Objective To quantify the mRNA of 5-lipoxygenase (5-LO) in the rat peritoneal macrophages cultured in vitro by capillary electrophoresis (CE). Methods The rat peritoneal macrophages were isolated and cultured for indicated time. The mRNA of 5-LO was detected by RT-PCR, and the products of RT-PCR were quantified by CE. Results The DNA fragments in the 100 bp DNA marker and the products of the RT-PCR were separated successfully by CE with the sieving buffer containing 1.8% hydroxypropylmethylcellulose (HPMC). It proved that there were no significant changes on the expressions of 5-LO mRNA when the cells were cultured for 72 h quantified by CE. The mRNA of 5-LO significantly decreased by almost 80% by CE with the cells cultured for 120 h in vitro.Conclusions The products of RT-PCR could be separated and quantified by CE directly.The 5-LO mRNA could express normally in the rat peritoneal macrophages for 72 h in vitro.

Objective To investigate the effect of activation of protein kinase B(PKB) and Caspase-9 signal transduction pathway of human gastric cancer cells on the cell growth and chemosensitivity to etoposide.Methods The gastric cancer cells SGC7901 were treated with etoposide or etoposide plus PKB inhibitor Wortmannin at different time.The growth rates of gastric cancer cells SGC7901 and their sensitivity to etoposide were examined by 3-(4,5-dimethylthiazol-2,1)-2,5 diphanytetrazolium(bromide) assay.Apoptosis of gastric cancer cells was(detected) by flow cytometry.PKB activity was measured by(immunoprecipitation.) Caspase-3 expression and Caspase-9 activity were determined by Western bolt analysis.Results Etoposide induced apoptosis of SGC7901 cells and inhibited its survival effectively,which was much(weaker) 12 h after treatment.PKB(activity) became higher gradually,and Caspase-3 expression,Caspase-9(activity) significantly reduced at 12 h treated with etoposide.(Conversely),after pretreated with Wortmannin,PKB activity remarkably(reduced,) and Caspase-3(expression),Caspase-9 activity markedly increased.(Wortmannin) suppressed growth and potentiated (apoptosis) caused by etoposide.Potentiation of apoptosis by Wortmannin(correlated) with etoposide-induced PKB and Caspase-9 phosphorylation.Conclusions PKB and Caspase-9 signal transduction pathway promotes(human) gastric cancer cells survival and resistance to(chemotherapy.) PKB inhibitor can enhance sensitivity of gastric cancer cells to chemotherapy.

Under the patterns of net-teaching,the construction of net-teaching database is an important tache to develop teaching favourably and improve the quality and efficiency of teaching quickly.This paper introduces the process of constructing a net-teaching database for medical haematology,the course learning of clinical hematology for the undergraduates and graduate students and the applications of the clinician in learning.

Objective To understand the roles of JAK STAT pathway in the process of differentiation of human cord blood CD34 + hematopoietic stem cells into dendritic cells (DCs) by detecting the expressions and activation of JAK2 and STAT5. Methods CD34 + hematopoietic stem cells isolated from human umbilical cord blood and cultured for two weeks were induced to differentiate into DCs in vitro . Total cellular JAK2 and STAT5 and tyrosine phosphorylated protein stimulated by granulocyte/macrophage colony stimulating factor (GM CSF) at different time points (0, 7, and 14) during DC differentiation were detected by Western blotting. Results The amount of JAK2 protein was similar at 0, 7, and 14 d without GM CSF stimulation. With the differentiation of cells into DCs, the amount of tyrosine phospho JAK2 induced by GM CSF increased markedly. Both total cellular and tyrosine phospho STAT5 expression increased markedly during DC differentiation. Maximal tyrosine phospho STAT5 expression was later than JAK2. Conclusion JAK STAT pathway may take part in the signal mechanism of DCs differentiation from CD34 + hematopoietic stem cells stimulated by GM CSF.

Objective:To compare the effects between electroacupuncture (EA) at Chize (LU 5, the He-Sea point of the Lung Meridian) and Shangjuxu (ST 37, the lower He-Sea point of the large intestine) in rats with ulcerative colitis (UC) on the variations of mesenteric microcirculation and vasoactive intestinal peptide (VIP) in the colon, lung, and hypothalamus. The relative specificity of acupoints was also explored.
Methods: A total of 28 male Wistar rats were randomized into a normal group, a model group, a Chize (LU 5) group and a Shangjuxu (ST 37) group, 7 rats in each group. The UC model was established by enema with acetic acid. Since the third day after modeling, rats in the Chize (LU 5) group and Shangjuxu (ST 37) group respectively received EA at Chize (LU 5) and Shangjuxu (ST 37), 15 min each time for successive 7 d. The variations of mesenteric microvascular calibers and blood flow status were observed by a microcirculation microscopic tester; VIP in the colon, lung and hypothalamus was measured by radioimmunoassay.
Results:Compared with the normal group, the mesenteric microvascular calibers were significantly expanded in the model group (P<0.05); there was no significant difference between the model group and Chize (LU 5) group (P>0.05); compared with the model group and Chize (LU 5) group, the calibers were obviously shrunk in Shangjuxu (ST 37) group (P<0.05). The four groups showed no significant inter-group differences in comparing blood flow status (P>0.05). The colonic VIP levels in the model group and Chize (LU 5) group were significantly higher than that in the normal group (P<0.01,P<0.05); the VIP level in Shangjuxu (ST 37) group was markedly lower than that in the model group (P<0.01). There were no significant differences among the four groups in comparing VIP level in lung and hypothalamus (P>0.05).
Conclusion:The effects of Chize (LU 5) and Shangjuxu (ST 37) were different in treating UC. Shangjuxu (ST 37) showed a more significant efficacy in down-regulating VIP in the colon and regulating mesenteric microcirculation, while the effects of Chize (LU 5) were not obvious.

AIM:To establish an HPLC-ELSD method for determing dulcitol and astragaloside in Compound Fufangteng Capsules(Radix et Rhizoma Ginseng rubra,Radix Astragali,Herba Euonymi,etc).METHOD:Chro-matographic conditions included Lichrospher 5-NH_2 column(250 mm?4.6 mm,5 ?m)and the mobile phase consisting of a mixture of acetonitrile-water(91 ∶9).The flow rate of mobile phase was 1.0 mL/min.The column tempe-rature was at 28 ℃.Detector:PL-ELS2100 ELSD(Eva:65 ℃,Ned:50 ℃,gas flow:0.9 L/min).RESULTS:The linear ranges of dulcitol and astragaloside were 2.91-29.1 ?g(r= 0.999 8)and 1.03-10.3 ?g(r= 0.999 1),respectively.The average recoveries of dulcitol and astragaloside were 99.24% and 103.17% with corresponding RSD of 2.6 % and 1.8%(n=6),respectively.CONCLUSION:The method is steady and with good repeatability,and can be used to determine the content of dulcitol and astragaloside in Compound Fufangteng Capsules.

Objective To investigate the effect of methylation of the APC gene on expression and the correlation with clinical data in pancreatic cancer.Methods Sixty postoperative tissue samples with pancreatic cancer were collected in the First Affiliated Hospital of Zhengzhou University from August 2010 to January 2011,20 benign pancreatic disease tissues were collected as control groups.APC promoter methylation and gene expression levels were detected by Methylation Specific PCR (MSP),Real Time PCR (RT-PCR) and Western blot in 60 pancreatic carcinoma,42 metastasis and 20 benign pancreatic disease tissues,then analyze the relation between methylation of the APC gene and the clinical data.Results APC promoter methlation was observed 48.53％,46.67％ and 1.16％ in pancreatic carcinoma,metastasis and benign pancreatic disease tissue,respectively.Methylation of APC in pancreatic carcinoma and metastasis increased significantly compared with control tissues (x2 =12.903,14.402; P ＜ 0.05).There were no statistically significant differences of APC expression in these tissues (P ＞ 0.05).There was a significant correlation between methylation of APC and clinicopathological stage (x2 =6.801,P ＜ 0.05),but no correlation with gender,age,tumor size,histological grade and metastasis (x2 =0.727,1.311,0.372,0.148,0.017 ; P ＞ 0.05).Conclusion The methylation of APC gene is closely related with pancreatic carcinoma inogenesis and the clinicopathological stage,but do not effect the expression of APC in tissues.

Objective To further research the biological functions of PES1,the ovarian SKOV3 cell line with inducible stable PES1 expression is established by using Tet-on system.Methods PES1 was cloned into pTRE-Tight vector via PCR and its expression was identified. After transfected the regulating plasmid pTet-on,SKOV3 cells were screened with G418 and re-transfected pTRE-Tight-PES1.The positive cell clones were screened out with hygromycin and were induced by doxycycline (Dox) to definite the best induction concentration.Growth velocity of SKOV3 cells stably expressing PES1 induced by Dox was detected with viola crystallina.Results The SKOV3 cells with inducible PES1 expression were screened out after the cells were transfected pTRE-Tight-PES1 constructed.Dox could dose-dependently induce the PES1 expression with the concentration under 2 mg/L,and 2 mg/L of Dox induced the highest PES1 expression.Growth velocity of SKOV3 cells transfected pTRE-Tight has no significant difference between the SKOV3 cells transfected nothing induced with Dox.However,the SKOV3 cells transfected pTRE-Tight-PES1 grew faster than the cells transfected pTRE-Tight or without transfection in the fourth day (P =0.001 ).Conclusion The inducible stable PES1 expression SKOV3 cells are successfully established and could be used to be an effective cell model to research the biological functions of PES1.The expression of PES1 could promote the growth of SKOV3 cells.

BACKGROUND:Few reports are found about the effect of ovariectomized rats' bone histomorphometry parameters using non-destructive dynamic loading system.OBJECTIVE:To observe the effect of different loads situations on the bone histomorphometry parameters in ovariectomized rats.DESIGN,TIME AND SETTING:A controlled observation on the bone histomorphometry was performed in the Biomedical Engineering Laboratory of Jilin University from April 2007 to August 2007.MATERIALS:A total of 35 9-month-old Sprague-Dawley female rats were randomly divided into five groups,including sham operation group,ovariectomized (OVX) control group,OVX loading 1 N group,OVX loading 2 N group,OVX loading 4 N group.There were 7 rats in each group.METHODS:Rats in OVX control group and castration load group were processed into bilateral OVX on the back. The sham operation group only underwent the excision of fat tissues on back,and then sutured. After castration for 1 week,rats were loaded with non-destructive dynamic loading system in the two sides of the tibia,15 minutes a day. The mechanical loads would continue for 4 weeks and the loads were 1N,2N and 4N.MAIN OUTCOME MEASURES:Changes of proximal tibia bone histomorphometry parameters.RESULTS:The area,number and thickness of trabecular bone in OVX loading group were all higher than OVX control group (P＜0.05,P＜0.01).The trabecular bone area and thickness in OVX 4 N and OVX 2 N groups were significantly higher than OVX control group (P ＜ 0.001).There was a downward trend of trabecular separation in OVX 4 N group compared with OVX control group (P ＜ 0.05). With the increasing loads,there was an increasing trend of the area,number and thickness of trabecular bone,which were close to sham-operated group. The trabecular separation was declined. Single fluorescent labeled surface and double fluorescent labeled surfaces in sham operated group were all lower than that in OVX control group. With the increase in loads,the single fluorescent labeled surface,double fluorescent labeled surface,interlabeled width and mineral apposition rate had been shown to increase. The OVX 2 N and OVX 4 N groups exhibited a remarkably higher level of mineral apposition rate than OVX control group (P ＜ 0.05).CONCLUSION:With the increase in load at the range of 1-4 N,all parameters of bone histomorphometry improve in the OVX rats,the bone microstrcture is greatly ameliorated,bone mass loss is reduced and the process of osteoporosis is delayed.

Physiology experiment can effectively stimulate the enthusiasm of the students to explore the scientific truth by providing the opportunity to observe real physiological phenomena,analyze,discuss and solve practical problems.Lab-based learning (LBL) and theory teaching have complementary advantages,promoting each other and can maximize the use of teaching resources.During the process of physiological LBL,training of students' thinking ability by creating problem situation,inspiration and guidance,carrying out exploring experiments can develop students' intelligence,promote innovation,improve the teaching effect.