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Objective To explore the mechanism of micro ribonucleic acid(miR)-3197 in diabetic retinopathy(DR)on the basis of the nuclear factor κB(NF-κB)signaling pathway.Methods A total of 47 DR patients admitted to Heng-shui People's Hospital from January 2021 to December 2021 were selected as the DR group,and 47 healthy individuals in the same period were collected as the control group.Their information in gender,age,fasting blood glucose(FBG),fast-ing insulin(FINS),triglycerides(TG),total cholesterol(TC)and miR-3197 were compared.The correlation between miR-3197 in DR patients and laboratory data was analyzed,and the receiver operating characteristic(ROC)curve of miR-3197 for DR diagnosis was drawn.The human retinal microvascular endothelial cells(hRMECs)were cultured in vitro and treated with 5.5 mmol·L-1 glucose[low glucose(NG)group]and 30 mmol·L-1 glucose[high glucose(HG)group],respectively.After transfecting with anti-miR-NC and anti-miR-3197,the cells were treated with 30 mmol·L-1 glucose(HG+anti-miR-NC group and HG+anti-miR-3197 group).Real-time fluorescence quantitative PCR was used to detect the relative expression level of miR-3197,flow cytometry was used to detect the apoptosis rate of hRMECs,enzyme-linked im-munosorbent assay was used for detecting tumor necrosis factor-a(TNF-a)and interleukin-6(IL-6),and Western blot was adopted to detect the expressions of aspartic protease 3 containing cysteine(cleaved caspase-3)protein,Bax protein and NF-κB signaling pathway-related proteins[phospho-NF-KB p65(p-p65),p65,phospho-NF-KB inhibited protein(p-IκBα),and NF-κB inhibited protein(IκBα)].Results The levels of FBG,FINS,TC and TG in the DR group were higher than those in the control group,and the differences were statistically significant(all P＜0.001).The relative expression level of miR-3197 in the peripheral blood of patients in the DR group(2.76±0.67)was higher than that of the control group(1.03±0.34),and the difference was statistically significant(P＜0.05).The miR-3197 level of patients in the DR group was positively correlated with FBG,FINS,TC and TG levels(r=0.672,0.587,0.511 and 0.423;all P＜0.05).The ROC curve graph showed that the area under the curve was 0.919,with sensitivity and specificity of 85.11％and 89.36％,respectively.Compared with the NG group,the HG group showed a significant increase in cell apoptosis rate and the pro-tein expressions of cleaved caspase-3,Bax,TNF-a,IL-6,p-IκBa and p-p65(all P＜0.05);compared with the HG+anti-miR-NC group,the HG+anti-miR-3197 group showed a significant decrease in cell apoptosis rate and the protein expres-sions of cleaved caspase-3,Bax,TNF-a,IL-6,p-IκBa and p-p65(allP＜0.05).Conclusion The miR-3197 is highly ex-pressed in the peripheral blood of DR patients and high glucose-induced hRMECs.Down-regulation of miR-3197 can allevi-ate high glucose-induced hRMEC apoptosis and inflammatory injury,and its mechanism of action may be related to the inhi-bition of the NF-κB signaling pathway.
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Objective To investigate the expression of circular ribonucleic acid carnitine palmitoyltrans-ferase 1A(circRNA CPT1A)in patients with diabetic retinopathy(DR)and its mechanism of action on neuropathy.Methods To-tally 80 patients(102 eyes)with type 2 diabetes admitted to our hospital from January 2019 to January 2022 were selected,including 22 patients(28 eyes)with no DR(NDR group),38 patients(48 eyes)with non-proliferative DR(NPDR group),and 20 patients(26 eyes)with proliferative DR(PDR group).Optical coherence tomography angiography was used to measure the thickness of the peripapillary retinal nerve fiber layer(pRNFL)and macular ganglion cell complex(GCC),the level of phosphatase and tensin homolog(PTEN)in peripheral blood was detected by Western blot,and the circRNA CPT1A level in peripheral blood was detected by fluorescence quantitative polymerase chain reaction.The levels of circRNA CPT1A and PTEN in peripheral blood,as well as the thickness of pRNFL and GCC,were compared among three groups,and Pearson correlation analysis was used to investigate the correlation between circRNA CPT1A and PTEN,pRNFL and GCC thickness.Results The circRNA CPT1A in the peripheral blood of the PDR group was higher than that in the NDR group and NPDR group;the PTEN in the peripheral blood of the PDR group was lower than that in the NDR group and NP-DR group(all P＜0.05).There was no statistically significant difference in the expression of circRNA CPT1A and PTEN be-tween NDPR group and NDR group(all P＞0.05).The Pearson correlation analysis showed that the expression level of cir-cRNA CPT1A in peripheral blood was negatively correlated with PTEN(P＜0.05).With the increase in disease severity,the thickness of pRNFL and GCC showed a decreasing trend;the thickness of pRNFL and GCC in the whole,upper and lower parts of eyes in the NDR group were higher than those in the NPDR group(all P＜0.05),while those in the NPDR group were higher than the PDR group(all P＜0.05).Pearson correlation analysis showed that the expression level of cir-cRNA CPT1A in peripheral blood was negatively correlated with the thickness of pRNFL and GCC in the whole,upper and lower parts of the observed eyes(all P＜0.05).Conclusion With the increase in the severity of DR,circRNA CPT1A in the peripheral blood of DR patients shows an increasing trend and is negatively correlated with peripheral blood PTEN lev-el,as well as macular pRNFL and GCC thickness.The mechanism of action may be that circRNA CPT1A negatively regu-lates the expression of PTEN and thus participates in the occurrence and development of DR.
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Objective:To evaluate the assistant role of manifestations under tracheoscopy in the diagnosis of invasive pulmonary aspergillosis (IPA) in severe patients.Methods:A retrospective study was conducted. The patients with suspected IPA admitted to intensive care unit (ICU) of Affiliated Hospital of Binzhou Medical College from January 2015 to December 2019 were enrolled. The diagnosis, clinical diagnosis and suspected diagnosis were made according to the grading criteria of Guidelines for the diagnosis and treatment of invasive fungal infection in severe patients (2007). Those who met the criteria were enrolled in the IPA group, and those who did not meet the criteria or other pathogens were enrolled in the non-IPA group. The general data of the patients were collected, and the changes of tracheal and bronchial mucosa under tracheal microscope before and after treatment were recorded, as well as the results of galactomannan (GM) test and aetiology culture of bronchoalveolar lavage fluid (BALF). The baseline, bronchoscopy and pulmonary CT manifestations and their dynamic changes were compared in each group. Results:A total of 142 patients with suspected IPA were finally enrolled. Among them, 12 were pathologically proven IPA, 77 were probable IPA, 22 were possible IPA, and 31 were undefined IPA. Of the 142 patients, 60 had typical manifestations of mucosal injury under bronchoscopy, including 7 proven IPA patients (58.3%), 52 probable IPA patients (67.5%), and 1 possible IPA patient (4.5%), but none undefined IPA patient. The patients undergoing lung CT scan were 12 proven IPA patients (100%), 73 probable IPA patients (94.8%), and 21 possible IPA patients (95.5%), respectively. Most of the Chest CT showed patchy or strip density increasing and other non-specific manifestations. There were 3 proven IPA patients (25.0%), 7 probable IPA patients (9.0%), and 0 possible IPA patient (0%) who had typical IPA CT manifestations (halo sign and cavity or crescent sign). Among the patients of proven IPA and probable IPA (89 cases), there were a total of 35 cases with endoscopic airway mucosal injury and tracheoscopy reexamination ≥ 3 times. All the 35 patients received anti-aspergillus treatment, among which 16 survived and 19 died. Among the 16 patients who survived, the microscopic appearance of mucosal injury was gradually reduced and the clinical manifestations were gradually improved. Of the 19 patients who died, 16 had deteriorated endoscopic airway mucosal injury.Conclusions:The specific manifestations of severe patients with bronchial mucosal injury are of great significance in the diagnosis of IPA. In the case of severe patients who cannot receive pathological examination or chest CT in time, dynamic observation of the changes of airway mucosal injury is a simple auxiliary method to discover the changes of patients' condition in time, evaluate the effect of antifungal therapy and the prognosis of IPA.
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Objective Detecting antibodies against RBCs,platelets,lymphocytes,and neutrophils by a solid-phase antibodies screening system.Methods mono-layer blood cell immobilized in the bottom of U-microplate respectively to prepare the solid-phase antibody screening system.Then detecting the antibodies exist or not in 2 150 random blood donor and 440transfusion patients' samples.Analyze the influence of antibodies against blood cells to the transfusion effect.Results 53RBC antibodies,22 platelet antibodies,13 lymphocyte antibodies,55 neutrophil antibodies were detected in random blood donor samples;31 RBC antibodies,38 platelet antibodies,24 lymphocyte antibodies,43 neutrophil antibodies and 115 mixed (two or more antibodies against RBC,platelets,lymphocytes or neutrophils exist at the same time) were detected in transfusion patients' samples after detection.233 suspected adverse reaction happened after transfusion,which antibodies were detected in 133 samples.Conclusion The antibodies against blood cells solid-phase screening system can applied in pre-and post-transfusion detection.The solid-phase screening method is more sensitivity than serologic tube test and flow cytometry.Existing of antibodies influence the transfusion effect.
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AIM:To study the effect of Hand2 (one of basic helix-loop-helix proteins’ transcription factors) expression on the development of the cardiac tissues in the fetal rats from gestational diabetes mellitus ( GDM) parents, and to investigate the potential pathogenesis of GDM-induced congenital cardiac defects in rats.METHODS: The adult Spra-gue-Dawley female rats were randomly divided into blank control group (n=24), GDM group (n=30), negative control group ( n=30) and insulin intervened group ( n=30) .The GDM model was established by intraperitoneal injection of 2%streptozotocin (STZ, 40 mg/kg body weight) to the pregnant rats on the successive day.The rats in insulin intervened group were injected with intermediate-acting insulin in order to keep the fasting blood glucose in the normal range.The rats in negative control group were injected with citric acid-sodium citrate buffer solution in the same position.Blood glucose and body weight were examined every day 72 h after STZ injection.On E12, E15 and E19, the rats were anesthetized and the embryonic cardiac tissues were collected after caesarean section.The histopathological changes of the cardiac tissues were observed under microscope with HE staining.The expression of Hand2 was analyzed by the method of immunohisto-chemistry.The expression of Hand2 in the cardiac tissues at mRNA and protein levels was determined by real-time fluores-cence quantitative PCR and Western blotting.RESULTS:During the development of embryonic heart, the protein expres-sion of Hand2 in the cardiac tissues was showed dynamic changes.Observed on E12, obviously increased on E15, and at the highest level on E19.Compared with the other 3 groups, the protein and mRNA expression of Hand2 in GDM group was decreased at the time points of E12 and E15.CONCLUSION:The morbidity of fetal cardiac malformation is significantly increased in GDM group, suggesting that Hand2 may be involved in the development of cardiac malformation in GDM.