Objective To construct the replication-deficient recombinant adenovirus coding ARP2 gene before the studies of gene transfection to ischemic myocardium.Methods From Dec.2004 to Oct.2005,in the Department of Cardiology,Changhai Hospital of the Second Military Medical Univesity,the ARP2-pAxCAwt was constructed by inserting the cDNA of interest into the SwaI site of pAxCAwt.Results The right direction of the insert was confirmed by restriction.Conclusion The direction of the insert must be confirmed by restriction analysis,and the DNA package protein is much helpful during the transfection of the recombinant cosmid to E.coli.

Objective To study the transfection efficiency, protein expression, and effect of adenovirus-mediated transfection on microvascular endothelial cells transfected by angiopoietin-related protein 2(Ad. ARP2)gene. Methods Mice coronary microvascular endothelial cells(CMECs) were isolated, cultured and transferred by Ad-ARP2. The transfection efficiency and cellular toxicity of adenovirus vector to CMECs were detected by immunofluorescence staining. Expression of Ad. ARP2 in CMECs and the secreted materials in culture medium were measured by Western blot and ELISA and compared among groups of Ad. ARP2, Ad. null, and PBS control. Vascular endothelial cells were incubated with conditional medium containing secreted ARP2, and effects on cells sprouting were observed in matrigel. Results Adenovirus-transfected CMECs showed a very high efficiency. When multiplicity of infection (MOD was 200, the transfection efficiency was 93. 5% ,and no harmful effect on CMECs growth was found. When CMECs were transfected with Ad. ARP2, there was a high ARP2 expression, which was significantly different from that with Ad. Null or PBS. The conditional medium containing ARP2 had an excellent ability to stimulate sprout of CMECs which phenomenon could not be seen in the control groups. Conclusions Adenovirus vector can be transferred into CMECs efficiently and safely. Ad. ARP2 gene transfection allows a high transient expression, and the expression products can stimulate the sprout of microvascular endothelial cells in vitro very well.

Objective To investigate the clinical efficacy of esmolol combined with atorvastatin on se-vere sepsis complicated with cardiac insufficiency.Methods This study was a prospective,double-blind,ran-domized controlled clinical trial.A total of 153 patients with severe sepsis complicated with cardiac insufficien-cy admitted to this hospital from January 2021 to December 2022 were selected and divided into groups A,B,and C by random number table method,with 51 cases in each.Patients in group A were given routine symp-tomatic supportive treatment after admission.On this basis,patients in group B and group C were given esmo-lol,esmolol+atorvastatin,respectively.The hemodynamic indexes,serological indexes and clinical prognosis of the three groups before and after intervention were compared.Results There was no significant difference in baseline data,and hemodynamic and serological indexes of three groups before intervention(P＞0.05).Compared with before intervention,after five days of intervention,heart rate,systemic vascular resistance in-dex(SVRI),blood levels of creatine kinase-MB(CK-MB),cardiac troponin Ⅰ(cTn Ⅰ),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and high sensitive C-reactive protein(hs-CRP)in three groups were de-creased,while the values of cardiac index(CI)were increased,and the differences were statistically significant(P＜0.05).After five days of intervention,the heart rate,SVRI,blood levels of CK-MB,cTn Ⅰ,TNF-α,IL-6,and hs-CRP in group C were lower than those in group A and group B,and the levels in group B were lower than those in group A;the value of CI in group C was higher than that in group A and group B,and group B was higher than that in group A,the differences were statistically significant(P＜0.05).After intervention,the length of stay in intensive care unit(ICU)in group C was the shortest,and that in group B was shorter than that in group A,the difference was statistically significant(P＜0.05).There was no significant difference in 28 d mortality among the three groups(P＞0.05).Conclusion Esmolol combined with atorvastatin can signif-icantly inhibit the inflammatory response in patients with severe sepsis complicated with cardiac insufficiency,relieve myocardial injury and promote rehabilitation,and the therapeutic effect is better than esmolol alone.

OBJECTIVETo explore the expression of estrogen receptors in the human periodontal ligament fibroblasts in vitro.

METHODSHuman periodontal ligament fibroblasts were cultured in vitro. The estrogen receptors (ER)-alpha, ER-beta were detected with immunocytochemistry staining; The mRNA of ER-alpha, ER-beta were measured by reverse transcription polymerase chain reaction; The expression of ER-alpha, ER-beta protein were detected by Western blot.

RESULTSThe mRNA and protein expression of two subtype of ER were observed on human periodontal ligament fibroblasts (HPLF). The intensity of the ER-beta bands were stronger than those of ER-alpha, indicating that ER-beta expression in HPLF was higher.

CONCLUSIONSER may play an important role in the function of human periodontal ligament fibroblasts.

Adolescent ; Blotting, Western ; Cells, Cultured ; Estrogen Receptor alpha ; genetics ; metabolism ; Estrogen Receptor beta ; genetics ; metabolism ; Female ; Fibroblasts ; metabolism ; Humans ; Immunohistochemistry ; Periodontal Ligament ; cytology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Thyroid cancer is the one of the most common endocrine tumors. The biological behaviors and prognoses of the thyroid cancer of different histological types remarkably differ. The highly invasive thyroid cancer responds poorly to traditional therapies. Recent research advances in the molecular mechanisms of the pathogenesis of thyroid cancer have revealed the roles of many genetic and epigenetic variations such as gene mutation, abnormal gene amplification, and abnormal gene methylation in the development of thyroid cancer, which provides new insights in the molecular diagnosis, prognosis, and target therapy of the thyroid cancer.
Carcinoma ; genetics ; Humans ; Mutation ; Signal Transduction ; Thyroid Neoplasms ; genetics

Objective To investigate the 7 kinds of respirovirus etiology of children with acute respiratory infection(ARI) in Wenzhou area from 2005 to 2006.Methods Three thousand nine hundred and seventy children with ARI visited the Yuying children's hospital were chosen,including 308 children with acute upper respiratory infection(URI) and 3 662 children with lower respiratory infection(LRI).Direct immunofluorescence(DIF) was used to detect the respiratory syncytial virus(RSV),adenovirus(ADV),influenza virus(IV) A and B,parainf-luenza virus(PIV) type 1,2,3 from nasopharyngeal secretions(NPS) collected from these patients.Results Among the 3 970 samples,1 773(44.7%) positive results were determined and the positive rate of RSV(36.2%) was the highest.The isolating rate of respirovirus were all conspicuous difference in sex(?2=9.2 P

Adolescent ; Common Variable Immunodeficiency ; complications ; Female ; Humans ; Lung Diseases, Interstitial ; etiology ; pathology ; physiopathology ; Pneumonia ; etiology ; pathology ; physiopathology

Objective Little is known about the effect of RNAi on mitochondrial apoptotic pathways. This study aims to explore the effects of the Survivin shRNA-APC double-gene on colon cancer mitochondrial apoptosis pathway-related factors survivin,cytochrome C (Cytc),second mitochondria-derived activator of caspases (Smac),and cysteine aspartic acid specific protease 9 (Caspase-9) as well as on the apoptosis of colon cancer transplanted tumor (CCTT) cells. Methods Thirty nude mice were randomly divided into five groups of equal number,Survivin shRNA-APC double-gene,survivin shRNA,APC,empty vector and blank transfection. The CCTT model was established in the nude mice by subcutaneous injection of the colon cancer cell strains stably transfected with the Survivin shRNA-APC double-gene,survivin shRNA,APC,an empty vector and HT-29,respectively,into the mid-posterior part of the left armpit of the nude mice. The rate of tumor growth inhibition was calculated by measuring the volume and weight of the CCTTs in the nude mice. The mRNA and protein expressions of survivin,Cytc,Smac and Caspase-9 in the tumor tissue were detected by real time PCR and immunohistochemistry,respectively,and the apoptosis rate of the CCTT cells was detected by TUNEL. Results The model of CCTT was successfully established in the nude mice. Com-pared with the empty vector and blank transfection groups,the mice in the double-gene,survivin shRNA and APC groups showed sig-nificantly decreased average volume and weight of the tumor tissue (P<0.05) but increased inhibition rate of its volume and weight (P<0.05). In comparison with the survivin shRNA and APC groups,the double-gene group exhibited remarkably decreased average volume and weight of the tumor tissue (P<0.05) but increased inhibition rate of its volume and weight (P<0.05). The mRNA and pro-tein expressions of survivin were significantly lower while those of Cytc,Smac and Caspase-9 markedly higher in the double-gene,sur-vivin shRNA and APC groups than in the empty vector and blank transfection groups (P<0.05),the former even lower (P<0.05) and the latter even higher in the double-gene than in the survivin shRNA and APC groups (P<0.05). The apoptosis rate of the CCTT cells was significantly increased in the double-gene ([56.78±3.04]%),survivin shRNA ([33.61±2.02]%) and APC groups ([30.16± 1.72]%) as compared with the empty vector ([10.05±0.42]%) and blank transfection groups ([9.87±0.30])% (P<0.05),even higher in the double-gene group than in the survivin shRNA and APC groups (P<0.05). Conclusion The Survivin shRNA-APC double-gene may induce apoptosis of colon cancer transplanted tumor cells by down-regulating the expression of the apoptosis inhibitor survivin,upregulating the expressions of Cytc,Smac and Caspase-9,and suppressing the growth of the colon transplanted tumor,with more significant abilities than a single gene in regulating apoptosis-related factors,inducing cell apoptosis and inhibiting the growth of the transplanted tumor.

Idiopathic pulmonary fibrosis(IPF) is a specific form of interstitial lung diseases(ILDs) with unknown causes. In 2018, the international expert panel of interstitial lung diseases updated the diagnostic criteria of IPF based on the imaging and histopathology published in 2011. We will interpret the new 2018 version of diagnostic guidelines for IPF.

OBJECTIVE: To observe the effects of immune complexes (IC) prepared from human oxidized density lipoprotein (oxLDL) antibodies and human oxLDL on the foam cell forming and the macrophage activation, and to further uncover the possible mechanisms of immune complexes contributing to the atherosclerosis occurrence. METHODS: The immune complexes of human oxLDL and purified human oxLDL antibodies were added to culture U937 cells by protocols: polyethylene glycol-precipitated insoluble IC (PEG-IC) and IC immobilized by absorption to red blood cells (RBC-IC). With oxLDL as controls and heat-aggregated gamma globulin as an inhibitor of Fc gamma receptor, we measured the cholesterol ester, total cholesterol of the cellular extracts, and quantified the secreted MMP-1 of supernatants from U937 cells. RESULTS: A significant increase of MMP-1 release [(0.769 +/- 0.030) ng/ml vs (0.513 +/- 0.034) ng/ml, P < 0.01] and a higher level of cholesterol ester accumulation [(20.271 +/- 1.668) microg/mg protein vs (17. 226 +/- 1.298 ) microg/mg protein, P < 0.05] in U937 cells incubated with RBC-IC were observed, compared with those incubated with RBC-oxLDL. However, the above quantative difference between the cholesterol ester accumulation induced by oxLDL and insoluble PEG-IC was even more striking, and cholesterol ester accumulation was dosage-dependent. Heat-aggregated gamma globulin (10 mg/ml) as an inhibitor of Fc gamma receptors competitively inhibited cholesterol ester accumulation and decreased PEG-IC stimulating MMP-1 secretion to 71%. CONCLUSION Immune complexe of ox-LDL can transform macrophages into foam cells and activted macrophages. The immunological function of oxLDL is involved in the process of atherosclerosis occurrence.
Antibodies ; pharmacology ; Atherosclerosis ; etiology ; metabolism ; Cholesterol Esters ; metabolism ; Foam Cells ; drug effects ; Humans ; Lipoproteins, LDL ; immunology ; pharmacology ; Macrophage Activation ; drug effects ; Matrix Metalloproteinase 1 ; biosynthesis ; U937 Cells