1.The effect of microRNA-155 on the proliferation,invasion,migration and apoptosis of hepatocellular carcinoma cells
Huanrong QIN ; Xiangkai WU ; Zheyu JIANG ; Yun ZHANG ; Liyun LIN ; Lizhou WANG ; Shi ZHOU
Journal of Interventional Radiology 2024;33(1):44-51
Objective To discuss the effect of PI3K-AKT signaling pathway regulated by microRNA-155(miRNA-155)targeted protein tyrosine phosphatase non-receptor type 21(PTPN21)on the proliferation,migration and invasion of hepatocellular carcinoma(HCC)cells.Methods Lentivirus transfection was used to silence the expression of miRNA-155 in human Huh7 HCC cells,and real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)was used to detect the silencing effect of miR-155.After obtaining stable cell lines,the cell lines were randomly divided into Blank group(normal Huh7 cells),shNC group(Huh7 cells+empty miR-155 vector),sh-miR-155(Huh7 cells+miR-155 silencing),sh-miR-155+Recilisib group(Huh7 cells+miR-155 silencing+PI3K-AKT agonist),shNC+Recilisib group(Huh7 cells+empty miR-155 vector+PI3K-AKT agonist).Dual luciferase assay was used to determine whether PTPN21 was the downstream of miR-155.The cell proliferation ability of cells in each group was detected by MTT assay.The apoptosis level of each group was tested by flow cytometry.The invasion and migration ability of cells was assessed by Transwell assay.Western blot analysis was used to observe the differences in protein expression of PTPN21,PI3K,P-PI3K,AKT,P-AKT,and apoptosis-related proteins including BAX,BCL-2 and caspase-3 in all groups.Results The expression level of miR-155 in sh-miR-155 group was lower than that in Blank group and shNC group(P<0.000 1),and the difference in miR-155 expression level between Blank group and shNC group was not statistically significant(P>0.05).MTT results showed that A values of Huh7 cells at 2,3,4 and 5 day in sh-miR-155 group were lower than those in Blank group and shNC group(P<0.000 1),while these differences between Blank group and shNC group were not statistically significant(P>0.05).In sh-miR-155 group the A values at 2,3,4 and 5 day were lower than those in sh-miR-155+Recilisib group and shNC+Recilisib group(P=0.0052 and P<0.0001,respectively),while the A values at 2,3,4 and 5 day in sh-miR-155+Recilisib were lower than those in shNC+Recilisib group(P<0.000 1).There was no significant differences in cell migration and number of invasion cells between the Blank group and shNC group(P>0.05).After activation of PI3K-AKT signaling pathway,the migration and invasion capacity of HCC cells in the shNC+Recilisib group were significantly enhanced when compared with the Blank group(P<0.000 1).In contrast,the number of migrated and invaded Huh7 cells after miR-155 silencing was significantly lower than that in the Blank group and shNC group(P<0.000 1)and this phenomenon became reversed by PI3K agonist.Compared with the sh-miR-155 group,in the sh-miR-155+Recilisib group the migration and invasion ability of HCC cells was enhanced(P=0.000 2).Lentiviral transfection of Huh7 human HCC cells to silence miR-155 and downregulate miR-155 inhibiting PTPN21 regulation of the PI3K-AKT signaling pathway,thus inhibiting the invasion,migration and proliferation ability of HCC cells and promoting the apoptosis of HCC cells.Conclusion miR-155 inhibits the migration,invasion and proliferation of HCC cells through targeting PTPN21 regulation of PI3K-AKT signaling pathway.The miR-155 may be a potential therapeutic target for HCC in the future.(J Intervent Radiol,2024,32:44-51)
2.Advance on surgical treatment of hepatolithiasis
Peng CHEN ; Zheyu ZHU ; Feifan WU ; Siyu WANG ; Yiyu HU ; Weimin WANG ; Chunmu MIAO ; Yunbing WANG ; Xiong DING
The Journal of Practical Medicine 2023;39(21):2857-2860
Hepatolithiasis is a common biliary tract disease in China and other Asian countries.It has insidious onset,recurrent disease,and may lead to biliary cirrhosis or even cancer in the late stage without inter-vention.At present,surgery is considered to be the best choice for treatment.In the 21st century of minimally inva-sive surgery,traditional procedures such as partial hepatectomy,choledochotomy,cholangioenterostomy and liver transplantation can be performed with the assistance of laparoscopy and da Vinci robots,and emerging endoscopic techniques such as endoscopic retrograde cholangiopancreatography(ERCP)and percutaneous transhepatic cho-ledochoscopic lithotripsy(PTCSL)have also been developed,bringing better treatment for patients with hepatoli-thiasis.As treatment options are becoming more varied,there are also some clinical problems that need to be addressed.In this article,we would like to briefly review the current surgical treatment modalities in order to provide a theoretical basis for optimizing the treatment modalities.
3.Construction of Hsf1 gene-knockout mouse model using CRISPR/Cas9
Zheyu WU ; Zhenhua ZHOU ; Zhipeng WU ; Yufeng ZHANG ; Zhiqiang YANG ; Yufan ZHU ; Yuanlong XIE ; Jianru XIAO ; Lin CAI
Chinese Journal of Orthopaedics 2021;41(8):515-524
Objective:To explore the efficient construction of HSF1 gene knockout mouse model using CRISPR/Cas9 gene editing technology, and to establish the early basis for the mouse model of primary osteosarcoma.Methods:According to exon 9 of HSF1 gene structure, the corresponding GRNA (guideRNA) was selected and screened. Then the transcription template of sgRNA (small guide RNA) was amplified by PCR, and four up stream primers were obtained. Subsequently, sgRNA was transcribed in vitro and screened by Tube Screen platform to screen the sgRNA with effective cutting, and the sgRNA with the highest cutting efficiency was selected from the screening results for subsequent experiments. The transcription template of SPCas9mRNA was amplified by PCR, and then Cas9mRNA was transcribed in vitro. The sgRNA transcribed in vitro and Cas9mRNA were injected into the fertilized eggs of healthy C57BL/6 mice, and the tissue was extracted from the tail of the born mice and identified by PCR sequencing. Heterozygous female mice of F0 generation were selected to mate with wild-type male mice too btain F1 generation off spring. The mutation of gene bases of F1 generation mice was detected by AGAR gel electrophoresis and gene sequencing. The heterozygous male mice of the F1 generation and female mice of the F0 generation were back crossed to obtain the F2 generation daughter mice. The tail tissues were cut and sequenced to obtain the F2 generation homozygous knockout mice. PCR was used to observe the cutting efficiency of sgRNA and the sequencing of rat tail tissue, and SNAPGene software was used for gene sequence alignment to determine the deletion of base fragments.Results:The up stream primers sgRNA-1 Primer-f, sgRNA-2 Primer-f, sgRNA-3 Primer-f, sgRNA-4 Primer-f and down stream primers sgRNA-4 Primer -r were obtained by PCR amplification. After in vitro tran scription and screening of sgrRNA, sgrRNA-1, sgrRNA-2 and sgrRNA-4 had high cleavage efficiency and were selected for subsequent experiments. T7 promoter was added to the 5 'end of Cas9 mRNA, and Cas9 mRNA was obtained by PCR and in vitro transcription kit. Mixed Cas9-sgRNA solution was injected into the fertilized eggs of mice and cultured. The cultured two-cell fertilized eggs were injected into the ampulla of the pseudo pregnant female mice, and the F0 generation mice were obtained successfully. A total of 8 heterozygous mice of F0 generation were obtained by Agar gel electrophoresis. Three heterozygous knockout mice of F1 generation were obtained by breeding the female heterozygous mice of F0 generation with healthy wild-type male mice and PCR and sequencing. Three heterozygous male mice of F1 generation were back crossed with female mice of F0 generation 3 to obtain F2 generation mice. Through the observation of electrophoresis and sequencing results of F2 generation mice, it was confirmed that 7 mice were missing HSF1 base sequence, and the electrophoresis results showed mutant bands and no wild-type bands, which were identified as homozygous. The F2 generation homozygous mice were able to breed stably. As eries of results proved that the HSF1 gene knockout mouse model was successfully established in this experiment.Conclusion:CRISPR/Cas9 technology was successfully used to construct HSF1 gene knockout mouse model, with strong stability and high reproducibility, which laida foundation for further study of HSF1 gene expression products and establishment of mouse model of primary osteosarcoma.
4.Analysis of correlation between serum NSE, hs-CRP levels and NIHSS score in patients with cerebral infarction
Chinese Journal of Primary Medicine and Pharmacy 2020;27(4):412-416
Objective To investigate the correlation between the changes of serum neuron specific enolase(NSE) and hypersensitive C-reactive protein (hs-CRP) levels and the degree of neurological deficit (NIHSS)score in patients with cerebral infarction.Methods From January 2017 to January 2019,63 patients with cerebral infarction admitted to Lishui Central Hospital were selected.According to NIHSS score,they were divided into 13 mild cases,30 moderate cases and 20 severe cases.According to infarction area,they were divided into large area group(16 cases),small area group (27 cases) and lacunar infarction group (20 cases).Another 60 cases underwent health examination in our hospital from January 2017 to January 2019 were selected as the control group.Enzyme-linked immunosorbent assay (ELISA) was used to determine the content of NSE,and immunoturbidimetric assay was used to determine the content of hs-CRP.The changes of serum NSE and hs-CRP levels in the cerebral infarction group and control group,serum NSE,hs-CRP levels and NIHSS scores in different severity and infarction area,and the correlation between serum NSE and hs-CRP changes and NIHSS scores were compared.Results The serum levels of NSE [(21.34 ± 3.27) ng/mL] and hs-CRP [(10.48 ± 2.14) mg/L] in the cerebral infarction group were significantly higher than those in the control group [(6.23 ± 1.08) ng/mL,(2.83 ± 0.46) mg/L] (t =34.061,27.095,all P < 0.05).The serum levels of NSE [(26.98 ± 3.64) ng/mL],hs-CRP [(15.36 ± 2.57) mg/L] and NIHSS score[(38.49 ±3.25) points] in the severe group were higher than those in the moderate group and mild group,which in the moderate group [(20.98 ± 3.21) ng/mL,(10.25 ± 2.09) mg/L and (22.18 ± 3.48) points]were higher than those in the mild group [(12.64 ± 2.78) ng/mL,(5.47 ± 1.40) mg/L and (7.38 ± 2.56)],the differences were statistically significant (F =14.975,9.132,15.873,all P < 0.05).The serum levels of NSE[(25.43 ± 3.35) ng/mL],hs-CRP [(16.54 ± 2.71) mg/L] and NIHSS score [(37.34 ± 3.75) points] in the large area group were higher than those in the small area group and lacunar infarction group,which in the small area group [(21.67 ± 3.12) ng/mL,(10.86 ± 2.21) mg/L and (21.25 ± 3.26) points] were higher than those in the lacunar infarction group [(13.45 ± 2.97) ng/mL,(4.79 ± 1.35) mg/L and (8.49 ± 2.15) points],the differences were statistically significant (F =13.241,9.893,17.482,all P < 0.05).The serum levels of NSE and hs-CRP were positively correlated with NIHSS score (r =0.829,0.713,all P < 0.05).Conclusion The levels of serum NSE and hs-CRP in patients with cerebral infarction increase with the progression of the disease,and there is a linear positive correlation between NSE and hs-CRP and NIHSS score.It is considered that NSE and hs-CRP are of great value in evaluating the degree of neurological impairment,the severity of the disease and the size of the infarct.
5.Diagnosis and treatment of easily misdiagnosed hepatic sclerosing hemangioma
Qi WU ; Xie SONG ; Zheyu NIU ; Bingqiang LIU ; Fangfeng LIU ; Jun LU ; Hong CHANG
Journal of Clinical Hepatology 2020;36(12):2843-2846
Hepatic sclerosing hemangioma (HSH) is a rare benign tumor that is considered fibrosis and hyaline change caused by degenerative changes of cavernous angioma, and changes in pathological features cause the changes in imaging features, making this atypical hemangioma easily misdiagnosed as primary or metastatic malignant tumor. Although there are many studies on the imaging findings of this disease, it is still difficult to diagnose and most patients underwent resection since it is misdiagnosed as malignant tumor. There is still a low rate of confirmed diagnosis before surgery. This article elaborates on the etiology, clinical manifestations and pathological features, imaging findings, diagnosis, and treatment of HSH, in order to provide a reference for the diagnosis and treatment of this disease.
6.Establishment of prognostic scoring system for single large hepatocellular carcinoma after hepatectomy
Junyi SHEN ; Chuan LI ; Tianfu WEN ; Lvnan YAN ; Jiayin YANG ; Yong ZENG ; Hong WU ; Wentao WANG ; Mingqing XU ; Zheyu CHEN ; Yonggang WEI ; Li JIANG ; Jiwei HUANG
Chinese Journal of Hepatic Surgery(Electronic Edition) 2018;7(3):197-201
Objective To establish a clinical scoring system for the prognosis of patients with single large hepatocellular carcinoma (HCC) after hepatectomy.Methods 268 patients with single large HCC who underwent hepatectomy in West China Hospital of Sichuan University from January 2009 to December 2013 were included in this prospective study. There were 227 males and 41 females, of which 198 cases aged≤60 years old, 70 aged>60 years old. The informed consents of all patients were obtained and the local ethical committee approval was received. The patients' survival were observed. The independent risk factors for postoperative prognosis of patients with single large HCC were selected by Cox proportional risk regression mode. Based on the risk factors, the prognostic scoring system for single large HCC wasestablished. The scoring system was tested through survival analysis by Kaplan-Meier and Log-rank test. Results The median overall postoperative survival was 45 months, and the tumor-free survival was 31 months. Platelet-to-lymphocyte ratio (PLR)≥107, tumor diameter≥6.8 cm and positive microvascular invasion (MVI) were the independent risk factors for postoperative overall survival and tumor-free survival in patients with single large HCC (HR=1.004, 1.092, 2.233 and 1.003, 1.062, 1.534; P<0.05). Every independent risk factor was assigned 1 point. All patients were divided into low risk group (0 point), moderate risk group (1-2 points) and high risk group (3 points). The 5-year survival rate of high risk group was 25.4%, and that of moderate and low risk group was 33.2% and 52.1% respectively, where significant difference was observed (χ2=23.1, P<0.05). Similar Results were observed when the scoring system was used in patients with or without cirrhosis.Conclusions PLR≥107, tumor diameter≥6.8 cm and positive MVI are the independent risk factors for the prognosis of patients with single large HCC after resection. The prognostic scoring system established in this study can be used to predict the postoperative long-term survival of patients.
7.Effect of Hydroxysafflor Yellow A on Pro/Anti-Inflammatory Cytokines in Peripheral Blood with Sepsis in Mice
Jinping WANG ; Ping WANG ; Runhua CHEN ; Peiyan ZHAO ; Zheyu HUANG ; Hanwei WU ; Jianlong WU
Journal of Sun Yat-sen University(Medical Sciences) 2017;38(5):665-669
[Objective]To explore the effect of HSYA on pro/anti-inflammatory cytokine′s levels and mRNA expression in peripheral blood of mice.with sepsis.[Methods]Dividing NIH mice into four groups,as normal group,sham group,CLP group and HSYA group,24 mice in each group. The CLP sepsis mouse model was established. HSYA(120 mg/kg)were injected intravenously at 12 h before the operation ,and 0 h and 12 h following CLP ,and other groups were given normal saline. observed the animals behavior changes,measured the levels WBC,PLT,ALT,AST,BUN in serum,detected the levels and mRNA expression of IL-6, IL-10 ,TNF-α in peripheral blood. cultured of peripheral blood bacteria loads.[Results]24 h after surgery ,mice in CLP group appeared furring,feces residues on anus etc. compared to normal group,sham group and HSYA group,WBC,PLT,ALT,AST, BUN,levels and mRNA expression of IL-6,IL-10,TNF-αshowed significant increases,it was also found that bacterial load was significant increased in model group.[Conclusions]HSYA has a therapeutic effect in mice with sepsis ,can reduce bacteria into the blood,and inhibit inflammatory mediators which caused tissue damage.
8.Research progress on application of assessment tools of humanistic nursing care
Bing WU ; Yang GAO ; Zheyu YAN ; Jinhua ZHANG
Chinese Journal of Practical Nursing 2016;32(12):950-953
Based on collecting, arranging, analyzing the literature, the most widely used assessment tools in caring behavior, caring ability and nursing caring characters were introduced. The problems were found out from its application at home and abroad. The aim was to provide the reference that developing and revising the assessment tools of nurses′humanistic that fit for the Chinese local culture.
9.Surgical Management for Giant liver Hemangiomas Greater Than 20 cm in Size.
Hui JIANG ; Zheyu CHEN ; Pankaj PRASOON ; Hong WU ; Yong ZENG
Gut and Liver 2011;5(2):228-233
BACKGROUND/AIMS: The aim of this study was to investigate the primary management experience for giant liver hemangiomas greater than 20 cm in size. METHODS: Records of patients referred for evaluation of radiologically and/or histopathologically proven giant liver hemangiomas between January 2007 and March 2010 were retrospectively analyzed. The reasons for referral, results of imaging studies, preoperative and surgical treatments, and outcome were reviewed. RESULTS: A retrospective analysis was performed for 14 patients diagnosed with a giant hemangioma on the basis of an imaging study and/or a histopathological examination. All cases were diagnosed as giant liver hemangioma with at least one lesion greater than 20 cm in size. Abdominal discomfort was the main presenting complaint for the referral in 9 patients (64.2%). Abdominal ultrasound established the diagnosis in 12 patients (85.7%). Twelve patients underwent liver resection, 2 of whom underwent staged resection. Enucleation was performed in 2 patients. Selective transcatheter arterial embolization was implemented in 9 patients. Postoperative morbidity occurred in 3 patients (21.4%). No complications related to the hemangiomas occurred during follow up. CONCLUSIONS: Liver resection is indicated for giant liver hemangiomas with abdominal discomfort, especially for lesions greater than 20 cm in size. Staged operations are performed for patients with multiple lesions. Preoperative selective transcatheter arterial embolization alleviates progressive abdominal pain.
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10.A comparative study on continuous hemihepatic with intermittent total hepatic inflow occlusion in hepatectomy for liver tumors
Zhixue LIAO ; Tianfu WEN ; Zheyu CHEN ; Lunan YAN ; Jian YANG ; Bo Lü ; Guochang WU ; Yu ZHANG
Chinese Journal of General Surgery 2009;24(4):295-299
Objective To evaluate if continuous hemihepatic inflow occlusion(HH)during hepatectomy can be as safe and effective as intermittent total hepatic inflow occlusion(TH)in reducing blood loss during hepatectomy.Methods From November 2001 to March 2006.eighty patients undergoing liver resections were included in a prospective randomized study comparning the intra-and postoperative course underTH(n=40)or HH(n=40).TH was performed with periods of 20 minutes of occlusion and 5 minutes of releasing,while HH with continuous occlusion.The surface area of liver transection was measured and blood loss was calculated.The amount of blood loss,levels of alanine aminotransferuse (ALT)and aspartate aminotransferase(AST),and postoperative course were recorded. Results The total ischemic time of the HH groups was longer than in the TH group[(42±13)min,(31±13)min,P=0.37],and the operative time in the HH group was longer than in the TH group[(236 ±49)min,(204±38)min,P=0.02 ].No signincant difierenee was found between HH and TH group in blood loss during liver parenchyma transection[(500 ±269)ml,(416 ±235)ml,P=0.14]and in the changes of ALT and AST on the first postoperative day[ALT:(677±572)IU/L,(577 ±327)IU/L,P=0.12;AST:(591 ±468)IU/L,(512±301)IU/L,P=0.66].There were no difierences on postoperative morbidity between the two groups(22.5%versus 20.0%,P=0.35).Conclusion The technique of continuous hemihepatic inflow occlusion is as safe and effective as intermittent total hepatic inflow occlusion.

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