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.

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 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

Objective To study the effect of community health education on lactation mastitis and breast feeding.Method Ninety parturients were given community health education for 6 months and then the rates of mastitis and breast feeding were recorded.Results The mastitis morbidity was 3.3%and the rate of breast feeding was 97.8%.Conclusion The implementation of the community health education can reduce the morbidity of maternal mastitis and raise the rate of breast feeding.

Heparin and low molecular weight heparin have been widely used in clinical therapy as anticoagulants in cardiovascular disease and in hemodialysis. Crude heparin is usually prepared from porcine intestinal mucosa. Purified heparin is a mixture of polysaccharides consisting mainly of repeating GlcNS(6S)-IdoA2S disaccharides and other disaccharides with different GlcNAc/GlcNS±3S±6S-GlcA/IdoA±2S residues. Heparin injections are drugs prepared from heparin active pharmaceutical ingredient ( API ) that is prepared from crude heparin. Low molecular weight heparins are dominant heparin-based drugs used clinically, which are prepared by degrading heparin into smaller sizes. As a result, low molecular weight heparins are sharing the same major disaccharides but have different reducing and non-reducing ends. In current study, we focused on the disaccharide compositional analysis of clinically used heparin and heparin-based drugs. HeparinaseⅠ,II, and Ⅲ were used to degrade all heparin and heparin-based drugs including heparin sodium injection, Enoxaparin sodium injection, Nadroparin calcium injection, Dalteparin sodium injection, Fondaparinux sodium into disaccharides. All the degraded products were analyzed by strong anion high perforance liquid chromatography ( SAX-HPLC) coupled with an UV-detector. Commercially available unsaturated disaccharide standards were then used for structral identification. Furthermore, unusual disaccharides present in Nadroparin, Dalteparin, and Fondaparinux were confirmed by reversed-phase ion pair HPLC coupled with mass spectrometry. The developed method produced detailed structural information, which should be useful for quality control of heparin and heparin-based drugs.

Objective:To study the positive expression rate of M2 subtype of macrophage cell surface molecules and the inflammatory factors of PPS in IL-4-induced M2 macrophage.Methods:The experiment was divided into 5 groups:blank control group, Model group,PPS groups(50 μg/ml,100 μg/ml and 200 μg/ml).The expression of CD206 and CD23 was used as bio-maker to confirm IL-4 induced macrophages by treating RAW264.7 with 20ng/ml of IL-4.IL-4 induced RAW264.7 cells were treated with PPS of 50μg/ml,100μg/ml and 200μg/ml for 24 h.Then the expression of CD206,CD16/32 and CD40 were analyzed by flow cytometry, and the mRNA expression of IL-1β,TNF-α,IL-10 and iNOS were detect by qRT-PCR.Results: After treated with IL-4,the positive rate of CD206 of RAW264.7 were high.After treated with PPS ,the rate of CD16/32 and CD40 in IL-4 induced RAW264.7 cells were high ,the expression of CD206 decreased,and the mRNA level of IL-1βand TNF-αincreased.Conclusion:RAW264.7 cells can be polarlized to M2 subtype macrophage by using 20 ng/ml IL-4.PPS enhances the mRNA of IL-1β,TNF-αand the expression of CD40, CD16/32 in IL-4-induced RAW264.7 cells .These results indicate that PPS can induce the M2 subtype to become M1 macrophages, can improve immune function of macrophages.

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 assess the left ventricular(LV)segmental function by velocity vector imaging(VVI)in uremic patients with diffferent quantity of parathyroid hormone(PTH).Methods Sixty patients with uremia were divided into 2 groups according to the value of PTH:group A with PTH less than 3 times of upper normal limit,group B with PTH more than 3 times of upper normal limit.Normal control group was 30 healthy subj ects.All people were evaluated by VVI.The parameters,including the longitudinal systolic strain rate(SSR),diastolic strain rate(DSR),were analysed among the three groups.Results Compared with normal group,in group A and B,SSR,DSR in all segments were lower.Compared with group A,in group B,DSR were all significantly lower(P＜0.05),SSR in apex of laternal wall,medium of anterior septum wall,posterior wall,medium and apex of posterior septum wall and superior wall,base,medium and apex of anterior wall were significantly lower(P＜0.05).There was no statistically significant difference of left ventricular ejection fraction between the two groups(P＞0.05).Conclusions VVI is a viable objective tool to quantitatively assess LV segmental function damaged by different quantity of PTH.