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Objective:To detect the mRNA expression and methylation status of leucine rich repeat containing 55(LRRC55) gene in pancreatic carcinoma tissues, and discuss the clinical value.Methods:Resected pancreatic ductal adenocarcinoma and normal adjacent specimens from 37 patients admitted in General Surgery of First Affiliated Hospital of Naval Medical University were collected from May 2019 to May 2021. Another two normal pancreas specimens and two blood samples from healthy adults were also collected. All patients′ age, gender, tumor location, tumor size, tumor differentiation, TNM staging, lymphatic metastasis, CEA and CA19-9 level were recorded. Bisulfite treatment of genomic DNA and sequencing analysis was used to study methylation patterns in CpG islands of the promoter for LRRC55 gene in fresh tissues from 2 pancreatic adenocarcinoma and adjacent tissues, 2 normal pancreatic tissues, 2 pancreatic cancer cell lines (PaTu8988 and ASPC1). LRRC55 mRNA in 35 pancreatic adenocarcinoma and adjacent tissues was detected by real-time quantitative PCR and the correlations with clinical parameters were analyzed.Results:CpG islands of LRRC55 in pancreatic adenocarcinoma tissues and pancreatic cancer cell lines was highly methylated and the mean methylation rate was 53% and 71%, respectively; while LRRC55 gene in pancreatic adjacent tissues and normal pancreatic tissues was lowly methylated, and the mean methylation rate was 8% and 11%. The relative expression in the pancreatic adenocarcinoma tissues and the paired adjacent normal tissues was 0.21 (0.02, 1.00 ) and 0.98 (0.33, 3.66 ), respectively; the former was significantly lower than the later and the difference was statistically significant ( P=0.003). Correlation analysis showed that LRRC55 mRNA expression level was related to tumor differentiation and CEA, but not correlated with patients′ age, gender, tumor location and size, CA19-9 level, lymphatic metastasis and TNM staging. Conclusions:Pancreatic cancer tissue and cell lines had abnormal methylation of LRRC55 gene; LRRC55 gene hypermethylation was related with its lower mRNA expression level in pancreatic cancer, which was correlated with the tumor differentiation and CEA level. LRRC55 may be a potential suppressor gene for pancreatic cancer.
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Objective:To investigate the clinical outcomes of patients with intrahepatic cholangiocarcinoma (ICC) undergoing surgical resection.Methods:Patients who undergoing radical surgical resection for ICC from Jan 2015 to Apr 2021 at the Department of General Surgery, the First Affiliated Hospital of Anhui Medical University were included in this retrospective cohort study.Results:There were 67 patients in the final analysis, The median follow-up duration was 14 months (range: 1-60 months). Firty three patients (79.1%) had tumor recurrence, 52 patients (77.6%) died, Among them, 49 patients (73.1%) died from tumor recurrence. The 1-、2-、and 3-year accumulated disease-free and overall survival rate were 35.6%, 19.6%, 16.8% and 53.7%, 32.4%, 20.8%. respectively. The overall survival rate of the group without microvascular invasion was significantly better than those of the group with microvascular invasion ( χ2=5.916, P=0.015). CA19-9≥1 000 U/ml was the only independent risk factor for the disease-free survival. CA19-9≥1 000 U/ml、blood loss≥600 ml、microvascular invasion and tumor recurrence were the independent risk factors for the overall survival. Conclusion:For ICC patients with single tumor, when the tumor diameter is less than 5 cm and has no microvascular invasion, surgical resection is recommended, and a satisfactory prognosis could be achieved.
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Objective To investigate the early predictive value of several commonly used biochemical markers for predicting persistent organ failure ( POF ) in patients with hyperlipidemic acute pancreatitis ( HLAP) . Methods Clinical data of 157 patients with HLAP within 72 hours after the onset of first attack who were admitted to the Dept. of Gastroenterology in Changhai Hospital from January 2015 to December 2017 were retrospectively analyzed, including 106 cases without POF ( non POF group ) and 51 cases with POF ( POF group) . Hct, BUN, Cr, APACHEⅡand BISAP were recorded within 24 hours after admission. Receiver-operating characteristic ( ROC) curve was drawn to calculate area under the ROC curve ( AUC) and evaluate the performance of Hct, BUN, Cr, APACHEⅡand BISAP scores in predicting HLAP complicated with POF, which was compared by DeLong test. Results Values of BUN, Cr, APACHEⅡand BISAP were significantly higher in HLAP patients with POF than those without POF [(10. 30 ± 7. 43) vs (5. 34 ± 2. 26) mmol/L, (165. 31 ± 123. 93) vs (65. 61 ± 20. 82)μmol/L, (10. 22 ± 6. 22) vs (4. 61 ± 2. 99) points, (2. 61 ± 0. 87) vs (1. 42 ± 1.07) points], and the differences were all statistically significant (all P<0.05), whereas Hct was not significantly different between the two groups. The AUC of Cr and BUN for predicting POF was 0. 77(95％ CI, 0. 69-0. 86) and 0. 71 (95％ CI, 0. 61-0. 81), respectively, and the optimum predictive Cut-off values were 130 μmol/L and 8. 95 mmol/L, respectively. The sensitivity was 53％, and the specificity was 99％ and 94％;the accuracy was 84％ and 81％;negative predictive value was 81％, and positive predictive value was 96％ and 82％. DeLong test showed that predictive performance of BUN and Cr was not statistically different from that of APACHEⅡand BISAP. Conclusions Cr≥130 μmol/L and BUN≥8. 95 mmol/L can be used clinically to predict the presence of POF in HLAP, and the predictive efficacy were comparable to APACHEⅡand BISAP.
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Objective To explore the effectiveness of endoscopic ultrasound( EUS)-guided holmium laser ablation for primary pancreatic implantation tumor in nude mice. Methods Pancreatic cancer cell line SW1990 were implanted into 204-6-week-old male balb/c nude mice to establish primary pancreatic implantation tumor in situ models. Then the nude mice were randomly divided into two groups, the treatment group(n=10) and the control group(n=10). The treatment group underwent EUS-guided holmium laser ablation in the pancreatic tumor. And no interventions were given to the control group. The volume of tumors of the two groups were measured under EUS at time points of 7 d, 14 d and 28 d after ablation. The activities, appetites and psychosis of all nude mice were evaluated in the meantime. At 28 d after ablation, lesions of pancreas were dissected and sliced for H&E staining. Results There were no complications in the treatment group, and all nude mice could tolerate the procedure. The mental state, activities and appetites of nude mice in the experimental group were better than those in the control group. Tumors of the control group enlarged. There was significant difference in the tumor size between the two groups at 28 d after ablation. HE staining showed coagulation necrosis in the ablation area. Conclusion EUS-guided holmium laser, producing coagulative necrosis in the ablation area, is effective for primary pancreatic implantation carcinoma in nude mice for about 28 days.
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Objective To investigate the effect of CYP3A5 on the proliferation of pancreatic cancer cells and its underlying mechanisms.Methods The protein expression of CYP3A5 in five pancreatic cancer cell lines BxPC-3,FG,MDA28,8902 and PANC1 was detected by Western blotting.The PANC1 cells with the lowest protein expression of CYP3A5 and the BxPC-3 cells with highest expression of CYP3A5 were transfected with CYP3A5 overexpression plasmid and CYP3A5 targeted-siRNA (siRNA-CYP3A5),respectively.CCK-8 and cloning formation assay were used to investigate the role of CYP3A5 overexpression and knockdown in the proliferation of pancreatic cancer cells.The changes of the protein and mRNA expression of cell cycle regulating gene cyclin E,cyclin D1 and apoptosis related gene Bcl-2 were detected by Western blotting and PCR,respectively.Results CYP3A5 protein expression in PANC1 cells increased significantly after the transfection of CYP3A5 overexpression plasmid (1.66 ± 0.14 to 1,P =0.0021),which greatly decreased in BxPC-3 cells transfected with siRNA CYP3A5 (0.18 ± 0.02 to 1,P ＜0.0001).A450 values of the CYP3A5 overexpression group and the empty plasmid group in PANC1 cells cultured for 48 and 72 h were 1.36 ±0.05 vs 1.15 ± 0.03,2.1 ± 0.09 vs 1.42 ± 0.03,respectively,which were significantly higher in CYP3A5 overexpression group than empty plasmid group,and the differences were statistically significant (P value ＜ 0.005 or 0.001).The A450 values of BxPC-3 cells in CYP3A5-siRNA transfected group and siRNA-NC transfected group were 0.62 ±0.01 vs 0.77 ± 0.03、0.83 ± 0.01 vs 1.18 ± 0.02,respectively,which in The CYP3A5-siRNA transfection group was significantly lower than that of siRNA-NC transfection group,and the difference was statistically significant (P ＜ 0.05 or ＜ 0.001).The clone formation rate of PANC1 cells in the overexpression group was (19.33 ± 0.58)％,which was significantly higher than that in the empty plasmid group (9.67±0.63) ％,and the clone formation rate in CYP3A5-siRNA group was (8.5± 0.8)％,which was significantly lower than that of group siRNA-NC (16± 0.6)％,and the differences were statistically significant (P ＜0.01).The protein expression of cyclin D1 in CYP3A5 overexpression PANC1 cells was 2.00 ± 0.11,which was obviously higher than 1.00 in empty plasmid group (P ＜0.01).The protein expression of cyclin D1 in siRNA CYP3A5 BxPC cells was 0.45 ±0.04,which was obviously lower than 1.00 in siRNA NC group,and the difference was statistically significant (P＜0.01).However,CYP3A5 overexpression or inhibition did not influence the relative expression of cyclin D1 mRNA and cyclin E,Bcl-2 pretein expression.Conclusions CYP3A5 can promote the proliferation of pancreatic cancer cells by up-regulating cyclin D1 protein expression.
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Objective To explore the effects of ApoC3 gene on the severity of hypertriglyceridemiainduced acute pancreatitis (AP).Methods ApoC3 transgenetic mice and C57BL/6J mice AP model was induced by cerulein intraperitoneal injection,and ApoC3 transgenetic mice and C57BL/6J mice injected by normal saline solution in equal volume served as control group.Serum triglyceride and cholesterol were detected,and the pathological changes of the pancreas were observed.RT PCR method was used to examine the changes of the inflammatory factor including IL-1β,IL-6,α-SMA and TNF-α mRNA levels,which reflected the severity of the inflammation.Results Serum triglyceride and cholesterol were higher in ApoC3 transgenetic mice than in C57BL/6J mice [(3.434 ± 0.931) mmol/L vs (0.766 ± 0.120) mmol/L,(2.553 ±0.178) mmol/L vs (1.996 ± 0.080) mmol/L],and the differences were statistically different (P ＜ 0.05).The pathological changes of the pancreas were more severe in ApoC3 transgenetic AP mice than in C57BL/6J AP mice,and the IL-1β,IL-6 and α-SMA mRNA levels in the pancreatic tissue were obviously higher in ApoC3 transgenetic AP mice than in C57BL/6J mice (1.72 ± 0.07vs 0.78 ± 0.09,1.58 ± 0.09vs 0.87 ±0.04,0.83 ± 0.05vs 0.44 ± 0.04),and the differences were statistically significant (P ＜ 0.05),while there was no statistical difference on TNF-αmRNA level (0.70 ± 0.09vs 0.65 ± 0.08,P ＞ 0.05).Conclusions ApoC3 gene could aggravate the severity of the inflammation in hypertriglyceridemia-induced AP.
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Objective To compare the effects of two kinds of lidocaine gel indwelling catheter on male patients with general anesthesia during recovery period. Methods A total of 90 cases of male patients with general anesthesia were selected and divided into experimental group and control group by random digits table method,45 cases in each group. The experimental group was given lidocaine gel injection indwelling catheter urethral surface anesthesia, the control group will lidocaine gel evenly to the catheter indwelling catheter. The incidence of catheter related bladder irritation and restlessness were observed during the recovery period of general anesthesia. Results The incidence of catheter related bladder irritation in grade 0, I, II, III during the recovery period of general anesthesia were 28, 14, 3, 0 cases in the experimental group and 7, 23, 12, 3 cases in the control group, there was significant difference between 2 groups (Z=1500, P<0.01). The incidence of restlessness in grade 0, I, II and III during the recovery period of general anesthesia were 27, 18, 0 and 0 cases, respectively in the experimental group and 7, 20, 9, and 9 cases in the control group, respectively. The difference between the 2 groups was statistically significant (Z=1435.5, P<0.01). Conclusions Lidocaine gel injection into the urethra surface anesthesia can effectively improve the comfort of male patients during the recovery period of general anesthesia.
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Objective To investigate several related problems possibly affecting the measurement results in the experiment of 125I-FSH in vitro cells uptake by domestic GC-1200 gamma RIA counter.Methods Before entering the measuring room,the sample was performed the background measurement.CPM measured in the different locations of same measurement frame and the different locations in unmeasured area were performed the statistical comparison.Results In high count,the influence of single sample reaching 1.9 ×106CPM on the adjacent low counting tube count was 7%;its influence on low counting tube count in adjacent detector was 7.33%;all samples were arranged from high to low order and the high count sample holder was placed on the measured location close to the detector,its influence on low counting tube count was 5.33%.Conclusion The domestic GC-1200 γ RIA counter is suitable for the measurement of the in vitro cell uptake experiment of 125I nuclear labeling.
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Objective To observe the effect of silencing Snail gene on the invasion and proliferation ability of human pancreatic cancer cell line PANC1.Methods Lentiviral vectors that can express small hairpin RNA(shRNA) targeting human Snail gene(shRNA-Snail) or shRNA sequence that did not match any known mRNA(shRNA-NC) were constructed,and transfected into PANC1 cells.Untransfected cells served as control.mRNA and protein expression of Snail,α-smooth muscle actin (α-SMA) and E-cadherin was determined by real time quantitative PCR and Western blotting,respectively.In vitro invasion ability was tested by Transwell model.Proliferation ability was measured by CCK-8 assay.Results Compared with those in shRNA-NC group,Snail mRNA (0.27 ± 0.02 vs 0.92 ± 0.03) and protein level (0.26 ± 0.02 vs 0.80 ± 0.02),and α-SMA mRNA (0.33 ±0.04 vs 0.97 ±0.07) and protein level (0.31 ±0.04 vs 0.74 ±0.06) in shRNA-Snail group were obviously decreased,but E-cadherin mRNA (1.57 ± 0.45 vs 0.95 ± 0.08) and protein level (0.86 ± 0.03 vs 0.20 ± 0.03) were greatly increased.The number of cells permeating the septum of transwell [(6.80 ± 0.73)/400 magnification vs (26.80 ± 2.52)/400 magnification,P ＜0.01] was significantly decreased,and cell proliferation was inhibited (0.74 ± 0.05 vs 1.47 ± 0.04,P ＜ 0.01).All the differences above were statistically significant (all P ＜ 0.01).No significant differences were observed between shRNA-NC and normal control group.Conclusions Silencing Snail gene may restrain the invasion and proliferation ability of PANC1 cells to a certain degree.
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Objective To observe the relationship between Toll-like receptor 4 (TLR4) and the sensitivity of PANC1 cells to gemcitabine (GEM),and to analyze the potential mechanism.Methods PANC1 cells were divided into GEM group,lipopolysaccharide (LPS) + GEM group and TLR4-siRNA + GEM group.GEM group was treated by GEM alone.LPS + GEM group was pretreated with 1 mg/L LPS for 4 h and then treated by GEM.TLR4-siRNA + GEM group was transfected with 100 pmol/mL TLR4-siRNA for 4 h and then treated by GEM.The untreated cells were used as the control group.MTT method was used to detect the cell proliferation.Morphological changes and apoptosis rate of the cells were examined by Hoechst33258 staining and flow cytometry,respectively.The protein expression of TLR4,phosphorylated AKT (p-AKT) and activated Caspase-3 were detected by Western blot.Results The median inhibition concentration (ICs0) of GEM in the GEM group,LPS + GEM group and TLR4-siRNA + GEM group was (8.9 ± 0.32),(14.21 ±0.95),(3.96 ± 0.27) mg/L,respectively.The IC50 in LPS + GEM group was significantly higher than that in GEM group (P ＜ 0.01),and the IC50 of GEM in TLR4-siRNA + GEM group was significantly lower than that in GEM group (P ＜0.01).Compared with that in GEM group,the cells with typical apoptotic morphological changes were decreased in LPS + GEM group,which was increased in TLR4-siRNA + GEM group.The apoptotic rate in control group,GEM group,LPS + GEM group,TLR4-siRNA + GEM group was (2.1 ± 0.3) ％,(15.1 ± 2.3) ％,(9.8 ± 1.5) ％,(22.9 ± 3.1) ％,respectively.Compared with that in GEM group,the cells apoptotic rate was significantly reduced in LPS + GEM group (P ＜0.01),which was significantly increased in TLR4-siRNA + GEM group (P ＜0.01).TLR4 protein level in the 4 groups was 0.83 ±0.08,0.81 ±0.07,0.85 ±0.07 and 0.16 ±0.03;p-AKT protein level 0.61 ±0.05,0.36 ±0.03,0.73 ± 0.07 and 0.21 ± 0.02;activated Caspase-3 protein level was 0.66 ± 0.05,0.73 ± 0.07,0.45 ± 0.04 and 0.91 ± 0.07,respectively.The expression of TLR4 and p-AKT in TLR4-siRNA + GEM group was significantly lower than that in GEM group (P ＜0.01),while the expression of activated Caspase-3 protein was increased significantly (P ＜ 0.05).Compared with the GEM group,the expression of p-AKT protein in LPS + GEM group was significantly increased (P＜0.01),and the expression of activated Caspase-3 protein was significantly decreased (P＜0.01).Conclusions TLR4 can inhibit the sensitivity of pancreatic cancer PNAC1 cells to GEM,and the mechanism is related to the activation of PI3K/AKT pathway and downregulation of activated Caspase-3.
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Objective To synthesize and identify artificial antigens of lung elastin degradation peptide and for the purpose of preparation of COPD test.Methods The artificial antigens were synthesized by Sulfo-SMCC and KLH.The complete antigens were identified by ultraviolet spectrum and SDS-PAGE.Immunize Balb/c mice was used to prepare antibody.The antiserum activity was evaluated by indirect competitive ELISA.Results The artificial antigens were identified by ultraviolet spectrum and SDS-PAGE. The protein concentration was 1.181 mg/mL.The titer of antiserum was 1∶64 000,and IC50 was 13.7 ng/mL.The antiserum had no cross-reaction with nonsense peptide.Conclusion The artificial antigens were acquired successfully,which had good immunoge-nicity.The results have laid basis for COPD test.
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Objective To determine the lower limit of detection (LLOD) and cut off values of K-ras mutation detection by peptide nucleic acid (PNA) clamping-PCR.Methods The genomic DNA of pancreatic cancer cell lines (PANC1 and SW1990) with codonl2,13 mutation and the genomic DNA of placenta with K-ras wild type were mixed and diluted serially into samples with different mutation rate (0,0.1％,0.2％,0.4％,0.8％,1.6％,3.1％,6.25％,12.5％,25％,50％),PANC1 cells with 1％ mutation rate and SW1990 cells with 30％ mutation rate and 4 samples with the quantity of DNA was 50,20,5,1 ng and 50,10,5,1 ng was prepared.Codon 12,13 mutation of K-ras was determined by PNA-PCR,and the mutation Ct values,overall Ct values were collected,and the △Ct values (mutation Ct values-overall Ct values) were calculated,and the tests were repeated for 10 times.ROC curve was used to analyze the △Ct values and determine the best cut off values for K-ras mutation,and the positive diagnostic rate,LLOD was evaluated.Results The mutation Ct,△Ct values of codon 12 mutation of PANC1 and codon 13 mutation of SW1990 of all the different mutation rates were statistically significantly different (P ＜ 0.05) when compared with negative control group,but the overall Ct values were not statistically significantly different from that of negative control group.For detection of K-ras codon 12 mutation by ROC curve,the relevant area of ROC curve (AUC) was 0.926,the optimum cut off value of △CT was 11,the sensitivity and specificity were 84％ and 100％,respectively,and the LLOD was 0.4 ng.For detection of K-ras codon 13 mutation by ROC curve,the relevant AUC was 0.906,the optimum cut off value of △CT was 9.5,the sensitivity and specificity were 71％ and 100％,respectively,and the LLOD was 1.5 ng.The mutation detection results of fixed rate further confirmed the LLOD.Conclusions This study successfully defines LLOD and cut off value of PNA clamping-PCR/K-ras method in detection of K-ras 12 and 13 codon mutations.This method meets the requirement of clinical application.
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Objective To explore the change of serum thyroid hormone related indicators and the probability of occurrence of thyroid dysfunction abnormality in the patient with type 2 diabetes mellitus(T2DM).Methods 86 patients with T2DM and 61 age-matched and gender-matched individuals with healthy physical examination as controls were selected and detected serum FT3,FT4 and TSH by the electrochemiluminescence method Results The serum FT3,FT4 and TSH in the T2DM group were 5.09 pmol/L, 17.32 pmol/L and 2.81 mIU/L respectively;which in the normal control group were 4.99 pmol/L,17.24 pmol/L and 2.71 mIU/L respectively,the differences between the two groups had no statistical significance(P >0.05).Among 86 cases of T2DM,29 cases had the serum abnormal TSH with the abnormal rate of 33.7%,which in the control group was 14.8% with statistical difference between the two groups(P <0.05).Among T2DM patients,the TSH abnormal rate of in females was 42.1%,which was higher than 17.2% in males.Conclusion The serum thyroid hormone detection is necessary for the T2DM patients,especially female pa-tients,which is conducive to early screening,prevention and treatment.
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Objective To assess the role of health education in outcomes of diabetes mellitus among high-risk populations.Methods The community physicians who participated this investigation received standardized training,and 307 community residents at high risk of developing diabetes obtained three-month intense health education and nine-month follow-up study.Paired t-test,and Analysis of Variance were used for data analysis.Results After systematic health education,professional level of community physicians was improved.Cognitive level of health knowledge was also significantly improved (5.5 vs 12.6,t=-28.511,P＜0.05).In addition,health knowledge of variant age (F=4.036,P＜0.05),education level (F=15.27,P＜0.05) and occupation (F=9.80,P＜0.05) subgroups was significantly increased.In comparison with baselines,the scores of each age subgroups (F=0.204,P＞0.05) showed no significant differences,although scores of different education level (F=4.71,P＜0.05) and occupation (F=4.87,P＜0.05) subgroups were significantly different.The risk factors of diabetes were effectively controlled.Conclusions Health education should be the key to health management of diabetes,which plays important roles in improving cognitive level of health knowledge among populations at high risk of developing diabetes and reducing the incidence of this condition.
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Objective To observe the effect of triptolide combined with gemcitabine on proliferation and apoptosis of pancreatic cancer cells ,and to analyze the relevant mechanisms .Methods After treated with TPL ,GEM or TPL combined with GEM in vitro , PANC-1 cells proliferation was accessed by MTT assay and the interaction between the two drugs was calculated .Apoptotic mor-phological changes and apoptosis rate of the cells were investigated by Hoechst 33258 staining and flow cytometry ,respectively .The expression of signal transduction and transcription factor 3(STAT3) ,cysteine aspartate specific proteases-3(caspase-3) protein were detected by Western blot analysis .Results TPL ,GEM or TPL combined with GEM could significantly inhibit the prolifera-tion of PANC-1 cells ,and the combination of the two drugs had a synergistic effect .The cells of the TPL group ,GEM group ,as well as the combined group showed typical apoptotic morphological changes .Compared with the TPL group and GEM group ,the number of apoptotic cells of the combined group increased significantly .Compared with the control group ,the cells apoptosis rate of the TPL group ,GEM group and combined group was significantly increased (P< 0 .05) ,and the apoptosis rate of the the combined group was significantly higher than that of the monotherapy group(P<0 .05) .TPL combined with GEM synergistically inhibited p-STAT3 protein expression and activated caspase-3 protein expression .Conclusion TPL combined with GEM can synergistically in-hibit proliferation and induce apoptosis of pancreatic cancer PANC-1 cells ,its mechanism is related to the inhibition of STAT 3 sig-naling pathway ,promotion the expression of caspase-3 protein .
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Objective To determine the expression of TM4SF1 mRNA in 5 human pancreatic cancer cell lines,and investigate its effect on the proliferation,migration and invasion of pancreatic cancer cells.Methods The expression of TM4SF1 mRNA in MPanc96,MiaPaCa-2,PANC1,AsPC-1,HPAC cells was determined by qRT-PCR,and the results were compared with that of human pancreatic ductal epithelial (HPDE) cells.RNA interference method was used to transiently transfect siRNA targeting at TM4SF1 and negative control siRNA into MPanc96,MiaPaCa-2 cells.The proliferation of cells were measured by MTS method,and migration and invasion of cells were determined by Transwell.Results The expression levels of TM4SF1 mRNA in pancreatic cancer cell lines MPanc96,MiaPaCa-2,PANC1,AsPC-1 and HPAC were 1.205 ± 0.073,1.096 ± 0.260,1.382 ± 0.075,1.374 ± 0.363 and 0.744 ± 0.096,which were significantly highly than that in HPDE (0.020 ± 0.003,P ＜ 0.01).Compared with cells transfected with negative control siRNA,the proliferation of MPanc96 and MiaPaCa-2 cells transfected with siRNA targeting at TM4SF1 was not significantly changed,but the migration abilitiy was decreased by (62.5 ± 7.6) ％ and (72.8 ± 4.0) ％,and invasion abilitiy was decreased by (69.5 ± 5.7) ％ and (78.6 ± 6.3) ％.Conclusions TM4SF1 is highly expressed in pancreatic cancer cells and appears to promote the migration and invasion abilities of the cancer cells.
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Objective To investigate the effects of microRNA-196a(miR-196a) inhibitory sequences transfection on HOXB8 expression in PANC1 cells.Methods PANC1 cells were divided into control group,miR-196a inhibitory sequences group and siRNA control group.Liposomal transfection method was applied to transfect miR 196a inhibitory sequences and siRNA control into PANC1 cells.RT-PCR and Western blot were used to detect the expressions of miR-196a and HOXB8 mRNA and protein.Results After miR-196a inhibitory sequences transfection,when compared with that of siRNA control group,the expression of miR-196a was significantly decreased (0.05 ± 0.054 vs.0.839 ± 0.025,t =3.12,P ＜0.05) ; and the expression of HOXB8 mRNA was significantly increased by 1.57 folds (2.20 ± 0.07 vs.1.29 ± 0.10,t =3.86,P ＜ 0.05),the expression of HOXB8 protein was also obviously increased (0.90 ± 0.03 vs.0.40 ± 0.10,t =3.11,P ＜ 0.05).Conclusions MicroRNA-196a down-regulates the expression of HOXB8.
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Objective: To analyze the constitution of hospitalization expenses and provide evidence for controlling unreasonable increase of medical costs. Methods: To compare the medical expense constitution of cerebral infarction patients with and without medical insurance; interview the doctors, patients and the medical insurance managers of medical insurance, analyze the influencing factors of the expense through combining the result of interview and quantitative data. Results: The patients with medical insurance spent higher expense than their counterparts, the per capita cost of medical insurance increase by year. Through analyzing the constitution of hospitalization cost, medicine fee and examination cost are the major medical costs. Conclusion: The costs of patients with medical insurance have over-treatment problem, which could be controlled by changing payment method and strengthening management.
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Objective To investigate the effect of Iodine 125 seeds short time low dose rate irradiation on perineural invasion (PNI) in pancreatic cancer Capan-2 cells,and explore its molecular mechanism.Methods The co-culture model was established by co-culturing the dorsal root ganglion (DRG) of SD rat and Capzn-2 cells line,while Capan-2 culture model and DRG culture model was also established.Iodine 125 seeds short time low dose rate irradiation tablet was used for the 3 models,and the model without irradiation was used as control.Cancer cell and DRG growth was observed under inverted microscopy,surface of neurite and cell colony growth was determined by image analysis software.The concentration of nerve growth factor (NGF),transforming growth factor-α (TGF-α) in cell culture supernatant and matrigel solution was tested by ELISA,and the expression of neurotrophin-3 (NT-3) mRNA was detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR).Results In the co-culture model,neurite of DRG showed a direction to cancer cells and had a concentrated growth towards cancer cells.And Capan-2 cells formed more colonies towards neurite.However,in irradiation groups,the symbiotic phenomenon was inhibited to some degree.Increased surface of neurite in co-culture model at 5th day was 290.15 ± 12.08,which was significantly higher than that in DRG group (124.83 ± 6.96,P ＜ 0.01),but the surface of neurite was decreased to 201.53 ± 12.20 after irradiation (P ＜0.01).Increased surface of Capan-2 cell was 300.47 ± 12.99,which was significantly higher than that in Capan-2 group (199.30 ± 8.60,P ＜ 0.01),but the surface of Capan-2 was decreased to 202.35 ± 7.97 after irradiation (P ＜ 0.01).NT-3 mRNA was seldom or not expressed in supernatant of co-culture model,but it was strongly expressed (0.68 ± 0.04) after irradiation (P ＜ 0.05).The concentration of NGF and TGF-α in supernatant of co-culture model were (27.56 ± 13.73),(40.86 ± 20.73) ng/ml,after irradiation they were increased to (94.98 ± 33.80),(157.54 ± 83.76) ng,/ml,and the difference between the two groups was statistically significant (P＜0.05 or ＜0.01).The concentration of NGF and TGF-α in matrigel lysate of co-culture model were (60.42 ± 33.03),(64.39 ± 21.52)ng/ml,after irradiation they were increased to (132.52 ±53.01),(138.38 ±83.58)ng/ml,and the difference of NGF concentration between the two groups was statistically significant (P ＜ 0.05).Conclusions Iodine-125 seeds short-time low-dose rate irradiation could inhibit interactions between nerve and Capan-2 cells,and the mechanism may be related to up-regulation of cancer cells perineural invasion promoter NGF,TGF-α and NT-3.
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Objective To investigate the correlation of K-ras gene mutations and activation of Hedgehog signaling pathway in pancreatic cancer cell lines.Methods Real-time PCR was used to detect K-ras gene mutations at codon 12 and 13 in pancreatic cancer cell lines of SW1990,PaTu8988,CFPAC-1,PANC1,AsPC-1,Capanc-2,BxPC-3 and the mRNA expression of Glil,Smo in these cell lines.Results The K-ras mutation of PANC1,CFPAC-1,AsPC-1,PaTu8988 was at codon 12,while SW1990 was at codon 13,BxPC-3,Capanc-2 were wild type.The expressions of Glil mRNA of AsPC-1,CFPAC-1,PANC1,PaTu8988,SW1990,BxPC-3,Capanc-2 were 7.84 ± 8.92,1.82 ± 3.45,1.00± 0.00,0.07 ± 0.10,0.88 ± 1.48,0.52 ± 0.98,0.15 ± 0.19,and the expressions of Smo mRNA were 144.00 ± 58.33,3.48 ±3.77,1.00 ±0.00,81.68 ±28.26,0.72 ±0.87,0.34 ±0.60,0.02 ±0.03.Glil,Smo mRNA expressions of cells with wild tpye K-ras were significantly lower than those with mutant type,and wild type BxPC-3's Glil,Smo mRNA expressions were significantly lower than that of mutant AxPC-1 (P ＜ 0.05).Conclusions Activation of Hedgehog signaling pathway may be related to K-ras gene mutations.