With the method of specific binding of DAPI and DNA, the mycoplasma contamination cell cultures are sensitively detected. In this way, the monolayer cells can be determinated if a certain cell culture has been contaminated by some mycoplasma. The process is simple and can be done within thirty minutes. This method can be widely applied as a routine determination for detecting mycoplasma contamination in laboratory cell cultures.

Objective To study the effects of tri-ortho-cresyl phosphate(TOCP) on mitochondrial membrane permeability in hen's nerve tissues and investigate the mechanism of the organophosphorus ester-induced delayed neurotoxicity(OPIDN).Methods Adult Roman hens were randomly divided into four groups,including three treated groups and one control group(24 in each group).The hens in the experimental groups were treated with TOCP by gavage at the single dosages of 185,375 and 750 mg/kg respectively.TOCP was dissolved in corn oil and administered at 0.65 ml/kg.The control hens received an equivalent volume of corn oil by gavage.The hens were sacrificed on the 1st,5th,15th and 21st day after treatment and the cerebrum,cerebellum,spinal cord were dissected and homogenized in ice bath.The mitochondria in these nerve tissues were extracted to determine the changes of the membrane permeability and membrane potential.Results Compared with the control group,no significant increase of the mitochondrial membrane permeability in the cerebrum was observed in treated groups.In the cerebellum,the membrane permeability in the 185 mg/kg group had no significant changes,while in the 375 and 750 mg/kg groups it increased significantly on the 1st and 5th day after TOCP treatment(P

Objective:To investigate relationships between the single nucleotide polymorphisms(SNPs) of interleukin-1B(IL-1B) and variable number of tandem repeat (VNTR)of interleukin-1RN(IL-1RN) promoter genes and susceptibility of the patients with gastric adenocarcinoma or the patients with gastric adenocarcinoma to helicobacter pylori (Hp) infection.Methods:The SNPs of the IL-1B(-31C/T and -511C/T) were determined by gene chip and The VNTR of IL-1RN were determined by agrose gel electrophoresis with a total of 130 cases of gastric adenocarcinoma and 142 healthy controls.The sera concentrations of IgG,IgM and IgA of Hp antibodies were measured by ELISA in all cases and controls.Results:H.pylori infection was detected in 69.2% of 130 patients and 46.5% of 142 controls (P=0.007,odds ratio [OR]=2.53).Frequencies of IL-1B-31TT genotype in patients with gastric adenocarcinoma or in patients with pooly-differentiated gastric adenocarcinoma were significantly higher than those in healthy controls or in well-differentiated groups,(P

To explore indications regularity of analogous decoctions of Erchen Decoction.323 analogous decoctions of Erchen Decoction were found in Concise Thesaurus of TCM Formula.Diseases and symptoms of TCM were analyzed.22 diseases and 118 symptoms of TCM were found.The main indication of analogous decoctions of Erchen Decoction included spleen-stomach and lung system,other symptoms were broadly involved.Erchen Decoction was the most appropriate for phlegm.

Objective To explore the effect of lysophosphatidic acid(LPA)on blood-brain barrier(BBB) permeability and its possible mechanism.Methods LPA or LPA+suramin(L+S)were stereotaxically injected into the right eaudate nucleus in SD rats in vivo.Evans blue(EB)was used to quantitatively measure the permeability of BBB at different time points.The expression of matrix metalloproteinase-9 was detected by immunohistochemistry technique.The pathological ultrastruetural changes of BBB were assessed by transmission electron microscopy.Results The BBB permeability began to increase after LPA administered into ipsilateral eaudate nucleus,and reached the peak at 24h.Then the permeability of BBB gradually lowered after 48h.In comparison with the same time points of control group,there were quite significant differences(P＜0.01).After L+S was injected,the change of BBB permeability had differences in comparison with those of LPA group in the same time points,(P＜0.05).MMP-9 positive cells were mainly vascular endothelial cells.The numbers of MMP-9 positive blood vessels grew at 6h in LPA group,and the expression of it reached maximum at 24h,then the number of it decreased at 48h,showing significant statistical differences in comparison with the L+S group(P＜0.01),It was observed microscopically that ultrastrueture of BBB of the LPA group was changed sharply,such as basement membrane roughed and fragmented,astroeyte end-feet swolled markedly and perivaseular space enlarged obviously.But there were no remarkable changes in BBB in L+S group.Conclusion LPA can induce increase of BBB permeability and its possible mechanism is the strong expression of MMP-9 protein produeted by endothelial cells through the mediation of LPA receptor,leading to degradation of basement membrane.

Objective To evaluate the outcome of unconstrained shoulder arthroplasty for severe injury of proximal part of the humerus.Methods Twelve eases were all treated with unconstrained shoul- der arthroplasty.Six patients with complex fractures of proximal humerus and four with bone tumor in the proximal part of the humerus used hemiarthroplasty and two patients with osteoarthritis were managed with total shoulder arthroplasty.Cemented prostheses were used in all the cases.Results The average age was 65 years and the follow-up was 2.3 years.Two cases of complex fractures had light pain with limited external rotation of the shoulder.No pain or prosthetic stems loosening were found,and the range of motion and the function of the shoulder were satisfactory in other cases.Conclusion Unconstrained shoulder arthroplasty is a satisfactory and safe technique for severe injury of proximal humerus.

To modify the splicing pattern of Bcl-x and compare the effect of this approach with that of the antisense gene therapy in BIU-87 cell line of bladder cancer, by using 5'-Bcl-x AS to target downstream alternative 5'-Bcl-x splice site to shift splicing from Bcl-xL to Bcl-xS and 3'-Bcl-x AS antisense to the 3'-splice site of exon III in Bcl-x pre-mRNA to down regulation of Bcl-xL expression, the inhibitory effects on cancer cells by modification of alternative splicing and antisense gene therapy were observed and compared by microscopy, MTT Assay, RT-PCR, FACS, Westhern bloting and clone formation. The growth of cells BIU-87 was inhibited in a dose- and time-dependent manner. Its inhibitory effect began 12 h after the exposure, reaching a maximum value after 72h. The number of cells decreased in S phase and the number increased in G1 phase. The ability to form foci was reduced and the antisense gene therapy was approximately half as efficient as modification of alternative splicing in inducing apoptosis. It is concluded that modification of splicing pattern of Bcl-x pre-mRNA in bladder cancer cell BIU-87 is better than antisense gene therapy in terms of tumor inhibition.

Aim To isolate cancer stem cells from human pan-creatic cancer cell line L3. 6pl and to identify their biological characteristics. Methods L3. 6pl cells were cultivated in com-mercial low adhesion plate with serum-free stem cell culture me-dium ( MEM/F12 1:1 ) supplemented with B27. The cancer stem cells reformed into floating spheres were isolated. The method of tumor sphere formation was used to isolate/enrich and characterize the cancer stem cells in pancreatic carcinoma cell line L3. 6pl. Cancer stem cell spheres were collected and sorted using magnetic cell sorting ( magnetic activated cell sorting, MACS) technology, with the cell surface markers of CD24 +CD44 + ESA+. Self-renewal and EMT-related oncogene expres-sion were measured with Western blot. Cancer stem cells differ-entiation potential and the expression of cancer stem cell related signs were checked with Immunofluorescence assay. To deter-mine tumorigenesis in vivo, Xenograft assay in NOD-SCID mice were performed respectively, then immunohistochemistry proto-oncogene c-Met and RON expression were also checked. West-ern blot was used to detect the changes of stemness relative tran-scriptional factors and epithelial markers expressed in spheres before and after differentiation. Drug resistance of pancreatic cancer stem cells to gemcitabine or paclitaxel was measured with MTT assay. Results CD24 + CD44 + ESA+ cells were signifi-cantly tumorigenic, and cultured in serum-free conditions to form spheroids, which had the characteristics of stem cells with self-renewal, EMT and drug-resistant capabilities, and had a posi-tive correlation with the c-Met, RON protein expression. Con-clusion Human pancreatic stem cells are successfully isolated, which provides a useful model for individualized therapy and e-valuation of the therapeutic efficacy for pancreatic cancer pa-tients.

Objective To investigate the effect of aurora kinase inhibitor VX-680 on cell apoptosis and Bcl-2 expressions in human bladder cancer T24 cells. Methods T24 cells were cultured and treated with VX-680 at various concentrations and time points in vitro. VX-680-induced apoptosis of T24 cells was calculated by flow cytometry. The morphological change of treated cells was observed by microscopy;Bcl-2 mRNA and protein expression in T24 cells was detected by RT-PCR and Western blot assay , respectively. Results The apoptosis rate of VX-680-induced T24 cells increased in a dose-and time-dependent manner. The increase of apoptosis rate and decrease of Bcl-2 mRNA and protein expression in VX-680-induced T24 cells were in a dose-dependent manner. Conclusion VX-680 can significantly induce the apoptosis of T24 cells by down-regulating Bcl-2expression in a dose-dependent manner.

Objective To analyze the perceived control of lung cancer patients undergoing chemotherapy and to explore its influencing factors. Methods A total of 185 patients with lung cancer were recruited and assessed by homemade general information questionnaire and the Cancer Experience and Efficacy Scale. Results The total score of cancer experience during chemotherapy in patients with lung cancer was (62.50±6.99) points. The total score of efficacy during chemotherapy in patients with lung cancer was (42.52±8.08) points. Single factor analysis showed the influencing factors of perceived control were personal income, sex, education, metastasis, surgical treatment and stage of disease. Conclusions The cancer-related experience and efficacy of patients with lung cancer is at middle level. Health-care worker should pay attention to the impact of lung cancer patients and take effective measures to mitigate the negative cancer-related experience, thereby improving the perceived control of the lung cancer patients.