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BACKGROUND:It has been suggested that CT multiplanar reconstruction should be performed prior to the placement of axial pedicle screws to determine the anatomy of the C2 pedicle in each patient,to design the appropriate screw locus and diameter,and to evaluate the feasibility of screw placement to reduce the incidence of surgery-related complications. OBJECTIVE:To evaluate the feasibility of axis pedicle screw placement by morphologic classification of pediculoisthmic component with CT multiplanar reconstruction. METHODS:The CT data of 200 patients(400 axial pedicle screws)with cervical spine were retrospectively studied by using Siemens Syngo.Via software.According to the direction of the axis of the pedicle,the CT multiplanar reconstruction positioning line was adjusted to reconstruct the sectional image of the narrowest part of the pediculoisthmic component.According to its morphological characteristics,the narrowest part of the pediculoisthmic component was divided into three types:type 1,"hook"type:Type 1a outer diameter width(a1)>0.4 cm,type 1b outer diameter width(a1)≤0.4 cm;type 2,"like circle/ellipse"type;type 3,"horizontal ellipse"type.The outer diameter width of the narrowest part of pediculoisthmic component(d1),medullary cavity width(d2),outer diameter height(a1),and medullary cavity height(a2)were compared among the three types,and the feasibility of pedicle screw placement of the three types was evaluated. RESULTS AND CONCLUSION:(1)A total of 400 axial pedicles included 269 cases of type 1,130 cases of type 2,and 1 case of type 3.(2)The mean external diameter height between types 1 and 2 was not significantly different(P>0.05).The mean medullary cavity height,mean outer diameter widths,and mean medullary cavity width were significantly different(P<0.001).There were 42 cases(15.6%)of type 1 and 0 cases(0.00%)of type 2 with mean external diameter width≤0.4 cm,and the difference was significant(P<0.001).There was only one case of type 3,whose external diameter height,medullary cavity height,outer diameter width and medullary cavity width were 1.20 cm,0.84 cm,0.64 cm and 0.31 cm,respectively.(3)These results confirm that axial pedicle screws can be safely inserted in patients with types 1a,2 and 3,which requires no further measurement and assessment.Pedicle screw insertion should be performed with caution in type 1b patients.Therefore,in type 1 patients,the width of the narrowest outer diameter of the pediculoisthmic component should be further measured to evaluate the feasibility of axial pedicle screw placement.
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Objective:By comparing the changes of metabolic parameters before and after laparoscopic sleeve gastectomy (LSG) in patients with type 2 diabetes mellitus (T2DM) and obesity, the insulin resistance index (HOMA-IR) and atherogenic index of plasma (AIP) were calculated to evaluate the effect of metabolic surgery on insulin resistance and atherosclerosis in patients with type 2 diabetes mellitus (T2DM) and obesity.Methods:LSG treatment were retrospectively analyzed in 54 patients with type 2 diabetes mellitus and obesity, detection of preoperative and postoperative 1 month, 6 month of fasting plasma glucose (FPG), fasting insulin (FINS), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), measuring blood pressure, body weight, calculating body mass index, and according to the steady state evaluation model and the formula for calculating HOMA-IR and AIP. Before and after surgery, paired t test was used, and Pearson correlation analysis and multiple stepwise regression analysis.Results:FPG, FINS, TG, HOMA-IR and AIP were (6.38±2.03) mmol/L and (5.36±1.33) mmol/L, (20.42±25.77) uU/mLand (11.22±3.62) uU/mL, (1.94±2.81) mmol/Land (1.70±2.33) mmol/L, (5.60±7.52) and (2.58±0.80), (0.15±0.27) and (0.08±0.25) ,which were significantly lower than those before surgery ( P<0.05) ,HDL-C was (1.04±0.20) mmol/L and (1.10±0.18) mmol/L at 1 and 6 months after operation, respectively, which was higher than that before operation ( P<0.05) .Preoperative correlation analysis showed that AIP was positively correlated with FPG, TG and HOMA-IR ( P<0.05), and negatively correlated with HDL-C ( P<0.05) .The results of multiple stepwise regression analysis showed that FPG, TG and HDL-C were independent influencing factors of AIP ( P<0.05) . Conclusion:LSG surgery can effectively reduce the blood glucose and lipid levels in patients with type 2 diabetes complicated with obesity, improve insulin resistance and reduce the plasma atherosclerosis index.
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Objective:To explore the effect of long-chain fat emulsion in parenteral nutrition therapy on the perioperative nutritional status of patients with low rectal cancer.Methods:A total of 204 patients who underwent rectal cancer surgery in the Third Hospital of Shanxi Medical University from January 2017 to June 2020 were retrospectively analyzed. The patients were divided into two groups according to the specific nutritional treatment methods, 100 cases in the study group used long-chain fat emulsion for parenteral nutrition support, and 104 cases in the control group used medium- and long-chain fat emulsion injection. After admission, the nutritional status of patients were evaluated according to the results of Scored Patient-Generated Subjective Global Assessment (PG-SGA) and related laboratory tests. At 7th day before the operation, the patients were treated with nutrition and electrolyte support. Parenteral nutrition and enteral nutrition combined treatment and early enteral nutrition were given after the operation. The albumin, prealbumin, retinol-binding protein, total cholesterol and body mass index (BMI) at 7th day before the operation, 1st day after the operation and 7th day after the operation and the patient's first exhaust time after surgery, occurrence of postoperative complications, postoperative fever and total hospital stay were recorded and compared between the two groups.Results:Postoperative first exhaust time [(42±11) h vs. (54±10) h], fever time [(48±8) h vs. (57±7) h], total hospital stay [(16.0±0.7) d vs. (18.0±0.9) d)], resting energy expenditure at the 7th day after surgery [(5 326±589) kJ/d vs. (5 840±599) kJ/d] and total cholesterol at the 7th day after surgery [(4.8±0.3) mmol/L vs. (5.0± 0.4) mmol/L] in the study group were lower than those in the control group, and albumin [(33±3) g/L vs. (28± 3) g/L], prealbumin [(0.189±0.041) g/L vs. (0.164±0.037) g/L] and retinol-binding protein [(0.039±0.016) g/L vs. (0.032±0.013) g/L] at the 7th day after surgery in the study group were higher than those in the control group, and the differences between the two groups were statistically significant (all P < 0.05). There was no statistical difference in other detection indexes between the two groups (all P > 0.05). Conclusion:The use of long-chain fat emulsion in low rectal cancer patients with malnutrition during the perioperative period may be more conducive to the recovery of the body.
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The clinical data of 15 patients with duodenal trauma who were admitted to Shanxi Bethune Hospital from January 2012 to June 2019 were retrospectively analyzed. There were 13 patients with blunt injury and 2 with penetrating injury. The surgical procedure was selected by patient′s condition and extent of injury combined with the clinical symptoms, imaging examination and the Organ Injury Scale grading system of the American Association for the Surgery of Trauma (AAST-OIS). All patients were followed up through outpatient examination and telephone interview till February 2020. Ten patients were diagnosed as duodenal trauma by CT scan before operation, and 5 patients were diagnosed during the operation. According to the AAST-OIS, 1 patient was with grade Ⅰ injury, 6 in grade Ⅱ, 5 in grade Ⅲ, 2 in grade Ⅳ and 1 in grade Ⅴ. All 15 patients received surgical treatment, including 1 with simple suture, 5 with break suture and duodenal diverticularization, 6 with break suture and biliary drainage (3 with hepatocystic duct drainage and 3 with cholecystostomy), 2 with pancreaticoduodenectomy. Postoperative complications occurred in 3 patients with Clavien system classification of Ⅲ b, Ⅱ and Ⅱ. One patient with duodenal stricture and severe abdominal infection was cured after gastrectomy and Billroth Ⅱ gastrojejunostomy 6 months after operation, and 2 cases with duodenal fistula were cured after conservative treatment. One patient who underwent pancreaticoduodenectomy was followed up for 6 months in the outpatient department, and 14 patients were followed up for 6-24 months. For emergency abdominal trauma patients with suspected duodenal injury, surgical exploration should be carried out actively. The site and range of intestinal wall injury should be considered in order to select a reasonable operation. Effective duodenal decompression and complete peritoneal drainage are important for the success of surgery. Early postoperative enteral nutrition support is one of the key measures for successful wound healing.
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Objective To study the effect of human CCR4-NOT transcription complex subunit 7 (CNOT7) gene knockdown on the immune microenvironment of HepG2 cells and explore its significance. Methods We designed a cell transfection protocol and performed the experiment with three groups:CNOT7-targeted knockdown group, control group, and CNOT7 overexpression group. The transfection efficiency was assessed using inverted fluorescence microscopy, and the expression level of CNOT7, transforming growth factor-β1 (TGF-β1), and nuclear factor-kappa B (NF-κB) p65 proteins was determined by Western blotting. The concentration of TGF-β1 secreted in the cell culture supernatant was measured by ELISA. The sensitivity of tumor cells to the killing function of natural killer (NK) cells was detected by flow cytometry. Results Compared with the control group, the expression level of TGF-β1 and NF-κB p65 proteins was significantly decreased in the CNOT7-targeted knockdown group, and the TGF-β1 concentration in the culture supernatant was also significantly reduced. However, in the CNOT7 overexpression group, the expression level of the two proteins and TGF-β1 concentration were significantly increased. NK cells were co-cultured with tumor cells, and the apoptosis rate of HepG2 cells transfected with CNOT7-specific shRNA was significantly increased. However, in the CNOT7 overexpression group, the apoptosis rate was significantly decreased. Conclusion CNOT7 forms the immune microenvironment of hepatocellular carcinoma. Targeted knockdown of CNOT7 can reduce TGF-β1 secretion and enhance the killing function of NK cells toward HepG2 cells.
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Objective To analyze tumor immune microenvironment and related mechanisms in liver cancer.Methods We included 10 cases of hepatocellular carcinoma,hepatitis B patients and healthy volunteers from January 2015 to December 2017 in Shanxi Grand Hospital.We first detected the peripheral and local GM-CSF level in each group,detected myeloid-derived suppressor cells (MDSCs) GM-CSF and pathway-related protein expression.from liver cancer,tumor margin and normal liver tissue through flow cytometry and immunohistochemistry,Finally,we transfected the CCR4-NOT transcriptional complex subunit 7 (CNOT7) recombinant plasmid in the hepatoma cell line,and then detected the related protein expression.Results There was no significant difference for peripheral blood GM-CSF level between liver cancer group,hepatitis group and control group (P＞0.05).The level of local GM-CSF was (32.2±8.9) ng/L,which was higher than that of hepatocellular carcinoma (9.7±2.7) ng/L and normal liver tissue (11.6±2.9) ng/L.The difference was statistically significant (P＜0.05).The proportion of MDSCs at the edge of the tumor was (9.9 ±3.6) ％,which was higher than that of liver cancer (4.0± 1.5) ％ and normal liver tissue (6.3±2.3) ％,and the difference was statistically significant (P＜0.05).Immunohistochemistrydata was consistent with previous data.Compared with normal liver tissue,CNOT7 and STAT3 were highly expressed in liver cancer tissues,while STAT1 was lowly expressed.HepG2 human hepatoma cells were selected for transfection.Compared with the empty plasmid group,CNOT7 expression was decreased in the knocking out group at the same time STAT1 expression was increased,STAT3 and GM-CSF expression was decreased.Conclusion In hepatocellular carcinoma,the secretion of GM-CSF increased and the number of MDSCs increased.Knocking out CNOT7 reduced GM-CSF secretion and activate the JAK/STAT signaling pathway.
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Objective To observe the changes in cytoskeleton (CSK) and glycosaminoglycan (GAG) synthesis following passage culture of articular chondrocytes and the correlation between CSK and GAG.Methods Eight male New Zealand White rabbits (8-month-old) were sacrificed by air embolism.After the chondrocytes from their knee joints were isolated by enzymolysis method,monolayer culture was performed.The chondrocytes of primary passage (P0) and passages 1 & 2 (P1,P2) were inoculated into 24-well plates with round cover slips put at the bottoms.Cell climbing slices were fixed after attachment of chondrocytes.The CSK proteins,actin,vimentin,tubulin and vinculin were stained by immuuofluorescence antibody on P0,P1 and P2 cell climbing slices,respectively.The CSK morphology was observed by laser confocal scanning microscopy and the fluorescence intensities of CSK proteins were detected by the fluorescence intensity software.The medium was changed for each generation after cell fusion and the GAG concentrations in the supernatants were measured at 24,36,48,60 h after medium change by alcian blue method.Results The intermediate filament networks became loosen and the dense distributions surrounding the nucleus decreased;more microtubule processes formed at the cell periphery with passage.The fluorescence intensity of actin of P1 chondrocytes was significantly increased than that of P0 (P ＜ 0.05),but there were no such significant differences between P0 and P2 or between P1 and P2 (P ＞ 0.05).The fluorescence intensities of vimentin and tubulin were significantly decreased with passage respectively,and there were such significant differences between any two of P0,P1 and P2 (P ＜ 0.05).The GAG concentrations in the supernatants were significantly decreased with passage at each time point,and there were such significant differences between any two of P0,P1 and P2 (P ＜ 0.05).Conclusions Passage culture of articular chondrocytes may lead to changes in morphology and protein expression intensity of the main components of CSK,and accordingly to decreased synthesis amount of GAG,one of the extracellular matrix of chondrocytes,indicating the changed characteristics of chondrocytes after passage and a certain correlation between CSK and GAG.
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Objective To explore the effect of lncRNA of growth arrest-specific 5 (lncRNA GAS5) on the radiosensitivity of colon cancer cells by targeting miR-223.Methods The expressions of lncRNA GAS5 in a few of colon cancer cell lines were detected by real-time quantitative PCR (qPCR).The cell lines with low expression level of lncRNA GAS5 were selected for subsequent study.The effect of overexpression of lncRNA GAS5 on the radiosensitivity of colon cancer SW480 cells was detected by cell cloning experiments.The target gene miR-223 of lncRNA GAS5 was predicted and validated by the bioinformatics database starBase and dual luciferase reporter assays.qPCR was used to detect the expression of miR-223 in various colon cancer cell lines and the influence of lncRNA GAS5 overexpression on the expression of miR-223 in SW480 cells.Results Compared with normal human colonic epithelial cells (NCM460),the expressions of lncRNA GAS5 in the colon cancer SW480,LOVO,HT-29 and SW620 cell lines were significantly lower(t =15.25,8.69,14.42,11.62,P ＜ 0.05),with the lowest level in SW480 cells.Both overexpression of lncRNA GAS5 and down-regulation of miR-223 significantly increased the radiosensitivity of colon cancer cells by decreasing cell survival fraction (at 8 Gy,lncRNA GAS5,t =13.51,P ＜ 0.05;anti-miR-223,t =14.93,P ＜ 0.05)and promoting apoptosis (lncRNA GAS5,t =8.30,P ＜ 0.05;anti-miR-223,t =7.32,P ＜ 0.05).Bioinformatics analysis showed that the 3'sequence of lncRNA GAS5 contained the binding sites with miR-223.After overexpression or downregulation of lncRNA GAS5,the expression of miR-223 was enhanced or reduced.Conclusions The lncRNA GAS5 promotes the apoptosis of colon cancer cells and inhibits its survival by targeting miR-223 expression,thereby increases the radiosensitivity of colon cancer cells.
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Objective To investigate whether astragaloside Ⅳ can regulate the multidrug resistance of HepG2/GCS-resistant cell lines,restore the sensitivity of drag-resistant cell lines to adriamycin (ADM) and its mechanism.Methods We used recombinant GCS (shRNAS) and control recombinant plasmids and did the transfection with HepG2 cells.RT-PCR and Western blot were used to analyze the expression of GCS.Astragaloside Ⅳ cytotoxicity experiments and ADM were performed in experimental and control groups.Hoechst 33258 was detected in two groups,apoptosis was detected by flow cytometry,and protein expression of caspase 9,3,Bax,and bcl-2 were detected by Western blot.Results RT-PCR and fluorescence observation showed that GCS was highly expressed after the transfection.Western blot showed that compared with control group,and HepG2GCS group,GCS and MDR1 expression were higher than;Astragaloside Ⅳ cytotoxicity experiment showed tumor proliferation was not regulated by GCS(FHepG2GCS=0.308,FHepG2EV =0.216,FHepG2 =0.153,P＞ 0.05),ADM in vitro tumor cell inhibition experiments showed that HepG2GCS cells were resistant to ADM (50％ cell transplantation concentration were 7.5,7.5,15 μg/ml);Hoechst 33258 and flow cytometry showed that Astragaloside Ⅳ can restore ADM tumor inhibition;Western blot showed that compared to untreated HepG2EV and HepG2GCS cells,protein level of caspase 9,caspase 3 were increased in Ast+HepG2EV and Ast+HepG2GCS groups (t=7.17,P＜0.05).At the same time,Bax and Bcl-2 were significantly different in each group (P＜ 0.05).Conclusions Astragaloside Ⅳ reverses multidrug resistance in HepG2/GCS cell lines,restores its sensitivity to ADM,promotes apoptosis in tumor cells through the caspase pathway and Bax pathway,and thus plays an important role in cancer chemotherapy.
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Objective To study the regulation of dendritic cells by recombinant glycated polylysine-coupled MIP-3α-FL double-gene targeting expression vector in liver cancer immune microenvironment.Methods H22 hepatocarcinoma cells were transfected with recombinant plasmid of MIP-3α-FL (shMIP-3α-FL) and injected into hepatoma model mice.The survival time,tumor size were compared.Flow cytometry was used to measure the number and phenotype of tumor infiltrating DCs.Results Western blot and ELISA demonstrated that the secretion of MIP-3α and FL in H22 cells was significantly increased after transfection with MIP-3α-FL.The survival time of the mice in the experimental group was significantly prolonged,the tumor size decreased.Flow cytometry showed that the number of tumor-infiltrating DCs in the experimental group was significantly higher than that in the control group;the expression of CD80 and CD86 in the infiltrating dendritic cells (TIDCs) was significantly higher than that of the control group.Conclusions The co-action of MIP-3α and FL can significantly promote DC accumulation,maturation,and conjugate glycosylated polylysine carriers increase the precision of targeting and enhance the antigenpresentation of the DCs.
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Objective To study the action of CNOT 7 (CCR4-NOT transcription complex subunit 7 human)gene and its mechanisms in the process of Vγ 9Vδ2T cell immunologic tolerance of HepG2 cells (Hepatoblastoma G2 Cell Line).Methods The shCNOT 7 (Recombinant plasmid of CNOT 7) and control vector shRNA were transfected into HepG2 cells.Vγ9Vδ2T cytokine stimulated each group before and after cell transfection,Cell apoptosis was detected by flow cytometry (FCM),CNOT 7 protein and STAT1,STAT3 expression level was detected by Western blot.CNOT 7,STAT1 and STAT3 protein expression levels of HepG2 liver cancer cell lines and L02 normal liver cell line was assayed by Western blot.Results When stimulated by Vγ9Vδ2T cytokine,the apoptosis rate of gene-knockdown group significantly improved from (7.55％ ±2.63％) to (20.59％ ±3.12％).Compared with L02 cells,the CNOT7 protein expression of HepG2 cells increased (F =28.76,P ＜ 0.01),STAT3 protein expression increased (F =110.29,P ＜ 0.01),while STAT1 protein expression was down-regulated (F =35.67,P ＜ 0.01).CNOT 7 knockout could induce HepG2 cells STAT1 expression (t =6.69,P ＜ 0.05).Conclusions CNOT 7 gene could induce HepG2 cells Vγ 9Vδ2T cellular immune tolerance.CNOT 7 knockout could reverse the Vγ 9Vδ2T cell immunologic tolerance of HCC.
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Objective To investigate the percentage,mature classification and Immune killing function of Vγ9Vδ2T cell in peripheral blood of HCC patients.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from HCC patients (n =25) and healthy donors (n =20) by discontinuous density gradient centrifugation.Proportion,mature and differentiate subtypes and IFN-γ and CD107a expressing of the delta 2 T cells were detect by using flow cytometry,δ2Tcell were selectively cultured with zoledronate and human IL-2.After 12-14 days cells were collected and tested for the second time.Results While the percentage of Vγ9Vδ2Tcell of total T cell in peripheral blood of HCC patients is lower than healthy people before culture (t =4.505,P ＜ 0.001),after augmentation in vitro the proportion increased significantly (t =8.782,P ＜ 0.001),to a level similar to healthy group (t =1.644,P =0.109).There was no statistically significant difference when differentiation subtypes of patient's Vγ9Vδ2Tcell were compared with healthy group before culturing (all P ＞ 0.05),after culture the proportion of Tn,Tcm and Temra decreased [t(Tn) =2.081,t(Tcm) =2.478,t(Temra) =2.953,all P ＜ 0.05],and the proportion of Tem,Tem+ Temra increased [t(Tem) =12.6,t (Tem + Temra) =9.843,all P ＜ 0.001].Cell culture did not alter the proportion of IFN-γ and CD107a secreting Vγ9Vδ2T cells in the peripheral blood in both HCC patients and healthy people (all P ＞ 0.05).Conclusions While the percentage of Vγ9Vδ2T cell of HCC patients in peripheral blood was lower than healthy people,its matured subtypes are similar to those of healthy people,and functions of expressing IFN-γ and CD107a are not different with healthy people.Applying ZOL + IL-2 can amplifyVγ9Vδ2T cells of patients with HCC.
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Objective To observe and compare the changes of weight,blood pressure,glycemia, blood lipid,serum uric acid,insulin sensitivity and insulin function of laparoscopic sleeve gastrectomy (LSG)for the treatment of type 2 diabetes mellitus(T2DM)with obesity,and analysed mechanism of meta-bolic surgery.Methods Changes of weight,blood pressure,glycemia,blood lipid ,serum uric acid,in-sulin sensitivity and insulin function of 8 type 2 diabetes mellitus(T2DM)with obesity patients undergoing LSG were retrospectively analyzed.Weight,BMI,EWL%,average systolic and diastolic pressure were measured 1,2,3,6 months and fasting plasma glucose (FPG),glycosylated hemoglobin (HbAl),insulin sensitivity parameter:insulin resistance index(HOMA-IR),insulin sensitivity index(ISI),insulin area un-der the curve (AUCI),insulin function parameter:glucose disposition index(DI)were measured 1,6 months after operation and compared with preoperative levels.Meanwhile medication change was observed af-ter operation and compared with preoperation.Results ⑴Weight ,BMI at 1,2,3,and 6 months after surgery showed a decreasing trend and EWL% showed a decreasing trend over time .⑵ FBG、HbA1c showed a decreasing trend after operation.Blood lipid,uric acid of 3 patients with dyslipidemia and hyperu-ricemia decreased to normal 1 month after operation.⑶ HOMA-IR,AUCI except for 1 patient showed a de-creasing trend and ISI was contrary after operation.DI ,trends were different.⑷7 patients stopped all an-tidiabetic drugs 6 months after operation,1 patient injected insulin with good glycemic control.Blood pres-sure of 5 patients with hypertension was normal without antihypertensive drugs 6 months after operation. Conclusions LSG procedure can significantly increase insulin sensitivity of T2DM with obesity,thereby improve glycemia,cholesterol,uric acid and further promot weight loss.
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Objective To study the relationship between Treg cell numbers and level of TGF-β1,IL-10 in the immune microenvironment of rat liver cancer.Method 40 male Wistar rats were randomly divided into three groups,liver cancer model group (n =20),saline control group (n =10) and blank control group (n =10).Liver cancer model was established by intraperitoneal injection of DEN (100 mg/kg).Percentage of Treg cells in cancer tissues was detected by fiow cytometry and the expression level of TGF-β1 and IL-10 by immunohistochemical SABC.Results The ratio of Treg cells in CD4 + T cells was 14.32％ ± 4.84％ in liver cancer tissues,significantly higher than that in the blank control group 5.64％ ±6.10％ and the saline control group 7.95％ ±3.55％.The difference was statistically significant (F =211.279,P ＜ 0.05).The expression level of TGF-β1 and IL-10 in cancer tissues was significantly higher than the blank control group and the saline control group (FTGF-β1 =250.740,FIL-10 =152.744,all P ＜0.05).The number of Treg cells was positively correlated with TGF-β1,IL-10 level (P ＜ 0.05).Conclusions TGF-β1,IL-10 and Treg cells significantly increased in rat experimental liver cancer tissues,and there was positive correlation between Treg cells,and TGF-β1 and IL-10 in liver cancer tissues.
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Objective To investigate the effect of sodium demethylcantharidate injection on adherence,invasion and metastasis and to investigate the related mechanism in human hepatocarcinoma cell line HepG2.Methods Adherence ability,migration and invasion of HepG2 cells inhibited by sodium demethylcantharidate injection were assessed by MTT and Transwell techniques.Expression levels of MMP-9 protein in HepG2 cells were determined by immunohistochemistry.Results The number of adhesion,migration and invasion of HepG2 cells were significantly lower in sodium demethylcantharidate injection group than those in the control group (P ＜ 0.05).HepG2 cells co-incubated with sodium demethylcantharidate injection in the concentration of 0.25 μg/ml for 30,60,90 and 120 min showed higher cell adhesion than the control group.The adhesion inhibition ratios were 48.11％,33.81％,28.97 ％ and 16.83 ％,respectively.The migration and invasion inhibition rates were 64.19 ％ and 58.19 ％.With concentration of sodium demethylcantharidate injection to increasing,expression levels of MMP-9 protein in HepG2 cells more and more lower than control group.Conclusion The adherence,migration and invasion abilities of HepG2 cells are markedly inhibited by sodium demethylcantharidate injection,the mechanisms is possible related to the expression levels of MMP-9 protein.
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Objective To summarize the clinical experiences of liver transplantation.Methods Of the nine patients, four operation was standard orthotopic liver transplantation,the latter five were the piggyback liver transplantation.The immunosuppressive protocols included methylprednisolone FK506 and mycophenolatemofeti. Meanwhile intravenous antihepatitis B immunoglobulin and Lamivudine were used to prevent hepatitis B recurrence.Results All patients were cured.Conclusion Liver transplantation can be employed for liver disease both cirrhosis and carcinoma as a conventional surgery.It is an effective way for the treatment of no metastatic liver carcinoma.The immunosuppressive protocols included methylprednisolone FK506 and mycophenolatemofeti,it can prevent immune rejection.
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Objective To explore the effect of FTY720 on human hepatocellular carcinoma cell line HepG-2 of invasion ability. Methods HepG-2 cells were cultured,and divided into 4 groups,namely control and FTY720 (0.05 g/ml,0.10 g/ml,0.15 g/ml) groups.The cell invasion ability was observed by the cell invasion assay.mRNA expression of MMP-2,MMP-9 was measured by RT-PCR.MMP-2,MMP-9 protein expression was measured by ELISA method.Results The invasive ability of cancer cells in each group significantly weaken,with the FTY720 concentration increased.The number of control group was (110±3)(P<0.05),the number of O.05 g/ml FRY720 treatment group cells was (74±4),while the number of cells of O.10 g/ml,0.15 g/ml the drug treated group,Was (42±3),(25±5)(P<0.01).Compared with control group,the MMP-2,MMP-9 expression of different concentrations of FTY720 groups decreased significantly (P<0.05).As the expression of FTY720 concentration increased,the MMP-2,MMP-9 expression decreased (P<0.01).Compared with control group,the MMP-2,MMP-9 protein expression of different concentrations of FTY720 groups decreased significantly (P<0.05), but the protein expression and concentration of FTY720 is inversely proportional in a certain range. Conclusion The invasive ability of HepG-2 cells can be inhibited by FTY720,and with FTY720 concentration increased,the invasive ability decreased gradually.One of the mechanisms may be related to reduce the MMP-2,MMP-9 protein and gene expression.
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Objective To study the effect of somatostatin on the Proliferation in Human HepG2 Cells and the Level of Gene p16. Methods The cell line HepG2 were divided into 3 groups: control groupA, low concentration group B and high concentration group C. The flow cytometry (FCM) was used to detect phase distribution of the cycles and detect the expression of p16 gene by using RT-PCR. Results Human HepG2 cells affected 24 hours later by different density SST, using FCM, we discovered that the G1 cell cycles of each A group, B group, C group occupied the proportion respectively is (51.41±3.27)%, (65.43±3.41)%, (69.02±3.75)%, that the S cell cycles is (36.86±2.31)%, (20.34±2.52)%, (18.50±3.09)%, that the G2 cell cycles is (11.46±1.96)%, (14.23±3.83)%, (12.48±3.18)%. Compare to the control groupA, the G1 cell cycles of groupB and group C were increased obviously. But the S cell cycles relatively reduced. The levels of p16 gene were expressed in each group detected by RT-PCR, but B group and C group expressed obviously. Conclusion The inhibited mechanism of somatostatin for HepG2 was expressed probably by improving levels of p16.
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Objective To investigate the effect of the lack of intestinal bile on liver regeneration after hepatectomy.Methods The model of interference with intestinal bile acid metabolism in rats was established by feeding rats with 0.2% cholic acid(cholic acid loading group), 2% cholestyramine resin(lack of bile group)and feeding the standard diet as the control group.Liver regeneration was compared among the 3 groups at 0, 1, 2, 3, and 7 d after 70% partial hepatectomy(PH)in rats and mRNA expression of the rate-limiting enzyme of bile acid biosynthesis(CYP7a1)and farnesoid X receptor (FXR)were detected.Results The rate of liver regeneration was significantly lower on days 3 and 7after PH in the lack of bile group than in the other groups(P＜0.05).On day 1, the labeling indices of PCNA and Ki-67 in the lack of bile group(22.21% ±2.31%、 17.25 % ± 6.50 %)were lower than those in the cholic acid loading group(44.4%±4.92%、 30.83% ± 3.91%)and control group (38.74% ±6.42% 、27.04% ±7.22%)and the peaking of labeling indices was delayed.After PH, the mRNA expression of FXR was significantly lower in the lack of bile group than in other groups.However, CYP7al mRNA had a trend towards increase after PH and was higher than that in other groups.Conclusion Lack of intestinal bile results in delayed liver regeneration of normal rat liver accompanied by decreased expression of FXR mRNA after hepatectomy.
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The liver regeneration is closely related to the bile acids. To avoid the toxic effects of bile acids on hepatocyte, the state of bile secretion, the rate-limiting enzyme of the bile acid synthesis, bile acids composition as well as the transporter changes at the process of liver regeneration. In vivo and in vitro experiments show that bile acids can promote liver cell proliferation, liver regeneration may be related to the signal which is released under the bile acid imbalance. The relationship between the liver regeneration and bile acid metabolism has an important practical significance in liver regeneration.