Objective:To investigate the inhibitory effects of melanoma differentiation associated gene-7(mda-7/IL- 24)on lymphoma cell line Namalwa in vitro and in vivo.Methods:Using RT-PCR,the expression of mda-7/IL-24 was examined in 10 malignant hematopoietic cell lines,including Namalwa,Raji,K562,NB4,U937,Ramous,CEM,KG1a, HL60,J6-1,etc.The coding region of mda-7/IL-24 was cloned from LPS-treated human peripheral blood mononuclear cells(PBMC)by RT-PCR,and the eukaryotic expression vector pTarget-IL-24 was constructed.The recombinant vector, after sequenced,was transfected into Namalwa cell line via lipofectamine reagent.The stable expression transfectants were selected by G418.The expression of mda-7/IL-24 mRNA and protein was verified by RT-PCR and Western blotting.MTT assay,colony forming assay,apoptosis detection,and tumorigenesis in nude mice were used to assess the effects of mda- 7/IL-24 on tumor proliferation,growth characteristics,colony forming,apoptosis,and tumorigenesis.Results:Expression of mda-7/IL-24 mRNA was not found in any of the 10 malignant hematopoietic cell lines and the expression of mda-7/IL- 24 mRNA and protein was found in Namalwa cells transfected with recombinant plasmid pTarget-IL-24.Significant de- crease in tumor cell viability was observed in Namalwa cells stably transfected with mda-7/IL-24,compared with control cells transfected with empty plasmid pTarget(P

AIMTo study the modulation of extracellular pH on the voltage-gated potassium currents (I(Kv)) in isolated pulmonary artery smooth muscle cells (PASMCs).

METHODSI(Kv) was recorded using whole-cell patch clamp technique under the external solutions with different pH. The electrophysiological characteristics of I(Kv) were then analyzed.

RESULTS(1) As compared to the normoxic group, I(K), decreased under acidic condition. When the extracellular pH were 7.0, 6.5, 6.0, the peak currents at a potential of +60 mV were inhibited by 16.93% +/- 2.47% (P < 0.01), 33.03% +/- 2.13% (P < 0.01), 41.59% +/- 6.53% (P < 0.01) respectively, and the current-voltage relationship (I/V) curve shifted to the right. (2) When the extracellular pH was 7.0, 6.5, 6.0, the voltage-depended Gk-Em was shifted to the direction of positive and the activation was sped up.

CONCLUSIONThe results suggest that with the development of hypoxic pulmonary vasoconstriction (HPV), extracellular pH may take part in the modulation of Kv channels partly, then make the cell depolarized and decrease the Kv currents, this will lead to open the L-type calcium channel and contract the pulmonary artery smooth muscle. It may be one of the mechanisms that hypoxic leads to HPV and finally accelerate the development of HPV.

Animals ; Hydrogen-Ion Concentration ; Male ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; metabolism ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Voltage-Gated ; physiology ; Pulmonary Artery ; cytology ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley

Objective To analyze factors influencing prognosis and to develop a prognosis predicting model for patients with chronic severe hepatitis. Methods Clinical data from 408 patients suffering from chronic severe hepatitis were divided into improved group and deteriorated group. The main clinical and laboratory variables were analyzed as predictive factors of prognosis with logistic regression analysis. Results It was shown that age, sex, Tbil, Dbil, [Cl-], [Na+], WBC, MCV, PT, NH3, PIA, and BUN were different between the two groups with statistical significance (P ＜ 0.05). The prognosis predicting model was P=1/(1+e-y), Y=- 4.636+0.022X1+0.034X2+0.096X3+0.047X4-0.042X5, (X1-age, X2-Tbil, X3-BUN, X4-MCV, X5-PTA) .Conclusion Age, Tbil, BUN, MCV and PTA are the independent risk factors related to prognosis of chronic severe hepatitis.

OBJECTIVE: To study the application value of surface electromyography in children with dysphagia. METHODS: A total of 20 children with dysphagia were enrolled as the observation group, and 20 healthy children, matched for sex and age, were enrolled as the control group. Surface electromyography was used to record the electromyography integral values of the submental and infrahyoid muscle groups in the resting state and the state after water swallowing. The two groups were compared in terms of the electromyography integral values of the submental and infrahyoid muscle groups in the resting state and the state after swallowing 5 mL water. The observation group was observed in terms of the changes in the electromyography integral values of the submental and infrahyoid muscle groups after 1 month of rehabilitation treatment. A Spearman correlation analysis was used to evaluate the correlation of the degree of dysphagia with the electromyography integral values of the submental and infrahyoid muscle groups in the observation group. RESULTS: There was no significant difference between the two groups in the electromyography integral values of the submental and infrahyoid muscle groups in the resting state (P>0.05), while after water swallowing, the observation group had significantly higher electromyography integral values than the control group (P<0.05). The observation group had significant improvements in the clinical symptoms of dysphagia after treatment, with significant reductions in the electromyography integral values of the submental and infrahyoid muscle groups (P<0.05). The severity of dysphagia was positively correlated with the electromyography integral values of the submental and infrahyoid muscle groups (P<0.01). CONCLUSIONS Surface electromyography is useful in the diagnosis and therapeutic effect evaluation for dysphagia in children.
Child ; Deglutition ; Deglutition Disorders ; Electromyography ; Humans

OBJECTIVETo investigate whether low molecular weight heparin (LMWH) may suppress the expression and secretion of vascular endothelial growth factor (VEGF) from tumor cells in vitro and inhibit the VEGF-induced proliferation of human tumor vascular endothelial cells.

METHODSHuman lung cancer cell line A549, human liver cancer cell line HepG2, human colon carcinoma cell lines HCT116 and HCT8 were used in this study. The expression levels of VEGF and TNF-alpha (tumor necrosis factor-alpha) in the tumor cells with or without pretreatment of LMWH/heparin were measured by standard sandwich ELISA technique. The VEGF mRNA level of HepG2 cells cultured with or without LMWH/heparin was determined by RT-PCR and real time PCR. Human umbilical vein endothelial cells (HUVEC) were cultured in tissue culture medium (TCM) with or without LMWH/heparin for 3 days. Then non-radioactive cell proliferation assay (MTS) kit and cell cycle assay by flow cytometry were performed to measure the proliferation of HUVEC.

RESULTSThe VEGF levels in the control, LMWH, and heparin groups of the pulmonary adenocarcinoma cell line A549 were (1045.89 +/- 165.30) pg/ml, (782.45 +/- 67.17) pg/ml and (916.54 +/- 71.25) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups of the colon adenocarcinoma cell line HCT116 were (955.76 +/- 51.14) pg/ml, (822.89 +/- 142.39) pg/ml and (951.77 +/- 188.22) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups in the colon adenocarcinoma cell line HCT8 were (1290.62 +/- 41.23) pg/ml, (1063.34 +/- 63.82) pg/ml and (1257.14 +/- 11.40) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups in the liver cancer cell line HepG2 were (1083.00 +/- 134.35) pg/ml, (758.00 +/- 84.85) pg/ml and (874.00 +/- 22.62) pg/ml, respectively. The VEGF expression levels in the above mentioned cell lines cultured in TCM were significantly reduced in the LMWH-treated groups compared with that of the control group (P < 0.05). But the level of TNF-alpha in TCM-cultured cells was unaffected by LMWH. The VEGF mRNA was reduced in the LMWH-treated HepG2 cell line. Moreover, TCM exhibited stimulating effect on proliferation of HUVEC and the effect was significantly impaired by LMWH treatment. Flow cytometric analysis revealed that LMWH treatment arrested HUVECs at the G1 phase of cell cycle.

CONCLUSIONLMWH can suppress the expression and secretion of VEGF by tumor cell lines and therefore have a potential inhibiting effect on angiogenesis induced by VEGF.

Adenocarcinoma ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media, Conditioned ; Endothelial Cells ; cytology ; HCT116 Cells ; Hep G2 Cells ; Heparin ; pharmacology ; Heparin, Low-Molecular-Weight ; pharmacology ; Humans ; Lung Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; secretion

OBJECTIVE: To investigate the risk factors for poor prognosis of neonatal bacterial meningitis. METHODS: A retrospective analysis was performed for the clinical data of 152 children with neonatal bacterial meningitis. According to their prognosis, they were divided into a good prognosis group with 122 children and a poor prognosis group with 30 children. The two groups were compared in terms of general status, initial symptoms, and laboratory findings, and the risk factors for poor prognosis were analyzed. RESULTS: Compared with the good prognosis group, the poor prognosis group had a significantly higher proportion of children with a very low birth weight, a peripheral blood white blood cell count (WBC) of <5×10/L or >20×10/L, a C-reactive protein level of >50 mg/L, a cerebrospinal fluid (CSF) WBC of >500×10/L, a CSF glucose level of <1 mmol/L, or a CSF protein level of >2 g/L, as well as significantly higher positive rates of blood culture and/or CSF culture, Gram-positive bacteria, and Streptococcus agalactiae (P<0.05). The multivariate logistic regression analysis showed that a CSF glucose level of <1 mmol/L and a CSF protein level of >2 g/L were independent risk factors for poor prognosis of neonatal bacterial meningitis. CONCLUSIONS A CSF glucose level of <1 mmol/L and a CSF protein level of >2 g/L are risk factors for poor prognosis of neonatal bacterial meningitis.
Child ; Humans ; Infant, Newborn ; Leukocyte Count ; Meningitis, Bacterial ; Prognosis ; Retrospective Studies ; Risk Factors

Plumbagin (Plumbago zeylanica L.) has a wide spectrum of anticancer activity with a relatively lower toxicity. The molecular mechanisms of proliferation inhibition and apoptosis induction by plumbagin on esophageal squamous carcinoma cell lines may be important for the structure modification and clinical application of plumbagin. After treatment of KYSE-30, KYSE-70 and KYSE-140 cells with 0-20 μmol·L^-1 of plumbagin for 24, 48, 72 h, CCK8 was used to examine the proliferation, Annexin V and PI immunofluorescence staining for apoptosis, real-time PCR and Western blot for FoxM1 mRNA and protein expression, dual-luciferase reporter gene assay for the transcriptional activity of FoxM1, respectively. In addition, the relationship between anti- tumor effect of plumbagin and FoxM1 was investigated in vivo. Plumbagin significantly inhibited proliferation and induced apoptosis of esophageal squamous carcinoma cell in vitro and in vivo. Moreover, plumbagin down-regulated the expression of FoxM1 through suppression of its gene transcription. Our findings suggest that plumbagin may inhibit the proliferation of esophageal squamous carcinoma cell in vivo and in vitro through down-regulating the expression of FoxM1.

Objective To investigate the measures of prevention and care on perioperative complications of sacral tumor resection and reconstruction. Methods Enhanced the awareness of the common complications before surgery and strengthen measures of prevention and care including psychological care, intraoperative bleeding prevention, bowel sphincter function training and skin preparation. The postoperative care including closely monitoring the vital signs, observing the spinal cord nerve function, strengthen nursing care containing the posture care, wound and drainage tube care, early stage rehabilitation guidance and limb functional training were carried out. Results During the perioperative period, without complications occurred. Conclusions Emphasis the prevention and care on complications and carrying out the proper treatment and nursing measures are essential to successful surgeries and the recovery of patients.

Objective To investigate the effects of liver X receptor (LXR) agonist on adiposederived mesenchymal stem cells (AD-MSCs) implantation into infarcted hearts of mice.Method AD-MSCFluc+ which stably expressed firefly luciferase (Fluc) were isolated from β-actin-Fluc transgenic mice and characterized by flowcytometry.Male FVB mice were randomly allocated into the following four groups (n=10each):(1) sham group; (2) MI + PBSgroup; (3) MI + AD-MSCFluc+ group; (4) MI + AD-MSCFluc + LXR agonist (T0901317) group.AD-MSCFluc+ or PBS were injected intramyocardially into periinfarcted region of mice heart after permanent left anterior descending ( LAD ) artery ligation.Bioluminensence imaging (BLI)was performed for quantification of injected cells retention and survival.Cardiac function was evaluated by echocardiography.Results The AD-MSCFluc+ were positive for CD44 and CD90 by flowcytometry.BLI evidenced the firefly luciferase expression of AD-MSCFluc+ which was positively correlated with cell numbers ( r2 =0.98 ).The results of BLI in vivo revealed that LXR agonist could improve the survival of AD-MSCFluc+ at day 7,14 and 21 after transplantation compared with AD-MSCFluc+ alone group.Cardiac function was further improved in combination therapy group compared with AD-MSCFluc+ alone group (P＜0.05).Conclusions LXR agonist T0901317 can improve the retention and survival of intramyocardial injected AD-MSCFluc+ post-MI,and the combination therapy of T0901317 and AD-MSCFluc+ has a synergetic effect on improving cardiae funetion in this model.

Objective To understand the epidemiologic characteristics of endemic typhus in Baoshan city. Methods Epidemiological data were collected and characteristics were analyzed. IgG antibody(Ab) of Rickettsia mooseri and Orientia tsutsuganushi in serum of patients were tested using both Weil-Felix and IFA method. The Rickettsia mooseri gltA gene, Rickettsia prowazekii gltA gene,Orientia tsutsugamushi 56 kDa protein gene, SFGR ompA gene, Ehrlichia sp. 16S rRNA gene and Anaplasma sp. 16S rRNA gene in spleen of mice were examined by PCR. Results Fifty- eight endemic typhus cases were found in Longyang district of Baoshan city, during July to August, 2009.Among them, 48 cases were confirmed by clinical diagnosis and 10 cases by laboratory tests. The Ab of Orientia tsutsugamushi Karp serotype was detected in 3 cases from laboratory diagnosis. The spleen samples from 85 Rattns flavipectus were tested using PCR. Of them, 3 samples for Rickettsia mooseri gltA gene showed positive (positive rate was 3.5% ), and the homology of 3 Rickettsia mooseri and Rickettsia mooseri Wilmington strain (GenBank U59714.1) was 100% through comparing gene sequence. The results of PCR for detecting Rickettsia prowazekii, Orientia tsutsugamushi, SFGR,Anaplasma sp. and Ehrlichia. sp were all negative. Conclusion The outbreak of endemic typhus was confirmed in Longyang district of Baoshan city through epidemiological data, clinical diagnosis and laboratory tests. Rickettsia mooseri DNA was detected in the dominant Raw flavipectus, suggesting that endemic typhus did exist in the local areas.