Objective To establish an acute rejection model of cervical heart transplantation in mice and evaluate the survival and dynamic rejection process post-transplantation. Methods Mice were randomly divided into sham operation group (n=10), syngeneic transplantation group (n=21), and allogeneic transplantation group (n=65). Sham operation, syngeneic cervical heart transplantation, and allogeneic cervical heart transplantation were performed respectively. The survival of recipient mice and grafts, histopathological changes of graft tissues, subpopulations of splenic lymphocytes, and expression of inflammatory factors in serum and grafts were observed. Results The survival rate and graft survival rate of the sham operation group and syngeneic transplantation group were 100% at 7 days after surgery. In the allogeneic transplantation group, 5 cases failed and died on the first day after surgery. The survival rate at 7 days after surgery was 86%, and all surviving mice had grafts that stopped beating at 7 days after surgery. The allogeneic transplantation group showed significant rejection at 7 days after surgery, accompanied by tissue damage and CD8⁺ T cell infiltration. The proportion of CD8⁺ T cells in the spleen continued to rise post-operation, while the proportion of CD4⁺ T cells showed a downward trend. The expression of interferon-γ in serum and grafts peaked at 5 days after surgery, while the expression of tumor necrosis factor-α showed no statistical significance. Conclusions Acute rejection following heart transplantation in mice intensifies between 5 to 7 days after surgery, which may be a critical time window for immunological intervention.

Objective:To explore the effects and mechanism of Salvianolic acid B(SalB)on Parkinson's disease(PD).Methods:Forty-eight C57BL/6 male mice were randomly separated into control group(control)withoutdrugs,model group(MPTP)with intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrah-ydropyridine(MPTP),SalB control group with intraperitoneal injection of SalB,and SalB treatment group(MPTP+SalB).Construction of PD mouse model by intraperitoneal injection of MPTP,and treatment with intraperitoneal injection of SalB.Pole climbing test was applied to assess behavior differences.The time of first black stool excretion and water content of feces were measured to evaluate intestinal dysfunctions.The number of tyrosine hydroxylase(TH)positive cells in substantia nigra and the level of Toll like receptor 4(TLR4)in colon were analyzed by immunohistochemistry.The pathological changes of colonic mucosa were observed by HE staining.The levels of calprotectin(CP)and tumor necrosis factor-α(TNF-α)in colon were determined by ELISA.Western blot was used to determine the level of TH in midbrain,the protein level of TH,tight junction protein(ZO-1),and protein level of TLR4/MyD88/NF-κB signaling pathways which express in colon.Results:Com-pared with the Control group,the climbing time,T-turn time and the first black stool excretion time in MPTP group increased while the fecal water content and the number of TH positive cells in substantia nigra were decreased.Accompanied by TLR4 positive cells in colon,pathological injury score of colonic mucosa,levels of CP and TNF-α in colon increased,expression of TH in midbrain and expression of ZO-1 in colon decreased.Expressions of TLR4,MyD88,Nuclear NF-κB p65 and p-NF-κB p65 in colon increased.Com-pared with MPTP group,SalB treatment shortened the climbing time,T-turn time and the first black stool excretion time in SalB treat-ment group,increased the fecal water content and the number of TH positive cells in substantia nigra,lowered TLR4 positive cells in colon,enhanced expression of TH in midbrain and colon,reduced the pathological injury score of colonic mucosa,significantly decreased levels of CP and TNF-α in colon,enhanced expression of ZO-1 in colon,inhibited expressions of TLR4,MyD88,Nuclear NF-κB p65 and p-NF-κB p65 in colon.Conclusion:SalB can protect the nerves and intestines and alleviate the intestinal inflamma-tion of PD mice,which may be related to the inhibition of TLR4/MyD88/NF-κB signal pathway.

BACKGROUND:Current studies have confirmed that Ganoderma lucidum polysaccharides can promote nerve regeneration in neurodegeneration-related diseases.The occurrence of neurodegenerative diseases is closely related to mitochondrial dysfunction,but the role of Ganoderma lucidum polysaccharides on the regulation of apoptosis and mitochondrial function in neurodegenerative diseases is not yet clarified. OBJECTIVE:To explore the regulatory effects and mechanisms of Ganoderma lucidum polysaccharides on apoptosis and mitochondrial dysfunction in H2O2-induced SH-SY5Y cells. METHODS:SH-SY5Y cells were divided into three groups:control group,H2O2 group,and Ganoderma lucidum polysaccharides group.Cells in the control group were normally cultured.Cells in the H2O2 group were treated with 300 μmol/L H2O2 for 24 hours.In the Ganoderma lucidum polysaccharides group,the intervention with 300 μg/L Ganoderma lucidum polysaccharides was conducted first for 1-2 hours,followed by the addition of 300 μmol/L H2O2 for 24 hours.The mitochondrial membrane potential was detected by JC-1 kit.Apoptosis was detected by TUNEL staining kit.The activities of malondialdehyde and superoxide dismutase were detected by malondialdehyde test kit and superoxide dismutase test kit,respectively.The apoptosis and expression of mitochondrial dynamics-related proteins were detected by immunofluorescence staining and western blot assay. RESULTS AND CONCLUSION:(1)Compared with the control group,the mitochondrial membrane potential and superoxide dismutase activity were significantly reduced,as well as apoptotic rate and malondialdehyde levels were significantly increased in the H2O2 group(P<0.05).After treatment with Ganoderma lucidum polysaccharides,the membrane potential and superoxide dismutase activities were significantly increased,and apoptotic rate and malondialdehyde levels were significantly reduced compared with the H2O2 group(P<0.05).(2)The expression levels of pro-apoptotic proteins Bax and Caspase-3 were significantly increased,but the expression of anti-apoptotic protein Bcl-2 was significantly decreased in the H2O2 group compared with the control group(P<0.05).Compared with the H2O2 group,the levels of Bax and Caspase-3 were significantly decreased,but the expression of anti-apoptotic protein Bcl-2 was significantly increased in the Ganoderma lucidum polysaccharides group(P<0.05).(3)Compared with the control group,the expression of mitochondrial splitting proteins Fis1 and p-Drp1 was significantly increased,but the expression of mitochondrial fusion proteins OPA1,Mfn1,and Mfn2 was decreased in the H2O2 group(P<0.05).Compared with the H2O2 group,Fis1 and p-Drp1 expression was significantly reduced,but the expression levels of OPA1,Mfn1,and Mfn2 were significantly increased in the Ganoderma lucidum polysaccharides group(P<0.05).(4)The above results confirm that Ganoderma lucidum polysaccharides can attenuate H2O2-induced oxidative stress damage and apoptosis in SH-SY5Y cells by ameliorating mitochondrial dysfunction.

Purpose To investigate the clinical pathologic features of five SMARCA4-deficient non-small lung cancers(SMARCA4-dNSCLCs).Methods Five cases of SMARCA4-dNSCLC was underwent by HE,immunohistochemical staining,and molecular detection,analyzed their clinicopathological char-acteristics and reviewed relevant literatures.Results All 5 ca-ses were male,and mean age was 66 years.Five patients had a history of smoking,three patients were treated with cough and blood in sputum as the first symptom,one was treated with a history of pulmonary tuberculosis combined with limb mobility disorder,and one was diagnosed with pulmonary nodules by physical examination.Under microscopic observation,tumor cells were poorly differentiated,with solid nest sheet distribu-tion,some with glandular structure,tumor cells had abundant e-osinophilic or transparent cytoplasm,vacuolar nuclear chroma-tin,nucleoli was visible,and nuclear mitosis was common.In-flammatory cell infiltration and sheet of necrosis were seen in the stroma.Immunohistochemical staining showed 5/5 diffuse ex-pression of CK(AE1/AE3)and CK7,5/5 loss expression of BRG1,1/5 diffuse expression of p40 and CK5/6,and Ki67 proliferating index ranged from 20％to 90％.FISH tests showed that 4/4 SMARCA4 genes missed.Five patients were followed up for 2-15 months,3 patients died and 2 patients survived.Conclusions SMARCA4-dNSCLC can have extensive morphologi-cal features,high degree of malignancy,and complicated treat-ment.BRG1 deficiency is helpful for diagnosis.Deepening the understanding of SMARCA4-dNSCLC can help the clinical cor-rect choice of treatment strategies and accurately evaluate patient prognosis.

Objective:To evaluate the application of intraoperative optical coherence tomography (iOCT) in vitreoretinal surgery.Methods:An observational case series study was conducted.A total of 316 patients (316 eyes) who received vitreoretinal surgery and iOCT from January 2015 to December 2017 in Hangzhou Branch of Eye Hospital, Wenzhou Medical University were enrolled.The iOCT scanning time and result, the consistency between iOCT result and observation under the operating microscope as well as the influence of iOCT on surgical strategy were recorded.The postoperative adverse reactions and complications were observed.The study protocol was approved by an Ethics Committee of Eye Hospital, Wenzhou Medical University(No.2019-168-K-160). Written informed consent was obtained from all patients prior to any medical examination.Results:There were 85.8% (271/316) of patients who successfully completed iOCT scanning, with an average scanning time of (3.54±2.30)minutes.Among the 271 successful eyes, there were 51 with idiopathic macular hole (IMH), 95 with epiretinal membrane, 50 with dense vitreous hemorrhage (VH), 30 with macular lamellar hole, 12 with vitreous macular traction syndrome, 33 with myopic maculopathy.There were 45 eyes, accounting for 16.6%, the iOCT results of which were inconsistent with the observation of operator under the operating microscope.There were 27 eyes, accounting for 10.0%, the surgical strategy of which was changed.The iOCT results of IMH showed that intra-retinal bridge connection appeared in 2 eyes after ILM peeling, then air tamponade was used.High-reflection strips on the edge of the MH after ILM peeling, called the Hole-door phenomenon, were found in 15 eyes, accounting for 32.6%.In MH with a diameter >400 μm, the postoperative best corrected visual acuity and continuity of outer limiting membrane were better in eyes with Hole-door phenomenon than eyes without Hole-door phenomenon.There were 8 eyes, accounting for 8.4%, showing residual membrane, 3 eyes of which received extra ERM peeling.There were 56 eyes, accounting for 58.9%, showing the secondary changes after membrane peeling.For 50 eyes with dense VH, there were 17 eyes showing the normal macular structure, accounting for 34%, and 33 eyes showing the abnormal macular structure, accounting for 66.0%.In addition, the macular structures of 14 eyes, accounting for 28.0%, observed in iOCT image were inconsistent with the intraocular microscope finding, and 11 of them received extra ERM peeling.Conclusions:The application of iOCT in vitreoretinal surgery can guide the selection of reasonable surgical methods during operation, predict postoperative recovery as well as improve postoperative outcomes.

Objective:To study a deep learning-based dual-modality fundus camera which was used to study retinal blood oxygen saturation and vascular morphology changes in eyes with branch retinal vein occlusion (BRVO).Methods:A prospective study. From May to October 2020, 31 patients (31 eyes) of BRVO (BRVO group) and 20 healthy volunteers (20 eyes) with matched gender and age (control group) were included in the study. Among 31 patients (31 eyes) in BRVO group, 20 patients (20 eyes) received one intravitreal injection of anti-vascular endothelial growth factor drugs before, and 11 patients (11 eyes) did not receive any treatment. They were divided into treatment group and untreated group accordingly. Retinal images were collected with a dual-modality fundus camera; arterial and vein segments were segmented in the macular region of interest (MROI) using deep learning; the optical density ratio was used to calculate retinal blood oxygen saturation (SO 2) on the affected and non-involved sides of the eyes in the control group and patients in the BRVO group, and calculated the diameter, curvature, fractal dimension and density of arteriovenous in MROI. Quantitative data were compared between groups using one-way analysis of variance. Results:There was a statistically significant difference in arterial SO 2 (SO 2 -A) in the MROI between the affected eyes, the fellow eyes in the BRVO group and the control group ( F=4.925, P<0.001), but there was no difference in the venous SO 2 (SO 2-V) ( F=0.607, P=0.178). Compared with the control group, the SO 2-A in the MROI of the affected side and the non-involved side of the untreated group was increased, and the difference was statistically significant ( F=4.925, P=0.012); there was no significant difference in SO 2-V ( F=0.607, P=0.550). There was no significant difference in SO 2-A and SO 2-V in the MROI between the affected side, the non-involved side in the treatment group and the control group ( F=0.159, 1.701; P=0.854, 0.197). There was no significant difference in SO 2-A and SO 2-V in MROI between the affected side of the treatment group, the untreated group and the control group ( F=2.553, 0.265; P=0.088, 0.546). The ophthalmic artery diameter, arterial curvature, arterial fractal dimension, vein fractal dimension, arterial density, and vein density were compared in the untreated group, the treatment group, and the control group, and the differences were statistically significant ( F=3.527, 3.322, 7.251, 26.128, 4.782, 5.612; P=0.047, 0.044, 0.002, <0.001, 0.013, 0.006); there was no significant difference in vein diameter and vein curvature ( F=2.132, 1.199; P=0.143, 0.321). Conclusion:Arterial SO 2 in BRVO patients is higher than that in healthy eyes, it decreases after anti-anti-vascular endothelial growth factor drugs treatment, SO 2-V is unchanged.

Corona Virus Disease 2019 (Corona Virus Disease 2019,COVID-19) has become a public health emergency that has attracted global attention because of its large-scale outbreak resulting in numerous human infections and deaths. COVID-19 is a highly contagious respiratory disease caused by novel coronavirus 2019-nCoV. Due to a large number of infections and fast transmission speed, it's significant to diagnose the infected people quickly and detect the asymptomatic infected people as soon as possible. At present, the preliminary screening is judged by the clinical manifestations of the patients, mainly involving the respiratory system, but recent studies have found that the patients infected with COVID-19 have unique oral manifestations, such as taste disturbance, xerostomia, halitosis, inflammation of salivary glands, necrotizing periodontal disease and some of them are earlier than typical symptoms such as dry cough, fever, etc. Paying attention to the oral manifestations of patients can further improve the COVID-19 screening procedure. At present, symptomatic treatment is mainly used for these oral symptoms.

High myopia (HM) is one of the main causes of vision loss. In recent years, optical coherence tomography and other techniques have shown a variety of vitreoretinal interface abnormalities (VRIA) in highly myopic eyes. Posterior vitreous detachment and paravascular abnormality are the relatively common manifestations of VRIA. Posterior vitreous detachment is classified in several different ways in HM eyes, the onset age of which is earlier in HM. Paravascular abnormality mainly includes paravascular microfold, paravascular cyst, paravascular lamellar hole, and paravascular retinoschisis. The former two are early-stage lesions, the latter two are advanced lesions. VRIA is closely related to many HM's fundus complications, such as myopic retinoschisis, macular hole, retinal detachment and so on. VRIA may develop into myopic retinoschisis, which in turn develop into full-thickness macular hole, and even retinal detachment. Therefore, the examination and judgment of VRIA in HM patients are of great significance for the early prevention and treatment of clinical retina diseases.

Objective:To observe the incidence and risk factors of microcystic macular edema (MME) in patients with idiopathic macular epiretinal membrane (IMEM) preoperatively and postoperatively.Methods:A retrospective case series study. From January 2017 to May 2021, 72 eyes of 72 patients with IMEM in Eye Hospital of Wenzhou Medical University at Hangzhou were included. There were 18 male and 54 female. Average age was 64.8±7.8 years. Eyes were all monocular. All patients received the examination of best corrected visual acuity (BCVA) by standard logarithmic visual acuity chart, which was represented logarithmic of minimum angle of resolution (logMAR). Optical coherence tomography was used to measure central macular thickness (CMT). MME was defined as small, vertically bounded cystic space located outside the fovea in the inner layer of the retina. According to the presence or absence of MME before surgery, the affected eyes were divided into two groups: non-MME group (35 eyes) and MME group (37 eyes). The difference of logMAR BCVA and CMT was statistically significant between Group A and B ( t=3.117, 2.589; P=0.003, 0.012). All patients with IMEM were treated with 23G three-channels pars plana vitrectomy (PPV) with epiretinal membrane and inner limiting membrane (ILM) peeling. The two groups were further divided into four groups according to whether there was MME in the postoperative follow-up time. The group A1 was without MME before and after surgery, group A2 was without MME before surgery and with MME after surgery. The MME group was subdivided into the group with MME before surgery and without MME after surgery (group B1) and the group with MME before and after surgery (group B2). The mean follow-up time was 8.8±7.7 months. The same equipment and methods were used to exam the patients during the follow-up. Paired t test was used to compare the changes of MME, BCVA and CMT before and after surgery. The differences of CMT and BCVA among groups before and after surgery were compared by independent sample t test and one-way ANOVA. Logistic regression was used to analyze the influencing factors of MME before and after surgery, and multiple linear regression was used to analyze the influencing factors of postoperative BCVA. Results:There were 35 eyes in the non-MME group, 18 eyes (51.43%, 18/35) in the A1 group and 17 eyes (48.57%, 17/35) in the A2 group, respectively. There were 37 eyes in MME group, 6 eyes (16.22%, 6/37) in group B1 and 31 eyes (83.78%, 31/37) in group B2, respectively. At last follow-up, the logMAR BCVA was 0.10±0.12, 0.25±0.17, 0.09±0.11, 0.30±0.26 in group A1, A2, B1, and B2, respectively. Compared with the logMAR BCVA before surgery, the differences were statistically significant ( t=3.779, 4.253, 7.869, 6.668; P<0.01). There was significant difference in logMAR BCVA among the four groups ( F=4.460, P<0.01). There was a significant difference in logMAR BCVA between group A1 and group A2 ( t=-2.930, P=0.006). There was no significant difference between group B1 and group B2 ( t=-1.921, P=0.063). The CMT of group A1, A2, B1 and B2 were 371.83±73.24, 431.24±83.13, 407.00±28.07 and 425.19±70.97 μm, respectively. Compared with those before operation, the differences were statistically significant ( t=5.197, 2.465, 3.055, 6.078; P<0.05). There was no significant difference in CMT among the four groups ( F=2.597, P=0.059). Logistic regression analysis showed that pre-operation MME was correlated with pre-operation IMEM stage ( β=1.494, P=0.004). New MME after surgery was correlated with age ( β=0.153, P=0.013). Multiple linear regression analysis showed that postoperative visual acuity was significantly correlated with CMT before surgery and MME after surgery ( β=0.001, 0.134; P=0.015, 0.019). Conclusions:PPV combined with epiretinal membrane and ILM peeling surgery for IMEM can improve visual acuity and decrease CMT. MME regress or regenerate after surgery. Age is an independent predictor of the risk of newly formed MME after surgery.

Objective:To explore the positive rate of scanning laser ophthalmoscope in the retromode (RM-SLO) in different types of diabetic macular edema (DME), and to analyze its correlation with foveal thickness (CMT) and macular volume.Methods:From March to May 2021, 40 patients (65 eyes) were diagnosed as DME by fundus examination combined with optical coherence tomography (OCT) in Affiliated Eye Hospital of Wenzhou Medical University at Hangzhou were included in the study. All eyes underwent best corrected visual acuity (BCVA), OCT and RM-SLO fundus imaging examinations, 47 eyes underwent fluorescein fundus angiography (FFA) examination. RM-SLO fundus imaging examinations were performed with Mirante SLO, including retro mode illumination deviated right (RMDR) and retro mode illumination deviated left (RMDL). If one or more of the RMDR and RMDL of the examined patient can identify macular edema, RM-SLO was considered to be able to identify macular edema. The macular volume at CMT and 6 mm from the fovea was measured by OCT software. DME were divided into 3 types based on OCT images: diffuse retinal thinkening (DRT) type; cystoid macular edema(CME) type; serous retinal detachment (SRD) type, focal leakage type, diffuse leakage type and diffuse cystic leakage type. The consistency of RMDR and RMDL in the diagnosis of DME in RM-SLO fundus imaging was evaluated, as well as their positive rate in different classifications of DME. The correlation between the detection of macular edema by RM-SLO and the DME type, CMT and foveal volume, and the correlation between BCVA and edema type, CMT and macular volume were analyzed.Results:Among 65 eyes, the positive rates of RMDR and RMDL fundus imaging to detect DME were 46 (70.77%, 46/65) and 48 (73.85%, 48/65), respectively. There was good consistency in identifying DME (Kappa value=0.770; P<0.001). The positive rates of RMDR and RMDL fundus imaging DRT, CME and SRD type of DME were 42.11% (8/19), 57.89% (11/19), 77.78% (28/36), 77.78% (28/36), 100.00% (10/10), 90.00% (9/10), respectively. In the FFA classification of them, the positive rates of focal leakage, diffuse leakage and diffuse cystic leakage were 68.75% (11/16), 62.50% (10/16), 68.00% (17/25), 76.00% (19/25), 100.00% (6/6), 100.00% (6/6), respectively. The results of Spearman correlation analysis showed that whether RM-SLO could identify DME was associated with CMT and OCT classification ( r=0.310, 0.365; P=0.120, 0.003); there was no correlation between FFA classification and macular volume ( r=0.113, 0.117; P=0.449, 0.352). BCVA was correlated with CMT and macular volume ( r=0.307, 0.269; P=0.013, 0.030), however, there was no significant correlation with OCT type, angiographic type ( r=0.051, 0.175; P=0.684, 0.240). Conclusion:The diagnostic agreement of DME are good between RMDR, RMDL of RM-SLO image. DME of DRT type and patients with smaller CMT in OCT are difficult to identified by RM-SLO fundus imaging.