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.

Objective To explore the molecular mechanism of hypophrenia induced by Down syndrome (DS).Methods Ts65Dn mice were used as DS animal model.Three female mice and three male mice of three to twelve weeks old were mated.Among the 17 first-generation mice alive,five mice remained Ts65Dn trisomy and 12 mice were normal.Five Ts65Dn mice and five normal mice were selected randomly as Ts65Dn group and control group,and bred till 16 to 18 weeks old for experiments.Differential proteins in hippocampus of mice were tested by isobaric tags for relative and absolute quantitation (iTRAQ).Expressions of the differential proteins in Ts65Dn group were detected compared with those in control group.Results A total of 2805 proteins were identified in hippocampus of Ts65Dn group and control group,and significant differences were observed in the expressions of 374 proteins.Compared with those in control group,expressions of 195 proteins increased and 179 reduced in Ts65Dn group.Sorted by P value from low to high,the seven proteins with the lowest P value were uncharacterized protein C2orf47 homolog,isoform 2 of filamin A-interacting protein 1-like,zinc finger protein,isoform 1 of pericentriolar material 1 protein,SEC23 interacting protein,BAG family molecular chaperone regulator 3 and serpin H1.Conclusions Differential proteins are observed in hippocampus of Ts65Dn mice,perhaps closely correlating to neurological defects.The new technology of iTRAQ helps to screen and identify differential proteins in hippocampus.

A secretive expression vector of Pichia pastoris system which can be used for the direct cloning of PCR products was constructed,and was verified through the expression of recombinant cellobiohydrolase II in Pichia pastoris.A randomly selected fragment was amplified with properly designed primers by PCR.The XhoI and Eam1105Ⅰ restriction sites were included in the 5'end of the fragment,and the Eam1105Ⅰ and XbaI restriction sites were included in its 3'end.The PCR amplified product was inserted into the P.pastoris expression plasmid pPICZ?A through XhoI and XbaI restriction sites and the resultant plasmid was digested with Eam1105Ⅰ,and lastly the big fragment was recovered,generating the P.pastoris expressive Tvector pPICZ?T.Then the cellobiohydrolase II of T.reesei was successfully expressed in P.pastoris with this expressive Tvector.Such results indicated that the constructed expression Tvector was convenient for PCR product cloning,and was effective for heterologous protein expression in P.pastoris.On the other hand,the application of the expression Tvector avoided the introduction of additional amino acids at the Nterminus of the expressed protein,which generally occurred when normal expression vectors were used in secretive expression system.

Objective To evaluate the efficacy of three-dimensional conformal radiotherapy (3DCRT) combined with concurrent chemotherapy for loco-regionally recurrent or mastastatie rectal cancer. Methods Between June 2004 and January 2007,47 patients with loco-regionally recurrent or nmatastatic rectal cancer were treated by 3DCRT of 55-65 Gy in 1.8-2.0 Gy fractions. Chemotherapy was given concurrenfly using oxaliplatin(100 mg/m2 ,iv drop,d1 ) and capecitabine(1500 mg/m2,orally,dl-14,21 days per cycle). Results After the follow-up of 12-35 months, the total response rate, complete response rate and partial response rate were 79% (37/47) ,19% (9/47) and 60% (28/47) ,respectively. The pain-alleviation rate and the mean pain-alleviation time were 85% and 6 months. The 1- and 2-year survival rates were 83% and 51%. Quality of life was improved without any treatment related death. Conclusions 3DCRT combined with concurrent chemotherapy is effective and well-tolerated in patients with post-operatively locoregionally recurrent or mastastatic rectal cancer.

Adult ; Amides ; adverse effects ; Benzene ; adverse effects ; Burns, Chemical ; pathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Nitro Compounds ; adverse effects ; Young Adult

Objective To study the value and clinical significance of diffusion tensor imaging(DTI)in differentiating high-grade astrocytomas and solitary metastatic tumors of brain.Methods 16 patients with intracranial solitary metastasis and 25 patients with high-grade astrocytoma confirmed pathologically were examined with MR DTI.Mean diffusivity(MD)and fractional anisotropy(FA)were measured in the regions of interest(ROI)of peritumor and cerebral parenchyma of the normal side on DTL Fiber tractography was reconstructed and the encountered patterns of tumor-related alteration to cerebral white matter were observed.Results The peritumor were all depicted as hyperintense or isointense signals on DTI.The FA values were 0.227±0.05 and 0.169±0.07 in the peritumor of high-grade astrocytoma and metastasis respectively,which was of significant difference(P＜0.05).Furthermore,fiber tractography could display the course of white matter tracts in three-dimensional space.Conclusion DTI may be help to differential diagnosis of solitary metastasis and high-grade astrocytoma in clinical practice.

OBJECTIVE To report a case of AIDS associated with penicilliosis marneffei and discriminate the morphology in bone marrow examination and cultural characteristic of Histoplasma capsulatum and Leishmania donovani.METHODS The three pathogens in bone marrow morphology and cultural characteristic were studied.RESULTS AIDS patient was often infected with P.marneffei,but we could differentiated easily from other pathogens because of its breeding by splitting up and the characteristis structure in bone marrow examination.CONCLUSIONS Isolation of the P.marneffei is the golden diagnosis standard,and the examination of bone marrow is important to diagnose P.marneffei infection.

Objective To investigate the application of vascularized free fibula flaps in reconstruction of mandibular defect,and to evaluate the survival rate and repair effect of the method.Methods 16 patients with mandibular tumor,having a desire to reconstruct mandible,and their systemic state could tolerate the long time operation, were selected to reconstruct mandibular defect by vascularized fibula flaps,of which 12 were male,4 female,aged 24-66 years old,average 45.2 years old.The resection of primary tumor and free fibula flaps were simultaneously conducted,then the fibular flaps were shaped according to the defect location and size of the mandible,and were fixed with reconstructive titanium plate for repair and reconstruction of mandible. The survival of flaps was determined by skin color, texture, and skin temperature of the flaps. The reconstruction effects were evaluated through the patients’surface like photos and X-ray picture of mandible before and after operation.Results All of flaps were survived and no serious complications were found. The complications of the supplied sites were skin tension and wound dehiscence, which were healed by dressing. The mandibular reconstruction effects were good through 2 or more persons’double blind evaluation.Conclusion Free fibula flaps have high survival rate and good results in mandibular reconstruction,and they can meet the needs of various types of mandibular defect repair.

Fibrinolytic Agents ; adverse effects ; Humans ; Male ; Middle Aged ; Thrombocytopenia ; chemically induced

Objective To investigate precision error between Varian auri-point localization system and Tosplane digital-bed localization system. Methods Varian Eclipse localization method and Topslane localization method were used separately to locate one target area for one same patient. Then deviations on the tumor center coordinate were measured under the simu- lation localization machine. Results Under two localization methods, the length error: 1.7mm ?0.3mm; the width error: 0.5mm?0.4mm; the altitude error: 1.2mm?0.4mm. Conclusion The curative effect of radiotherapy would be improved by applying the Topslane localization bed to the Varian Eclipse plan system.