Objective:To analyze the optical coherence tomography (OCT) imaging characteristics in patients with Coats disease and their value in predicting macular fibrosis.Methods:A nested case-control study was performed.A total of 43 patients (43 eyes) diagnosed with Coats disease through color fundus photography, ocular B-scan ultrasonography, fundus fluorescein angiography, and spectral-domain OCT examination were enrolled from January 2008 to October 2021 at the Xijing Hospital.Among them, there were 40 males and 3 females, aged from 2 to 60 years old, with a median age of 13 years.Macular fibrosis was used as an indicator of poor prognosis, and patients were divided into two groups based on whether macular fibrosis occurred at the end of follow-up.The differences in OCT characteristics between two groups were compared and logistic regression analysis was used to identify the risk factors for macular fibrosis.This study adhered to the Declaration of Helsinki and was approved by the Ethics Committee of Xijing Hospital of Fourth Military Medical University (No.KY20202009-C-1).Results:The OCT clinical features of 43 cases of Coats disease included intraretinal hard exudates in 43 eyes (100%), subretinal fluid in 21 eyes (48.8%), macular cysts in 17 eyes (27.9%), subretinal exudates in 9 eyes (20.9%), anterior retinal hyperreflective dots in 7 eyes (16.3%), epiretinal membrane in 21 eyes (48.8%), and intraretinal fluid in 22 eyes (51.2%).In color fundus photos of 41 eyes, 38 eyes (93.0%) had hard exudates distributed in the posterior pole and 27 eyes (65.9%) had the mid-peripheral region.OCT examination showed that hard exudates were distributed in the inner nuclear layer in 35 eyes (81.4%) and the outer nuclear layer in 33 eyes (76.7%).Among 21 eyes with exudative retinal detachment detected by OCT, 9 eyes (42.9%) were detected by fundus photography and 18 eyes (85.7%) were detected by B-scan ultrasonography.The proportions of eyes with subretinal fluid and subretinal exudates were higher in the macular fibrosis group than in the non-macular fibrosis group, and the differences were statistically significant ( χ2=20.755, P<0.001; χ2=6.133, P=0.013).Logistic regression analysis showed that the presence of subretinal fluid was a risk factor for macular fibrosis (odds ratio=48.345, 95% confidence interval: 4.272-547.066, P=0.002). Conclusions:OCT examination can detect subretinal fluid, subretinal exudates, macular cysts, macular exudates, and hyperreflective spots in the retina of patients with Coats disease.Subretinal fluid is a risk factor for macular fibrosis.

Objective:To investigate the molecular expression and pathological features of endothelial cell (EC) in a murine model of choroidal neovascularization (CNV) based on single-cell RNA sequencing (scRNA-seq).Methods:Six C57BL/6 mice aged 6-8 weeks were randomly divided into two groups, with 3 mice in each group.Bilateral eyeballs were enucleated.The choroidal tissues from the two groups were isolated by shearing the complex and scraping the choroid, respectively.Single-cell suspension was prepared by continuous digestion with trypsin/type Ⅰ collagenase at 37 ℃, and the cell viability and EC ratio were detected by flow cytometry to determine the preparation method of single-cell suspension.Another 6 mice were randomly assigned into the control group and the CNV group, with 3 mice in each group.The CNV model was induced by laser photocoagulation and single-cell suspensions were prepared 7 days after modeling.Gene expression library construction was performed using the Chromi-um (10x Genomics) instrument.High throughput sequencing was performed using the Illumina Novaseq6000 to obtain the expression matrix.The EC subpopulations were classified according to previous researches and the Cellmarker database.Pseudo-time analysis was performed in EC, revealing the gene expression matrix of different states.CNV-EC were further selected with preliminary analysis of the expression characteristics.Another 6 mice were selected to establish the CNV model and eyeball frozen sections were prepared 7 days after modeling.Expression and distribution as well as the area percentage of EC marker Pecam1, mitochondrial outer membrane proteins Tomm20 and mt-Co1, and capillary markers Kdr and Plvap were observed by immunofluorescence staining, and the vascular diameter was calculated.The use and care of animals followed the ARVO statement.This study protocol was approved by the Experimental Animal Welfare and Ethics Committee of Air Force Military Medical University (No.20200181).Results:The cell viability of the single-cell suspension prepared from choroidal-scleral fragments and choroidal scrapings was 99.4% and 99.1%, respectively, both of which met the sequencing requirements.The percentage of EC detected by flow cytometry was approximately 1.58%.The scRNA-seq result revealed that both the normal control and CNV groups contained 13 choroidal cell clusters.Compared with the normal control group, the proportions of rod/cone photoreceptor cells, EC and hematopoietic cells all increased, while the retinal pigment epithelium (RPE) and Schwan cells reduced in the CNV group.Among all clusters, EC constituted 18.4%.The pseudo-time analysis demonstrated that EC could be further divided into 4 states.The percentage of state 2 EC was 29.1% in the CNV group, which was significantly higher than 9.5% in the normal control group.Differentially expressed gene analysis showed that the expression of mitochondrion-related genes, including mt-Nd4 and mt-Atp6, were upregulated in state 2 EC, while capillary-related genes, including Kdr and Esm1, were downregulated.Immunofluorescent staining revealed that the area of Tomm20 and mt-Co1 in Pecam1-positive EC in the CNV area was (19.50±4.68)% and (4.64±2.82)%, respectively, which were both higher than (3.00±2.09)% and (0.18±0.34)% in normal area ( t=7.88, 3.84; both at P<0.01). The area of Kdr and Plvap in Pecam1-positive EC in the CNV area was (1.50±0.29)% and (0.79±0.97)%, respectively, which were both lower than (31.30±5.44)% and (10.43±2.28)% in the normal area ( t=13.40, 9.48; both at P<0.01). The vascular diameter in the CNV area was (5.52±1.85)μm, which was larger than (4.21±1.84)μm in the normal area ( t=9.57, P<0.001). Conclusions:When CNV occurs, the proportion of EC in choroid increases, and CNV-EC shows pathologic features of mitochondrial metabolic activation and loss of capillary properties, suggesting the mitochondrial activation of EC may play a role in the formation of CNV.

AIM: To explore the molecular mechanism of sequoiaflavone affecting gastric cancer cells. MEHTODS: Gastric cancer cell line AGS cells were treated with gradient concentrations of sequoiaflavone, and then induced by PI3K/AKT signaling pathway activator. The optimal inhibitory concentration and time of semi-inhibitory concentration of red cedar flavonoid on AGS cells were detected by CCK-8, and the changes of cell proliferation, migration and invasion ability were detected by colony formation assay, transwell assay and wound healing assay. PI3K/AKT signal pathway related proteins p-PI3K, PI3K, p-AKT and AKT were detected by western blot. RESULTS: Sequoiaflavone inhibited AGS cells in a concentration-dependent manner. The half inhibitory concentration was 0.5 mmol/L, the optimal treatment time was 48 h. The protein expression of p-PI3K and p-AKT was down regulated. The proliferation, migration and invasion of AGS cells were decreased after treated with sequoiaflavone. After treated with PI3K / AKT signal pathway activator, the protein expression level of pPI3K and p-AKT was partially reversed, and the ability of cell viability, proliferation, migration and invasion was also partially improved. CONCLUSION: Inactivation of PI3K/AKT signaling pathway caused by sequoiaflavone inhibited gastric cancer cells proliferation, migration and invasion ability.

Objective:To investigate the fundus fluorescein angiography (FFA) characteristics of spontaneous regression in retinopathy of prematurity (ROP) and the range of retinal vascularization.Methods:A clinical retrospective study. A total of 82 eyes of 41 infants with ROP, who underwent FFA from January 2019 to December 2021 in Department of Ophthalmology of Xijing Hospital after completion of ROP regression, were included. There were 25 males (50 eyes) and 16 females (32 eyes). ROP was diagnosed in Zone Ⅱ in 44 eyes, with 38 eyes in stage 2 and 6 eyes in stage 3, and in zone Ⅲ in 38 eyes of stage 2. All patients underwent FFA examination under general anesthesia, at postmenstrual age of 70.70±12.25 weeks, after the natural regression of ROP was completed. Focus on the retinal vascular development, as well as choroid circulation and macular abnormalities, and compare and observe the differences between zone Ⅱ and Ⅲ after spontaneous regression. The extent of retinal vascularization was determined by the ratio between the distance of the center of the disc to the border of the vascularized zone (DB) and the center of the disc to the fovea distance (DF). The width of avascular area, recorded as the distance from the ora serrata to the vascular termination, was counted by disc diameters (DD). The measurement data between zone Ⅱ and zone Ⅲ ROP were compared by the independent sample t-test, and the count data were compared by χ2 test or Fisher exact probability test. Results:The linear choroidal pattern was present in 9 eyes (21.95%, 9/41), and the tortuous arteries in the posterior retina were detected in 32 eyes (39.02%, 32/82). It was noted that increased branching of vessels presented in 45 eyes (54.88%, 45/82), straight shape of vessels shown in 27 eyes (32.93%, 27/82), circumferential vessels arisen in 45 eyes (54.88%, 45/82), arteriovenous shunt appeared in 18 eyes (21.95%, 18/82), and capillary bed lost in 46 eyes (56.10%, 46/82) in areas from initial ridge to vascular termini. Punctate or linear dye leakage was observed in 23 eyes (28.05%, 23/82) during the late phase of FFA. Macular abnormalities, such as the absence of foveal avascular zone and hypoperfusion, were observed in 28 eyes (34.15%, 28/82), of which the macular ectopia presented in 1 eye. The mean DB/DF ratio of all the 82 eyes on the temporal side was 4.63±0.29 and 3.77±0.23 in the nasal. The mean avascular area on the temporal retina was 1.74±1.00 DD. Compared with ROP in zone Ⅲ, increased branching of vessels and dye leakage were more common ( χ2=9.303, 10.774; P=0.002, 0.001), the extent of temporal retinal vascularization was smaller ( t=-2.285, P=0.026), and the avascular area of the retina was more significant ( t=5.491, P<0.001) in zone Ⅱ ROP. Conclusions:Even after completion of spontaneous regression in ROP, incomplete retinal vascularization and vascular abnormalities may exist permanently in FFA, including those such as the tortuous arteries in the posterior retina, increased branching and straight shape of vessels, circumferential vessels, capillary bed lost and macular abnormality. Further appropriate follow-up is needed for long-term safety.

Drug optimization, which improves drug potency/specificity by structure‒activity relationship (SAR) and drug-like properties, is rigorously performed to select drug candidates for clinical trials. However, the current drug optimization may overlook the structure‒tissue exposure/selectivity-relationship (STR) in disease-targeted tissues vs. normal tissues, which may mislead the drug candidate selection and impact the balance of clinical efficacy/toxicity. In this study, we investigated the STR in correlation with observed clinical efficacy/toxicity using seven selective estrogen receptor modulators (SERMs) that have similar structures, same molecular target, and similar/different pharmacokinetics. The results showed that drug's plasma exposure was not correlated with drug's exposures in the target tissues (tumor, fat pad, bone, uterus), while tissue exposure/selectivity of SERMs was correlated with clinical efficacy/safety. Slight structure modifications of four SERMs did not change drug's plasma exposure but altered drug's tissue exposure/selectivity. Seven SERMs with high protein binding showed higher accumulation in tumors compared to surrounding normal tissues, which is likely due to tumor EPR effect of protein-bound drugs. These suggest that STR alters drug's tissue exposure/selectivity in disease-targeted tissues vs. normal tissues impacting clinical efficacy/toxicity. Drug optimization needs to balance the SAR and STR in selecting drug candidate for clinical trial to improve success of clinical drug development.

Objective:To investigate the ocular clinical manifestations in pediatric patients with incontinentia pigmenti (IP).Methods:A case series study was carried out and a retrospective analysis was performed.Clinical data of 13 pediatric patients with IP treated from January 2013 to December 2019 in Xijing Hospital were collected.All the patients underwent regular ophthalmologic examination.Three patients accepted fundus fluorescein angiography and six eyes of five patients were treated with retinal photocoagulation or anti-vascular endothelial growth factor (VEGF) intravitreal injection according to severity of the condition.The follow-up period ranged from 6 months to 6 years.The medical history, family history, systemic manifestations, ocular characteristics, diagnosis, treatment as well as ocular and systemic changes during follow-up were recorded and analyzed.This study followed the Declaration of Helsinki and the study protocol was approved by the Ethics Committee of Xijing Hospital, Fourth Military Medical University (No.KY20203287-1).Results:All the 13 patients were female aged from 5 days to 42 months at first visit, with the average age of 2.0 (1.0, 8.5) months.As for the main skin lesions at first visit, there were 4 cases in erythematous vesicle stage, 3 cases in verrucous exanthema stage, and 6 cases in hyperpigmented stage.There were 7 cases in shrinkage stage during follow-up.Among the 26 eyes of 13 patients, 18 eyes of 10 patients showed ocular anomalies, accounting for 76.9% of total patients (69.2% of total eyes). Among the 13 patients, 8 patients presented bilateral ocular involvement, 2 patients showed unilateral anomalies, and 3 patients had no ocular lesions.The retina was involved in all patients with ocular manifestations.The typical retinal lesions included avascular zone of peripheral retina in 13 eyes, tortuous and dilated retinal vessels in 10 eyes, increased vascular branch in 7 eyes, white linear retinal arteries and partial vascular occlusion in 4 eyes, retinal neovascularization in 3 eyes, total retinal detachment in 2 eyes, and retinal fold with macular lamellar hole in 1 eye.In addition, there was retinal hemorrhage in 11 eyes, retinal pigment changes in 4 eyes, grey ridge lesions in 3 eyes, macular dysplasia in 2 eyes, choroidal atrophy in 1 eye, optic gliosis in 1 eye and yellowish-white retinal exudate in 1 eye.There were also 4 patients with other ocular manifestations, such as strabismus and eyeball atrophy.Retinal photocoagulation was performed in 4 eyes of 3 patients and anti-VEGF intravitreal injection in 2 eyes of 2 patients.The retinal lesions regressed and the condition of patients kept stable during follow-up.Conclusions:The ocular clinical manifestations in patients with IP are usually typical and diverse, and the retinal vascular lesion is the main type.Early diagnosis and timely treatment are of great significance.

Mesenchymal stem cells (MSCs) are capable of self-replication and multi-directional differentiation, which are very important for the development and reconstruction of mesenchymal tissue. Bone tissue damage repair involves the participation of various cells and molecules. The recovery of bone mass requires sufficiently many MSCs to migrate to the damaged site to perform the reconstruction function. The local inflammatory response at the injury site can recruit MSCs and promote new bone formation. Simultaneously, niche changes during the migration of MSCs will affect their biological performance and initiate the phase of directed differentiation. This article explores the relevant mechanisms that mediate the migration of MSCs in the process of bone injury repair, including the regulation of immune cells and chemotactic signaling molecules in the inflammatory response in the bone repair stage through signaling pathways such as BMP/Smads. Then, it summarizes the mechanism by which the high matrix stiffness upregulates the expression of the integrin and focal adhesions to promote the MSCs migration and osteogenic differentiation. Simultaneously, the migration ability of MSCs can be regulated through drugs or genetic modification to promote the bone injury repair. The improvement of MSCs migration ability can shorten the time of bone tissue damage repair and improve the bone quality. This article reviews the role of the MSCs migration ability in bone tissue injury repair to provide a reference for the application of MSCs with high migration ability in the fields of stem cell therapy for bone related diseases and bone tissue engineering.

Objective: To investigate the correlation of functional connectivity (FC) and the integrity of connective fibres between hippocampus and thalamus in Alzheimer′s disease(AD) and amnestic mild cognitive impairment (aMCI). Methods: Both resting-state functional magnetic resonance imaging (rs-fMRI) and diffusion tensor imaging (DTI) data of 40 AD patients, 37 aMCI patients and 41 normal control subjects matching with age and educational level were collected. These subjects were all recruited from outpatient Department of Neurology in the Second Medical Center of Chinese PLA General Hospital, as well as poster, from May 2016 to January 2018. The FC strength between bilateral hippocampus and thalamus, as well as the parameters representing integrity of connective fibres, including fractional anisotropy (FA) and mean diffusivity(MD),were analyzed. Also, the correlations between FC strength and FA or MD strength were analyzed in the study. Results: Compared to that of normal control subjects, the FC strength between billateral hippocampus and thalamus in patients with AD, aMCI were not significantly different(P>0.05). The integrity of bilateral connective fibres between hippocampus and thalamus were damaged in AD patients when compared to normal control subjects(P<0.01). A positive correlation of connective fibres integrity with FC strength between hippocampus and thalamus was found in the left side(r=0.25,P<0.05) but rather in the right side. Conclusion In AD and aMCI patients, structural connectivity between left hippocampus and thalamus affects the functional connectivity between them.

Objective To establish and assess the new method of APTT assay based on the combinations of Mg2+and Ca2+for lupus anticoagulants(LA)measurements.Methods This prospective study included 309 trisodium citrate anticoagulated plasma samples from 244 random patients and 65 patients with different autoimmune diseases(AID)to establish and assess the method of LA measurement, respectively.Final concentrations of 0,2.0, 4.0, 8.0,16.0 mmol/L Mg2+were added into 25 mmol/L Ca2+solution, and Actin reagent was used to measured plasma APTT of 94 patients.The applied concentration of Mg2+-Ca2+solution was confirmed through the special and significant alteration of APTT from LA-positive and -negative plasma observed in the presence of Mg 2+(test solution).Based on Actin reagent use,the test solution and 25 mmol/L Ca2+solution were applied to measure APTT of patients and normal individuals, respectively, and the ratio of Mg2+-Ca2+-APTT to Ca2+-APTT(Mg2+-Ca2+-APTT indices)and normalized Mg2+-Ca2+-APTT indices(NAR)were calculated, respectively.Mixed plasma NAR was measured,and CV%was calculated to evaluate the repeatability and stability of Mg 2+-Ca2+-APTT method.APTT of 150 patients was measured with the test solution and Actin reagent to calculate Mg 2+-Ca2+-APTT indices, and normalized LA ratio was determined with dRVVT method.The applicability of Mg2+-Ca2+-APTT assay was assessed through comparisons of the results from the two methods.Finally, NAR and NLR of 65 patients with AID(including 26 SLE patients)were measured with Mg2+-Ca2+-APTT assay and dRVVT method, respectively, and ROC curve was also used to assess the efficacy of the two methods for LA measurements.Results In all LA-negative plasma,APTT increased from 28.1 ±4.5 s to 61.2 ±7.9 s in normal APTT group,47.2 ±8.9 s to 97.5 ±10.3 s in increased APTT group,and 27.6 ± 5.1 s to 61.2 ±7.9 s in ACA-positive group when Mg2+increased from 0 to 8 mmol/L in Mg2+-Ca2+solution(F=34.12, 38.9 and 28.35,P<0.01).Following increased Mg2+concentration, APTT shortened from 0 to 4.0 mmol/L, but simultaneously prolonged from 4.0 to 16.0 mmol/L in LA-positive plasma with prolonged or normal APTT(F=31.55 and 39.51, P<0.01), and APTT was significantly higher in 8.0 mmol/L than that in 4.0 mmol/L(P<0.001).The test concentration of Mg 2+/Ca2+solution was 4.0 mmol/L.The within, inter-day CV% of NAR was 1.39%,2.30%, and 3.44%, respectively. According to the judging criteria of <0.966 and >1.034 of Mg2+/Ca2+indices, there was 141 patients with increased indices and NLR <1.20, and 9 patients with decreased ones and NLR≥1.20 in all 150 patients.The area under ROC curve of NAR and NLR for LA detection was 0.913(95%CI:0.848-0.978) and 0.892(95%CI:0.817-0.966), respectively, and the cut-off value was 0.87 and 1.13, respectively. The sensitivity and specificity of NAR(85% and 77%)was higher than that of NLR(81% and 74%), respectively.The accordant rate of positive,negative,and total results between NAR and NLR was 94.4%, 98.5%,and 98%,respectively.Conclusion The method of APTT assay based on Mg2+combining Ca2+for LA measurements is feasible,and can be used to detect plasma LA of patients.

Objective Investigate the effects of early application of noninvasive respiratory support on very low birth weight infant(VLBWI). Method A total of 65 VLBWI(born during September 2015 to September 2016 with 28-32 weeks gestational age;1000 g ≤ birth weight < 1500 g;exclusion of combination with congenital deformity)were randomly divided into the early application of noninvasive respiratory support as observation group (n = 33) and the application of endotracheal intubation with gen respiratory support as control group (n = 32). Comparison of two groups was carried out by SPSS in terms of incidence of endotracheal intubation,BPD,pulmonary infection,pneumothorax,and necrotizing enterocolitis,together with rescue ratio,total oxygenation time and hospitalization. Results No significant difference was found on incidence of pneumothorax,necrotizing enterocolitis and rescue ratio between two groups. The incidence of endotracheal intubation,BPD,pulmonary infection and total oxygenation time was markly decreased in observation group. Conclusion Early application of noninvasive respiratory support benefits VLBWI via reducing incidence of endotracheal intubation,BPD,pulmonary infection, total oxygenation time.