To construct the eukaryotic vector that expresses the fusion protein of Axud1 and influenza virus hemagglutin HA epitope tag, the total RNA was isolated from the peripheral blood lymphocytes, and reverse transcription reaction was used to amplify the full length of human Axud1 cDNA. PCR product of Axud1 was then amplified using specific primers containing HA epitope sequence, and inserted into eukaryotic expression plasmid pcDNA3.1(+)digested with BamH Ⅰand Xba Ⅰ. The recombinant plasmid was identified by polymerase chain reaction, restriction endonuclease mapping and sequencing, and then transfected into human lung adenocarcinoma SPC-A1 cell lines.The fusion HA-Axud1 protein expression in anti-G418 clones was verified by Western blot. This study might be instrumental in further study of the function of Axud1 protein in tumor cells.

Objective: To determine the potential diagnostic value of miRNA-29 (miR-29) for malignant tumor. Methods: A systematic search of literature regarding miR-29 was performed in three English databases (PubMed, Web of Science, and Embase) and two Chinese databases (Chinese National Knowledge Infrastructure [CNKI] and WanFang). The retrieval was ended until September 15, 2018. Search terms included miRNA-29 (miR-29), tumor, cancer, serum, plasma, diagnosis, etc. Quality Assessment of Diagnostic Accuracy Studies-2 (QUADAS-2) was carried out to evaluate the quality of the selected articles. STATA12.0 was used to calculate the combined sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR) and diagnostic odds ratio (DOR). Subgroup analysis and Meta-regression analysis were carried out to explore the origin of heterogeneity. Results: Twenty eligible articles were selected from 1 172 literatures related to tumors and miR-29. The combined sensitivity was 0.76 (95%CI: 0.68-0.83), combined specificity was 0.83 (95%CI: 0.74-0.89), combined PLR was 4.5 (95%CI: 2.7-7.4), combined NLR was 0.28 (95%CI: 0.20-0.41), DOR was 16 (95%CI: 7-35), and theAUC was 0.86 (95%CI: 0.83-0.89). The combined specificity of plasma samples was higher than that of serum samples, and the difference was statistically significant (P<0.01). There was a higher diagnostic value of miR-29 for breast cancer and pancreatic cancer (DOR=101.52, 11.22), but lower diagnostic value for colorectal cancer and non-small cell lung cancer (DOR=5.05, 6.57); miR-29b showed a high diagnostic value for cancer (DOR=60.91). The publication bias was not obvious in this study (P>0.05). Conclusion: This systematic review and Meta-analysis suggests that miR-29 family is a potential biomarker in the diagnosis of cancers with great sensitivity and specificity.

Objective To investigate the expression profiles of microRNA in early esophageal squamous cancer and normal esophageal tissues and verify the significantly different expression miRNAs , further to study the effects on proliferation of EC109. Methods The microarray assay was performed to analyze miRNA expression profiles in three pairs of early esophageal squamous cancer and the corresponding normal esophageal tissues. Quantitative real-time polymerase chain reaction (qRT-PCR) was used in another 38 pairs samples to further verify the differentially expressed miRNAs . Three verified miRNAs ( miR-29a, miR-221 and miR-222) mimics were transfected into EC109 respectively and CCK8 method was used to study the effect of cell proliferation in each miRNA. Results Microarray technique selected 53 miRNAs that differentially expressed in early esophageal squamous cancer and normal esophageal tissues , 32 miRNAs were up-regulated and 21 miRNAs were down-regulated. qRT-PCR verified that miR-29a was significantly down-regulated (P < 0.05) and miR-221, miR-222 were significantly up-regulated (P < 0.05) in early esophageal squamous cancer tissue. Over-expression of miR-29a could significantly inhibit the proliferation of EC109 (P < 0.05) whereas over-expression of miR-221 or miR-222 could both significantly promote the proliferation of EC109 (P < 0.05). Conclusion There was significant difference of miRNAs expression between early esophageal squamous cancer and normal esophageal tissues, and the differentially expressed miRNAs could be used as new biomarkers for early diagnosis of esophageal squamous cancer.

Objective To investigate the safety and feasibility of modified hand-assisted laparoscopic surgery (MHALS) in the abdominal surgery.Methods The clinical data of 8 patients who underwent long-sleeved MHALS at the First People's Hospital of Foshan between September 2014 and January 2015 were retrospectively analyzed.Among the 8 patients,right liver cancer with intrahepatic metastasis was found in 2 patients,left liver cancer in 1 patient,hepatic peripheral nerve sheath tumor in 1 patient,left retroperitoneal leiomyosarcoma in 1 patient,extra-and intra-hepatic cholangiolithiasis in 1 patient,choledochocyst in 1 patient and ampulla cancer in 1 patient.Laparoscopic protection sleeve went through the middle of incision-retractor,and then wrapped around it about 10 cm.Incision-retractor was fixed at the abdominal incision firstly,laparoscopic procedures were performed when the wrist of assisted hand was bound and fixed by the distal of sleeve.The patients were followed up by outpatient examination and telephone interview till March 2015.Results All the 8 patients underwent successful MHALS,including 1 of right hemihepatectomy in situ,1 of ligation of right portal vein + left liver split (the patient gave up two-stage operation due to intractable ascites and elevated bilirubin),1 of hepatic left lateral lobectomy (the patient underwent hemostatic sutures in open surgery due to hemorrhage of liver's cutting surface),1 of hepatic peripheral nerve sheath tumor resection,1 of left retroperitoneal leiomyosarcoma resection,1 of choledocholithotomy + left hepatectomy + cholecystectomy + T tube drainage,1 of choledochocyst + biliary enteric drainage and 1 of pancreaticoduodenectomy.Four patients had assisted incision of 4 cm,and another 4 patients of 7 cm.Eight patients were followed up for a median time of 3 months (range,2-7 months).The patient who received ligation of right portal vein + left liver split died at postoperative month 3,and the others didn't have recurrence of tumor or lithiasis.Conclusion The MHALS is safe and feasible in the abdominal surgery.

Objective To evaluate the diagnostic efficacy of magnetic resonance(MR)-guided prostate biopsy based on abnormal diffusion weighted imaging for prostate cancer in elder men.Methods From July 2014 to Dec 2016,56 patients (age≥ 65 years) with diffusion weighted imaging (DWI)abnormalities undergoing MR-guided prostate biopsy were retrospectively evaluated.According to pathological diagnosis,patients were divided into prostate cancer and non-prostate cancer groups.The clinical data of two groups were analyzed.The correlation of Gleason score,prostate specific antigen(PSA),and the location and number of biopsy were compared.Results Among 56 cases,32 (57.1％)were diagnosed as prostate cancer,and 24 (42.9％)as chronic prostatitis and prostatic hyperplasia (non-prostate cancer).There were statistically significant differences between prostate cancer and non-prostate cancer in age[(73.0±5.7)vs.(70.1±4.3)year]and PSA[(9.3±6.0)μg/L vs.(6.0± 3.9)μg/L] (both P＜0.05),and no statistical differences in biopsy location and biopsy number(t =2.08,2.37,P＞ 0.05).Gleason score had no correlation with PSA level and biopsy location and biopsy number(r=0.189、-0.183、0.082,P＞0.05).Conclusions MR-guided prostate biopsy based on MR-DWI has some merits,such as accurate positioning,fewer numbers of biopsy,and lower false-negative rate.

Objective The metabolism and excretion of isochlorogenic acid B in rats were investigated by UHPLC-MS.Methods Feces,urine and plasma were individually collected before and at different time points after administration of 20 mg·kg-1.Post-prepared samples were analyzed by UHPLC-MS.Results According to the retention times,m/z,characteristic fragment ions and related literature,a total of 22 metabolites were detected,of which 18 metabolites were present in rat feces,3 metabolites in urine and one metabolite in plasma.The main metabolic pathways contain hydrolysis,hydrogenation,methylation,sulfation and so on.The cumulative excretion of isochlorogenic acid B and its main metabolite chlorogenic acid in feces and urine was further performed.Conclusion The metabolism and excretion of isochlorogenic acid B in rats were explored to provide experimental basis for its further research and development.

Objective:To evaluate the efficacy of ultrasound-guided adductor canal approach to nerve block using one-puncture four-point technique for analgesia after total knee arthroplasty (TKA).Methods:Sixty patients of both sexes, aged 65-80 yr, with body mass index of 18-27 kg/m 2, of American Society of Anesthesiologists physical status Ⅱor Ⅲ, scheduled for elective unilateral TKA under general anesthesia, were enrolled in this study.Peripheral nerve block was performed for postoperative analgesia under ultrasound guidance.All the patients were allocated into 2 groups ( n=30 each) using a random number table method: adductor canal block combined with posterior approach to sciatic nerve block group (group ASB) and adductor canal approach to nerve block using one-puncture four-point technique including sciatic nerve, medial vastus muscle nerve, saphenous nerve, anterior femoral cutaneous nerve at adductor canal level group (group ANSB). When visual analogue scale score ≥4 during postoperative movement, ketorolac tromethamine 0.6 mg/kg was intravenously injected as rescue analgesic. The muscle strength of quadriceps on the affected side, range of knee joint motion and rate of the ability to walk for 10 m using the walker were recorded at 2, 8, 24 and 48 h after surgery.The consumption of analgesics for rescue analgesia, manipulation completion time, length of hospital stay and patients′ satisfaction score were recorded within 48 h after surgery. Results:Compared with group ASB, the consumption of ketorolac tromethamine was significantly reduced, manipulation completion time and length of hospital stay were shortened, patients′ satisfaction scores were increased, and the range of knee joint motion and rate of the ability to walk for 10 m using the walker were increased at 2, 8, 24 and 48 h after surgery in group ANSB ( P<0.05). There was no significant difference in muscle strength of quadriceps between two groups ( P>0.05). Conclusion:Ultrasound-guided adductor canal approach to nerve block using one-puncture four-point technique provides better efficacy than adductor canal block combined with posterior approach to sciatic nerve block when used for analgesia after TKA.

OBJECTIVETo investigate the effects of different anesthesia depth on stress response in elderly patients undergoing elective laparoscopic surgery for colorectal cancer.

METHODSA total of 105 ASA I-III patients aged 60-91 years undergoing elective laparoscopic surgery for colorectal cancer with general anesthesia were randomized into 3 groups, namely group A with a target Narcotrend index (NI) maintained at D0 level, group B with a NI at D2 level, and group C with a NI at E1 level. The anesthetics (profopol and remifentanil) were adjusted according to Narcotrend monitoring results to maintain the specified anesthesia depth. The patients' heart rate (HR) and mean artery pressure (MAP) were recorded before anesthesia (T0), before intubation (T1), immediately after intubation (T2), at 2 min before pneumoperitoneum (T3), 2 min after pneumoperitoneum (T4), at the end of the surgery (T5) and extubation (T6). Serum levels of cortisol, adrenocorticotropic hormone (ACTH), endothelin-1 (ET-1), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and C-reactive protein (CRP) were measured by standard ELISA and radioimmunoassay before anesthesia (Ta), at the end of the surgery (Tb) and 1 day after the surgery (Tc).

RESULTSHR and MAP in group A increased significantly at T2, T4, and T6 compared to those at T0 (P<0.05), and were higher than those in group B and group C (P<0.05). The MAP in all the 3 groups all decreased at T1 and T3 (P<0.05 or P<0.01), and was markedly lower in group C than in groups A and B (P<0.05). The incidence of hypertension was significantly higher in group A than in groups B and C (P<0.05), while the incidence of hypotension was much higher in group C (P<0.01). There were no obvious differences in serum levels of cortisol, ACTH, CRP, IL-6, TNF-a, or ET-1 among the groups at Ta (P>0.05). The serum levels of ACTH in the 3 groups all significantly increased at Tb and Tc (P<0.01). CRP, IL-6 and TNF-a levels in group A were increased at Tb and Tc (P<0.05 or P<0.01) and significantly higher than those in groups B and C (P<0.05 or P<0.01). Cortisol in groups A and B increased at Tb and Tc (P<0.05) to a significantly higher level than that in group C (P<0.01). ET-1 level in group C at Tb and Tc was lower than those in groups A and B (P<0.05 or P<0.01).

CONCLUSIONSMaintaining the anesthesia depth for a NI at the D2 and E1 level can both attenuate the stress response in elderly patients undergoing laparoscopic surgery for colorectal cancer, but the hemodynamic stability can be better at a D2 level.

Adrenocorticotropic Hormone ; blood ; Aged ; Aged, 80 and over ; Anesthesia, General ; methods ; Blood Pressure ; C-Reactive Protein ; metabolism ; Colorectal Neoplasms ; surgery ; Elective Surgical Procedures ; Endothelin-1 ; blood ; Heart Rate ; Humans ; Hydrocortisone ; blood ; Interleukin-6 ; blood ; Laparoscopy ; Middle Aged ; Piperidines ; administration & dosage ; Propofol ; administration & dosage ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo investigate AXIN1-related CSRNP1 gene expression and the mechanism of its transcriptional regulation in TGF-β1-induced tumor cells.

METHODSHuman lung carcinoma A549 cells or human prostate cancer PC3 cells were treated with TGF-β1 at different doses (0, 20, 40, and 80 ng/ml) or at 20 ng/ml for 0, 8, 12, or 24 h, and the dose and time effect of TGF-β1 on CSRNP1 mRNA expression in the tumor cells were evaluated with real-time RT-PCR. A549 cells were also treated with TGF-β1 and cycloheximide to clarify whether CSRNP1 expression induced by TGF-β1 required de novo protein synthesis. A549 cells transfected with pcDNA3.1, flag-SMAD3, or flag-SMAD3-mu, after serum starvation, were treated with or without TGF-β1 (20 ng/mL) for 24 h, and the overexpression of wild-type SMAD3 and dominant negative SMAD3-mu mutant were confirmed by Western blotting. The effect of SMAD3 or SMAD3-mu overexpression on CSRNP1 mRNA expression was also measured by real-time RT-PCR.

RESULTSIn both A549 and PC3 cells, TGF-β1 dose- and time-dependently stimulated CSRNP1 expression, which required de novo protein synthesis in A549 cells. Overexpression of wild-type SMAD3 significantly increased the expression of CSRNP1 mRNA induced by TGF-β1, while overexpression of dominant negative SMAD3 mutant remarkably reduced CSRNP1 mRNA expression in response to TGF-β1 in A549 cells.

CONCLUSIONTGF-β1 may contribute to CSRNP1 expression through SMAD3 activation and downstream signaling in tumor cells.

Apoptosis Regulatory Proteins ; genetics ; metabolism ; Axin Protein ; genetics ; metabolism ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Humans ; RNA, Messenger ; genetics ; Signal Transduction ; Smad3 Protein ; genetics ; metabolism ; Transfection ; Transforming Growth Factor beta1 ; pharmacology

OBJECTIVETo investigate the dynamic changes in angiogenesis within the tumor tissue of mice bearing S180 tumor at different day-points of oral administration with a Chinese medicine compound "Yiliuyin" (YLY) and to explore the anti-tumor mechanisms of YLY in vivo.

METHODFifty-six BALB/c mice were divided into YLY group and control group (28 mice/group) and each group was divided into four subgroups (7 mice/subgroup), randomly. After 24 hrs of inoculation with S180 tumor cells subcutaneously in the right axilla, YLY in the mice of YLY group and equal volume of cold boiled-water in the mice of control group were administered orally twice every day, 0.5 mL each time. The mice of one subgroup from the two groups apiece were killed at 10, 20, 30 th and 40 th day-point of oral administration, respectively. The tumors were isolated and were made into paraffin embedded sections. The dynamic changes of the angiogenesis (CD34 staining), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor-2 (VEGFR-2) and endostatin (ES) in tumor tissue were detected by immunohistochemistry staining, and the results were shown as PED (positive enzyme dot).

RESULTYLY could remarkably decrease the angiogenesis within tumor tissues. The PED of CD34 in control group at 10, 20, 30 th and 40 th day-point was 392.86+/-42.01, 481.49+/-58.34, 386.31+/-54.91 and 376.69+/-28.71, and that in YLY group was 334.46+/-33.38, 289.34+/-39.63, 257.09+/-40.00 and 246.57+/-36.78, respectively. The PED of CD34 in YLY group at each day-point was lower than that in control group (P<0.05, P<0.01, P<0.01 and P<0.01, respectively). The PED of VEGF in control group at 10, 20, 30 th and 40 th day-point was 852.63+/-81.65, 1168.40+/-96.69, 1292.60+/-147.54 and 1124.74+/-139.64, and that inYLY group was 718.40+/-94.94, 866.54+/-72.40, 859.31+/-74.02 and 753.34+/-72.95, respectively. The PED of VEGF in YLY group at each day-point was lower than that in control group (P <0.05, P <0.01, P <0.01 and P <0.01, respectively). The PED of VEGFR-2 in control group at 10th, 20th, 30th and 40th day-point was 618.63+/-59.08, 750.09+/-56.72, 684.91+/-72.86 and 644.06+/-60.25, and that in YLY group was 523.91+/-64.66, 449.03+/-46.85, 400.06+/-60.12 and 339.89+/-45.39, respectively. The PED of VEGFR-2 in YLY group at each day-point was lower than that in control group (P <0.05, P <0.01, P <0.01 and P <0.01, respectively). The PED of ES in control group at 10th, 20th, 30th and 40th day-point was 250.26+/-36.27, 298.60+/-44.41, 450.86+/-38.95 and 398.43+/-34.19, and that in YLY group was 249.57+/-40.23, 350.03+/-40.92, 499.40+/-40.29 and 497.94+/-42.76, respectively. There was no difference between the two groups at 10th day-point.The PED of ES in YLY group was higher than that in control group at 20, 30, 40 th day-point (P <0.05, P <0.01 and P <0.01, respectively) .

CONCLUSIONYLY could exert the anti- tumor role by down-regulating the expression of VEGF and VEGFR-2, up-regulating the expression of ES and inhibiting the angiogenesis within tumor tissue.

Administration, Oral ; Animals ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Endostatins ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Neoplasms ; blood supply ; drug therapy ; genetics ; pathology ; Neovascularization, Pathologic ; drug therapy ; pathology ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism