OBJECTIVETo investigate the effects of prostate stromal cells from different zones of normal prostate tissue on the growth of prostate cancer cells and their action mechanisms.

METHODSWe extracted stromal cells in the fresh normal prostatic tissue derived from the peripheral zone (PZ) or transitional zone (TZ), amplified them in vitro, and used the supernatants of the cells as conditioned media to culture hormone-resistant prostate cancer DU145 cells. We measured the growth curve of the tumor cells using the CCK8 method, determined the number and viability of the cells by trypan blue staining, evaluated their invasiveness by scratch test, and detected the effects of the stromal cells on the key enzymes in the glycolysis of the tumor cells by Western blot.

RESULTSThe conditioned medium with the PZ-derived stromal cells promoted, while that with the TZ-derived stromal cells inhibited the growth of the tumor cells. The former significantly increased, while the latter markedly decreased the expressions of the key enzymes hexokinase 2 (HK-2), pyruvate kinase 2 (PKM-2), lactate dehydrogenase (LDHA), and pyruvate dehydrogenase (PDH) in the glycolysis of the tumor cells.

CONCLUSIONProstate stromal cells from different zones exert different influences on the growth of tumor cells, which may be associated with their different effects on the glycolysis of tumor cells.

Blotting, Western ; Cell Culture Techniques ; Cell Proliferation ; Culture Media, Conditioned ; Glycolysis ; Humans ; Male ; Prostate ; cytology ; Prostatic Neoplasms ; pathology ; Stromal Cells ; physiology ; Tumor Cells, Cultured

ObjectiveTo evaluate prognostic factors and the surgical results of pulmonary carcinoid tumors.Methods We retrospectively reviewed the medical records of 62 patients who were diagnosed as pulmonary carcinoid tumors between January 2000 and October 2010 at Department of Thoracic Surgery,Shanghai Chest Hospital.The following information was available for each of the 62 patients:age,sex,pathological type,and TNM stage.ResultsThere were no operative death.The 3-year and 5-year survival rates after surgery were 92.1％ and 77.8％,respectively.Of the 62 patients,42 were diagnosed as typical carcinoid tumor,and among them,4 patients (8.3％) had lymph node metastases.Their 3-year and 5-year survival rates were 97.8％ and 94.7％,respectively.The remaining 20 patients were diagnosed as atypical carcinoid tumor,and among them,6 patients (37.5％) had lymph node metastases.Their 3-year and 5-year survival rates were 84.4％ and 58.8％,which were statistically significant compared with typical carcinoid tumor( P =0.0047 ).There was significant difference in survival rate between the patients with lymph node metastases and the patients without lymph node metastases (P =0.0048).CondusionThe main risk factors affecting survival rate of those patients who were diagnosed as pulmonary carcinoid tumors were pathological types and lymph node metastases.

ObjectiveTo determine whether the inhibition of caveolin-1 tyrosine residues 14 (Cav-1-Y14) phosphorylation with protein tyrosine kinase inhibitors (PP2) will upregulate heme oxygenase-1 (HO-1) activity to protect against ventilation induced lung injury in vivo of an animal model.Methods Fifty-four male Sprague-Dawley (SD) rats were randomly divided into nine groups (eachn = 6). Group A served as normal control group, in which rats did not receive ventilation but tracheotomy. Groups B1 and B2 received lung protective ventilation respectively for 1 hour or 2 hours. Groups C1 and C2 received high tidal volume (40 mL/kg) ventilation for 1 hour or 2 hours, respectively. The group D1 or D2 also received high tidal volume ventilation for 1 hour or 2 hour respectively, but they were given PP2 1 hour before high tidal volume ventilation. The groups E1 and E2 also received high tidal volume ventilation respectively for 1 hour or 2 hours, but tyrosine kinase inhibitor PP2 and HO-1 inhibitor zinc protoporphyrinⅨ(ZnPPⅨ) were given to animals 18 hours before high tidal volume ventilation. All the animals were sacrificed after ventilation, and the specimens of lung tissues and bronchoalveolar lavage fluid (BALF) were harvested. Then the changes in pathology of lung tissue was observed, and diffuse alveolar damage scores (DAD) were calculated, myeloperoxidase (MPO) activity was measured by colorimetric analysis, lung wet/dry ratio (W/D) was estimated. The expressions of phosphorylated caveolin-1 (P-Cav-1-Y14), caveolin-1 (Cav-1) and HO-1 were determined by Western Blot. The expressions of high mobility group B1 (HMGB1) and advanced glycation end product receptor (RAGE) in lung tissues were assayed with immunohistochemistry staining. The levels of tumor necrosis factor-α(TNF-α) in BALF were measured by enzyme linked immunosorbent assay (ELISA).Results There was no significant difference in all the parameters between group A and groups B. Compared with group B1, DAD score, W/D ratio, the activity of MPO and the concentration of TNF-α in BALF in group C1 were significantly increased [DAD score:7.97±0.59 vs. 0.55±0.13, W/D ratio: 5.70±1.61 vs. 5.04±0.63, MPO (U/g): 1.82±0.14 vs. 0.77±0.26, TNF-α(ng/L): 370.10±29.61 vs. 54.38±8.18, allP< 0.05], and the injury in ventilation 2 hours group was more serious than that in ventilation 1 hour group. Compared with groups C, all the parameters in groups D were significantly decreased. The parameters in groups E were significantly higher than those in groups A, B, and D, but no significant difference was found as compared with groups C. Compared with groups B, the protein expressions of Cav-1 and P-Cav-1-Y14 (gray value) in groups C were significantly increased (1 hour: 1.49±0.02 vs. 1.26±0.13, 1.34±0.02 vs. 0.87±0.04;2 hours: 1.58±0.02 vs. 1.27±0.27, 1.31±0.01 vs. 0.95±0.02, allP< 0.05), and the expression of HO-1 protein (gray value) was significantly decreased (1 hour: 0.59±0.02 vs. 1.10±0.01, 2 hours: 0.49±0.01 vs. 1.20±0.02, both P< 0.05). No significant difference in Cav-1 protein expression between groups D as well as groups E and groups C. The protein expression of P-Cav-1-Y14 in groups D and E was significantly lower than that in groups C. The protein expression of HO-1 in groups D was significantly higher than that in groups C, but the phenomenon was not found in groups E as compared with groups C. Compared with group A, the positive expression of HMGB1 and RAGE in lung tissue in groups C and E was significantly increased, but no significant difference was found between groups B as well as groups D and group A.Conclusion Cav-1-Y14 phosphorylation is the key factor for ventilator induced lung injury, which can not only lead to a decrease in vascular barrier function, but also inhibit the activity of HO-1 enzyme, thus further aggravates inflammatory injury of the lung as induced by mechanical ventilation.

Objective To investigate the clinical effect of heart failure recombinant human brain natriuretic peptide (rhBNP) in treatment of chronic pulmonary heart disease (CPHD).Methods Fifty-six CPHD patients with heart failure were randomly divided into control and research group who were hospitalized from January 2010 to December 2011.Patient in two groups were given oxygen,anti-infection,nutritional support and complications treatment.In addition patients in the treatment group was treated with rhBNP.Clinical symptoms,signs and cardiac,pulmonary function of two groups were recorded.Results The pulmonary artery pressure in treatment group were (39.7 ± 6.2) mm Hg and (26.5 ± 3.8) mm Hg before and after treatment,and the difference was significant(t =14.992,P =0.000).The pulmonary artery pressure in control group were (38.4 ±5.1) mm Hg and (31.5 ±4.5) mm Hg before and after treatment,and significant difference were seen (9.378,P =0.000).In addition,pulmonary artery pressure were different between in treatment and control group(t =-9.742,P =0.000).The level of BNP in treatment group was (873.0 ± 12.9) ng/L and (382.0 ± 11.4) ng/L,there was significant difference(t =353.627,P =0.000) ;While in control group,the level of BNP was (862.0 ± 12.3) ng/L and (568.0 ± 12.6) ng/L before and after treatment,and the difference was significant(t =156.135,P =0.000).And there was sinificant difference between the two groups after treatment (t =-103.490,P =0.000).The left ventricular ejection fraction before and after treatment in treatment group was (38 ±9)％ and (65 ±8)％,and the difference was significant(t =-23.056,P =0.000) ;While in control group,the Left ventricular ejection fraction was (32 ± 7) ％ and (47 ± 5) ％ before and after treatment,and the difference was significant (t =-16.485,P =0.000).And the difference between two groups was significant(t =18.308,P ＜ 0.01).24 h urine volume in treatment group was (0.9 ± 0.4) L and (1.6 ± 0.3) L before and after treatment,and the difference was significant(t =-17.320,P =0.000) ;While in control group,24 h urine volume was(0.9 ± 0.2) L and (1.0 ± 0.6) L before and after treatment,and the difference was significant (t =-5.250,P =0.000).And the difference between two groups was significant (t =6.592,P =0.000).The total effective rate in treatment was 82.2％ (23/28),higher than that in the control group (57.1％ (16/28),and the difference was significant(x2 =4.139,P ＜ 0.05).Conclusion rhBNP can improve heart function of CPHD patients with heart failure.

Objective To investigate the protective effects and molecular mechanism of peroxisome proliferate-activated receptor α(PPARα) activation on acute myocardial damage induced by isoproterenol (Iso) in rats. Methods Thirty male Wistar rats, weighting 160～180 g, were randomly divided into control group, Iso group, fenafibrate(FF) group(each n=10) according to physique quantity. Acute myocardial injury caused by Iso abdomen cavity injection induced ischemia was established and the protective effects of peroxisome proliferate-activated receptor α activation were accessed by the level of ereatine kinase(CK), lactic dehydrogenase(LDH) in serum as well as the activities of myoperoxidase(MPO) in myocardium, and the protein expressions of PPABα in myocardium by Western blot. Results The level of serum CK in control group, lso group and FF group, was (62.41±9.47),(101.71±11.05),(75.64±11.73)kU/L, respectively(F= 34.34, P<0.01). Whereas the level of serum CK in Iso group and FF group was higher than that in control group(P<0.01 or<0.05), the level of serum CK in FF group was lower than that in Iso group(P<0.01). The levels of LDH in these three groups were (5912.20±204.44), (6365.78±137.10), (6089.76±169.60) U/L, respectively(F= 17.54, P<0.01). Compared with the control group, the levels of LDH in Iso and Fir groups were significantly increased(P<0.01 or<0.05). But the level of LDH in FIr group was decreased compared with that in Iso group(P<0.01). The activities of myocardial MPO in these three groups were (1.95±0.10),(3.89±0.17),(2.49±0.19)U/g, espectively(F=391.68,P< 0.01). The activities of myocardial MPO in Iso and FF groups were higher than that in the control group (all P< 0.01), while the activities of myocardial MPO in FIr group were lower than that in lso group(P<0.01). The protein expressions of PPARα in myocardium of these three groups were 251.57±10.95,191.97±10.74,215.08±9.61, respectively(F=82.69, P<0.01). Conclusion PPARα activation by its actor FF can exert protective effects on the acute myocardial ischemia injury induced by lso in rats through inhibiting the release of inflammatory cell factors.

Objective To establish a gene diagnosis assay for spinal muscular atrophy(SMA) in children. Method Analysis of the survival motor neuron (SMN) gene in 19 SMA patients and in 21 normal controls were performed by using polymerase chain reaction - fragment length polymorphism (PCR - RFLP) method. Result Deletion of exon 7and 8 in SMNt gene were found in all 19 SMA patients, while no such changes were found in normal controls. Conclusion The SMNt gene exon 7 and 8 examine can be applied to SMA gene diagnosis, and the PCR- RFLP method have higher sensitivity and particularity to the SMA diagnosis.

A simple and rapid method was established for the determination of the content of L-malic acid in fermentation liquor. This procedure includes six steps: (1) Use a microi- njector to inject the fermentation liquor quantitively on the filter paper, then use the chromatography solution (n-butanol: formic acid: water=8:1.5:1) for paper chromatography. (2) Atomize the display reagent (10ml 0.02?10~(-2) bromocresol green: 0.2ml 10?10~(-2) sodium hydroxide) on the chromatography paper. (3) Scissor off L-malic acid spots from the paper and put in into 5 ml distilled water for three hours. (4) In 1ml eluent, add 6 ml 96?10~(-2) sulphuric acid and 0.1ml 1.0?10~(-2) ?-naphthol.(5) Heat the reaction system in 100℃ water for sixty minutes. (6) After cooling determine the optical density at the wavelength 476 am in the colorimeter.

OBJECTIVE To investigate the effect of furanodiene(FDE),a diterpene derived from the medicinal plant Zedoary,on apoptosis of human gastric cancer MGC-803 cells induced in vitro. METHODS MGC-803 cells were treated with FDE 46.29~740.74μmol·L-1 for 24,48 and 72 h,and the cell viability was detected with MTT assay. Cell morphology was observed by light microscopy and Hoechst33342 staining. Flow cytometry was used to detect cell apoptotic rate and cell cycle. Rh123 staining and fluorescence probe DCFH-DA were employed to detect the changes in mitochondrial membrane potential (MMP) and reactive oxygen species(ROS). RESULTS MTT Results showed that FDE 46.29-740.74μmol · L-1 exhibited significantly higher cytotoxicity to gastric cancer MGC-803 cells. IC50 for MGC-803 of 24,48 and 72 h treatment was 347.91,257.41 and 101.01μmol·L-1,respectively. Treatment with FDE 92.58-370.32μmol·L-1 for 24 h also caused significant morphological changes in MGC-803 cells. AnnexinⅤ-FITC/PI double staining showed that the apoptotic rate increased after FDE 92.58-370.32μmol·L-1 treatment for 24 h(P<0.05). FDE enabled MGC-803 cell cycle arrest in S phase. DCFH-DA staining showed that FDE resulted in an increase in intracellular ROS levels(P<0.05) when PDE concentration was 370.37μmol·L-1(P<0.05). MMP decreased after FDE treatment when PDE concen?tration was 370.37μmol·L-1(P<0.05). CONCLUSION FDE Possesses potent tumor selected toxicity and can induce apoptosis of MGC-803 cells through cell cycle arresting,which is related to inhibition of DNA biosynthesis.

AIM: To investigate the effect of human amniotic fluid on the release of thromboxane A 2 (TXA 2), prostaglandin I 2 (PGI 2) and Leukotriene C 4(LTC 4) from blood cells. METHODS: 1 mL human amniotic fluid and 10 mL oneself blood collected from 38-41 weeks with cesarean section were cultured at 37℃ for 30 min, and then centrifuged. The supernatants were taken and stored at -70℃. TXB 2 and 6-Keto-PGF 1? of the superntants were determined by radioimmunoassay and LTC 4 by enzyme immunoassay. RESULTS: It was found that the levels of TXB 2 and LTC 4 in blood were elevated from (63.5?52.0) ng/L and (40.1?39.2) ng/L to (189.1?102.0) ng/L and (293.5?206.1) ng/L respectively (P0.05).CONCLUSION: Amniotic fluid might stimulate the release of TXA 2 and LTC 4 from blood, it might affect the balance of TXA 2 and PGI 2 in blood, which might play an important role in the pathogenesis of amniotic fluid embolism.

Objective To investigate whether the inhibition of caveolin-1 (Cav-1) phosphorylation will regulate effectively nuclear factor-erythroid 2-related factor (Nrf2) signal pathway and downstream effector molecules and protest against ventilation induced lung injury (VILI) in an animal model in vivo. Methods Ninety male Sprague-Dawley (SD) rats were randomly divided into nine groups (each n = 10): sham group in which rats did not receive ventilation but received tracheotomy; lung protective ventilation (PV) for 1 hour or 2 hours group; mechanical ventilation (MV) at high volume tidal (VT, 40 mL/kg) for 1 hour or 2 hours group; protein tyrosine kinase inhibitor PP2 or rosiglitazone (Rsg) pretreatment + high VT ventilation for 1 hour or 2 hours groups. The two pretreatment groups were given intraperitoneal injection PP2 15 mg/kg or intragastric administration of Rsg 5 mg/kg 1 hour before ventilation respectively. The rats were sacrificed after model reproduction, and bronchoalveolar lavage fluid (BALF) was collected. Pulmonary vascular permeability was measured by Evans blue (EB). The levels of tumor necrosis factor-α (TNF-α), activator protein-1 (AP-1), nuclear factor-κB (NF-κB), and interleukin-8 (IL-8) in BALF were determined by enzyme linked immunosorbent assay (ELISA). Then the lung tissues were collected, the lung wet/dry ratio (W/D) was calculated, the changes in pathology was observed with light microscope, and myeloperoxidase (MPO) activity was determined by colorimetric analysis. Nrf2 mRNA was determined by reverse transcription-polymerase chain reaction (RT-PCR). The expressions of Cav-1 tyrosine residues 14 phosphorylation (pCav-1-Y14), Cav-1, peroxisome proliferators-activated receptor γ (PPARγ) and claudin-5 as well as Nrf2 in cytoplasm and nucleus were determined by Western Blot. The positive expressions of PPARγ and claudin-5 in lung tissues were assayed with immunohistochemistry staining. Results There were no obvious pathological changes in the lung tissue in sham group and PV groups, and there were no significant differences in all the parameters between the two groups either. However, the injury in lung tissue was severe in the high VT groups in which W/D ratio, EB contents, MPO activity, and TNF-α, AP-1, IL-8, NF-κB levels in BALF as well as the protein expressions of Cav-1 and pCav-1-Y14 were significantly higher than those of sham group and PV groups, and the protein expressions of PPARγ and claudin-5 were significant lower than those of sham group and PV groups with a dose-dependent manner; but Nrf2 expressions in cytoplasm and nucleus did not show a statistical increase. After pretreatment of PP2 or Rsg, W/D ratio, MPO activity, EB contents, TNF-α, AP-1, IL-8, and NF-κB in BALF were significantly decreased as compared with those of high VT group, and RT-PCR showed significant up-regulation of Nrf2 mRNA in lung tissues too. Moreover, there was a statistically significant increase in expressed Nrf2 proteins in nucleus in PP2 or Rsg groups as compared with those of high VT groups [Nrf2 in nucleus (gray value): 0.61±0.06, 0.56±0.06 vs. 0.31±0.02 at 1 hour, 0.38±0.06, 0.43±0.07 vs. 0.22±0.03 at 2 hours; all P < 0.05], but no significant difference was found in the expression of Nrf2 protein in the cytoplasm among all groups. The protein expressions of pCav-1-Y14 in PP2 pretreatment groups were significantly lower than those of high VT groups (gray value: 0.89±0.04 vs. 1.48±0.02 at 1 hour, 0.86±0.02 vs. 1.31±0.01 at 2 hours; both P < 0.05); but expressed PPARγ proteins and expressed claudin-5 proteins in PP2 or Rsg pretreatment groups were significantly higher than those of high VT groups [PPARγ (gray value): 0.34±0.07, 0.42±0.13 vs. 0.17±0.07 at 1 hour, 0.38±0.09, 0.33±0.07 vs. 0.16±0.03 at 2 hours; claudin-5 (gray value): 0.33±0.05, 0.38±0.07 vs. 0.14±0.03 at 1 hour; 0.30±0.06, 0.31±0.04 vs. 0.17±0.04 at 2 hours; all P < 0.05]. Conclusions The inhibition of Cav-1-Y14 phosphorylation can increase the expression of Nrf2 in the nucleus, then result in an increase in the protein expressions of PPARγ and claudin-5 of its effector molecules. This effect can reduce the inflammation and capillary permeability of lung tissue in the model of VILI.