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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.
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Objective:To compare the effectiveness and safety of laparoscopic hepatectomy (LH) versus radiofrequency ablation (RFA) in treatment of hepatocellular carcinoma (HCC).Methods:The medical literatures on LH and RFA for HCC were searched in PubMed, Web of Science, Embase, VIP, Wanfang, CNKI and other electronic databases. The retrieval date was from database construction to June 7, 2021. According to the inclusion and exclusion criteria, studies were extracted by two authors, and Revman 5.3 software was used to conduct a meta-analysis to compare differences in operation time, blood loss, length of hospital stay, total complications, overall survival and disease-free survival outcomes between the LH group and the RFA group.Results:Of 3 690 patients who were included in 32 studies, there were 1 708 patients in the LH group and 1982 patients in the RFA group. Meta-analysis showed that compared with the LH group, the RFA group had significantly shorter surgical duration ( MD=-86.41, 95% CI: -116.21--56.60), less blood loss ( MD=-213.22, 95% CI: -273.43--153.00), shorter hospital stay ( MD=-3.23, 95% CI: -4.13--2.32), and lower incidence of complications ( OR=0.33, 95% CI: 0.26-0.43). However, local recurrence rate was significantly higher ( OR=1.83, 95% CI: 1.38-2.41). (All P<0.05). The 5-year survival rate of the LH group was significantly better than the RFA group ( OR=0.68, 95% CI: 0.51-0.90, P=0.008). Conclusion:LH provided better overall survival outcomes and lower local recurrence rates than RFA in HCC patients.
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Objective To explore the curative effects of mesenchymal stem cells(MSC)that overexpress in murine type 1 diabetes nephropathy (DN).Methods Mice were randomly divided into normal control(NC) group,DN group,C3-treated group,C3-MIGR1-treated group and C3-MIGR1-ICAM-1-treated group.Mice were given streptozotocin until the DN model was set up.The murine DN model was treated with murine MSC(C3H10T1/2),transfection empty vector of murine MSCs(C3H10T1/2-MIGR1/MSC) and murine MSCs (C3H10T1/2-ICAM-1/MSC)that overexpressed ICAM-1.After transplantation, the pathological features of kidneys were observed by Masson staining and the number of homing MSC cells to the kidney was calculated on days 1,3,7 by frozen section, while qPCR was used to analyze the expression of signaling molecules for collagen1, TGF-β1 and SMAD2 after treatment with various MSCs.Results Compared with DN group, the renal fibrosis treated with MSCs overexpressing ICAM-1 was significantly decreased by Masson staining.Three and seven days after transplant, the homing cells of MSC in different groups displayed no difference using tissue freezing section method.Furthermore, TGF-β1/SMAD signaling was lowly activated after the treatment with MSCs that overexpressed ICAM-1 compared with model mice(P<0.01).Conclusion MSCs that overexpress ICAM-1 can protect kidneys in the DN model.
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Objective To explore the effect of telomerase reverse transcriptase(TERT) gene transfected bone marrow stem cell(BMSC)on the memory function and hippocampal CA1 region synaptic plasticity in vascular dementia rat.Methods A total of 60 rats were randomly divided into the negative control group(group A),model group(group B),conventional BMSC group(group C) and transfected BMSC group(group D).The related indicators in each group were detected by using the Morris maze test,RTPCR and Western blot respectively.Results The escape latency period in the group C and group D was significantly longer than that in the group B,which in the group D was significantly longer than that in the group C.Compared with the group A,the expressions of brain-derived neurotrophic factor(BDNF)mRNA,TERT mRNA,SYP mRNA and protein in the group B,group C and group D were significantly decreased.The synaptic cleft arrange in group A was clear with more SYN positive ceils.The synaptic cleft in the group D was clearer,and the number of SYN positive cells was close to that in group A.Conclusion TERT transfected BMSC has obvious therapeutic effect on vascular dementia rats and its mechanism may be related to the promotion of BDNF,TrkB expression and the improvement of synaptic plasticity.
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Objective To investigate the effects of cyclopamine (CYP) on endometrial carcinoma (HEC-1A) cell survival and on induction of cell apoptosis .Methods HEC-1A cells were treated with various doses of CYP (0, 5,10, 20 and 40 μmol/L) for 24 h respectively .Then,the inverted microscope was used to observe cell morphology .Cell proliferation and apoptosis were tested by CCK-8 assay and AO/EB bi-labelling assay.The apoptosis rate of HEC-1A was analyzed using flow cytometric analysis , and the key gene expression of Bax and Bcl-2 was detected by quantitative PCR .Results The HEC-1A cells exhibited dramatic morphological changes after treatment with CYP and in a dose-dependent manner .CYP significantly inhibited HEC-1A cell proliferation using CCK8 assays(P<0.05), and induced cell death by AO/EB bi-labelling assay.Moreover,flow cytometry analysis showed that CYP treatment resulted in HEC-1A cell apoptosis, and that a higher concentration of CYP induced severer cell apoptosis (P<0.05).Meanwhile, CYP treated HEC-1A cells exhibited up-regulated expression of Bax and down-regulated expression of Bcl-2 according to Q-PCR.Conclusion Our findings indicatee that CYP can inhibit HEC-1A cell proliferation and induce cell apoptosis .
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OBJECTIVE:To investigate therapeutic efficacy and safety of leosimendan in the treatment of acute left heart failure (ALHF) and its effects on left ventricular function indexes and serum markers.METHODS:A total of 110 patients with acute left ventricular function selected as research objects from No.251 Hospital of PLA during Jan.2014-Dec.2015,and then were divided into control group (53 cases) and observation group (57 cases) according to random number table.Both groups received routine therapy.Control group was additionally given Dopamine hydrochloride injection or Dobutamine hydrochloride injection as cardiotonic on the basis of routine therapy.Observation group was additionally given Levosimendan injection with initial dose of 10 μg/kg+0.9％ Sodium chloride injection 50 mL,ivgtt (10 min),and then with micro pump infusion at the rate of 0.1 g/(kg· min) for continuous 24 h.Both groups were treated for continuous 7 d.Clinical efficacies of 2 groups were observed,and the levels of left ventricular function indexes (PER,PFR,LVEF,LVSF) level,serum marker (NT-proBNP) and galectin-3 (Gal-3) before and after treatment,the occurrence of ADR was recorded.RESULTS:Two cases were withdrawn from the study due to death (one case in each group).Finally,a total of 108 cases were included,involving 52 cases in control group and 56 cases in observation group.Clinical total response rate of observation group (94.64％) was higher than that of control group (86.54％),but without statistical significance (P＞0.05).Before treatment,there was no statistical significance in left ventricular function indexes or serum markers levels between 2 groups (P＞ 0.05).After treatment,the levels of left ventricular function indexes were improved significantly in 2 groups,and LVEF and LVSF of observation group were significantly higher than those of control group,with statistical significance (P＜0.05).NT-proBNP and Gal-3 of 2 groups were decreased significantly,and the observation group was significantly lower than the control group,with statistical significance (P＜0.05).No obvious ADR was found in 2 groups during treatment.CONCLUSIONS:Leosimendan in the treatment of ALHF have the similor clinical efficacy with dopamine,but helps to strengthen the left heart pump function,reduce heart failure markers levels with good safety.
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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.
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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 .
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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.
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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.
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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 .
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<p><b>OBJECTIVE</b>To study the prevention of colorectal cancer liver metastasis by adenoviral transduction of the endostatin gene.</p><p><b>METHODS</b>The 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.</p><p><b>RESULTS</b>HUVEC 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.</p><p><b>CONCLUSIONS</b>The 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.</p>
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Animals , Mice , Adenoviridae , Genetics , Cell Division , Collagen , Genetics , Therapeutic Uses , Colorectal Neoplasms , Pathology , Disease Models, Animal , Endostatins , Endothelium, Vascular , Pathology , Genetic Therapy , Genetic Vectors , Liver Neoplasms , Mice, Inbred BALB C , Neoplasm Metastasis , Neoplasm Transplantation , Peptide Fragments , Genetics , Therapeutic Uses , Transduction, GeneticABSTRACT
<p><b>OBJECTIVE</b>To construct a retroviral-mediated vector of FLT3 ligand (FL) and express it in human bone marrow stromal cells.</p><p><b>METHODS</b>FL cDNA was inserted into the retroviral vector pLXIN by gene recombination technology. The recombinant plasmid pLFIN was transferred into retrovirus packaging cell line PA317 by lipofectamine, and the positive clones were selected by G418. The mRNA expression in human stromal cells and integration of genome DNA were assayed by reverse transcriptase-polymerase chain reaction (RT-PCR) and genomic DNA-PCR. The expression of FL protein and its biological activities in the culture were investigated by ELISA and mouse bone marrow CFU-GM assay.</p><p><b>RESULTS</b>The recombinant plasmid pLFIN was successfully constructed. In genome of these transfected target cells, Neo gene and FL gene were integrated, FL mRNA was transcripted and FL protein was expressed at 4.35 ng/ml/24 h. The specific activities of FL in the culture indicated that human bone marrow stromal cells transfected with FL could significantly express FL in vitro.</p><p><b>CONCLUSION</b>The retroviral-mediated FL gene was expressed in bone marrow stromal cells and the biological activities of FL were detectable in the supernatant of the transfected cells. These results provide a basis for studies on hematopoietic regulation by gene transfected bone marrow stromal cells.</p>
Subject(s)
Animals , Humans , Mice , 3T3 Cells , Bone Marrow Cells , Cell Biology , Metabolism , Cell Line , Colony-Forming Units Assay , Enzyme-Linked Immunosorbent Assay , Gene Expression , Genetic Vectors , Genetics , Membrane Proteins , Genetics , Metabolism , RNA, Messenger , Genetics , Metabolism , Retroviridae , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Stromal Cells , Cell Biology , Metabolism , TransfectionABSTRACT
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.
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Transfer of drug resistance genes into hematopoietic cells is an attractive approach to protect hematopoietic system from the toxic effects by chemotherapeutic agents in cancer patients. In this study, transduction of mdr-1 in combination with dihydrofolate reductase (dhfr) gene was performed, and the expression of exogenous genes and chemoprotection capacity in mouse bone marrow cells were observed. The results showed that approximately 15% of bone marrow cells transfected with the retroviral vector expressed mdr-1 as assayed by flow cytometry. Gene transfer resulted in about 0.9 - 13 fold and 0.5 - 2.6 fold increase in the resistance of CFU-GM to taxol and methotrexate in vitro, respectively (P < 0.05). Moreover, seven months after transplantation to syngeneic mice with mdr-1 and dhfr-transfected bone marrow cells, peripheral blood cells in recipients were still positive for gp170 as evaluated by FACS as well as for mdr-1 and dhfr by PCR amplification. These results indicate that hematopoietic progenitors can be transfected by retrovirus containing mdr-1 and dhfr genes, and that functional drug resistance accompanies their expressions. Furthermore, genetic chimerism might exist in hematopoietic stem cells. In conclusion, transfer and expression of mdr-1 and dhfr genes in bone marrow cells might be applicable in gene therapy research in cancer patients.
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The generation of large quantities of novel human T cell clones ex vivo would make a wide range of gene-and immuno-therapies for tumor and AIDS possibly. Although it is well established that T cells are derived from CD34(+) cells, the involvement of thymic fragments from either human or murine fetus makes the in vitro T cell perliferation very cumbersome. In this report, cord blood mononuclear cells were used as accessory cells to support the differentiation of CD34(+) cells into naive T cells stimulated with SCF and IL-2. CD4(+) and CD8(+) T cells, under the cultural conditions, were continuously produced in vitro at least over a period of 3 weeks and their ratios changed gradually. CD4/CD8 double positive T cells and RAG-2 gene were existed, and RAG-2 gene, reponsible for TCR rearrangement, was expressed during the cell proliferation. Our study presents a simple culture system in vitro to acquire large quantities of naive T cell clones.
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Objective:In order to determine the nature of P 200 autoantigen Methods:12 cases of anti-P 200 pemphigoid sera were collected The skin sections from 6 cases of recessive dystrophic epidermolysis bullosa were studied with these sera by indirect immunofluorescence Results:All the 12 anti-P 200 pemphigoid sera could react with basement membrane zone (BMZ)of 5 cases of recessive dystrophic epidermolysis bullosa, while epidermolysis bullosa acquisita sera were negative in these skins In addition, in a case of recessive dystrophic epidermolysis bullosa, epidermolysis bullosa acquisita sera react with both BMZ and intracytoplasmic deposition of type Ⅶ collagen, while no anti-P 200 pemphigoid sera showed this reactivity Conclusion:These results suggested that the 200 kD antigen is not a component of type Ⅶ collagen, but a specific autoantigen