Objective　To investigate clinical features of neonatal ventilator associated pneumonia.Methods　Retrospective analysis of clinical features was performed in twenty-two newborns with ventilator associated pneumonia.Results　Ventilator associated pneumonia were found in 22 ou t of 36 neonates with mechanical ventilation.Conclusion　The incidence of VAP is significantly increased if the time of mechanical ventilation in longer than 96 hours.

Objective To evaluate the clinical significance of cytokeratin19(CK19) mRNA in the peripheral blood of patients with non-small cell lung cancer (NSCLC). Methods The expression of CK19 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR) in the peripheral blood from 98 NSCLC patients (NSCLC group), 20 benign pulmonary lesion patients (benign pulmonary lesion group) and 20 healthy adults(control group). Results The positive rate of CK19 mRNA in peripheral blood of NSCLC group, benign pulmonary lesion group and control group was 77.55 % (76/98), 3.57% (1/28),0 (0/20) respectively,and there was significant difference among three groups ( x2 = 73.680,P ＜ 0.01 ). The positive rate of CK19 mRNA in different pathological types of NSCLC had no significant difference (x2 =0.414, P ＞ 0.05 ), but had significant difference in different lymphy node metastasis status( x2 = 17.523, P ＜0.01 ) and different clinical stage ( x2 =13.453,P ＜ 0.01 ). Conclusions The higher expression of CK19mRNA in peripheral blood of NSCLC may relate to cancer metastasis. Detection of CK19 mRNA has significance in early prediction of its prognosis.

Objective To investigate the expression and significance of endothelin-1 ET-1 in urothelial transitional cell carcinoma. Methods Immunohistochemistry was applied to detect ET-1 expression in 30 cases of urothelial transitional cell carcinoma and 20 cases of normal urothelial transitional cell samples, the results were analyzed in combination with clinical data. Results ET-1 expression in urothelial transitional cell carcinoma was 40.5%, which was significantly higher than that in normal urothelial transitional cell samples(P ＜0.05). ET-1 expression was correlated with grade and stage of urothelial transitional cell carcinoma (P ＜0.05). Conclusion The over expression of ET-1 in urothelial transitional cell carcinoma tissues correlates closely with cancer genesis progression and invasiveness, suggesting that ET-1 might be a biomarker and as a therapeutic target in human urothelial transitional cell carcinoma.

Objective To evaluate the clinical outcome of external bracket fixation in the treat- ment of complex tibia diaphysis fracture involving intra-articular fractures.Methods Forty-two cases of complex tibia diaphysis fracture with proximal and distal intra-articular fractures treated surgically in our hospital from January 1999 to January 2004 were analyzed.The complex tibia diaphysis fractures were categorized according to the AO classification as type C2 (multiple segments fracture) and type C3 (ir- regular fracture),proximal and distal intra-articular fractures in 23 and 19 cases,respectively.Definite operation was done within one week.Twenty-two cases were treated with simple external fixator,and 20 cases treated with screws and external fixator.Results All the 42 cases were followed-up regularly. According to AO evaluation of the knee and ankle joint movement,83% (35/42 cases) of the cases gained satisfactory functional outcome,14% (6/42 cases) had quite satisfactory results and 2% (1/42 case) had unsatisfactory functional outcome.Conclusion External bracket fixation can obtain outcome of relative length of the tibia and fibula,tube structure reconstruction,smoothness of the articular surface and the parallel and symmetric relation of knees and ankles for complex tibia diaphysis fracture with proxi- mal and distal intra-articular fracture.The arthritis resulting in pain in movement and restriction of func- tion is considered to be the most important factor affecting the joint function.Early functional exercise is important for best recovery of knee and ankle function.

OBJECTIVETo observe the effect of Chinese herbal extract HuNan A-1 (HNA-1) on the thymic output function in Simian immunodeficiency virus (SIV) chronically infected rhesus macaques.

METHODSEight Chinese rhesus macaques had been infected by SIVmac239 for 16 to 21 months, and then they were randomly divided into the treatment group and the control group, 4 in each group. Monkeys in the treatment group were administered with HNA-1 by gastrogavage, once daily for 2 successive months, while those in the control group were administered with equal volume of normal saline by gastrogavage, once daily for 2 successive months. The general condition and body weight of monkeys were observed. Plasma viral loads were detected using real-time fluorescent quantitative PCR assay. CD4 percentages and counts, as well as naive CD subsets were detected using flow cytometry. T-cell receptor excision circles (TREC) were detected using real-time fluorescent quantitative PCR assay. The thymus tissue was pathologically observed using routine HE staining. The correlation between lesions of the thymus tissue, CD4 counts, naive CD counts, and TREC were analyzed.

RESULTSThere was no statistical difference in body weight, viral loads, absolute CD ratios between the two groups after treatment (P > 0.05). The altered TREC multiple showed an obvious decreasing tendency in the control group, while it showed an increasing tendency in the treatment group (P < 0.05). In both groups, destroyed structures of the thymus tissue could be seen, filled with pink unstructured material. Increased connective tissues, lowered connective cell density, and confused arrangement could also be seen in the two groups, with no obvious difference. TREC contents were positively correlated with naive CD4 counts after removing extremum (r = 0.926, P = 0.001). Naive CD4 counts were positively correlated with CD4 counts (r = 0.961, P = 0.005).

CONCLUSIONSTREC content determination, as a marker of newly thymic emigrants, could be taken as a testing method for evaluating the thymic output function. Besides, HNA-1 treatment increased the thymic output significantly in SIV chronically infected monkeys. Correlation existed among TREC contents, naive CD4 counts, and pathologies of thymus tissues, especially in late infection stage.

Animals ; CD4 Lymphocyte Count ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Macaca mulatta ; Plant Extracts ; pharmacology ; Random Allocation ; Simian Acquired Immunodeficiency Syndrome ; drug therapy ; Simian Immunodeficiency Virus ; Thymus Gland ; drug effects ; Viral Load

Objective To evaluate the capability and accuracy of multi-shce spiral computed tomography(MSCT)in detecting atherosclerotic plaques in nonstenotic coronary arteries with reference to the findings of intravascular ultrasound(IVUS)in a segment analysis.Methods Both IVUS exams and 16-row MSCT scans were performed on 35 consecutive patients among whom 30 patients had successful MSCT scans.A total of 94 coronary segments without significant coronary stenoses were paired-analyzed both on IVUS and MSCT segment by segment.The plaques were classified as calcified,fibrotic and soft types according to the echogeneity on IVUS.Plaque attenuation on MSCT was measured and expressed by Hounsfield units(HU).Results When referred to IVUS,MSCT had a sensitivity of 82.1%(46/56)and specificity of 89.5% (34/38),respectively in detectiong any plaques.For the detection of calcified plaques,the sensitivity and specificity were 92.1%(35/38)and 96.4%(54/56),respectively.For the detection of mixed and noncalcified plaques,MSCT had sensitivity of 73.2%(30/41)and specificity of 88.7%(47/53).But for the detection of the noncalcified plaque,the sensitivity was 66.7%(12/18). According to the findings On IVUS,the plaques were classified as calcified(n=19),fibrotic(n=19)and soft(n=16).The CT attenuation of calcified plaques was(489?169)HU(196 to 817 HU),fibrotic plaques(69?21)HU(25 to 117 HU)and soft plaques(23?18)HU(-12 to 47 HU).Nonparametric Kruskal-Wallis test revealed a significant difference of plaque attenuation among the three groups(P

Objective To investigate the pathological distribution and clinical characteristics of pineal region tumors (PRTs) with variant pathology.Methods We retrospectively analyzed the clinical and pathological data of 133 patients with PRTs that were performed surgical removal from January 2000 to January 2009 in our hospital.Results Sixty-one patients (45.9%) were diagnosed as having germ cell tumors,17 (12.8%) pineal parenchymal tumors,28 (21.1%) gliomas and 27 (20.2%)other resource tumors.Sex ratio of patients with germ cell tumors,pineal parenchymal tumors and gliomas were 14.25:1 (male:female),2.4:1 and 1.15:1,respectively; their median ages were 15.3,24.7 and 28.1,respectively.The serum immunologic test showed abnormal results in 33 patients; except for 1 with metastatic tumors,the others were diagnosed as having germ cell tumors.Conclusion PRTs have many pathological types and patients with PRTs are mainly diagnosed as having germ cell tumors.Correct diagnosis can not be made by imaging,serum immunologic test or biopsy, thus,obtaining complete clinical specimen during the total tumor removal is very necessary in studying the pathology of PRTs.

OBJECTIVETo investigate the apoptosis effect of diffuse large B-cell lymphoma cell line (DLBCL) SUDHL-4 induced by IL-21 and its related mechanism.

METHODSSUDHL-4 cells were treated with IL-21 at different concentration (1000 ng/ml, 100 ng/ml, 10 ng/ml, 1 ng/ml) for 24 h, 48 h, 72 h, respectively. The inhibitory rate of cell proliferation was detected by CCK-8 assay. The cell growth curves were drawn and half inhibitory concentration (IC(50)) values were calculated. The cell apoptosis were detected by flow cytometry (FCM), the expression of the caspase-9, caspase-3, cleaved caspase-3, Bcl-2, Bcl-XL, Bid, Bax and c-myc protein in SUDHL-4 cells treated with IL-21 by western blot, the mRNA expression of Bcl-2, Bcl-XL, Bid, Bax, c-myc by Survivin gene with RT-PCR.

RESULTSIL-21 markedly inhibited SUDHL-4 cell growth in a time- and dose-dependent manner. The 48 hIC(50) was 140.9ng/ml; The FCM showed that the apoptosis proportion of SUDHL-4 cells treated with 100 ng/ml of IL-21 apoptosis (AnnexinV-FITC(+) positive cells) gradually increased (48 h: 19.7 ± 2.3%). The protein expression of caspase-9, caspase-3, Bcl-2 and Bcl-XL decreased in a time-dependent manner. The Bax and c-myc protein markedly increased, but the Bid protein level did not change. IL-21 up regulated c-myc and Bax gene expression, however down regulated Bcl-2 and BCL-XL gene expression, but the gene expression of Bid and Survivin hadn't been changed significantly.

CONCLUSIONSIL-21 can inhibit proliferation and induce apoptosis of SUDHL-4 cell. The mechanism may involve in endogenous mitochondrial pathway mediated by the c-myc and the Bcl-2 genes.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Humans ; Interleukins ; administration & dosage ; pharmacology ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-X Protein ; metabolism

OBJECTIVETo investigate the effects of lentivirus-mediated RNA interference targeting vascular endothelial growth factor receptor 1 (VEGFR1) gene on the proliferation, migration and apoptosis of leukemic cell line U937.

METHODSShort hairpin RNAs (shRNA) targeting VEGFR-1 was synthesized and cloned into pRNAT-U6.2 lentiviral vector. The expression vectors were transfected into 293T cell line to produce packaged lentivirus. After infected with the packaged lentivirus, the expression of VEGFR-1 gene of U937 cells at mRNA and protein level was detected by real-time PCR and Western blot. VEGF production by the cells was determined by ELISA. Cell proliferation and survival under regular culture and in the presence of cytarabine (Ara-C) was determined by CCK-8 assay. Migration assays were performed by 5 µm pore transwell inserts.

RESULTSThe lentiviral shRNA vector targeting VEGFR-1 was successfully constructed and transfected into U937 cells. The shRNA vector effectively inhibited the expression of VEGFR-1 gene in U937 cell line at mRNA and protein levels. As compared to that of the control, the proliferation rate of U937-shVEGFR-1 cells reduced; The VEGF production and migrated cell number of U937-shVEGFR-1 cells decreased dramatically. After treated with Ara-C, the inhibition rate and apoptotic rate of U937-shVEGFR-1 cells increased significantly. The number of migrated cells in the KD group under regular culture and in the presence of VEGF was markedly lower than that in the NC group and CON group. Bevacizumab could decrease the number of migrated cells in the NC group and CON group, but could not in the KD group.

CONCLUSIONSLentivirus-mediated RNA interference targeting VEGFR1 gene reduces the proliferation, migration of U937 cell line and enhances its sensitivity to Ara-C.

Apoptosis ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Gene Silencing ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; U937 Cells ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the effects of hypoxia on sumoylation of HIF-1α, as well as transcriptional activity and protein stability of HIF-1α in Jurkat cells, and explore its effect and significance on modulating the transcriptional activity of down stream target gene.

METHODSCoCl(2) was used as a chemical inducer to simulate the hypoxia environment. Real-time fluorescence quantitative PCR and western blot were used to evaluate transcriptional activity and protein stability of HIF-1α, levels of SUMO-1 and SENP1 protein, and gene transcripts of VEGF, mdr1, mdr3, Mcl-1 and survivin respectively.

RESULTSAfter 4 h, 8 h, 16 h, 24 h and 72 h after hypoxia induction, the gene transcripts of HIF-1α were 0.79 ± 0.19, 2.65 ± 2.05, 4.19 ± 4.72, 2.77 ± 3.37, 0.09 ± 0.05 and 0.69 ± 0.55-fold (P > 0.01) of that of 0h in Jurkat cells, respectively, while the protein stability increased first, then decreased (P < 0.01). SENP-1 protein up-regulated first, then down-regulated, and the SUMO-1 protein changed in an opposite trend. Excepting for survivin gene, transcriptional activities of VEGF, mdr1, mdr3, and Mcl-1 were affected by the stability of HIF-1α protein.

CONCLUSIONHypoxia induces changes in SENP-1 expression, which increases the stability of HIF-1α by decreasing the sumoylation of HIF-1α and affects biological process by regulating the transcriptional activities of VEGF, mdr1, mdr3 and Mcl-1 gene.

Cell Hypoxia ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; Jurkat Cells ; RNA, Messenger ; genetics ; Sumoylation ; Transcriptional Activation ; Vascular Endothelial Growth Factor A ; metabolism