1.LYN′s effects on the migration and invasion of gastric cancer cells through Wnt/ β-catenin signaling pathway
Rui SU ; Yan LU ; Xin ZHENG ; Jun ZHANG
International Journal of Surgery 2021;48(2):78-82,F3
Objective:To explore the effects of LYN on the biological behaviors of gastric cancer cells, such as migration and invasion, and analyze the regulatory mechanism of Wnt/ β-catenin signaling. Methods:Gastric and paracancerous tissue samples from gastric cancer patients admitted to the Affiliated Hospital of Chengde Medical University from Oct. 2017 to May 2018 were selected to detect the expression level of LYN in clinically collected gastric cancer and paracancerous tissue samples by qRT-PCR.Gastric cancer AGS cells were used as experimental subjects, and shRNA-LYN was transfected into gastric cancer AGS cells using Lipofectamine 2000 as experimental group (Sh-LYN group), and untransfected cells as control group (NC group). LYN mRNA expression in AGS cells before and after transfection was detected by qRT-PCR.Transwell detected cell migration and invasion. Cell apoptosis was detected by TUNEL.Western Blot analysis was performed to detect proteins related to the Wnt/β-catenin signaling pathway and to study its regulatory mechanism.Measurement data were expressed as ( Mean± SD), and independent sample t test was used for comparison between the two groups. P<0.05 was considered statistically significant. Results:LYN mRNA expression was significantly up-regulated in gastric cancer tissues ( t=11.04, P<0.001).Sh-LYN transfected gastric cancer cells could make LYN protein expression lower than that of the normal control group( F=961.26, P=0.016).After transfection, migration and invasion of cells in the sh-LYN group were inhibited, and the apoptosis rate of cells in the sh-LYN group was significantly higher than that in the normal control group ( t=3.437, 4.450, P=0.011, 0.026) .Western Blot results: sh-LYN reduced the expression of Wnt3 and β-catenin significantly when compared with the normal control group.The detection of downstream proteins showed that after LYN knockdown, the expression of EMT epithelial labeled E-cadherin was increased, and EMT mesenchymal labeled N-cardherin and vimentin were down-regulated. The other related proteins snail 1 and snail 2 were also down-regulated. Conclusion:LYN knockdown inhibits migration and invasion of gastric cancer cells and induces apoptosis in gastric cancer cells, a process associated with the Wnt/ β-catenin signaling pathway.
2.Clinical features of acute pancreatitis in pregnancy and risk factor analysis for fetal death
Rui ZHONG ; Huan XU ; Yongfeng YAN ; Xin JIANG ; Yan PENG ; Xiaowei TANG
Chinese Journal of Pancreatology 2021;21(1):45-50
Objective:To explore the clinical features of acute pancreatitis in pregnancy (APIP) and the risk factors for fetal death.Methods:The clinical data of 90 patients with APIP in the Affiliated Hospital of Southwest Medical University were retrospectively analyzed from January 2013 to June 2020. Based on the severity, the patients were classified into MAP groups ( n=41), MSAP groups ( n=33), SAP group ( n=16). According to the presence of fetal deaths, the patients were divided into fetal death group ( n=13) and fetal survival group ( n=77). The clinical characteristics and indicators of patients in each group were compared. Binary logistic regression analysis was performed on the variables with differences between groups to explore independent risk factors for fetal death. The receiver operating characteristic curves of laboratory indicators were drawn to evaluate their diagnostic efficacy. Results:Hyperlipidemia was the main cause in 90 APIP cases (42/90, 46.7%). The levels of LDH, CRP, blood glucose, D2 polymer, albumin and ApoA1, the 1-min and 5-min Apgar scores of neonates were statistically significant among MAP group, MSAP group and SAP group (all P<0.05). There were no maternal deaths in 90 cases and 13 fetal deaths (14.4%). Fetal mortality increased with the severity of APIP. APIP combined with hypertension ( OR=14.742, 95% CI 1.157-187.890, P=0.038), ketoacidosis ( OR=19.587, 95% CI 1.789-214.469, P=0.015) and CRP level ( OR=1.013, 95% CI 1.001-1.025, P=0.031) were risk factors for fetal death. ApoA1 level ( OR=0.118, 95% CI 0.021-0.664, P=0.015) was a protective factor for fetal death. The sensitivity and specificity of ApoA1 for predicting fetal death were 84.6% and 79.2%, the sensitivity and specificity of CRP for predicting fetal death were 76.9% and 84.4%, and the sensitivity and specificity of the combination of the two indicators for predicting fetal death were 100% and 67.5%. Conclusions:The severity of APIP was closely related to fetal death. Hypertension, ketoacidosis and blood level CRP were independent risk factors for fetal death, which should be paid special attention.
4.Clinical examples of professor LI Zhi-dao's "tonifying three qi" acupuncture method.
Rui-Chao LI ; Yan LI ; Yuan-Xin FU ; Xiang-Fei ZHAO ; Jing SUN ; Lan-Yuan LI
Chinese Acupuncture & Moxibustion 2014;34(8):810-812
Professor LI Zhi-dao, according to acupoint selection of syndrome differentiation in TCM basic theory, concluded a new therapy, namely "tonifying three qi" that is mainly based on three acupoints in the Conception Vessel. This method is consisted of Danzhong (CV 17), Zhongwan (CV 12) and Qihai (CV 6) in the Conception Vessel, which could successively nourish clear qi, stomach qi and original qi. In clinic, according to the severity of symptoms of three qi, the acupoints are selected flexibly, which could respectively treat deficiency of heart-lung qi, deficiency of stomach-spleen qi and deficiency of original qi. Some examples are also given in the article.
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5.Comparative study of MR diffusion weighted imaging for breast with different b values
Xin CHEN ; Rui YAN ; Huafeng KANG ; Youmin GUO ; Xian ZHAO ; Changan HE ; Yili ZHANG
Chinese Journal of Radiology 2009;43(4):356-359
Objective To explore the optimal b value in MR DWI for breast.Methods Forty patients with palpable masses of breasts underwent MR and DWI at b = 1000, 800 and 600 s/mm2 before surgery according prospective planning. Visual assessment, signal to noise ratio (SNR) of DWI and the ADC values of benign and malignant lesions were compared among three b values using one-way ANOVA test.The diagnostic value of ADC were analyzed by ROC curves.Results Of the 40 patients, there were 26 patients with malignant lesions and 14 patients with benign lesions verified by histopathology.36 patients with 42 lesions were examined by DWI, and detectabilities at three b values were alike.Most DWI at b = 1000, 800 and 600 s/mm2 were of high quality, and images of grade A were 75.0% (30/40), 77.5% (31/40) and 77.5% (31/40), respectively.Mean SNR at b = 1000, 800 and 600 s/mm2 were 63 ± 22,82 ± 27 and 96 ± 29 respectively.Compared with one another, there was statistically significant difference (P <0.01).Mean ADC of benign lesions at different b values were (1.44 ± 0.28) × 10-3, (1.50 ± 0.32) × 10-3 and (1.52±0.29) × 10-3 mm2/s respectively. Compared with one another, there was no statistically significant difference (P = 0.725).Mean ADC of malignant lesions were (1.00 ± 0.25) × 10-3, (0.98 ± 0.19) × 10-3 and (1.07 ± 0.22) × 10-3 mm2/s respectively.Compared with one another, there was also no statistically significant difference (P = 0.358).The area under the ROC curves at b = 1000,800 and 600 s/mm2 were 0.879,0.885 and 0.865 respectively.Threshold value to distinguish benign and malignant lesions were 1.295 × 10-3,1.435×10-3 and 1.335×10-3 mm2/s respectively.Sensitivity of diagnosing breast cancer were 80.0%, 92.0% and 84.0%, and specificity were all 90.0% . Positive predictive values were 95.2%, 95.8% and 95.5% at b = 1000, 800 and 600 s/mm2.Conclusion Combining SNR and the value of ADC in diagnosing benign and malignant breast lesions, the study indicated that b = 800 s/mm2 was the optimal b value in breast DWI.
6.Comparison of low-dose dobutamine stress echocardiography and single photon emission computed tomography and delayed contrast MRI in the diagnosis of myocardial viability: Meta-analysis
Wenling LI ; Xin CHEN ; Xiaojuan GUO ; Min LIU ; Rui YAN ; Youmin GUO
Chinese Journal of Radiology 2009;43(9):942-947
elayed contrast enhanced MRI showed significant different pooled sensitivity in comparison to low-dese dobutamine stress echocardiography.
7.Clinical characteristics of ocular toxocariasis patients on the first attendance
Xiaochun LI ; Qing CHANG ; Rui JIANG ; Xin HUANG ; Yanqiong ZHANG ; Qian CHEN ; Saimei YAN
Chinese Journal of Ocular Fundus Diseases 2016;32(1):40-43
Objective To investigate the clinical characteristics of 40 patients with ocular toxocariasis (OT) on the first attendance.Methods A total of 40 consecutive patients who were clinically and serologically diagnosed with OT were retrospectively reviewed.Results The mean age of patients was (12.12±10.42) years.There were 29 males and 11 females.29 cases presented with decreased vision,4 children with leukocoria,2 cases with strabismus and 5 cases was found abnormal during regular eye examination.Initially 8 eyes (20%) were misdiagnosed as retinoblastoma (1 eye),Coat' s disease (1 eye),cataract (2 eyes),iridocyclitis (2 eyes) and retinal detachment (2 eyes).23 eyes had retinal detachment,19 eyes had cataract.OT was the initial diagnosis for 15 patients (37.5%).The best corrected visual acuity (BCVA) were NLP to 0.7.Ultrasound biomicroscopy (UBM) were performed in 29 eyes,and identified peripheral granulomas in 23 eyes and adjacent tractional retinal detachment in 12 eyes.We also identified 17 cases (68.0%) with elevated IgE level among 25 patients with positive serological antibody test.Conclusions Tractional retinal detachment,vitreous opacities and cataract are the common clinical findings at the first attendance of OT patients.The adjunctive test of serum total IgE level may be helpful for the diagnosis.The application of UBM and specific IgG detection in serum and intraocular fluid,can also improve the diagnosis.
8.Expression of major histocompatibility complex in ovarian epithelial carcinoma
Li-Xin SUN ; Li-Juan YAN ; Li-Gong SUN ; Rui-Xia ZHAO ; Shao-Yun ZHANG ;
Cancer Research and Clinic 1997;0(03):-
Objective To investigate the numbers and the expression of MHC-Ⅰpositive cells in hu- man ovarian epithelial carcinoma tissues and provide the experimental data in the futher biological therapy of ovarian carcinoma.Methods Thirty one samples of ovarian epithelial carcinoma were analysed with flow cy- tometry for MHC classⅠexpression.Results The number of MHC classⅠcells(25.22?21.23,4.37?3.63)was fewer in ovarian epithelial carcinoma than that in normal ovarian tissues(43.56?18.47,7.43?5.87),and was related with pathologic grades(P
9.Construction of TK Gene-deleted PRV SH StrainContaining a Single LoxP Site
Min-Xiu WANG ; Xin-Ming SU ; Chun-Mei YU ; Rui-Bing CAO ; Pu-Yan CHEN ;
China Biotechnology 2006;0(10):-
Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.
10.Notch4 Inhibition Suppresses Invasion and Vasculogenic Mimicry Formation of Hepatocellular Carcinoma Cells
CHENG RUI ; CAI XIN-RAN ; KE KUN ; CHEN YAN-LING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(5):719-725
Vasculogenic mimicry (VM) is a process by which aggressive tumor cells generate non-endothelial cell-lined channels in malignant tumors including hepatocellular carcinoma (HCC).It has provided new insights into tumor behavior and has surfaced as a potential target for drug therapy.The molecular events underlying the process of VM formation are still poorly understood.In this study,we attempted to elucidate the relationship between Notch4 and VM formation in HCC.An effective siRNA lentiviral vector targeting Notch4 was constructed and transfected into Be17402,a HCC cell line.VM networks were observed with a microscope in a 3 dimensional cell culture system.Cell migration and invasion were evaluated using wound healing and transwell assays.Matrix metalloproteinases (MMPs) activity was detected by gelatin zymography.Furthermore,the role of Notch4 inhibition in Be17402 cells in vivo was examined in subcutaneous xenograft tumor model of mice.The results showed that downregulation of Notch4 destroyed VM network formation and inhibited migration and invasion of tumor cells in vitro (P<0.05).In vivo,tumor growth was also inhibited in subcutaneous xenograft model (P<0.05).The potential mechanisms might be related with down-regulation of MT1-MMP,MMP-2,MMP-9 expression and inhibition of the activation of MMP2 and MMP9.These results indicated that Notch4 may play an important role in VM formation and tumor invasion in HCC.Related molecular pathways may be used as novel therapeutic targets for HCC antiangiogenesis therapy.