To explore the function of human laryngeal carcinoma Hep-2 cell after down-regulating the expression of E-cadherin gene to provide theoretical rationales for gene therapy of laryngeal cancer.According to the GenBank database, 3 pairs of shRNA sequences of E-cadherin gene were designed and synthesized. shRNAs were transfected into the cell line Hep-2 by liposome. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the silencing effect of E-cadherin expression. The changed capacity of cell proliferation were detected in vitro by methyl thiazalyl tetrazolium (MTT) assay in the transfected Hep-2 cells and the cell proliferation rate (survival rate) was calculated. And Transwell was used to detect the migratory capacity of Hep-2 cells after siRNA transfection.The E-cadherin gene expression of RNAi transfected Hep-2 cells significantly decreased in interference group. And the proliferation of interference group became markedly enhanced. In Transwell test, the migrated cell numbers in interference group were significant higher than those in negative control group (262 ± 15, 288 ± 12, 292 ± 6 vs 74 ± 8, all P < 0.01).RNA interference silencing of E-cadherin gene expression can significantly enhance the proliferation and migratory capacity of Hep-2 cells. And E-cadherin may be considered as one of gene therapy targets for laryngeal cancer.

To explore the effects of phosphatidylinositol 3 kinase-protein kinase B (PI3K-AKT) signal pathway on the proliferation, apoptosis and invasiveness of colon cancer cell line SW480 and explore its possible mechanisms.SW480 cells were cultured with various concentrations (0, 5, 10, 20 and 40 µmol/L) of LY294002 and methyl thiazolyl tetrazolium (MTT) assay was conducted after 24, 48 and 72 hours. SW480 cells were cultured for 24 hours with various concentrations (0, 10, 20 and 40 µmol/L) of LY294002. The apoptotic rate was measured by flow cytometry (FCM). The difference of invasiveness was examined by Transwell invasion test. Western blotting was used to examine the expressions of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9). One-way ANOVA was used for statistical analysis.MTT assay showed that the proliferation rate of all SW480 cells except for the lowest concentration group (5 µmol/L) was lower than the control group (0 µmol/L) and a time-dosage dependence existed (all P < 0.05). The results of FCM demonstrated that the apoptotic rates of 10, 20 and 40 µmol/L groups were 13.3% ± 0.9%, 30.9% ± 2.5% and 41.2% ± 4.1% respectively, and were all significantly higher than control group (5.2% ± 1.8%, all P < 0.05). The number of cells penetrating through the membrane of 10, 20 and 40 µmol/L groups were 87 ± 6, 65 ± 7 and 46 ± 11 respectively. All invasiveness groups were all lower than control group (100 ± 10, all P < 0.05) except the 10 µmol/L concentration group (P = 0.096). Western blotting showed that the expressions of VEGF and MMP-9 were all lower than control group (all P < 0.05).PI3K-AKT signal pathway plays an important role in the proliferation, apoptosis and inhibition of colonic cancer cells. Its mechanism is probably related with the inhibitions of VEGF and MMP-9. PI3K-AKT signal pathway may become a potential target for the treatment of colon cancer.

To explore the mechanism of selective phosphodiesterase 4 (PDE4) inhibitor and observe its effects on the in vitro expression of aquaporin 5 (AQP5) in a rat model of airway mucus hypersecretion.Forty healthy male rats were randomized into 4 groups (n = 10 each). The normal control and drug control groups underwent normal saline aerosol inhalation or YM976 lavage respectively for 12 days while the model and treatment groups acrolein atomization inhalation or crolein atomization inhalation plus YM976 lavage respectively for 12 days. The middle lobe of right lung were fixed, embedded and stained by hematoxylin and eosin to observe the histological changes of small airway. The mucin secretion by goblet cells in lung tissue was tested by alcian blue staining. And the effects of selective PDE4 inhibitor on the expression of AQP5 were detected by immunohistochemical stain. Reverse transcription-polymerase chain reaction and Western blot were used to determine the levels of AQP5 mRNA and protein.In the treatment group, mucus gland hyperplasia and airway mucus hypersecretion and positive staining area ratio of alcian blue staining decreased significantly compared with the model group (10.23% ± 0.94% vs 14.74% ± 1.06%, P < 0.05). And the expression of AQP5 mRNA (1.26 ± 0.19) and protein (0.56 ± 0.13) also declined significantly versus the model group ((0.96 ± 0.17), (0.43 ± 0.15), P < 0.05).Selective PDE4 inhibitors alleviate the pathological damage of lung tissue and enhance the expression of AQP-5 in airway mucus hypersecretion.

To explore the effects of 3-methyladenine (3-MA) and rapamycin (Rapa) on autophagy and uridine 5'-diphospho-glucuronosyltransferase 1A1 (UGT1A1) induced by sulforaphane (SFN) in human colon cancer Caco-2 cells.Western blot was used to detect the expression of microtubule-associated protein 1 light chain 3 (LC3) and UGT1A1 proteins. And immunocytochemistry was employed to observe the intracellular distribution of LC3 and nuclear localization of NF-E2-related factor 2 (Nrf2). Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) was employed to examine the mRNA expression of UGT1A1 and human pregnane X receptor (hPXR).After the treatment of SFN, the LC3-II protein was induced in a dose and time-dependent manner. SFN-induced LC3-II protein could be attenuated and enhanced by 3-MA and Rapa respectively. In comparison with the control group, UGT1A1 mRNA levels increased significantly after the treatment of Rapa, SFN or their combination (2.4, 4.12 and 2.41 folds respectively, all P < 0.01). And the combination of SFN and Rapa possessed the highest level. UGT1A1 protein band intensity was also enhanced in three groups. There was no obvious nuclear staining of Nrf2 in control group while intense nuclear fluorescent labeling of Nrf2 could be observed in the SFN-treated groups, especially the combination group of SFN and Rapa. The hPXR mRNA levels increased significantly in the Rapa and combination groups (1.82 and 1.4 folds respectively, both P < 0.01).The treatment of 3-MA or Rapa may attenuate or enhance SFN-induced autophagy respectively. And Rapa also potentiates SFN-induced UGT1A1 expression. The mechanism for the synergic effect of Rapa and SFN on UGT1A1 induction may be a simultaneous activation of Nrf2 and hPXR signaling pathway.

To explore the effects of atovastatin and tinidazole on atherosclerosis and the levels of tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1) and matrix metalloproteinase-2 (MMP-2) in periodontitis.A total of 48 male New Zealand white rabbits were randomly divided into 4 groups (n = 12 each): control group (A), atovastatin group (B), tinidazole group (C) and combination group (D, atovastatin + tinidazole). All groups received interventions according to the experiment design. During Week 1-4, mandibular first premolars were used to establish periodontitis model. For Week 1, adaptive feeding was provided with 50% normal diet + 50% high-fat diet. Then a full high fat-diet was used to establish atherosclerosis model. During Week 16-20, experimental drug intervention was administered twice weekly: group A received the same volume of saline, group B atorvastatin tablets 1.5 mg/kg, group C tinidazole tablets 150 mg/kg and group D atorvastatin tablets 1.5 mg/kg + tinidazole tablets 150 mg/kg. At the end of 20-week intervention, the animals were sacrificed to take vascular and heart tissue samples. Immunohistochemistry and fluorescent polymerase chain reaction (PCR) were employed for quantitative determinations.The positive areas of MMP-2 expression in groups B, C and D were smaller than that of group A respectively (35% ± 17%, 69% ± 5%, 30% ± 7% vs 86% ± 9%, all P < 0.05). And the PCR results showed the levels of TNF-α and IL-1 in group D was the lowest of four groups (all P < 0.05).Atovastatin and tinidazole can reduce the expression levels of TNF-α, IL-1 and MMP-2 in the rabbits with atherosclerosis and periodontitis respectively. And the combination of both drugs may achieve a better efficacy.

To explore the cell signal transduction pathway of calcium-sensing receptor (CaSR) mediated hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells (PASMCs).The expressions of proliferating cell nuclear antigen (PCNA) and extracellular signal-regulated protein kinase 1, 2 (ERK1, 2) were analyzed by Western blot. Cell proliferation was tested by a 5-bromo-2-deoxyuridine (BrdU) incorporation assay. Cell cycle and proliferation index (PI) were analyzed by flow cytometry.Hypoxia significantly increased the expression of PCNA (0.528 ± 0.028), p-ERK1, 2 (1.12 ± 0.05, 0.91 ± 0.06), BrdU incorporation (143.3 ± 4.2) and cell proliferation index (12.5 ± 0.9) (all P < 0.05, versus control group, 0.243 ± 0.025, 0.47 ± 0.03, 0.40 ± 0.03, 100.0 ± 5.4, 7.5 ± 1.2). Gadolinium chloride (GdCl3, a CaSR agonist) amplified the effect of hypoxia (0.770 ± 0.039, 1.50 ± 0.06, 1.61 ± 0.05, 187.4 ± 3.9, 19.8 ± 0.6, all P < 0.05). PD98059 (a MEK1 inhibitor) decreased the up-regulation of PCNA expression, BrdU incorporation and the increase of cell proliferation index induced by hypoxia and GdCl3 in PASMCs (0.441 ± 0.020, 0.71 ± 0.07, 0.72 ± 0.06, 115.5 ± 4.0, 9.3 ± 1.1, all P < 0.05).Calcium-sensing receptor mediates hypoxia-induced proliferation of rat pulmonary artery smooth muscle cells through ERK1, 2 pathways.

To evaluate the diagnostic utility of bronchoscopy in solid organ transplant recipients with pulmonary opportunistic infections.From December 2006 to September 2011, 117 cases of bronchoscopy with protected specimen brush and bronchoalveolar lavage were performed in 114 solid organ transplant recipients at Tianjin First Central Hospital. The indication for bronchoscopy was suspected pulmonary infections. The bronchoscopic manifestations were described and the specimens analyzed with regards to bacteriology, cytomegalovirus, P carinii, mycobacterium tuberculosis and other fungal cultures.A definite infectious etiology was confirmed in 63 patients (53.8%). And opportunistic infections were the most frequent etiology (56/63, 88.9% including 7 cases with two mixed opportunistic infections). Among 63 pathogens, P carinii was demonstrated in 36 episodes, cytomegalovirus in 24 episodes and mycobacterium tuberculosis in 3 episodes. Bacterial infections (mainly Gram-negative) accounted for 16 of 63 episodes (25.9% including 9 cases with mixed opportunistic/bacterial infection). In accordance with the diagnostic results, antibiotic treatment was changed in 45 cases.As an extremely useful tool, bronchoscopy is of great value for pathogenic detection in transplant recipients with suspected pulmonary infections, especially for opportunistic infections. And the bronchoscopic findings may guide targeted antimicrobial therapy.

To explore the efficacies and safety of combined treatment of remifemin and paroxetine for perimenopausal depression.A total of 120 patients with perimenopausal depression were digital randomly divided into the treatment and control groups (n = 60 each). The treatment group received oral remifemin one tablet twice daily and paroxetine 20 mg once daily for 2 months while the control group oral paroxetine 20 mg once daily for 2 months. The Hamilton depression scale (HAMD) and Kupperman scale were used to assess the therapeutic efficacies. Blood and urine routine, electrocardiography, liver function, kidney function and blood pressure before and after treatment were examined to assess the side effects.For the improvement of perimenopausal depression on HAMD, the total effective rates of the treatment and control groups were 88.3% and 78.3% respectively. The therapeutic efficacy of the treatment group was significantly higher than that of the control group (P < 0.05). After 8-week treatment, Kupperman menopausal indices of the treatment and control groups were 9.89 ± 3.76 and 15.75 ± 5.84 respectively. There was also significant difference (P < 0.01). No significant changes existed in blood routine, urine routine, liver function, kidney function, blood pressure, ECG or blood pressure before and after treatment (P > 0.05).The combined treatment of remifemin and paroxetine for perimenopausal depression can improve the efficacies. It is easily accepted by patients for its higher safety and fewer side-effects. It is worthy of a wider application and further researches.

To explore the safety and efficiency of transurethral plasmakinetic enucleation of prostate (TUPKEP) and suprapubic small cut in the treatment of high-risk and senior patients with benign prostatic hyperplasia and bladder stones.A retrospective review was conducted for 68 high-risk and senior patients with benign prostatic hyperplasia and bladder stones. All of them were treated by TUPKEP and suprapubic small cut.Operation was successfully performed in all 68 cases. And there was no instance of transurethral resection syndrome, shock, myocardial infarct, cerebral infarction, cerebral hemorrhage, permanent urinary incontinence or surgical site infection. Seven patients with temporal urinary incontinence recovered at a mean time of (9.48 ± 1.52) days post-operation. The mean operative duration was (48.63 ± 4.14) min and the mean volume of blood loss (50.97 ± 5.33) ml. The changes of maximum flow rate (Qmax), international prostatic symptom score (I-PSS) and quality-of-life (QOL) were statistically significant before and after operation. Qmax increased from (4.56 ± 0.35) to (18.82 ± 1.65) ml/s (P < 0.001), I-PSS decreased form (21.96 ± 1.89) to (11.23 ± 0.86) (P = 0.018) and QOL decreased from (4.94 ± 0.35) to (1.95 ± 0.32) (P = 0.011).The approach of TUPKEP and suprapubic small cut is both safe and effective in the treatment of high-risk and senior patient with benign prostatic hyperplasia and bladder stones and should be widely applied.

To acquire the case constitution of bipolar disordered (BPD) in mood disordered (MD) outpatients at two local Beijing hospitals to understand the case constitution and medication intention of clinical psychiatrists in terms of differentiation and treatment of BPD.All psychiatrists at Anding Hospital and neurological physicians at Xuanwu Hospital were surveyed by self-rated questionnaires and the analysis focused on the composition of proportions of BPD cases out of MD patients. The items included estimated ratio of BPD cases, doctors' intention of drug prescription and clinical therapeutic regimen for treatment of BPD.(1) BPD ratio in MD outpatients of two hospitals were 41.79% and 12.24% respectively; (2) Doctor's ratio who estimated BPD ratio in MD cases < 40% were 60% and 100% respectively; (3) 100% psychiatrists at Beijing Anding Hospital and 72.22% neurological physicians at Beijing Xuanwu Hospital prescribed antidepressant to BPD cases; (4) Doctor's ratio who adopted therapeutic schedule of 'antidepressant + mood stabilizer' or 'antidepressant + mood stabilizer + antipsychotic' agents for BPD were 100% and 66.67% respectively.Regardless of mental specialized hospital or general hospital, the physicians should pay great attention to the differentiation of BPD and the rationality of drug prescription.