Objective To analyze the risk factors of type 2 diabetic peripheral neuropathy (DPN),in order to provide incidence for clinical treatment and prevention.Methods Two hundred and eighty-seven patients with type 2 diabetes were divided into DPN group(113 cases) and non-DPN group(174 cases)according to electrophysiological examination and diagnosis.The clinical information were collected including body mass index (BMI),blood pressure,fasting plasma glucose (FPG),2 h postprandial plasma glucose (2 h PG),fasting plasma insulin (FINS),C peptide,glycosylated hemoglobin (HbAl c),blood fat and cholesterin.The DPN prevalence in patients with type 2 diabetes was calculated,and t or x2 analysis and multivariate logistic regression analysis were applied.Results Among the 287 patients with type 2,the DPN prevalence was 39.4％ (113/287).The level of age,duration of diabetes and smoking in DPN group were significantly higher than those of non-DPN group,while exercise,income situation and educational background were significantly lower than those of non-DPN group (x2 =4.378,8.430,4.525,4.500,4.203,6.890,P ＜ 0.05 or ＜ 0.01).Systolic blood pressure((137.52 ± 16.10) mmHg),FPG ((11.42 ± 3.08) mmol/L),2 hPG ((18.70 ± 4.61) mmol/L),HbA1c ((10.21 ± 2.50)％) in DPN group were higher than those of non DPN group ((systolic pressure (132.67± 15.80) mmHg,FPG(9.96 ±3.76) mmol/L,2 hPG(15.38 ±5.26) mmol/L,HbA1c(9.54 ±2.83)％).In DPN group,Fasting insulin,2 h insulin,fasting C peptide were (13.52 ± 4.92) mmol/L,(36.20 ± 17.52) mmol/L,(1.44 ± 0.62) mmol/L,(3.89 ± 3.01) mmol/L,lower than those of non DPN group ((16.76 ± 5.24) mmol/L,(47.95 ± t5.04) mmol/L,(1.83 ± 0.57) mmol/L,(5.24 ± 3.45) mmol/L),and the differences were significant (t =2.512,3.592,5.635,2.105,5.312,5.863,5.372,3.502,P ＜0.05 or P ＜ 0.001).Multiple logistic regression analysis revealed that the level of duration of diabetes,HbA1c,2 hPG were positively correlated with DPN prevalence,and those were the independent risk factors of DPN (OR(95％CI) 1.040(1.018-1.062),1.331(1.032-1.717),1.366(1.044-1.787),P＜0.05).The level of FINS,Fasting C peptide,2 h C peptide were negatively correlated with DPN prevalence,and those were independent protective factors of DPN (OR (95 ％ CI) 0.803 (0.725-0.889),0.923 (0.731-0.954),0.863 (0.801-0.930),P ＜ 0.05).Conclusion The occurrence of DPN is common in patients with type 2 diabetes.The indices of duration of diabetes,HbA1c,2 hPG were risk factors of DPN,and there is more dangerous with the lower level of FINS,Fasting C peptide,2 h C peptide.

Objective To conduct mutation screening of SCN1A 3′ untranslated region (UTR) on Dravet syndrome (DS) patients without mutations in the SCN1A coding region and promoter region, and functional analysis of the variant from DS patients.Methods Twenty-eight DS patients without mutations in the SCN1A coding region and promoter region were screened for SCN1A 3′ UTR mutations using PCR and direct sequencing.Functional analysis of the detected mutation was done via luciferase assay, mRNA stability analysis and RNA electrophoretic mobility shift assay (RNA-EMSA).Results A novo variant (c.*20A>G) in SCN1A 3′ UTR was found in one DS patient.The variant (c.*20A>G) reduced the luciferase gene xpression by 30% through increasing the affinity of pluripotent embryonal carcinoma cell line NT2/cytoplasmic protein binding and reducing luciferase gene mRNA stability (t=8.5,P<0.01).Conclusions A functional variant was detected from one patient with DS.This variant negatively regulated the gene expression by increasing the affinity of pluripotent embryonal carcinoma cell line NT2/cytoplasmic protein binding and reducing mRNA stability.

Objective:To investigate the correlations of human leukocyte antigen (HLA)-A, B, C, and DRB1 genotypes with cross-reactivity caused by aromatic antiepileptic-drugs.Methods:A case-control association study was carried out on subjects who accepted treatments/physical examination in our hospitals from September 2016 and September 2020; 31 patients with aromatic antiepileptic drugs (carbamazepine, phenytoin, oxcarbazepine, lamotrigine and phenobarbital)-induced cross-reactivity were enrolled as patient group, 52 tolerant subjects who took the 5 antiepileptic drugs for more than 3 months without cross-reactivity were chosen as tolerant control group, and 500 healthy volunteers were recruited as normal control group. The ethnicity of all patients and controls was Han Chinese. High-resolution genotyping was performed to compare the HLA-A, B, C, and DRB1 genotypes in subjects of the 3 groups. χ2 test or Fisher's exact test were used to analyze the correlations of HLA genes with cross-reactivity caused by aromatic antiepileptic-drugs. Results:The presence of HLA-B*13:01 genotype in the patient group, the tolerant control group, and the normal control group was 45.2% (14/31), 15.4% (8/52) and 14.6% (73/500), respectively. The presence of HLA-B*13:01 genotype in the patient group was significantly higher as compared with that in the tolerant control group and normal control group ( Pc<0.017). No other HLA genotypes were found to be associated with cross-reactivity caused by aromatic antiepileptic-drugs. Conclusion:HLA-B*13:01 is the risk genotype for cross-reactivity caused by aromatic antiepileptic-drugs.

Objective:To establish an artificial intelligence robot-assisted diagnosis system for fundus diseases based on deep learning optical coherence tomography (OCT) and evaluate its application value.Methods:Diagnostic test studies. From 2016 to 2019, 25 000 OCT images of 25 000 patients treated at the Eye Center of the Second Affiliated Hospital of Zhejiang University School of Medicine were used as training sets and validation sets for the fundus intelligent assisted diagnosis system. Among them, macular epiretinal membrane (MERM), macular edema, macular hole, choroidal neovascularization (CNV), and age-related macular degeneration (AMD) were 5 000 sheets each. The training set and the verification set are 18 124 and 6 876 sheets, respectively. Through the transfer learning Attention ResNet structure algorithm, the OCT image was characterized by lesion identification, the disease feature was extracted by a specific procedure, and the given image was distinguished from other types of disease according to the statistical characteristics of the target lesion. The model algorithms of MERM, macular edema, macular hole, CNV and AMD were initially formed, and the fundus intelligent auxiliary diagnosis system of five models was established. The performance of each model-assisted diagnosis in the fundus intelligent auxiliary diagnostic system was evaluated by applying the subject working characteristic curve, area under the curve (AUC), sensitivity, and specificity.Results:With the intelligent auxiliary diagnosis system, the diagnostic sensitivity of the MERM was 93.5%, the specificity was 99.23%, and AUC was 0.983 7; the diagnostic sensitivity of macular edema was 99.02%, the specificity was 98.17%, and AUC was 0.994 6; the diagnostic sensitivity of macular hole was 98.91%, the specificity was 99.91%, AUC was 0.996 2; the diagnostic sensitivity of CNV was 97.54%, the specificity was 94.71%, AUC was 0.987 5; the diagnostic sensitivity of AMD was 95.12%, the specificity was 97.09%, AUC was 0.985 3.Conclusions:The artificial intelligence robot-assisted diagnosis system for fundus diseases based on deep learning for OCT images has accurate and efficient diagnostic performance for assisting the diagnosis of MERM, macular edema, macular hole, CNV, and AMD.

IL-34 can promote osteoclast differentiation and activation, which may contribute to steroidinduced osteonecrosis of the femoral head (ONFH). Animal model was constructed in both BALB/c and IL-34 deficient mice to detect the relative expression of inflammation cytokines. Micro-CT was utilized to reveal the internal structure. In vitro differentiated osteoclast was induced by culturing bone marrow-derived macrophages with IL-34 conditioned medium or M-CSF. The relative expression of pro-inflammation cytokines, osteoclast marker genes, and relevant pathways molecules was detected with quantitative real-time RT-PCR, ELISA, and Western blot. Up-regulated IL-34 expression could be detected in the serum of ONFH patients and femoral heads of ONFH mice. IL-34 deficient mice showed the resistance to ONFH induction with the up-regulated trabecular number, trabecular thickness, bone value fraction, and down-regulated trabecular separation. On the other hand, inflammatory cytokines, such as TNF-α, IFN-γ, IL-6, IL-12, IL-2, and IL-17A, showed diminished expression in IL-34 deficient ONFH induced mice. IL-34 alone or works in coordination with M-CSF to promote osteoclastogenesis and activate ERK, STAT3, and non-canonical NF-κB pathways. These data demonstrate that IL-34 can promote the differentiation of osteoclast through ERK, STAT3, and non-canonical NF-κB pathways to aggravate steroid-induced ONFH, and IL-34 can be considered as a treatment target.

ObjectiveExploring the role of microRNA126 （miRNA126） in chronic kidney disease combined with atherosclerosis （CKD AS） by regulating the phosphatidylinositol-3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway and the mechanism of Shenshuai Xiezhuo decoction in the intervention of CKD AS rats with 5/6 nephrectomy combined with high-fat feeding. MethodA total of 60 SD rats were randomly divided into sham operation group， model group， losartan group， and low， medium， and high dose groups of Shenshuai Xiezhuo decoction. The CKD AS rat model was established by 5/6 nephrectomy combined with high-fat feeding for 10 weeks. The low， medium， and high dose groups （6.0， 12.0， 24.0 g·kg^-1·d^-1） of Shenshuai Xiezhuo decoction and the losartan group （20 mg·kg^-1·d^-1） were gavaged， and the corresponding intervention was carried out for eight weeks. Then， the rats were killed， and samples were collected for corresponding detection. Fully automated biochemical analyzers were used to detect kidney function and blood lipids in rats： blood creatinine （SCr）， blood urea nitrogen （BUN）， total cholesterol （TC）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C） levels. Hematoxylin-eosin （HE） and Masson staining of aortic tissue and pathological observation under a light microscope were carried out， and autophagosomes and autophagy lysosomes were observed by transmission electron microscopy. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to determine the mRNA levels of miRNA126， PI3K， Akt， and mTOR in rats， and Western blot was used to determine the protein expression levels of phosphorylated （p）-PI3K， PI3K， p-Akt， Akt， p -mTOR， mTOR， benzyl chloride 1 （Beclin-1）， and microtubule-associated protein light chain 3Ⅱ/Ⅰ （LC3Ⅱ/LC3Ⅰ）. ResultCompared with the sham operation group， the serum SCr， BUN， TC， TG， and LDL-C in the model group were significantly increased （P<0.01）. Compared with the model group， the SCr， BUN， TC， TG， and LDL-C were decreased in the losartan group and low， medium， and high dose groups of Shenshuai Xiezhuo decoction （P<0.05）. Compared with the sham operation group， thickening plaques， infiltration of mononuclear macrophages， a small number of foam cells， disordered arrangement of smooth muscle fibers in the tunica media， and increased collagen fibers were observed in the model group， and the lesions in the losartan group and Shenshuai Xiezhuo decoction groups were alleviated compared with those in the model group. Compared with the model group， the number of autophagosomes and autophagy lysosomes increased in the medium and high dose groups of Shenshuai Xiezhuo decoction. Compared with the sham operation group， the expression of miRNA126 in the aortic tissue of the model group was significantly decreased （P<0.01）， and the mRNA expressions of PI3K， Akt， and mTOR were significantly increased （P<0.01）. Compared with the model group， the expression of miRNA126 in the aortic tissue of rats in high， medium， and low dose groups of Shenshuai Xiezhuo decoction and losartan group was significantly increased （P<0.01）， while the mRNA expressions of PI3K， Akt， and mTOR were significantly decreased （P<0.01）. Compared with the sham operation group， the protein expressions of p-PI3K， PI3K， p-Akt， Akt， p-mTOR， and mTOR in the model group were significantly increased （P<0.01）， while the protein levels of Beclin-1， LC3Ⅰ， and LC3Ⅱ were significantly decreased （P<0.01）. Compared with the model group， the protein expressions of p-PI3K， PI3K， p-Akt， Akt， p-mTOR， and mTOR in the losartan group and low， medium， and high dose groups of Shenshuai Xiezhuo decoction were decreased （P<0.05）， while the protein levels of Beclin-1 and LC3Ⅱ/LC3Ⅰ were increased （P<0.05）. ConclusionThe expression of miRNA126 is decreased in the aortic tissue of CKD AS rats， and the PI3K/Akt/mTOR pathway is activated to inhibit autophagy flux. Shenshuai Xiezhuo decoction regulates the PI3K/Akt/mTOR signaling pathway through miRNA126， restores the autophagy of aortic endothelial cells， protects the damage of CKD vessels， reduces the formation of As plaques， and slows the development of cardiovascular complications.