1.Relationship between the expression of E-cadherin and ki-67 in hepatocellular carcinoma tissues and prognosis of the patients after hepatectomy
Shugang SUN ; Zusen WANG ; Yujun LI ; Tianhua GUO ; Lianjun ZHAO
Chinese Journal of Current Advances in General Surgery 1999;0(02):-
Objective: To study the relationships between the expressions of E-cadherin and ki-67 in the tissues of heatocellular carcinoma(HCC)and the prognosis of HCC patients after hepatectomy as well as their clinical pathology. Methods: We examine the expressions of E-cadherin and ki-67 in 255 HCC tissues by tissue microarray and PV-6000 two-step method of immunohistochemistry and analyze the correlations between their expressions and clinical pathological data, 1-year recurrent rate and overall survival time after hepatectomy. Results: The expression of E-cadherin correlated with the tumor size and the 1-year recurrent rate of positive group was higher than that of the negative group. The expression of ki-67 correlated with vascular invasion and differentiation of the tumor, the positive group showed a higher 1-year recurrent rate and a shorter overall survival time. Multivariate Cox regression analysis indicated that the expression of ki-67 was an independent risky factor. Conclusions: The negative expression of E-cadherin and the positive expression of ki-67 predict a higher recurrent rate of early stage. The expression of ki-67 is an independent risky factor which can be used to evaluate the prognosis of patients with HCC after hepatectomy.
2.Neurobehavioral effects of long-term mild hypothermia combined with compound porcine cerebroside and ganglioside injection after traumatic brain injury in rats
Wanyong ZHAO ; Shugang XU ; Jingjing WANG ; Xiaohong LI ; Hongtao SUN
Acta Universitatis Medicinalis Anhui 2024;59(3):448-454
Objective To investigate the neurobehavioral effects of long-term mild hypothermia(MHT)combined with compound porcine cerebroside and ganglioside injection(CPCGI)after traumatic brain injury(TBI)in rats and its mechanism.Methods 36 healthy adult male SD rats were randomly divided into model group,MHT group,CPCGI group and MHT+CPCGI group.The TBI model was prepared using an electronically controlled cortical in-jury device.The rats in model group received an intraperitoneal injection of an equal amount of normal saline(NS,2 ml/kg)and were treated at room temperature(37℃)for 48 hours.The rats in MHT group received an intraper-itoneal injection of an equal amount of NS and were treated at a slightly low temperature(33.0±1.0)℃for 48 hours.The rats in CPCGI group received an intraperitoneal injection of an equal amount of CPCGI(0.6 ml/kg)and were treated at room temperature for 48 hours.The rats in MHT+CPCGI group received an intraperitoneal in-jection of an equal amount of CPCGI and were treated at a slightly low temperature for 48 hours.The sensorimotor function of rats was evaluated by modified Neurological Severity Score(mNSS).The motor and spatial memory a-bilities of rats were detected by Morris water maze test,and the motor function of rats was evaluated by beam walk-ing test(BWT)and inclined-grid climbing test.The number of neurons in hippocampus was observed by Nissl stai-ning and immunofluorescence was used to detect the expression of doublecortin(DCX)and neuronal nuclear anti-gen antibody(NeuN).Western blot was used to observe the protein expression of B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax)and cysteine proteinase-3(Caspase-3).Results Compared with MHT group and CPCGI group,MHT+CPCGI group had a lower mNNS score,shorter escape latency,higher times across the platform and the percentage of time in the target quadrant,higher BWT score and larger climbing angle,increased numbers of neurons,DCX and NeuN positive cells,increased Bcl-2 expression and decreased expression of Bax and Caspase-3.(P<0.05).Conclusion Long-term mild hypothermia combined with CPCGI can effectively im-prove the neurological deficits of TBI rats by promoting nerve regeneration and inhibiting cell apoptosis,and provide potential strategies and basis for the clinical treatment of TBI.
3.Two cases of epilepsy without periventricular nodules caused by the ARF1 gene mutation and literature review
Weiran ZHANG ; Na SUN ; Shugang WANG ; Liu LIU ; Lihua JIANG ; Congying ZHAO ; Feng GAO ; Peifang JIANG ; Zhefeng YUAN
Chinese Journal of Applied Clinical Pediatrics 2024;39(12):936-939
A retrospective analysis was made on the clinical data and gene mutation characteristics of 2 children admitted to the Children′s Hospital of Zhejiang University School of Medicine for epilepsy without periventricular nodules caused by the ARF1 gene mutation from August 2023 to February 2024, and relevant literature was reviewed.Both patients presented with seizures and psychomotor retardation, and 1 of them was diagnosed with West syndrome.Whole exome sequencing confirmed that the 2 patients carried a missense mutation in the ARF1 gene (c.55C>A, p.R19S).Brain magnetic resonance imaging (MRI) of 2 patients revealed no obvious abnormalities.A summary analysis of 5 cases of ARF1 gene mutations reported in three foreign literatures showed that patients with ARF1 gene mutations usually presented with seizures, developmental delay, hypotonia, mental retardation, and motor stereotypies.MRI showed periventricular nodular heterotopia, corpus callosum dysplasia, subcortical white matter abnormalities, and delayed myelination.This study found for the first time that ARF1-related disorders can occur without significant brain structural malformations, indicating that there are inconsistencies in neuroimaging findings, adding valuable phenotypic information to this gene.The differences in imaging findings may be the result of genetic background or variation in ARF1-interacting proteins, or may be caused by altered regulatory mechanisms of protein activity.
4.Effect of micro-ribonucleic acid-21 on the malignant biological behavior of cholangiocarcinoma cells by targeting the PTEN/PI3K/Akt pathway
Zhe SHI ; Liyuan ZHOU ; Guodong ZHAO ; Shugang SUN ; Liang XUE
Journal of Clinical Hepatology 2022;38(9):2091-2098
Objective To investigate the effect of micro-ribonucleic acid-21(miR-21) on the malignant biological behavior of human cholangiocarcinoma cell line QBC939 by targeting the protein tyrosine phosphatase (PTEN)/inositol phosphate 3-kinase (PI3K)/protein kinase B (Akt) pathway. Methods The cholangiocarcinoma cell line QBC939 in the logarithmic growth phase was divided into empty vector group, blank control group, overexpression group, and silencing group.An inverted fluorescence microscope was used to observe transfection efficiency; MTT assay, flow cytometry, Transwell assay, and wound healing assay were used to measure cell proliferative activity, apoptosis rate, invasion activity, and migration activity.Quantitative reverse transcription PCR was used to measure the mRNA expression levels of miR-21, PTEN, PI3K, Akt, and mammalian target of rapamycin (mTOR); Western blotting was used to measure the protein expression levels of PTEN, PI3K, Akt, phosphorylated Akt (p-Akt), and mTOR; dual-luciferase reporter assay was used to verify the effect of miR-21 on PTEN.A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the SNK- q test was used for further comparison between two groups. Results Transfection efficiency was 90.27%±18.03% in the overexpression group, 91.43%±18.26% in the silencing group, and 92.16%±18.41% in the empty vector group.Compared with the blank control group and the empty vector group, the overexpression group had a significant increase in the proliferative activity of QBC939 cells (both P < 0.05) and a significant reduction in apoptosis rate (both P < 0.01);compared with the blank control group, the empty vector group, and the overexpression group, the silencing group had a significant reduction in proliferative activity ( P < 0.01) and a significant increase in apoptosis rate ( P < 0.01).Compared with the blank control group and the empty vector group, the overexpression group had significant increases in the migration rate of QBC939 cells and number of cells penetrating the membrane (all P < 0.01);compared with the blank control group, the empty vector group, and the overexpression group, the silencing group had significant reductions in migration rate and number of cells penetrating the membrane (all P < 0.01).Compared with the blank control group and the empty vector group, the overexpression group had significant increases in the mRNA expression levels of miR-21, PI3K, Akt, and mTOR and a significant reduction in the mRNA expression level of PTEN (all P < 0.05);compared with the blank control group, the empty vector group, and the overexpression group, the silencing group had significant reductions in the mRNA expression levels of miR-21, PI3K, Akt, and mTOR and a significant increase in the mRNA expression level of PTEN (all P < 0.05).Compared with the blank control group and the empty vector group, the overexpression group had significant increases in the protein expression levels of PI3K, Akt, p-Akt, and mTOR and a significant reduction in the protein expression level of PTEN (all P < 0.01);compared with the blank control group, the empty vector group, and the overexpression group, the silencing group had significant reductions in the protein expression levels of PI3K, Akt, p-Akt, and mTOR and a significant increase in the protein expression level of PTEN (all P < 0.01).Furthermore, miR-21 showed targeted regulation of PTEN expression. Conclusion MiR-21 silencing may inhibit the malignant biological behavior of human cholangiocarcinoma cell line QBC939 by targeting the PTEN/PI3K/Akt signaling pathway to upregulate the expression of PTEN and downregulate the expression of PI3K, Akt, mTOR, and p-Akt.