Objective:To investigate the effect of dimethyl amiloride (DMA) on invasive activity of PGCL3 cells from a human highly-metastatic lung carcinoma cell line in vitro and elucidate its possible mechanism. Methods:The invasion and migration capacities of PGCL3 cells were measured by using Transwell chamber assay after pretreatment with DMA. The effects of DMA on the activity of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) secreted by PGCL3 cells were measured by chromogenic substrate assay. The effects of DMA on uPA, urokinase-type plasminogen activator receptor (uPAR), and PAI-1 mRNAs transcription were determined by RT-PCR. The expression levels of extracellular regulated protein kinases 2 (ERK2) and ras protein were assessed by Western blot. Results:DMA inhibited invasion and migration capabilities of PGCL3 cells in vitro, down-regulated the mRNA transcription of uPA, uPAR and PAI-1, as well as up-regulated the expression of ras protein. After 24-hour treatment, DMA reduced the activity of uPA at higher concentration, but DMA had no effects on the activity of secreted PAI-1 protein and expression of ERK2 protein. Conclusion:DMA inhibits the invasion and migration of highly-metastatic lung cancer PGCL3 cells. The mechanism might be associated with down-regulation of the expression of uPA system.

Objective: To investigate the effect of amiloride on in vitro invasive activity and uPA (urokinase-type plasminogen activator)system of human highly metastatic lung carcinoma cell line PGCL3. Methods: At 6 hours after treatment with amiloride at the concentrations of 25μmol/L，50μmol/L and 100μmol/L for PGCL3 cells，Transwell Chamber assay was performed to detect the effect of amiloride on the invasive and migratory capacity of PGCL3 cells.Effect of amiloride on the activity of uPA and PAI-1(plasminogen activator inhibitor-1)secreted by PGCL3 cells were measured by chromogenic substrate assay after PGCL3 cells were incubated with amiloride for 24 hours.RT-PCR was used to analyze the effect of amilorede on mRNA levels of uPA，uPAR(urokinase-type plasminogen activator receptor)and PAI-1.The expression levels of uPA，ERK2(extracellular regulated protein kinases 2)and ras protein were assessed by Western blot. Results: The number of cells through membrane was significantly decreased in invasion and migration test in vitro.The inhibitory rates of invasion and migration after treatment with amiloride of 100μmol/L were 37.7%±4.1%and 64.9%±4.9%.respectively,with a significant difference from those in the control group(P<0.01).At 24 hours after amiloride treatment，the chromogenic substrate assay showed direct inhibition of the activity of uPA and PAI-1 secreted by PGCL3 cells.No effect on the expression of uPAR in mRNA level was observed,but the expression of PAI-1 in mRNA level was significantly inhibited.Amiloride of 100μmol/L dramatically inhibited the expression of uPA mRNA.The expression level of uPA protein was decreased with the increase of the concentration of amiloride,but no effect was observed on the expression of ERK2 and ras in protein level.Conclusion: Amiloride can inhibit the invasion and migration of PGCL3 cells,through inhibiting the expression and activity of uPA and PAI-1.Amiloride is a potential agent to inhibit cancer invasion and metastasis.

This paper summarized the assessment model reform practiced in the postgraduate cell biology course for several years and successfully obtain a new testing method based on research papers re-porting and questioning. Before the assessment, each student searched and selected one required research paper to read and make a PowerPoint reporting document. During assessment , some students were ex-tracted to speak and comprehensive score, which was regarded as student's final examination results was made by teaching group. The rest of the students listened to speeches , questioned and filled the questions on the record sheet.Teaching group reported a score , which was regarded as part of student's final exami-nation result based on questioning record sheets and printed PowerPoint documents.This method can well evaluate students' scientific thinking and strengthen the training of scientific thinking of postgraduates.

Objective To establish a CLCN3/MMTV-PyMT double transgenic mouse model of spontaneous breast tumor with simultaneously overexpressing ClC-3.Method CLCN3 transgenic mice were crossed with MMTV-PyMT spon-taneous mammary tumor model mice.The genotype was determined by PCR.The expression of ClC-3 in tissues was detec-ted by immunofluorescence and Western blot.Results CLCN3 and MMTV-PyMT transgenic mice were bred and CLCN3/MMTV-PyMT hybrid mouse model was successfully established.The ClC-3 expression in CLCN3/MMTV-PyMT hybrid mice was higher than that in the MMTV-PyMT mice, assessed by immunofluorescence and Western blot analysis.Conclu-sions Transgenic mouse models of spontaneous breast cancer with simultaneously overexpressing ClC-3 are successfully es-tablished.The double transgenic mice provide a good animal model for further research of ClC-3 in tumor growth and metas-tasis.

Background and purpose:Checkpoint kinase 1 and 2 have been proposed to be potential therapeutic targets to sensitize cancers to radioor chemo-therapeutics. However, little is known about whether Chk1/2 is also a suitable target for sensitizing cancers to curcumin. In the present study, we investigated effects of Chk1/2 siRNA on curcumin-induced apopotosis in hepatoma cell line Huh7 and evaluated the effectiveness of Chk1/2 as a therapeutic target to potentiate human hepatoma to curcumin. Methods:Effect of curcumin on the cell cycle checkpoint-associated proteins was detected by Western blot. The knockdown efficacy of Chk1/2 siRNA was measured by RT-PCR and Western blot. Effect of Chk1/2 siRNA on curcumin-induced apoptosis in Huh7 cells was evaluated by DAPI staining. Effect of Chk1/2 siRNA on cell cycle distribution in curcumin-treated Huh7 cells was analyzed by flow cytometry. Results:Curcumin significantly inhibited phosphorylation of cell cycle checkpoint-associtaed proteins Chk1(S317), Cdc25C(S216) and Cdk1(Y15). Chk1 siRNA decreased Chk1 mRNA and protein by 95% and 92% and Chk2 siRNA decreased Chk2 mRNA and protein by 60% and 55% respectively as compared with negative control siRNA (P

Background and purpose: Checkpoint kinase 1 and 2 have been proposed to be potential therapeutic targets to sensitize cancers to radio- or chemo-therapeutics. However, little is known about whether Chk1/2 is also a suitable target for sensitizing cancers to curcumin. In the present study, we investigated effects of Chk1/2 siRNA on curcumin-induced apopotosis in hepatoma cell line Huh7 and evaluated the effectiveness of Chkl/2as a therapeutic target to potentiate human hepatoma to curcumin. Methods: Effect of curcumin on the cell cycle checkpoint-associated proteins was detected by Westem blot. The knockdown efficacy of Chk1/2 siRNA was measured by RT-PCR and Westem blot. Effect of Chk1/2 siRNA on curcumin-induced apoptosis in Huh7 cells was evaluated by DAPI staining. Effect of Chk1/2 siRNA on cell cycle distribution in curcumin-treated Huh7 cells was analyzed by flow cytometry. Results: Curcumin significantly inhibited phosphorylation of cell cycle checkpoint-associtaed proteins Chk1(S317), Cdc25C(S216) and Cdk1(Y15). Chk1 siRNA decreased Chk1 mRNA and protein by 95% and 92% and Chk2 siRNA decreased Chk2 mRNA and protein by 60% and 55% respectively as compared with negative control siRNA (P<0.01). Inhibition of Chk1, but not Chk2, increased apoptotic rate from (21.3±1.8)% to (29.5±2.6)% (P<0.05). Neither Chk1 nor Cbk2 siRNA had any impact on cell cycle distribution in Huh7 cells induced by curcumin. Conclusion: Chk1 siRNA sensitized Huh7 cells to curcumin-induced apoptosis, suggesting that Chk1 is a potential therapeutic target to sensitize human hepatoma to curcumin.

gluconate in both G1 phase and S phase cells.The permeability of G1 phase cells to I-was higher than that in S phase cells,but to gluconate was lower than that in S phase cells.CONCLUSIONS: The density of the volume-activated Cl-current,the anion permeability of the channel and the sensitivity of the current to tamoxifen were different between the CNE-2Z cells in G1 phase and those in S phase.The results suggest that the expression of tamoxifen-sensitive,volume-activated chloride channels is differentiated at different stages of the cell cycle.

Objective:To investigate the clinical characteristics of children with high-risk acute lymphoblastic leukemia (ALL) receiving lasting remission after severe infection.Methods:The data of 3 children with high-risk ALL who were treated in Children's Hospital Affiliated of Zhengzhou University in 2014, 2015 and 2017 were analyzed retrospectively. The clinical and laboratory characteristics of all patients were also analyzed, and the relevant literatures were reviewed.Results:All 3 children were clinically classified as high-risk ALL with severe infection. A variety of anti-infective drugs and blood products were used in the treatment, and all achieved lasting remission.Conclusions:Children with high-risk ALL after severe infection can acquire lasting remission, which may be related with the production of infection stimulating inflammatory factors and cytokines to activate certain immune pathways, or various kinds of antibiotics, blood products participating in the immune regulation to make the body regain the immune surveillance function of the tumor cells.

S. The migration of CNE-2Z cells was inhibited by chloride channel blockers (ATP, NPPB and tamoxifen), but the inhibitory effect of the blockers varied with cells at different stages. CONCLUSIONS: The migratory ability is associated with the cell cycle in CNE-2Z cells. Chloride channels play an important role in cell migration of CNE-2Z cells.

Objective To compare the safety and efficiency of laparoscopic transabdominal preperitoneal (TAPP) and totally extraperitoneal (TEP) procedure for herniorrhaphy. Methods Between Jan 1997 and Jan 2005, 235 patients (274 hernias) underwent laparoscopic inguinal hernia repair with 163 TAPP in 139 patients and 111 TEP procedures in 96 patients. Postoperative patients were followed-up for 6-24 months. Results No conversion was necessary. There were no significant differences between TAPP and TEP when considering operating time, postoperative hospital stay, VAS and time to return to daily activity. The recurrence rate was 1. 8% and 0. 9% respectively in TAPP and TEP( P = 0. 524). The total incidence of postoperative complications were 15. 3% and 10. 8% respectively ( P = 0. 282 ) , with seroma, transient neurapraxia and urinary retention being 5. 5% vs. 4. 5% ( P = 0. 707 ) , 5. 5% vs. 3. 6% ( P = 0.464) and 3. 1% vs. 2.7%(P = 1.000). The inhospital cost was significantly higher in TAPP than in TEP(P=0.000). Conclusion TAPP and TEP are both safe and efficient tension-free technique. Surgeons' experience was more important than choice of approach to laparoscopic repair for groin hernias.