Objective To explore the molecular effect and interaction among nephrin, podocin, CD2AP and ?-actinin-4. Methods Firstly, the recombinant RNA interference (RNAi) plasmid-psiRNA-hH1GFPzeo, specifically targeting to the mRNA of nephrin, podocin, CD2AP or ?-actinin-4, was respectively tansfected into the mouse podocyte clone (MPC5) to each knockdown (KD) the expression of nephrin, podocin, CD2AP or ?-actinin-4. Molecular distributions were revealed by confocal microscopy, and the mRNA and protein expressions were detected with semi-quantitative RT-PCR and Western blotting. Results (1)In podocin KD group (siPod966 and siPod54), the mRNAs of podocin and nephrin were not detected, their protein decreased 92% and 79%, 82% and 67%, respectively. The mRNA and protein level of CD2AP increased 62% and 42%, 71% and 46%, respectively, whereas ?-actinin-4 did not change. In nephrin KD group (siNep492), the mRNA expression and protein level of nephrin were not detected, CD2AP increased 35% and 48%, respectively; and whereas podocin and ?-actinin-4 did not change. In CD2AP KD group (siCda744 and siCda21), the mRNA of expression CD2AP was not detected, and its protein level decreased 92% and 83%, the mRNA and protein of nephrin decreased 60% and 48%, 76% and 72%, respectively; podocin increased 38% and 22%, 56% and 44%, respectively; whereas ?-actinin-4 did not change. In ?-actinin-4 KD group (siAct1790 and siAct319), the mRNAs expression of ?-actinin-4 and nephrin decreased 69% and 58%, 64% and 49%, respectively; their protein level decreased 81% and 55%, 71% and 64%, respectively. However, the mRNAs of podocin and CD2AP increased 50% and 34%, 45% and 28%, respectively; and their protein level increased 64% and 46%, 65% and 42%, respectively. (2) With their expression change, the distributions of nephrin, podocin and CD2AP shifted evidently from the cell membrane surface to the nucleus circumference, whereas ?-actinin-4 showed no change, which was still localized in the cytoplasm and further extended to foot processes. Conclusion (1) Nephrin might more independently play a crucial role in the slit diaphragm complex. (2) Alpha-actinin-4 might interact direcdy or indirectly with nephrin, podocin and CD2AP. (3) The relationship among these podocyte molecules might not be spontaneous, either a single-directional or bi-directional reaction. (4) The normal localization of these podocyte molecules might depend on their normal expression quantity.

Objective To dynamically observe the expression of slit diaphragm complex molecules, including nephrin, podocin, CD2AP, and cytoskeleton protein a-actinin-4, in adriamycin-induced nephrotic (ADN) rats, and to further explore the molecular behavior of podocyte proteins during the occurrence and development of proteinuria and their possible mechanisms. Methods Adriamycin nephropathy was induced by a single tail intravenous injection of adriamycin. Renal tissue samples were collected at day 3, 7, 14, and 28, respectively. The distribution, mRNA expression and protein expression of nephrin, podocin, CD2AP and a-actinin-4 were examined by indirect immunofluorescence, real-time PCR and Western blotting, respectively. Results (1) After the adriamycin injection, a significant increment of the 24-hour urinary protein was observed at day 14 and persisted up to day 28 (P

Objective To explore the molecular mechanisms underlying therapeutic responses of the anti-proteinuria drugs from the view of podocyte molecule. Methods Adriamycin (ADR) nephropathy was induced by a single tail intravenous injection of adriamycin. Lisinopril, prednisone and all-trans retinoic acid (ATRA) were administered once a day to the adriamycin-induced nephrotic rats at the first day after adriamycin injection respectively. Renal tissue samples were collected at day 3, 7, 14, and 28 after adriamycin injection respectively. The distribution, mRNA expression and protein expression of nephrin, podocin, CD2AP and ?-actinin-4 were examined by indirect immunofluorescence, real-time PCR and Western blotting, respectively. The interactions among nephrin and podocin, nephrin and CD2AP, as well as the nephrin phosphorylation were detected by immunoprecipitation, respectively. Results Compared to the control rats, 24 h urinary protein of the ADR rats increased significantly at day 14 (P

Objective To develop a simple high-resolution melting ( HRM) analysis method for differentiation of Pc and P2 variants in class 1 integron.Methods DNA fragments containing Pc and P2 variants were amplified from plasmids pACW ( PcW ) and pACWP2 ( PcW-P2 ) respectively , then these purified PCR products and P 2 promoters were analyed full-length amplicon by HRM .Eight DNA fragments containing different Pc promoters were amplified and site-specific mutated from plasmids pACS ( PcS ) , pACH2 ( PcH2 ) , pACH1 ( PcH1 ) , pACW ( PcW ) , genomic DNA of Klebsiellar pneumonia HS07-68 (PcWTGN-10)and HS05-1792(PcH2TGN-10)respectively.The purified PCR products and eight Pc variants were characterized by HRM analyses of an unlabeled probe and full-length amplicon.This assay was applied to the differentiate Pc and P2 variants in 109 class 1 integrons from 95 urine clinical Escherichia coli isolates in Huashan Hospital during 2004 -2007.The differentiation results were compared with that determined by direct sequencing .Results P2 promoter with a significant higher melting temperature ( Tm ) can be identified by HRM analysis clearly .P2 promoters were identified in 2 class 1 integrons and consistent with direct sequencing results .Eight Pc variants were classified into three groups: PcS, PcSTGN-10 , PcW, PcWTGN-10, PcH1, PcH1TGN-10.Using direct HRM analysis.PcH2, PcH2TGN-10 were classified into four groups:PcS, PcH1, PcH2, PcW, PcSTGN-10 , PcH1TGN-10 , PcH2TGN-10 , PcWTGN-10 according to the melting curves of the unlabeled probe .Combined the HRM analyses of the whole amplicon and unlabeled probe , the eight Pc variants can be differentiated from each other .Five different Pc variants, PcS, PcW, PcH1, PcH2TGN-10 and PcWTGN-10 , were identified and consistent with direct sequencing results .Conclusions This developed a simple Pc and P 2 variants differentiation method via simultaneous HRM analyses of an unlabeled probe and full-length amplicon .This method is cost-effective and accurate , could be used in differentiation of Pc and P2 variants of class 1 integrons in clinical isolates .

Objective To investigate the clinicopathologic significance of Y-box binding protein-1 (YB-1)expression in cervical cancer and its correlation with epithelial-mesenchymal transition. Methods A series of 202 samples,including 50 cases of normal cervical tissues,100 cases of cervical intraepithelial neo-plasia(CIN)and 52 cases of squamous cell carcinoma(SCC),were examined YB-1 and E-cadherin by immu-nohistochemical staining. Results The E-cadherin cell membrane immunoreactivity for normal/ CINⅠ,CINⅡ-Ⅲ,and SCC tissues were 100% ,64% and 3. 85% ,respectively( χ2 = 40. 909;χ2 = 119. 088;χ2 =25. 274;P < 0. 05). The negative and aberrant expression of E-cadherin was higher in metastatic SCC (100. 0% ,20 / 20)than that in non-metastatic SCC(68. 75% ,22 / 32)(χ2 = 5. 857,P = 0. 016). 94. 23%(49 / 52)cases of SCC exhibited strong YB-1 cytoplasmic immunoreactivity. The positive rates in normal/ CINⅠand CINⅡ-Ⅲ were 0(0 / 100)and 10. 00%(5 / 50)(χ2 = 72. 591;χ2 = 139. 059;χ2 = 5. 857;P < 0. 05). The cytoplasmic expression of YB-1 was higher in metastatic SCC(100. 0% ,20 / 20)than that in non-metastatic SCC(59. 38% ,19 / 32)(χ2 = 10. 833,P = 0. 001). The rates were 60. 71%(17 / 28)and 91. 67%(22 /24)in early stage SCC and late stage SCC(χ2 = 6. 603,P = 0. 01). Conclusion YB-1 over-expression is as-sociated with the malignant transformation of cervical epithelium,stage progression and metastasis of cervical cancer. The up-regulation of YB-1 is also associated with the down-regulation of E-cadherin,and it may predict the malignant transformation of CIN and distal metastasis of cervical cancer.

Abstract: Objective To understand the morphological characteristics and ultrastructure of the dominant species of Rhipicephalus sanguineus in Hainan at different developmental stages, and provide theoretical basis for the identification of the lineage and control of Rhipicephalus sanguineus. Methods The external morphology of different developmental stages of the dominant species of Rhipicephalus sanguineus, including larva, nymph and adult tick in Hainan were observed by scanning electron microscope. Results The division between each segment of larva pedipalps was not obvious, and setae was serrated; dental formula type 2 | 2; 3 pairs of podomere; a pair of setae on the anal valve; none of anal groove, spiracular plate, porous area and genital aperture. There was a clear boundary at the beginning of each segment of nymph pedipalps; dental formula type 2 | 2; 4 pairs of podomere; 3 pairs of setae on the anal valve; anal groove; none of porous area and genital aperture. The male adult tick's trichotheca are covered by the pedipalps, and the whole bristles are conical; dental formula type 3 | 3; 4 pairs of podomere; anal groove and paraprocts; 7 setae on the anal valve; genital aperture was oval. The female of adult tick can be distinguished by dental formula 3 | 3; pairs of podomere; porous areas with 3 short setae; anal groove; 4 pairs of setae and 2 pores on the anal valve; genital pore was broadly U-shaped. In addition, the male adult's scutum occupies almost the entire dorsal surface, the basis capituli of larva, nymph and adult tick all were hexagonal, and the existence of Haller's organ was found on the first pair of legs. Conclusions Scanning electron microscopy observation of the different developmental stages of R.sanguineus revealed clear morphological features, preliminarily suggesting that R.sanguineus in Hainan Province may belong to the tropical lineage, which provide a certain experimental basis for the identification of the tick and the comprehensive prevention and control of local tick-borne diseases.

To analyze the lesion distribution situations and relationship of lesions detected by gastroscopy and colonoscopy in asymptomatic population in Beijing. Data of 1 663 patients who received gastroscopy and colonoscopy in the physical examination center of Peking Union Medical College Hospital and Beijing Hospital between January 2016 and December 2018 were analyzed retrospectively. Statistical analysis was conducted on the detection rate and relationship of different lesions based on the information of gender and ages. Gastroscopy data showed that chronic non-atrophic gastritis and chronic atrophic gastritis accounted for 1 240 (74.6%)and 423 (25.4%)cases respectively. Chronic atrophic gastritis was more common in population aged over 40. Other common diseases included erosions of gastric body and/or antrum, fundic gland polyps, reflux esophagitis, duodenitis, bile regurgitation and so on. Upper gastrointestinal tumors including esophagus cancer and gastric cancer were both early lesions. Colonoscopy results showed that colonic polyps were common lesions, among which there were 382 (23.0%)cases of colonic adenoma and 217 (13.0%)hyperplastic polyps. Incidence of colonic polyps increased with age. Colorectal cancer accounted for 0.7%. Colon diverticulum and melanosis coli were more common in population aged over 40. Colon adenoma was more common in male and melanosis coli was more common in female. The positive rate of HP was 32.2%. There was no positive relationship between HP infection and fundic gland polyps( P=0.329). There was no positive relationship between fundic gland polyps and colon adenomas as well as colorectal cancer( P=0.152, P=0.616). Gastroscopy and colonoscopy play important roles in different kinds of digestive diseases, especially in the early detection of tumors. More attention should be paid to the application of endoscopy in asymptomatic population.

OBJECTIVETo investigate the clinicopathological significance of Twist and YB-1 up-regulation in cervical cancer, and to correlate the expression of the two genes with E-cadherin, a marker of epithelial-mesenchymal transition (EMT).

METHODSA total of 202 tissue samples were collected during January 2008 to December 2013, including 50 cases of normal cervical tissues, 100 cases of cervical intraepithelial neoplasia (CIN) and 52 cases of squamous cell carcinoma (SCC). Twist, YB-1 and E-cadherin expression was investigated by MaxVision.

RESULTSIncreased expression levels of Twist and YB-1 were found and correlated with the malignant transformation of cervical epithelium, histological progression and metastasis of cervical cancer. In addition, Twist and YB-1 overexpression was also associated with aberrant expression of E-cadherin. Regression analysis revealed that Twist expression was an independent factor for the histological progression of cervical cancer.

CONCLUSIONSIt is suggested that Twist and YB-1 overexpression is significantly linked to cervical cancer tumorigenesis and progression, likely related to EMT through (YB-1)-Twist-(E-cadherin) pathway. Twist and YB-1 may be markers for determining the metastatic potential of cervical cancer.

Biomarkers, Tumor ; genetics ; metabolism ; Cadherins ; genetics ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Transformation, Neoplastic ; Cervical Intraepithelial Neoplasia ; metabolism ; pathology ; Disease Progression ; Epithelial-Mesenchymal Transition ; Epithelium ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Nuclear Proteins ; genetics ; metabolism ; Twist-Related Protein 1 ; genetics ; metabolism ; Up-Regulation ; Uterine Cervical Neoplasms ; metabolism ; pathology ; Y-Box-Binding Protein 1 ; genetics ; metabolism

With the enhancement of aesthetic awareness of children's oral maxillofacial development, multi-disciplinary doctors pay attention to children's oral maxillofacial management. Artificial intelligence (AI) technology has been gradually applied to all fields of children's oral maxillofacial management because of its outstanding advantages in medical screening and auxiliary decision-making. This article reviews the application of AI technology in the screening, diagnosis, treatment and follow-up of oral maxillofacial management in children.
Child ; Humans ; Artificial Intelligence ; Administration, Oral

Ferroptosis is a novel iron-dependent mode of programmed cell death characterized by iron deposition and accumulation of lipid peroxidation. More and more studies have found that ferroptosis is closely related to the pathogenesis of type 2 diabetes mellitus （T2DM）. The yin-fire theory is an important part of LI Gao's spleen-stomach theory， and it is believed that qi-fire imblance and yin-fire internal generation is the main pathogenesis of T2DM. Abnormal iron metabolism may be an important prerequisite for T2DM yin-fire internal generation， while oxidative stress is the specific manifestation of T2DM qi-fire imbalance. Reactive oxygen species （ROS） is the end product of qi-fire imbalance， and lipid peroxide is the pathological products of T2DM yin-fire internal generation. This study intends to explore the pathological mechanism of qi-fire imbalance and yin-fire internal generation from the perspectives of iron metabolism， oxidative stress and lipid peroxidation， enriching the modern connotation of yin-fire theory， and benefiting traditional Chinese medicine to target against ferroptosis， and prevent and treat T2DM precisely.