ABSTRACT
Acting as a difficult and high risk procedure in the operation, laparoscopic central tumor anatomical hepatectomy possesses various technical points that shall be noted in the process of application. The choice of laparoscopic approach has been recognized to be one of the key technical links for this technique. According to the anatomical characteristics of the middle lobe of liver and "Easy first" strategy, the authors summarize the laparoscopic central tumor anatomical hepatectomy technology with left side hepatic parenchymal transection-first approach. The left side hepatic parenchymal transection-first approach is found to be simple, fast, safe and effective in operation, which overcomes the challenges of complicated target Glisson pedicle operation and high technical risk of laparoscopic anatomical hepatectomy. The approach also better complies with the "no-touch" principle for malignant tumors. Significantly, it exhibits clinical application value in laparoscopic central tumor anatomical hepatectomy.
ABSTRACT
Diabetes mellitus patients are at high risk of tuberculosis, and tuberculosis is one of the important reasons for inducing and aggravating acute complications of diabetes. At present, China is facing a serious dual epidemic of tuberculosis and diabetes, which has caused great harm to the health of our people, but also brought great burden to our social economy. In this paper, PubMed, Web of Science, CNKI and Wanfang data knowledge service platform were used to search the relevant literature. Research progress on the epidemiology of tuberculosis patients complicated with diabetes, the effect of diabetes and blood glucose on the treatment outcome of tuberculosis patients, and the mechanism of diabetes affecting the treatment outcome of tuberculosis patients were analyzed and summarized, and some references were provided for the treatment and management of tuberculosis patients in China in the future.
ABSTRACT
Objective To explore the 3-D anatomical structure of blood vessel around the pancreas and its clinical significance.Methods Fifty objects were scanned by 64-slice CT of GE,the artery,portal vein,spleen vein,superior mesenteric vein and inferior mesenteric vein were 3-D reconstructed by Myrian system,and the scientific data were recorded.Results The artery,portal vein,spleen vein,superior mesenteric vein and inferior mesenteric vein were all reconstructed;the length of portal vein was(44.28±10.23)mm and the length of post-pancreas trunk was(32.13±7.08)mm;the angle between portal vein and spleen vein were classified into three types:the angle of type Ⅰ was90 °.30％of the inferior mesenteric vein fed into the spleen vein,56％fell into the superior mesenteric vein and 14％came into the the meeting point of the spleen vein and superior mesenteric vein.Conclusion The 3-D reconstruction of blood vessels around the pancreas can provide the anatomical basis for surgeon and reduce the risk of pancreatic surgery before operation.
ABSTRACT
Objective To determine the effect of COMMD7 inhibition on invasion and migration in liver cancer stem cells (LCSCs),and investigate the possible mechanism.Methods After LCSCs were infected by shRNA lentiviral vectors of COMMD7,adhesion assay and Transwell assay were used to detect the invasion and migration,and phalloidin staining was employed to observe the morphological changes.Western blotting was adopted to measure the expression of E-cadherin,N-cadherin and Vimentin.Results COMMD7 knockdown significantly inhibited the invasion and migration of LCSCs.The relative cell quantity of adhesion was 1.00 ± 0.12 and 2.35 ± 0.20 respectively in control cells and infected cells,suggesting there were significantly more adhesive cells in the infected group (P ＜ 0.05).The relative cell quantity per visual field of migration was 1.00 ±0.04 and 0.24±0.03,and that of invasion was 1.00 ±0.05 and 0.24 ±0.04 respectively in the control cells and infected cells,and there were significantly less invasive and migrated cells in the infected group (P ＜0.05).What's more,COMMD7 knockdown also induced some morphological changes of cells corresponding to the weakened abilities of migration and invasion.All the changes above were associated with up-regulation of E-cadherin (P ＜ 0.05) and down-regulation of N-cadherin and Vimentin (P ＜0.05),the molecules related to mesenchymal-epithelial transition (MET).Conclusion COMMD7 knockdown inhibits the invasion and migration in LCSCs,which may be through its regulation on the MET course.
ABSTRACT
Objective To investigate the mechanism of COMMD7,a hepatocellular carcinoma gene,promoting human hepatocellular carcinoma cell line (HePG2 cells)proliferation and migration.Methods The specific siRNA (small interference RNA)was designed for COMMD7 gene,siRNA transfected HepG2 cells was set as Si-HePG2 group,and the PTEN inhibitor treating group (Si +BpV-HePG2),the control group (HePG2),the empty vector group (HePG2 was infected empty vector,N-HePG2 group)were also set up.qRT-PCR was per-formed to evaluate the mRNA expression changes of COMMD7 and PTEN,and Western blot was performed to test the expression changes of COMMD7,PTEN,p-AKT,and AKT.MSP method was performed to detect methylation level.CCK8 was performed to test cell proliferation a-bility.Transwell was performed to detect the invasion of cells.Results The results of qRT-PCR showed that the expression of COMMD7 gene in Si-HePG2 group was lower (0.101 times)than the control group and the N-HePG2 group,and the expression of PTEN gene was higher (2.841 times)than the control group and the N-HePG2 group,and all the results above were of statistically singificant difference (P <0.05). The results of Western blot showed that in Si-HePG2 group,the expression of COMMD7 reduced obviously,the expression of PTEN increased and the expression of p-AKT was significantly inhibited.In Si +Bpv-HePG2 group,the expression of PTEN was significantly inhibited and the expression of p-AKT was increased.The result of MSP showed that compared with the control group and the N-HePG2 group,Si-HePG2 group was more obviously demethylated,which indecated that the expression of COMMD7 gene could induce PTEN demethylation.The results of CCK8 showed that the proliferation in Si-HePG2 group was decreased compared with the control group,N-HePG2 group and Si +BpV-HePG2 group,and the difference was singificant (P <0.05).The results of Transwell showed that the numbers of cell permeating septum in Si-HePG2 group,N-HePG2 group,control group and PTEN group were (17.4 ±2.7),(36.2 ±3.2),(41.6 ±4.5)and (47.6 ±1.8)respec-tively,which were obviously decreased with a significant difference (P <0.05).Conclusion siRNA interference decreased the expression of COMMD7.It can induced the demethylation of PTEN gene in HePG2 and increase the expression of PTEN gene to inhibit PI3K/AKT signaling pathway,thus decreasing the proliferation,migration and invasion ability of HePG2 cells.
ABSTRACT
Objective To observe the changes of the cells of human hepatocellular carcinoma (HepG2)using RNA for silencing the expression of COMMD7 gene,and investigate related mechanism of COMMD7 gene promoting HepG2 proliferation.Methods COMMD7 gene shRNA was designed and constructed into COMMD7-shRNA plasmid.HepG2 cells were divided into the HepG2 group,control-shRNA group (empty vectors were infected) and COMMD7-shRNA group (positive vectors were infected).Cells shapes were observed by fluorescence microscope after infecting.The expression of COMMD7 and expression and phosphosylation of extracellular regulated protein kinase1/2 (ERK1/2) and MEK1/2 protein were measured by Western blot.The cell vitality was measured by cholecystokinin octapeptide (CCK-8),and the apoptosis of cell was detected by flow cytometry.The measurement data with normal distribution were presented as (x) ± s.The comparisons among groups were evaluated with the one-way ANOVA,and pairwise comparison was analyzed by the LSD-t test.Results The cells were oval or spindle shapes and displayed green fluorescent after infected successfully.The results of Western blot showed that the relative quantitative expression of COMMD7 protein in the HepG2 group,control-shRNA group and COMMD7-shRNA group were 0.90 ±0.18,1.03 ±0.05 and 0.23 ±0.03,respectively,with a significant difference among the 3 groups (F =152.08,P ＜ 0.05),and the expression of COMMD7 protein in the COMMD7-shRNA group was significantly lower than those in the other 2 groups (t =20.74,21.16,P ＜ 0.05).The results of CCK-8 showed that the scores of the HepG2 vitality in the HepG2 group,control-shRNA group and COMMD7-shRNA group were 1.193 ±0.024,1.225 ±0.034 and 1.147 ±0.021,respectively,with a significant difference among the 3 groups (F =6.90,P ＜ 0.05),and the HepG2 vitality in the COMMD7-shRNA group was significantly lower than those in the other 2 groups (t =3.53,3.69,P ＜ 0.05).The results of flow cytometry showed that the apoptosis rate of HepG2 in the HepG2 group,control-shRNA group and COMMD7-shRNA group were 6.1％ ± 0.3％,7.8％ ± 0.5％ and 20.9％ ± 1.4％,showing a significant difference among the 3 groups (F =270.80,P ＜0.05),and the apoptosis rate of HepG2 in the COMMD7-shRNA group was significant higher than those in the other 2 groups (t =21.77,19.36,P ＜0.05).The results of Western blot showed that the relative quantitative expression of phosphorylation (p)-ERK1/2 and p-MEK1/2 in the HepG2 group,control-shRNA group and COMMD7-shRNA group were 0.932 ±0.046,0.945 ±0.017,0.553 ±0.052 and 0.452 ±0.031,0.468±0.027,0.263 ± 0.022,respectively,showing significant differences among the 3 groups (F =93.61,49.16,P ＜ 0.05),and the relative quantitative expression of p-ERK1/2 and p-MEK1/2 in the COMMD7-shRNA group were significantly lower than those in the other 2 groups (t =11.94,12.17,9.33,8.65,P ＜ 0.05).Conclusions COMMD7 gene can promote HepG2 proliferation via activating ERK/mitogen-activated protein kinase (MAPK) signaling pathway,and its mechanism may be promoting the phosphorylation of expression of ERK1/2 and MEK1/2.
ABSTRACT
Cell therapy has a very promising potential for end-stage liver diseases (ESLD).Fetal liver stem/progenitor cells (FLSPCs) have advantages of safety,high survival and proliferation rates,and a small volume,all which make them ideal for liver disease stem cell therapy.During the early phase of our study,we applied a three-step separation method to enrich FLSPCs and obtained a separation efficiency similar to that of the flow-cell sorting method.Additionally,using a fulminant hepatic failure model in rats,we have demonstrated that FLSPCs can contribute to morphological and functional recovery of the liver.This manuscript will discuss how FLSPCs can be induced to accurately differentiate into hepatocytes and cholangiocytes and how FLSPCs maintain self-renewal.The Notch signaling plays a critical role in regulating the differentiation and self-renewal of many types of stem cells.Our previous findings have shown that the Notch signaling plays an important role in FLSPCs differentiation into hepatocytes.Therefore,the Notch signaling might be involved in the differentiation and self-renewal of FLSPCs.We conducted a study on the regulatory effects and relative molecular mechanisms of the Notch signaling on FLSPCs and found the corresponding interfering target,which might become an index for the clinical application of FLSPCs.
ABSTRACT
Objective To develop an ideal cultural method to amplify embryonic hepatic stem cells and inhibit their differentiation in vitro. Methods Suspension of ED 14 Fischer (F) 344 rat em-bryonic hepatic stem cells was prepared by collagenase digestion and mechanical disaggregation. Then cells were divided into two groups randomly. The cells in group 1 were seeded into type I collagen-coated plates by adherent culture while those in group 2 were seeded into soft agar medium by suspen-sion culture. After culture for 2 weeks, the morphology and ultrastructure of cells in both groups were observed and compared by inverted microscope and transmission electron microscope, respectivley.The expression of CD90. 1 and CD49F, the two specific stem cell surface markers, was tested by flow cytometry to manifest the establishment of embryonic hepatic stem cells. Alkaline phosphatase stai-ning was used to detect stem cell differentiation. Result Embryonic hepatic stem cells in group 2 were characterized by higher nucleus-cytoplasm ratio and less cell organelles, higher expression of CD90. 1 and CD49F, and stronger positive reaction for alkaline phosphatase staining compared with those in group 1. Moreover, the cells in group 1 showed significant differentiation features. Conclusion Em-bryonic hepatic stem cells cultured suspendedly in soft agar medium experience less differentiation than those adherently cultured in serum-added culture medium, and can proliferate and form clone ball with a specific stem cell feature.