ObjectiveTo explore the recurrence inhibition of colorectal cancer by adenoviral transducted endostatin gene. Methods The recombinant adenovirus expressing endostatin was constructed. Its biological activities were observed. The levels of endostatin in mice peripheral blood, tumor local recurrence and tumor cell apotosis were analyzed after the endostatin gene transduction by adenovirus. Results The infecting supernatant of recombinant adenovirus significantly inhibited HUVEC proliferation. After the injection of the recombinant adenovirus, persistent high serum levels of endostatin in peripheral blood was observed, local recurrence rate decreased, and apotosis of recurrence tumor cells increased.Conclusions The intraveneously injection of recombinant adenovirus mediated endostatin gene produces high concentration and stable expression of endostatin,which effects prevention of local recurrence after surgical resection of colorectal cancer.

Objective To evaluate the effect of SAHA or/and PTX on survival and apoptosis of human paclitaxel-resist-ant ovarian cancer OC3/P cells, and explore whether the combination of two drugs has a synergistic effect .Methods The morphology of OC3/P cells in different drug-groups was observed by inverted microscope .Cell viability was evaluated by MTT assay.The apoptosis rate was analyzed by Annexin V-FITC/PI assay.Results The morphology change of OC 3/P cells treated with different drug was observed by inverted microscope , and the change in combination group was more signif-icant than one drug alone group .The result of cell survival measured by MTT assay showed that inhibition rate of combina -tion group was more higher than one drug alone group (P<0.05).The analysis of factorial design and gold formula method all proved that the two drugs had synergy .Further the result of flow cytometry showed that apoptosis rate in combination group was significantly higher than SAHA or PTX alone group (P<0.05).Conclusion SAHA and PTX can inhibit the survival and induce apoptosis of OC 3/P cells, and two drugs have synergistic antitumor effects .

Objective To detect the expression of metastasis sappressor 1(MTSS1) gene in cervical cancer tissue and to clarify its association with cervical cancer.Methods Totally 103 cases of cervical tissue were collected between Dec 2011 and Dec 2014 and classified according to biopsy and stage .Q-PCR and Western blotting were used to detect the expression of MTSS1 in normal cervical tissue and in different clinical stages of cervical cancer tissue .Results The expression of MTSS1 inⅡB-Ⅳstages of cervical cancer tissue was significantly higher than that of normal tissue or Ⅰ-ⅡA stages through q-PCR (P=0.000).Western blotting results showed that MTSS1 was positively expressed in normal cervical tissue at a rate of 23.3% or 53.3% in cervical cancer tissue.Moreover, the expression of MTSS1 was poorly correlated with age, tumor differentiation and lymphnode metastasis in cervical cancer tissue (P>0.05).The protein level of MTSS1 expressed in ⅡB-Ⅳ stages was significantly higher than that ofⅠ-ⅡA stages(P=0.005).Conclusion The expression of MTSS1 indicates the clinical stage of cervical cancer , suggesting that MTSS1 may play an important role in the development of cervical cancer .

OBJECTIVE To investigate the effect of overexpression of vascular cell adhesion molecule-1(VCAM-1)on the migration in vitro of the murine mesenchymal stem cells(MSCs)and its possible mechanism. METHODS The migration ability of normal mouse MSC (C3) ,empty vector-transfected MSC(C3+N) and VCAM-1 transfected MSC(C3+VCAM-1)was assessed by Transwell culture system in vitro after incubation for 8 and 12 h,respectively. The fetal bovine serum (FBS) was used as the chemotactic agent to induce MSC migration. The transmigrated cells were detected with methylosaniliam chloride(crystal violet)as well as DAPI staining.Furthermore,the specific chemical inhibitors of mitogen-activation protein kinase (MAPK) pathway ( SB203580,PD98059 and JNK inhibitorⅡ)were added to the Transwell system for 12 h and the alteration of the MSC migration ability was evaluated. RESULTS After incubation with FBS for 8 and 12 h,the absolute migrated cell number(7467 ± 485 and 8795 ± 255)and migration rate〔(14.9 ± 1.0)% and(17.6 ± 0.5)%〕of MSC in C3+VCAM-1 group were significantly increased compared with C3 group〔2731±562 and 4779±224, (5.5 ± 1.1)%and(9.6 ± 0.4)%〕and C3+N group〔2539 ± 321 and 5645 ± 1080,(5.1 ± 0.6)%and(11.3 ± 1.1)%〕(P<0.05,P<0.01),but there was no significant difference between C3 and C3+N groups. Moreover,the MSC migration ability of C3+VCAM-1 group was partially suppressed by addition of JNK inhibitorⅡ. The transmigrated cell number(4843 ± 167)and migration rate〔(9.7 ± 0.3)%〕were decreased compared with those of C3+VCAM-1 group without JNK inhibitorⅡ(P<0.01). SB203580 and PD98059,as specific chemical inhibitors of MAPK pathway,had no effect on MSC migration. CONCLUSION VCAM-1 can enhance mouse MSC migration in vitro and th4e mechanism may be related to JNK/MAPK pathway activation.

OBJECTIVE To study the molecular mechanism of cisplatin(DDP)by which HeLa cell growth and proliferation are inhibited. METHODS Cultured HeLa cells were treated with DDP 0.02-75 μmol · L-1 for 24 or 48 h. CCK-8 assay was used to determine the cell proliferation. The wound scratch assay was used to detect the cell migration and invasion. Flow cytometry was used to detect the cell cycle arresting. q-PCR was used to test the expression of metastasis suppressor gene 1 (MTSS1)mRNA. Western blot was used to determine protein levels of MTSS1,phosphorylated-extra?cellular signal-regulated kinase(p-ERK) and phosphorylated-serine-threonine kinase(p-AKT). RESULTS Following the treatment with DDP for 24 or 48 h,the proliferation of HeLa cells was inhibited significantly (P<0.05),the value of the half inhibitory concentration (IC50) of cells was 4.14 and 11.82 μmol · L-1. Migration and invasion activity of HeLa cells were reduced according to the wound scratch assay(P<0.05). Flow cytometry results showed that the cell cycle was arrested at S phase. q-PCR results showed that MTSS1 mRNA expression changed with DDP in a concentration-dependent manner (r24 h=-0.965,P<0.01;r48 h=-0.953,P<0.01). Western blot showed that the protein levels of MTSS1,p-ERK and p-AKT expression declined significantly with the increase in DDP concentrations(p-ERK:r24 h=-0.875,P<0.01;r48 h=-0.966,P<0.01. p-AKT:r24 h=-0.831,P<0.01;r48 h=-0.863,P<0.01. MTSS1:r24 h=-0.969,P<0.01;r48 h=-0.988,P<0.01). CONCLUSION DDP treatment inhibits HeLa growth and proliferation by interfering with the MTSS1 expression and disturbing the activation of ERK and AKT signaling pathways.

Objective To investigate the effect of resveratrol on miRNA-106b in Alzheimer′s disease ( AD ) animal model.Methods Fifty Kunming male mice were divided into five groups by completely randomized block sampling.The five groups included three dosage resveratrol groups , an AD model group and a control group.The AD models were established in one month prior to treatments. Subsequently, from the 31st day various doses of resveratrol were provided intragastricly for 60 days.Then the memory function was observed by the step-down test.Meanwhile, the varying expressions of APP , P62, ApoA1, miRNA-106b, ABCA1 were tested in each group to determine whether there is the binding site for miRNA-106b in APP 3′UTR sequence.Results Compared with the control group by step-down test, the memory function of the AD model group mice decreased in different degree , which in the drug treatment group was higher than that in the model group (P<0.05).Compared with the AD group, the expression of APP (1.131 ±0.035) in the drug treatment group was higher than that in the model group (0.652 ± 0.026), while the P62 (0.412 ±0.022) and ApoA1 (0.534 ±0.032) were lower than the model group ( all P<0.05 ).High and medium dose groups of resveratrol treatment reduced varying degrees of APP (0.733 ±0.018,0.929 ±0.019,F=177.733) levels, and increased P62(0.954 ±0.035,0.633 ±0.015, F=434.5 ) and ApoA1 ( 1.042 ±0.051, 0.824 ±0.034, F=286.582 ) levels ( all P<0.05 ).The expression of miRNA-106b (0.464 ±0.313) and ABCA1(0.293 ±0.042) in the model group was lower than that in the control group (miRNA-106b 1.064 ±0.032, F=238.159; ABCA1 0.781 ±0.027,F=341.61;both P<0.05).The miRNA-106b (0.843 ±0.034, 0.601 ±0.012) and ABCA1 (0.882 ± 0.025, 0.624 ±0.036) levels in the high, medium dose resveratrol treatment groups increased to different extent ( both P<0.05 ).After the drug treatment , luciferase reporter vector experiments showed that the APP 3′UTR sequence contains the binding site of miRNA-106b.Conclusions APP is one of the target genes of miRNA-106b.Resveratrol is capable of improving AD by enhancing the expression of miRNA-106b and down-regulating the target genes including APP , P62 and ApoA1.This provides a new theoretical basis for the clinical treatment of AD.

Objective To evaluate the effect of suberoylanilide hydroxamic acid(SAHA) or/and paclitaxel(PTX) on lethality and autophagy of human ovarian cancer OC3 cells,and to explore whether the combination of the two drugs has a synergistic function.Methods The morphology of OC3 cells was treated with SAHA and/or PTX, and then the morphology of treated OC3 cells was observed under an inverted microscope, cell proliferation was detected by MTT assay and autoph-agy was analyzed by AO/EB double staining assay.The synergistic effect of SAHA and/or PTX was analyzed by factorial design and gold formula method.Results After treatment with SAHA and/or PTX, the morphology of OC3 cells in the combination group ( SAHA+PTX) displayed significant morphological changes.OC3 cells became less adherent and refrac-tive than in other groups.Cell proliferation by MTT assay demonstrated that the growth inhibition rate of the combination groups was higher than in groups treated with SAHA or PTX respectively( P<0.05) .Furthermore, the synergistic effect af-ter treatment with a combination of SAHA with PTX was proved by the factorial design and gold formula method.The auto-phagy rate of the combined groups was significantly higher than in single treatment groups (P<0.05) by AO/EB double staining.Conclusion SAHA and PTX can inhibit the survival of OC3 cells and induce its autophagy.The two drugs have synergistic antitumor effects.

OBJECTIVETo study the prevention of colorectal cancer liver metastasis by adenoviral transduction of the endostatin gene.

METHODSThe recombinant adenovirus expressing endostatin was constructed. Its biological activities were surveyed in vitro, as determined in human umbilicus vein endothelium cell (HUVEC) proliferation inhibition, and in vivo, by reduction of liver metastasis.

RESULTSHUVEC proliferation was obviously inhibited by the infecting supernatant of recombinant adenovirus. Persistent high serum levels of endostatin in peripheral blood, especially in the liver vein were observed. The production of liver metastasis was intervened.

CONCLUSIONSThe single injection in the vein of the recombinant adenovirus realizes the high effective and stable expression of endostatin in general body and liver, which brings about the ideal prevention of liver metastasis.

Adenoviridae ; genetics ; Animals ; Cell Division ; drug effects ; Collagen ; genetics ; therapeutic use ; Colorectal Neoplasms ; pathology ; Disease Models, Animal ; Endostatins ; Endothelium, Vascular ; drug effects ; pathology ; Genetic Therapy ; Genetic Vectors ; Liver Neoplasms ; prevention & control ; secondary ; Mice ; Mice, Inbred BALB C ; Neoplasm Metastasis ; prevention & control ; Neoplasm Transplantation ; Peptide Fragments ; genetics ; therapeutic use ; Transduction, Genetic

Objective To explore the value of tissue Doppler imaging(TDI)corrected diaphragm ultrasonic parameters for evaluating diaphragmatic function in dyspnea patients who underwent non-invasive mechanical ventilation.Methods Thirty-one acute dyspnea patients who underwent non-invasive mechanical ventilation less than 1 h(non-invasive ventilation group)and 31 healthy subjects(control group)were prospectively enrolled,and ultrasound of diaphragm was performed.Routine diaphragmatic parameters,including diaphragm displacement(DD),diaphragm thickness of end-expiratory(DTee),diaphragm thickness of end-inspiratory(DTei)and diaphragm thickening fraction(DTF)were measured and calculated,while peak systolic velocity of diaphragm(DPSV)and peak diastolic velocity of diaphragm(DPDV)were measured using TDI mode.Based on DD and DTF after rapid shallow breathing index(RSBI),DD-RSBI and DTF-RSBI were corrected,DPSV-RSBI and DPDV-RSBI were obtained by calculating product of DPSV and DPDV with respiratory rate,respectively.The index of compensatory work of diaphragm during systole(DD/DPSV and DTF/DPSV)and diastole(DD/DPDV and DTF/DPDV)were obtained by corrected DD and DTF with DPSV and DPDV,respectively.The ultrasonic parameters of diaphragm were compared between groups.Receiver operating characteristic(ROC)curve was drawn,and the area under the curve(AUC)was calculated to evaluate the efficacy of ultrasonic parameters of diaphragm for assessing diaphragmatic function in dyspnea patients who underwent non-invasive mechanical ventilation.Results DPSV,DPDV,DD-RSBI,DTF-RSBI,DPSV-RSBI and DPDV-RSBI in non-invasive ventilation group were all higher,while DD/DPSV,DD/DPDV,DTF/DPSV and DTF/DPDV were all lower than those in control group(all P＜0.05).No significant difference of DD,DTee,DTei nor DTF was found between groups(all P＞0.05).DPSV-RSBI and DPDV-RSBI had excellent efficacy for assessing diaphragmatic function in dyspnea patients after non-invasive mechanical ventilation(AUC=0.974,0.919),DPSV,DD-RSBI,DTF-RSBI,DD/DPSV and DTF/DPSV had good efficacy(AUC 0.760-0.881),while DD,DPDV,DTee,DTei,DTF,DD/DPDV and DTF/DPDV had bad or general efficacy(AUC 0.467-0.698).Conclusion TDI corrected diaphragm ultrasonic parameters could effectively evaluate diaphragmatic function in dyspnea patients who underwent non-invasive mechanical ventilation.

Objective:To analyze the cardio-pulmonary ultrasound features of cardiogenic pulmonary edema (CPE) and pneumonia in adults with acute dyspnea, and to construct a differential diagnosis model.Methods:Seven hundred and forty-three patients with sudden acute dyspnea admitted to Hebei General Hospital from November 2018 to May 2022 were retropectively included. Ultrasonographer A performed lung ultrasound with 12 zone method, and interpreted and recorded the ultrasonic signs (including A-lines area, B-lines area, consolidation area and pleural effusion area) together with ultrasonographer B. According to the ultrasonic characteristics of the whole lung, it was divided into A-profile and B-profile. According to the continuity and symmetry of the distribution of B-lines in bilateral lung fields, it could be divided into bilateral lung continuous and discontinuous B-profile, bilateral lung symmetric and asymmetric B-profile. Left ventricular ejection fraction (LVEF), left ventricular filling pressure (E/e′), right ventricular dilatation, tricuspid annular systolic displacement (TAPSE) and inferior vena cava diameter (IVCD) were evaluated by echocardiography, and all the indexes were transformed into binary variables. According to the final clinical diagnosis and treatment results, the disease was divided into CPE group and pneumonia group. Binary Logistic regression model was used to screen independent influencing factors, and partial regression coefficient β value was used as a weight to assign a score, and a differential diagnosis model was established based on the total score. The predictive value of the model was evaluated by the receiver operating characteristic curve (ROC) and area under curve (AUC). After the model was built, 30 patients with CPE or pneumonia were independently collected by ultrasonographer C as external validation data, which were included in the model to draw ROC curve and evaluate the differential diagnosis efficiency of the model. The consistencies between ultrasonographer A and B, A and C in observing lung ultrasound were explored.Results:A total of 743 patients from 43 clinical departments were included, including 246 cases in CPE group and 497 cases in pneumonia group. Multivariate logistic regression analysis showed that bilateral lung continuous B-profile, bilateral lung symmetric B-profile, ≥1 pleural effusion area, LVEF<50%, E/e′>14 were the risk factors for CPE (all OR>1, P<0.05), and ≥1 consolidation area and ≥1 pleural sliding disappearance area were the protective factors for CPE (all OR>1, P<0.05). The sensitivity, specificity and AUC of combined cardio-pulmonary ultrasound index β value weight score in the differential diagnosis of CPE and pneumonia were 0.939, 0.956 and 0.986, respectively. The AUC of external validation data was 0.904. Ultrasonographer A and B, A and C had good consistency in the interpretation of lung ultrasound signs ( P<0.05). Conclusions:The differential diagnosis model based on combined cardio-pulmonary ultrasound indexes has high differential diagnosis efficiency for CPE and pneumonia, and can be used in bedside cardio-pulmonary ultrasound practice.