomer and dimer were 86.0% and 14.0% respectively. By counting the steps of GFP fluorescence bleaching, the proportions of receptor monomer, dimer, trimer, and tetramer were 81.4%, 12.9%, 5.5%, and 0.3% respectively. Conclusion:In the absence of ligands, VEGFR2 coexists in the form of monomers and dimers on the surface of RF/6A cell membrane, and monomers are dominant.

Circulating exosomal inflammation-related protein S100A8 as a potential biomarker for the severity of diabetic retinopathy / 中华眼底病杂志

Objective:To observe the expression of S100A8 in plasma exosomes, microvesicles (MV), plasma and vitreous in patients with diabetic retinopathy (DR), and verify it in a diabetic rat model, and to preliminarily explore its role in the occurrence and development of DR.Methods:A case-control study. From September 2018 to December 2019, a total of 73 patients with type 2 diabetes, hospitalized patients undergoing vitrectomy, and healthy physical examinations in the Tianjin Medical University Eye Hospital were included in the study. Among them, plasma were collected from 32 patients and vitreous fluid were collected from 41 patients, which were divided into plasma sample research cohort and vitreous sample research cohort. The subjects were divided into simple diabetes group (DM group), non-proliferative DR group (NPDR group) and proliferative DR group (PDR group) without fundus changes; healthy subjects were regarded as normal control group (NC group). In the study cohort of vitreous samples, the control group was the vitreous humor of patients with epimacular membrane or macular hole. Plasma exosomes and microvesicles (MVs) were separated using ultracentrifugation. Transmission electron microscopy, nanometer particle size analyzer and Western blot (WB) were used to characterize exosomes and MVs. The mass concentration of S100A8 was determined by enzymelinked immunosorbent assay. Eighteen healthy male Brown Norway rats were divided into normal control group and diabetic group with 9 rats in each group by random number table method. The rats of diabetes group were induced by streptozotocin to establish diabetic model. Five months after modeling, immunohistochemical staining and WB were used to detect the expression of S100A8 in the retina of rats in the normal control group and the diabetes group. t test was used for the comparison of measurement data between the two groups. Single-factor analysis of variance were used for the comparison of multiple groups of measurement data.parison of measurement data between the two groups. Single-factor analysis of variance were used for the comparison of multiple groups of measurement data. Results:Exosomes and MVs with their own characteristics were successfully separated from plasma. The concentrations of plasma exosomes and vitreous S100A8 in the PDR group were higher than those in the NPDR group, DM group, NC group, and the difference was statistically significant ( P=0.039, 0.020, 0.002, 0.002, P<0.000,<0.000). In the plasma sample cohort study, It was not statistically significant that the overall comparison of the S100A8 mass concentrations of plasma and plasma MV in the four groups of subjects ( F=0.283, 0.015; P=0.836, 0.996). Immunohistochemical staining showed that retinal ganglion cells, bipolar cells, cone rod cells and vascular endothelial cells in the diabetic group all expressed S100A8 protein. Compared with the normal control group, the expression level of S100A8 in the retina of the diabetic group increased, and the difference was statistically significant ( t=8.028, P=0.001). Conclusions:The level of S100A8 protein in circulating exosomes increases significantly with the severity of DR in patients with type 2 diabetes. S100A8 may be an influential factor in the inflammatory environment of DR and a potential anti-inflammatory therapeutic target.

Effects of intravenous injection of human umbilical cord mesenchymal stem cells-derived small extracellular vesicles on experimental autoimmune uveitis in mice / 中华实验眼科杂志

Objective:To explore the therapeutic effects of intravenous injection of small extracellular vesicles (sEVs) derived from human umbilical cord mesenchymal stem cells (MSCs) on experimental autoimmune uveitis (EAU) in mice.Methods:MSCs from human umbilical cord were cultured and the supernatant was collected.The sEVs were isolated by ultracentrifugation method and a NanoSight instrument was used to analyze the particle size.The expression of surface markers sEVs, CD9, CD81 and CD63 was determined via Western blot.The morphology of sEVs was observed with a transmission electron microscope.Forty-eight 7-week-old female C57BL/6 mice were seclected to establish the EAU model through immunization with interphotoreceptor retinoid-binding protein peptide 651-670 (IRBP 651-670). The mice were divided into sEVs treatment group and phosphate buffer solution (PBS) control group using a random number table, with 24 mice in each group.The mice in the sEVs treatment group were injected with 50 μg of MSCs-derived sEVs via tail vein on the 11th day after modeling.In the PBS control group, the mice were injected with the same volume of PBS.Six mice in each group were randomly selected to observe the inflammation of the retina after mydriasis with an ophthalmoscope every other day from 8th day following modeling and the inflammation scores were evaluated.Six mice were randomly selected and sacrificed on the 14th day and 6 on the 18th day following modeling in each group, and both eyeballs of the mice were enucleated.Retinal tissue sections of the 6 mice sacrificed on the 18th day were stained with hematoxylin-eosin and the pathological scores were evaluated.The infiltration of helper T 1 (Th1) cells and Th17 cells in the eyeballs of the 6 mice sacrificed on the 18th day following modeling was detected by flow cytometry.T cells were isolated from spleen and lymph nodes of the 6 mice sacrificed on the 14th day, and the proliferation of T cells under different concentrations of IRBP 651-670 (0, 1, 10 and 20 μg/ml) was detected using a 5-bromodeoxyuridine (BrdU) method.To further study the effects of MSCs-derived sEVs on Th1/Th17 cells differentiation, naive T cells of spleen from another 3 normal mice were isolated by magnetic bead negative sorting and incubated with 10 μg/ml MSCs-derived sEVs or 10 μg/ml PBS, and then were cultured under Th1/Th17 cell differentiation conditions, respectively.Flow cytometry was used to measure the differentiation of naive T cells into Th1/Th17 cells.This study protocol complied with the regulations of the care and use of laboratory animals in China and was approved by an Ethics Committee of Tianjin Medical University (No.TJYY2019103022). Results:The isolated human MSCs-derived sEVs was with an average diameter of (102.4±33.6) nm and showed a double-layer membrane vesicle structure under the transmission electron microscope.The CD9, CD63 and CD81 proteins were highly expressed in sEVs.The inflammation scores of the sEVs treatment group were significantly lower than those in the control group on day 14, 16, 18, 20 and 22 after modeling (all at P<0.05). The pathological score of mice in the sEVs treatment group was significantly lower than that of PBS control group on the 18th day following modeling ( P<0.05). The flow cytometry results showed that on day 18 after modeling, the proportions of Th1 and Th17 cells in eyeballs in the sEVs treatment group were (15.55±2.03)% and (15.67±2.15)%, respectively, which were significantly lower than (21.35±0.72)% and (20.90±1.10)% in the PBS control group ( t=6.58, 5.31; both at P<0.01). BrdU results showed that when the IRBP 651-670 concentration was 20 μg/ml, the T cell proliferation ability in the sEVs treatment group was inhibited obviously compared with the control group ( P<0.05). The proportions of naive T cells differentiated into Th1 cells and Th17 cells in the sEVs treatment group were (28.15±1.32)% and (11.60±2.23)% respectively, which were significantly lower than (31.58±1.75)% and (23.52±1.76)% of the PBS control group, and the differences were statistically significant ( t=3.93, 10.26; both at P<0.05). Conclusions:Intravenous injection of human umbilical cord MSCs-derived sEVs can reduce the inflammation in EAU mice.The mechanism may be related to inhibiting the differentiation of naive T cells to Th1 and Th17 cells, and reducing the infiltration of Th1 and Th17 cells in the eyeballs.

Quantitative proteomic analysis of human umbilical cord mesenchymal stem cells with pigment epithelium-derived factor gene modification mediated by lentivirus / 中华实验眼科杂志

Objective:To investigate the protein expression changes of human umbilical cord mesenchymal stem cells (hUCMSCs) modified with pigment epithelium-derived factor ( PEDF) gene mediated by lentivirus. Methods:The hUCMSCs were divided into the control group and experimental group.Cells in the control group were normal hUCMSCs and the cells in experimental group were hUCMSCs with PEDF modification.The proteins from the two groups were collected and processed by FASP method.Samples were fractionated by liquid chromatography and analyzed by tandem mass spectrometry, and SWATH mode was applied.Differential protein analysis, Gene Ontology (GO) analysis and Reactome pathway enrichment analysis were performed. Results:A total of 5 361 quantified proteins were detected in this experiment, of which 432 proteins were differentially expressed (fold change>1.5, P<0.05). There were 219 of the 432 proteins up-regulated, including serpin family F member 1 (SERPINF1) (PEDF), DEAD-box helicase 59 (DDX59) and thrombospondin 1 (THBS1), etc., whereas 213 proteins were down-regulated, including collagen type Ⅰ alpha 1 (COL1A1), COL18A1, etc.GO analysis indicated that the differential proteins were mainly involved in fibrinolysis, extracellular structure organization, regulation of transporter activity, biosynthetic process of secondary alcohol and cholesterol, coenzyme metabolic process and regulation of peptidase activity, etc.Reactome pathway analysis showed that the differential proteins were mostly involved in regulation of insulin like growth factor (IGF) transport and uptake by IGF binding protein, post-translational protein phosphorylation, extracellular matrix organization, metabolism of steroids. Conclusions:After gene modification with PEDF mediated by lentivirus, the expression of many proteins in hUCMSCs were changed. PEDF gene modification can alter the structure of extracellular matrix and regulate the expression of proteins associated with cell proliferation, self-renewal and multipotency.

Current state and progress of intravitreal injection of chemotherapeutic agents for the treatment of vitreoretinal lymphoma / 中华眼底病杂志

Primary vitreoretinal lymphoma (PVRL) is one of the most common type of primary intraocular lymphoma. The current treatment options include local ocular radiotherapy (radiotherapy), systemic chemotherapy (chemotherapy), local ocular chemotherapy, and combination therapy. The treatment options are different at different stages of PVRL, however, there is no uniform treatment guideline. Local ocular chemotherapy can make the drug reach effective therapeutic concentration in the eye, and it can be repeated many times. At the same time, it can avoid the adverse reactions caused by systemic medication or radiotherapy. It is an ideal choice for relieving ocular symptoms. At present, the mainstream ocular local chemotherapeutics are methotrexate (MTX) and rituximab (RTX). The basic consensus about the intravitreal injection of MTX (IVM) is the induction-consolidation-maintenance model, however, the time of each stage and frequency of IVM are diverse. The time interval of intravitreal injection of RTX is also variable, ranging from 1 time/week to 1 time/months and so on. Corneal epithelial lesions caused by frequent MTX injections and the higher recurrence rate after RTX treatment are the main reasons for changing the treatment plan. For patients with primary central nervous system lymphoma and PVRL, combined treatment with neurology department is necessary to save patient's lives, ophthalmology treatment relieves ocular symptoms and improves the patient's quality of life. For patients with PVRL alone without central nervous system involvement, ophthalmology treatment is necessary to control patient's eye symptoms, and close follow-up should be followed to find the involvement of the central nervous system in time, and then combined with neurological treatment to save patient’s lives.

Current state and progress of rapamycin in the treatment of non-infectious uveitis / 中华眼底病杂志

Glucocorticoids (GCS) are the main treatment for non-infectious uveitis (NIU). However, long-term GCS treatment may induce systemic side effects including hypertension, hyperlipidemia and diabetes. Patients may develop cataract, ocular hypertension or glaucoma because of topical application of GCS. Rapamycin (RAPA) exhibits immunosuppressive, antiangiogenic and antiproliferative effects. Animal experiments and clinical trials have shown that RAPA has therapeutic potential for NIU, especially the treatment of intravitreal injection. In particular, intravitreal injection of RAPA can result in minimal systemic exposure and reduce adverse events. Meanwhile, systemic unwanted effects should be concerned about. In recent years, some studies have attempted to employ nanostructured carriers to improve penetrating abilities of RAPA and efficacy of treatment for ocular posterior segment diseases. These carriers include micelles, liposomes, nanocrystals, polymeric nanoparticles, magnetic nanoparticles and so on. Whether they can load RAPA for treating NIU deserves further study and exploration.

Effect of butorphanol mixed with ropivacaine for erector spinae plane block on postoperative outcomes in patients undergoing thoracoscopic pulmonary lobectomy / 中华麻醉学杂志

Objective:To evaluate the effect of butorphanol mixed with ropivacaine for erector spinae plane block (ESPB) on postoperative outcomes in the patients undergoing thoracoscopic pulmonary lobectomy.Methods:Eighty patients of either sex, aged 35-64 yr, with body mass index of 19-30 kg/m 2, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, undergoing elective thoracoscopic pulmonary lobectomy, were divided into 2 groups ( n=40 each) using a computer-generated random number table method: butorphanol mixed with ropivacaine for ESPB group (group EB) and ropivacaine for ESPB group (group E). ESPB was performed under ultrasound guidance in both groups.A mixture of 0.1% butorphano 1 ml and 0.375% ropivacaine 20 ml was injected in EB group, and 0.375% ropivacaine 20 ml was injected in E group.The other anesthesia methods were the same in the two groups.And target-controlled infusion was stopped and PCIA was performed at the end of skin suture in the two groups.The intraoperative consumption of remifentanil, first time to press an analgesia pump, requirement for rescue analgesia within 24 h after surgery, and occurrence of ESPB-related complications were recorded.Quality of Recovery-40 (QoR-40) scores were recorded at 7 days after surgery.Before induction of anesthesia (T 1) and at 24 h after operation (T 2), the peripheral venous blood samples were collected for determination of plasma interleukin-6 (IL-6) and IL-10 concentrations, and bedside pulmonary function test was performed, and FEV 1/FVC was calculated. Results:Compared with group E, the QoR-40 scores were significantly increased at 7 days after operation, FEV 1/FVC was increased at T 2, the plasma concentrations of IL-10 were decreased at T 2, the plasma concentrations of IL-10 were increased at T 2, the intraoperative consumption of remifentanil was reduced, the first time to press an analgesia pump was prolonged, and the requirement for rescue analgesia within 24 h after surgery was decreased in group EB ( P<0.05). No ESPB-related complications were found in either group. Conclusion:Butorphanol mixed with ropivacaine for ESPB can improve postoperative outcomes in the patients undergoing thoracoscopic pulmonary lobectomy.

Intravitreal injection of conbercept after operation in the treatment of proliferatived diabetic retinophathy complicated with vitreous hemorrhage / 中华眼底病杂志

Objective:To observe the efficacy and safety of intravitreal injection of conbercept in the treatment of proliferatived diabetic retinophathy (PDR) complicated with vitreous hemorrhage by minimally invasive vitreoretinal surgery.Methods:Prospective clinical study. A total of 50 patients with PDR complicated with vitreous hemorrhage clinically diagnosed in Tianjin Medical University Eye Hospital who needed vitrectomy were recruited in this study. According to the principle of informed consent, the patients were divided into two groups: postoperative injection group and the control group. Twenty-five eyes of 25 patients in each group were examined before operation. No significant proliferative changes in the posterior pole and traction retinal detachment were observed. There was significant difference of age between two groups ( t=-24.697, P=0.030), but no significant difference of sex ( χ2=0.330, P=0.564), duration of diabetes ( t=-1.144, P=0.258), logMAR BCVA ( t=-0.148, P=0.883), lens state ( χ2=0.397, P=0.529), panretinal laser photocoagulation ( χ2=1.333, P=0.248). The postoperative injection group was treated with intravitreous injection of 0.05 ml conbercept (10 mg/ml) immediately after 27G minimally invasive vitrectomy. The other treatment and follow-up were the same as those in the postoperative injection group except for conbercept injection. All patients underwent routine slit-lamp examination, indirect ophthalmoscope and B-ultrasound examination before operation. The main outcome measure included the time of operation, the incidence rate of iatrogenic retinal holes and silicone oil filling. The recurrence of vitreous hemorrhage, BCVA, intraocular pressure, central retinal thickness (CRT), postoperative complications and progression were recorded 1 week, 1 month, 3 months and 6 months after operation. Results:At 1 week and 1, 3, 6 months after operation, there was significant difference of logMAR BCVA between the two groups ( t=-4.980, -4.840, -4.892, -5.439; P<0.001). At 3 and 6 months after operation, the recurrence of vitreous hemorrhage in the postoperative injection group was lower than that in the control group, but there was no statistical difference between two groups ( χ2=3.030, 4.153; P=0.192, 0.103). At 1 week and 1, 3, 6 months after operation, the CRT in the postoperative injection group was lower than that in the control group, the difference was significant ( t=-2.622, -2.638, -3.613, -3.037; P=0.012, 0.010, 0.001, 0.004, 0.005). There was no complications such as choroid detachment, proliferative vitreoretinopathy, retinal detachment, iris redness and neovascular glaucoma in all the eyes after operation. Conclusions:Intravitreal injection of conbercept in the treatment of PDR after operation is safe and effective. It can reduce the recurrence of vitreous hemorrhage after vitrectomy, improve the BCVA.

Lung protection induced by goal-directed fluid therapy in patients undergoing pulmonary lobectomy / 中华麻醉学杂志

Objective:To evaluate the lung protection induced by goal-directed fluid therapy (GDFT) in the patients undergoing pulmonary lobectomy.Methods:Eighty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients of both sexes, aged 35-64 yr, with body mass index of 18-30 kg/m 2, scheduled for elective pulmonary lobectomy with general anesthesia, were divided into 2 groups ( n=40 each) using a random number table method: control group (group C) and GDFT group (group G). In group C, mean arterial pressure was maintained at 65-90 mmHg, central venous pressure at 8-12 cmH 2O, and urine volume>0.5 ml·kg -1·h -1.In group G, GDFT was performed, maintaining cardiac index>2.5 L·min -1·m -2 and stroke volume variation ≤11%.Peak airway pressure and plateau airway pressure were recorded immediately after intubation (T 1), 30 min of one-lung ventilation (T 2), 1 h of one-lung ventilation (T 3) and at the end of the surgery (T 4). Blood samples were collected from the radial artery at T 1-4 and 24 h after operation (T 5) for blood gas analysis to determine the alveolar-arterial difference of oxygen tension (A-aDO 2). Broncho-alveolar lavage fluid (BALF) and blood samples from the internal jugular vein were collected at T 1 and T 4 to detect the concentrations of interleukin-6 (IL-6) and IL-10 in serum and BALF, and the ratio of IL-6 to IL-10 was calculated.Ultrasound was used to measure the inferior vena cava respiratory variations index at T 1, 4.The intraoperative total volume of fluid infused, amount of colloid solution infused, urine volume and blood loss were recorded, and the hospitalization time was also recorded. Results:Compared with group C, peak airway pressure and plateau airway pressure at T 2-4, A-aDO 2 at T 2-5, concentration of IL-6 in BALF at T 4 and IL-6/IL-10 ratio were significantly decreased, the total volume of fluid infused and urine volume were reduced, and the amount of colloid solution infused was increased in group G ( P<0.05). Conclusion:GDFT can improve the intraoperative respiratory dynamics, inhibit inflammatory responses in lung tissues and improve lung function in the patients undergoing pulmonary lobectomy.

Single-port robot-assisted laparoscopic radical prostatectomy through different approaches: initial experience and outcomes / 中华泌尿外科杂志

Objective:To evaluate the feasibility and effectiveness of sing-port robot-assisted laparoscopic radical prostatectomy(sp-RALP) through different approaches.Methods:The data of 115 patients who underwent sp-RALP through different approaches from May 2018 to June 2020 were retrospectively reviewed. In our study, 92 cases through extraperitoneal approach, 10 through transperineal approach and 13 though transvesical approach. Preoperative, intraoperative and early postoperative outcomes after various approaches were collected and analyzed. The mean age was 67(52-84) years, mean preoperative body mass index was 24.44(19.52-32.33) kg/m 2. Patients were graded as cT 1-3aN 0M 0 with no signs of regional lymph node invasion or distal metastasis. The median preoperative PSA was 9.77(6.54, 15.32) ng/ml. Results:All 115 sp-RALP were successfully completed with no conversion to open surgery or additional ports being placed. The mean operative time was 91.8(40-200)min, with 88.0(40-200)min for extraperitoneal approach, 132.5 (90-190)min for transperineal approach and 87.3(60-150)min for transvesical approach. The mean estimated blood loss was 85.5 (45-400)ml, with 77.6(50-200)ml for extraperitoneal approach, 178.0(80-400)ml for transperineal approach and 70.4 (45-150)ml for transvesical approach. Of the 115 patients, 60.9% (70/115)were <pT 3a, 39.1%(45/115) were ≥pT 3a. Positive margin(PSM) rate was 17.4%(20/115), with 31.1%(14/45) for ≥T 3a group, 8.6%(6/70) for <T 3a group. Postoperative pathology revealed a Gleason score of 3 + 3 in 6 patients, 3 + 4 in 45 patients, 4+ 3 in 52 patients, and ≥8 in 2 patients. Mean hospital stay was 3(1-7)d. Median postoperative PSA was 0.03(0.01, 0.05)ng/ml at 1 month postoperatively, and 88.7% patients reported less than 1 pad per day at 3 month postoperatively. Conclusions:With several advantages, such as better cosmesis, postoperative fewer pain, and better recovery as well as comparable oncological outcomes in the short-term follow-up, sp-RALP through various approaches is a feasible and effective procedure for organ-confined prostate cancer.