Objective: To explore the effect of activation of peroxisome proliferator-activated receptor gamma (PPARγ) on cell cycle arrest of gastric carcinoma cell line MGC803. Methods: The inhibitory effect of pioglitazone (PGZ) on proliferation of MGC803 cells was analyzed by MTT assay. Cell cycle were detected by flow cytometry (FCM). The protein expression of PPARγ, cyclinD1 and cell cycle protein-dependent kinase CDK4 in MGC803 cells was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Treatment with 0.1-10 μmol/L, PGZ for 96 h significantly inhibited cell proliferation. The proportion of MGC803 cells at G1 phase was significantly increased when treated with 10 μmol/L PGZ for 48, 72 and 96 h, and showed an apparent G1 phase arrest. The expression of PPARγ was at a low level in MGC803 cells and significantly up-regulated when treated with 10 μmol/L, PGZ for 48 h (P < 0.01). The expression of CDK4 in MGC803 cells was significantly down-regulated when treated with 10 μmol/L, PGZ for 96 h (P < 0.01) and the expression of cyclinD1 was slightly down-regulated. Conclusion: Activation of PPARγ significantly induces G1 phase arrest, which is associated with down-regulation of the expression of CDK4 and cyclinD1.

Child ; Humans ; Myelodysplastic Syndromes ; diagnosis ; Prognosis

AIM: To explore the skills and characteristics of corneal neovascular model in rat induced by micropocket assay. ·METHODS: Nine eyes of nine Sprague-Dawley rats were studied. Pellets made of vascular endothelial growth factor (VEGF), poly-2-hydroxylethylmethacrylate and sucralfate were implanted into the corneal stroma nocloser than 1mm from the limbus. Biomicroscopic features of corneal neovascular were observed on 1,3, 5, 7th day after the implantation. ·RESULTS: On day 1 after operation, the limbal vessels were dilated, with no angiogenesis appeared. On day 3, angiogenesis began to invade peri-cornea with a brush shape, the area of CNV was (2.23±0.59) mm2. On day 5, new vessels reached the lower margin of pellet densely, the area of CNVwas (6.81±1.35)mm2. On day 7, new vessels continued to elongate, parts of them extended as loops toward the pellet, the area of CNV was (8.92± 1.79)mm2. Neither hyphema or other complications occurred.·CONCLUSION: Corneal neovascular induced by micropocket assay in rat grows steadily, with no complication, and is suitable for quantitative researches.

Background Microvessels are composed of two interacting cell types: endothelial cells and pericytes. Over the past decades, studies of corneal angiogenesis have concentrated mainly on endothelial cells, while interest in pericytes has lagged behind. Objective The present study aimed to investigate the recruitment of vascular endothelial cells and pericytes in rat corneas after alkali burn. Methods Corneal alkali burn models were established in the right eyes of 36 adult SPF SD rats by putting 4 mm medicators containing a 1% 1 mol/L NaOH solution at the central corneas for 30 seconds, and 3 matched normal rats were used as controls. Corneas were excised 1,2,3,5,7 and 10 days after surgery. Frozen sections that parallel with the corneoscleral limbus were constructed. Double immunofluorescence staining was used to observe the dynamic expression of CD31 and α-smooth muscle actin (α-SMA) in corneal tissue for the evaluation of the number of endothelial cells and pericytes. The pericyte coverage index (PCI) was calculated to quantify the recruitment of pericytes to neovascular sites. The use of experimental animals followed the Statement of Association for Research in Vision and Ophthalmology. Results CD31 was expressed in the superficial stromal layer of the cornea on the 1 st day, showing the presence of red fluoresence. The positive cell number for CD31 was gradually increased with the passage of time and proceeded into the deep stromal layer from days 2 through 5 but decreased after that. However,α-SMA was positively expressed on the 2nd day in the cornea after alkali burn with the presence of green fluorescence. The positive cell number for α-SMA was less than those of CD31 throughout the experimental period. The PCI was 0, 16.07%, 11.95%, 43.84%, 73.97% and 86. 21% , respectively, 1,2,3,5,7 and 10 days after surgery. Conclusion Pericytes recruitment to corneal new vessels may play a key role in the stabilization and maturation of angiogeneis.

Cultured human testicular tissues were grouped as follows:control group,group tracted with single dosc of human chorionic gonadotropin(hCG), group treated with double doses of hCG given at three-day-interval and group treated with multiple doses of hCG. The dose of hCG was 20kIU/L,and it was added into the medium and incubated for 24 hours. Testosterone was measured by RIA. The result indicated that all three hCG-treated groups had a testosterone secretion peak at 48-72 hours after the first dose of hCG. After the second hCG treatment given in three days interval the testosterone increased again. but its maximal rise reduced. Multiple doses of hCG given successively inhibited the response of Leydig cells to hCG stimulation.

objective To detect CK19 mRNA and CEA mRNA in blood of the peripheral vein and to investigate the effect of ligating pulmonary vein firstly or ligating pulmonary artery firstly during surgical operation on haematogenous dissemination of malignant cells.Methods Fifty six non-small cell lung cancer patients were collected and random assigned to two groups before operation (ligating pulmonary vein firstly or ligating pulmonary artery firstly).The patients were accepted radical operation and their operations were put in practice by doctors of one team.Vein blood was collected one day before operation and one week after operation.CK19 mRNA and CEA mRNA in blood were detected by nested reverse transcriptase-polymerase chain reaction. Result The positive rate of expression of CK19 mRNA and CEA mRNA after operation is lower than that before operation.The positive rate of expression of CK19 mRNA and CEA mRNA in ligating pulmonary vein firstly after operation is lower than that in the other group. Conclusions Surgical operation have effects on the dissemination of malignant cell and ligating palmonary vein firstly during operation can reduce the dissemination of malignant cell.

OBJECTIVETo observe the effect of the signal pathway of phosphoinositol-3-kinase (PI3K)/Akt-antypical protein kinase C(iotazeta) (aPKC(iotazeta))-Nuclear factor-E2 related factor (Nrf2) on gamma-glutamylcysteine synthetase (gamma-GCS) of the bronchial epithelial cells of rats after exposure to cigarette smoke extracts (CSE).

METHODSgamma-GCS, Nrf2, p-Akt and p-aPKC(iotazeta) proteins were semi-quantified by Western blot. gamma-GCS protein expression was assessed by immunocytochemistry. gamma-GCS mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). Nrf2 protein was observed by immunofluorescence. The rate of the cells expressed p-Akt were analyzed by flow cytometry. GSH content and gamma-GCS activity were measured.

RESULTSGSH content, Nrf2 protein of nucleus, p-aPKC(iotazeta) protein, p-Akt protein and positive cells, gamma-GCS protein and mRNA and activity were significantly increased after exposure to CSE for 3 hours. aPK(iotazeta) inhibitor RO813220 significantly reduced the expression of p-aPKC(iotazeta) protein, gamma-GCS protein and mRNA and activity, but enhanced Nrf2 protein of cytoplasm expression, had no effect on p-Akt. p-Akt inhibitor LY294002 and RO813220 + LY294002 decreased p-aPKC(iotazeta) protein, p-Akt protein and positive cells, gamma-GCS protein and mRNA and activity expression, increased Nrf2 protein of cytoplasm expression. The correlation analysises demonstrated that there were a positive correlation between Nrf2 and gamma-GCS, p-Akt, p-aPKC(iotazeta), between p-Akt and Nrf2, p-aPKC(iotazeta), gamma-GCS, between p-aPKC(iotazeta) and Nrf2, p-Akt, gamma-GCS.

CONCLUSIONCSE might upregulate gamma-GCS expression through PI3K/Akt-aPKC(iotazeta)-Nrf2 signaling pathway in the bronchial epithelial cells of rats.

Animals ; Bronchi ; cytology ; enzymology ; Environmental Exposure ; adverse effects ; Epithelial Cells ; enzymology ; GA-Binding Protein Transcription Factor ; metabolism ; Glutamate-Cysteine Ligase ; genetics ; metabolism ; Isoenzymes ; metabolism ; Male ; Oncogene Protein v-akt ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Protein Kinase C ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tobacco Smoke Pollution ; adverse effects

Background and purpose:Δ133p53 can promote tumor cell growth, but the exact mechanism is not clear. This study was aimed to observe the expression and signiifcant of the p53 isoformsΔ133p53 and p53 gene downstream molecules MDM2 and cyclin G1 genes by 5-FU-MKN45 gastric cancer cell line model. Methods:Af-ter using different concentrations of 5-FU (50μg/mL, 100μg/mL) to human gastric cancer cell line MKN45, inhibition rate should be detected by MTT assay, the changes ofΔ133p53 mRNA, MDM2 mRNA and cyclin G1 mRNA express-ing were detected by reverse transcription-polymerase chain reaction (RT-PCR). Differences between these groups were analyzed by ANOVA, comparisons within groups were analyzed by t-test, bivariate correlation was analyzed by Pearson linear correlation. Results:MTT results showed that with the increased concentration of 5-FU and the extension of time, the cell inhibition rates increased gradually. The inhibition rates of 50μg/mL 5-FU were 41.10%, 54.79%and 68.48%, for culturing 24, 48 and 72 hours. There were statistically signiifcant differences between the groups(F=45.52, P=0.00). The inhibition rates of 100μg/mL 5-FU were 69.53%, 78.21%and 86.92%, for culturing 24, 48 and 72 hours. There were statistically signiifcant differences between the groups(F=85.58,P=0.00). The inhibition rates of 50 and 100μg/mL were 41.10%and 69.53%, for culturing 24 hours. There were statistically signiifcant differences between the groups(F=51.29, P=0.00). The inhibition rates of 50 and 100μg/mL were 54.79%and 78.21%, for culturing 48 hours. There were statistically signiifcant differences between the groups(F=51.29, P=0.00). The inhibition rates of 50 and 100μg/mL were 68.48%and 86.82%, for culturing 72 hours. There were statistically signiifcant differences between the groups(104.91, P=0.00). RT-PCR results showed that with the increase of the concentration of 5-FU, theΔ133p53 mRNA, MDM2 mRNA and cyclin G1 mRNA expression gradually declined in gastric cancer cell line MKN45 cells, and there were statistically signiifcant differences between the groups(F=738.532, 1 396.607, 2 785.56,P=0.00). Cor-relation analysis showed that the expressions ofΔ133p53 mRNA and MDM2 mRNA in gastric cancer were positively correlated (r=0.871, P=0.01), while the expression of cyclin G1 mRNA and p53 mRNA had no obvious relevance (P=0.13). Conclusion:In the 5-FU-MKN45 gastric cancer cell line model, anti-tumor pathway ofΔ133p53 isomers is related with MDM2 but was not related with cyclin G1.

Background and purpose: Little about the function of p53 isoforms in gastric cancer was reported. This study was designed to explore the role ofΔ133p53 in the effect of recombinant mutant human tumor necrosis factor (rmhTNF) on gastric cancer cells, and provide a new basis for the diagnostics and therapeutics of gastric carcinoma. Methods: MKN45 (withΔ133p53 expression) or SGC7901 (withoutΔ133p53 expression) cells were treated with rmhTNF of different concentrations only or combined with 5-FU (a traditional gastric cancer cellular killer), and the growth inhibition rate and apoptosis was detected by CCK-8 and lfow cytometry. mRNA expressions ofΔ133p53, Gadd45αand CyclinB1 were measured by nested reverse transcription-polymerase chain reaction (nRT-PCR) or real-time polymerase chain reaction(RT-PCR). Results:On MKN-45 cells with positiveΔ133p53 expression, the inhibitory effect of rmhTNF was signiifcant, the inhibition rates of 50 and 500 IU/mL rmhTNF were 24.82%, 72.33%after culturing for 24 h (t=-9.558, P<0.01);also, the inhibitory effect of 5-FU was improved by rmhTNF remarkably in time-and dose-dependence, the inhibition rates of 5-FU (25 μg/mL), rmhTNF (50 IU/mL) combined with 5-FU (25 μg/mL), rmhTNF (500 IU/mL) combined with 5-FU (25 μg/mL) were 18.20%, 48.66%, 59.83%, separately, after culturing for 24 h (F=82.742, P<0.01);the inhibition rates of rmhTNF (50 IU/mL) combined with 5-FU (25 μg/mL) were 48.66%, 68.20%, 85.23%, separately, after culturing for 24 h, 48 h and 72 h (F=128.583, P<0.01). However, on SGC-7901 cells with negativeΔ133p53 expression, no growth inhibition was showed by rmhTNF only, the inhibition rates of 50 and 500 IU/mL were 2.74%, 3.25%after culturing for 24 h (t=-0.121, P>0.05). In apoptosis test, the apopto-sis-enhancing effect of rmhTNF was signiifcant on MKN45 cells, and the apoptosis-enhancing effect of 5-FU was fur-ther promoted signiifcantly by rmhTNF, the apoptosis of rmhTNF (50 IU/mL), rmhTNF (50 IU/mL) combined with 5-FU (25 μg/mL), rmhTNF (500 IU/mL) combined with 5-FU (25 μg/mL) were 18.20%, 48.66%, 59.83%, separately, after culturing for 24 h (F=123.931, P<0.05). In mRNA measurement, down-regulation ofΔ133p53 and CyclinB1, up-regula-tion of Gadd45αwere signiifcant in MKN45 cells treated by rmhTNF alone or combined with 5-FU. In nRT-PCR anal-ysis, the mRNA levels ofΔ133p53 were relatively 0.886, 0.499, 0.330, 0.161 (F=240.927, P<0.01);In real-time PCR analysis, the mRNA levels of Gadd45αwere 1.227, 1.694, 3.394, and the mRNA levels of CyclinB1 were 1.221, 0.722, 0.316, relatively. The expression ofΔ133p53 was positively related to CyclinB1 (r=0.977, P<0.01), but negatively re-lated to Gadd45α(r=-0.950, P<0.01). Conclusion:InΔ133p53 positively expressed MKN45 cells, rmhTNF showed as an effective tumor inhibitor and an enhancer of 5-FU as well, and this effect might be helped by two p53 down-stream molecules CyclinB1 and Gadd45α. The results suggest thatΔ133p53 might be a key target for the biological effect of rmhTNF against gastric cancer.

Influenza caused by influenza virus,seriously threaten human life and health.Drug treatment is one of the effective measurement. However, there are only two classes of drugs, one class is M2 blockers and another is neuraminidase (NA)inhibitors. The recent antiviral surveillance studies reported a global significant increase in M2 blocker resistance among influenza viruses, and the resistant virus strains against NA inhibitor are also reported in clinical treatment.Therefore thediscovery of new medicines with low resistance has become very urgent.As all known,traditional medicines with multi-target features and network mechanism often possess low resistance. Compound Yizhihao, which consists of radix isatidis,folium isatidis,Artemisia rupestris,is one of the famous traditional medicine for influenza treatment in China, however its mechanism of action against influenza is unclear. In this study, the multiple targets related with influenza disease and the known chemical constituents from Compound Yizhihao were collected, and multi-target QSAR (mt-QSAR) classification models were developed by Na?ve Bayesian algorithm and verified by various datasets. Then the classification models were applied to predict the effective constituents and their drug targets.Finally,the constituent-target-pathway network was constructed,which revealed the effective constituents and their network mechanism in Compound Yizhihao. This study will lay important basis for the clinical uses for influenza treatment and for the further research and development of the effective constituents.