Effects of mannan-oligosaccharide (MOS) on the growth of Lactobacillus were investigated by pure culture of three swine originated Lactobacillus strains or co-culture with swine pathogenic E. coli. The pure culture results revealed that OD values and lactic acid concentration of three Lactobacillus strains in the MOS supplemented cultures were higher than those of the control group without MOS supplementation, a lower pH was observed for MOS supplement compared with that of the control group. The co-culture results showed that MOS were utilized by both Lactobacillus strains and swine pathogenic E. coli strain. But Lac-tobacillus grew faster than the latter. A significant increase in lactic acid concentration (P

Objective To evaluate the clinical application of May-Grunwald-Giemsa staining followed by fluorescence in situ hybridization (MGG-FISH) technique in the differentiation diagnosis of Ph-chromosome positive acute lymphoid leukemia (Ph + ALL) from chronic myeloid leukemia in lymphoid blast crisis(CML-LBC). Methods The bone marrow smears of 4 patients with Ph+ ALL, 4 patients with CML-LBC, 1 patient with CML in myelocytic blast crisis complicated with lymphoma and 1 patient with CML in mixed blast crisis were assayed with the MGG-FISH technique in which the spectrum green labeled BCR and spectrum orange labeled ABL dual color dual fusion probes were used. Based on the morphological classification, the percentages of BCR-ABL positive cells were subsequently determined respectively in the erythroid, myeloid and lymphoid hneages for the 10 specimens. Results According to the MGG-FISH analysis, the erythroid lineage was not involved in the 4 Ph+ ALL specimens without BCR/ABL positive cells. While the BCR/ABL positive percentage of myeloid cells was 11% (1/9), 8% (1/12), 0% (0/8) and 10% (1/10) respectively and that of lymphoid cells was 97% (76/78), 98% (87/89), 98% (97/99) and 97% (75/77) respectively. On the other hand, the BCR/ABL positive percentage was 100% (8/8), 91% (10/11), 82% (9/11), 88% (7/8) in the erythroid lineage, 89% (8/9), 96% (94/98), 100% (47/47), 98% (40/41)in the myeloid lineage and 96% (78/81), 93% (52/56), 96% (68/71), 95% (58/61) in the lymphoid lineage respectively for the 4 CML-LBC specimens. The BCR/ABL positive percentages of the other 2 specimens were all above 80% and through MGG-FISH analysis we also identified the source of the malignant clones and ascertained the diagnosis of the 2 patients. Conclusions The MGG-FISH technique has proved useful in providing rapid and precise differentiation between Ph + ALL and CML-LBC. The source of the malignant clones can also be analyzed by this technique.

ObjectiveTo investigate the radiosensitizing effect of 3-(5'-hydroxy-2'-furyl)-1-benzyl indazole ( YC-1 ) on hypoxic human adenocarcinoma cell line A549.MethodsMTT assay was used to test the inhibitory effect of YC-1 on proliferation of A549 cells.Clonogenic assay was performed to determine the radiosensitizing effect of YC-1 on hopxic A549 cells.Single-hit multi-target model was used to plot survival curve and calculate sensitization enhancement ratio (SER).The cell cycle and apoptosis were measured by flow cytometry.ResultsThe proliferation of A549 cells was inhibited by YC-1 in a time-dose-dependent manner.In normoxic and hypoxic cells,the IC20 was 16.7 μmol/L and 39.2 μmol/L at 24 h,respectively.In the group of hypoxia plus YC-1,SERD0 and SERDq were 1.11 and 1.26,respectively.In hypoxia,YC-1combined with 2 Gy irradiation could induce cell apoptosis and prolong G2 + M phase arrest ( ( 30.17 ±1.21 )％ ∶ ( 15.44 ±0.96) ％,P =0.000; (21.56 ±0.47 )％ ∶ (6.16 ±0.16)％,P =0.000).Concinsions YC-1 could enhance the radiosensitivity of hypoxic A549 cells.

Advances in mechanism and application of the heating effect in breeding of microorganism are reviewed in this paper. Heat produces mutagenesis effect and screening effect. Heating mutagenesis effect is occurred through the substitution of G-C base pair induced by heat, and heating screening effect produces higher forward mutation rate induced by other mutagens.

A bacterial strain which could grow well on the substrate of PAEs as the sole source of carbon and energy was isolated from contaminated sludge in the river of WeiFang in ShangDong province and it was designated as JDC-3. Based on the morphology,biophysical and biochemical properties as well as molecular characteristics,this isolate was preliminarily identified as Delftia sp.. A fragment of phthalate dioxygenase gene was successfully amplified from the genus of Delftia for the first time using a set of degenerate primers. Meanwhile,the degradation capability of JDC-3 was determined by HPLC using DMP as test substrate. The results showed that the optimal pH and temperature were at 7.0～8.0 and 30?C～35?C respectively. The degradation kinetics of JDC-3 was studied in different initial DMP concentration under optimal conditions. The results indicated that the degradation dynamic equation was ln C =-0.06837 t + A when DMP concentration was lower than 300 mg/L,with half life of 12.48 h. The degradation rate decreased and half life of JDC-3 prolonged as the initial concentration kept on increasing.

OBJECTIVETo observe the electroacupuncture (EA) pretreatment at Baihui (GV20) on the concentration of adenosine deaminase (ADA) and adenosine, and to evaluate its effects on the neurologic function score and the infarction volume after middle cerebral artery occlusion (MCAO) ischemia/reperfusion (I/R), thus exploring its mechanisms for relieving the ischemia/reperfusion injury.

METHODSTotally 54 male SD rats were randomly divided into 3 groups, the sham-EA group, the EA group, and the control group, 18 in each group. Rats in the control group were not intervened after anesthesia. Rats in the EA group were needled at Baihui (GV20) for 30 min. Rats in the sham-EA group received the same procedure as those performed in the EA group without electricity connected. The changes of adenosine and ADA contents were detected at 30, 60, and 120 min after EA respectively. The I/R model was established. Totally 48 male SD rats were randomly divided into 6 groups, i.e., the model group (Group A), the EA group (Group B), the EA +8-Cyclopentyl-1,3-dipropylxanthine (DPCPX) group (Group C), the EA + DMSO group (Group D), the Deoxycoformycin (Deo) group (Group E), and the normal saline group (Group F). Rats in Group B, C, and D received EA for 30 min before modeling. Rats in Group C and D were peritoneally injected with DPCPX (1 mg/kg) and DMSO (1 mL/kg) at 30 min before EA. The neurologic function score was evaluated and the infarct volumes were detected after 24-h reperfusion.

RESULTSCompared with the sham-EA group, there was no statistical difference in the contents of the adenosine or ADA in the control group at each time point (P > 0.05). Compared with the control group at the same time point, the content of ADA significantly decreased at 60 min in the EA group [(315.0 +/- 22.9 U/L), P < 0.05], and restored to the normal level at 120 min after EA. The content of adenosine increased in the EA group at 120 min [(20.4 +/- 2.2) ng/microL, P < 0.05]. Compared with the model group, the neurologic function score decreased (P < 0.05) and the infarct volumes were obviously reduced (P < 0.01) in Group B, D and E. There was no statistical difference in the neurologic function score or the infarct volumes in other groups, when compared with the model group (P > 0.05)

CONCLUSIONEA at Baihui (GV20) showed protective effects on the cerebral I/R rats, which might be achieved through lowering the ADA concentration and elevating the adenosine content, and further activating adenosine A1 receptor.

Adenosine Deaminase ; metabolism ; Animals ; Brain Ischemia ; metabolism ; Electroacupuncture ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism

OBJECTIVETo investigate the feasibility of stable transfection of human hypoxia inducible factor-1alpha (HIF-1alpha) gene into fibroblasts cells and the effects of supernatant from the transfected cell culture on hair follicle cells.

METHODSPcDNA-HIF1alpha was stably transfected into fibroblasts cells with lipofectamine 2000. Expression of HIF-1alpha was observed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The supernatant was obtained to detect the expression of vascular endothelial growth factor (VEGF) by ELISA. The mRNA expression of basic fibroblast growth factor (bFGF) was detected by RT-PCR. MTT was used to detect the activity of fibroblasts cells and dermal sheath cells added with supernatant.

RESULTSPcDNA-HIF1alpha was successfully transfected into fibroblasts cells. HIF-1alpha could be detected by RT-PCR and Western blot. The expression of VEGF in the supernatant of cells transfected with PcDNA-HIF1alpha was detected. The mRNA expression of bFGF was significantly higher than in the control group (P < 0.01). MTT showed the activity of cells added with supernatant was enhanced (P < 0.05).

CONCLUSIONPcDNA-HIF1alpha can stably transfected into fibroblasts cells, and the expressed HIF-1alpha induces the expression of VEGF and bFGF, and the expressed VEGF enhances the activity of cells.

Cell Culture Techniques ; Fibroblast Growth Factor 2 ; biosynthesis ; Fibroblasts ; metabolism ; Hair Follicle ; cytology ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; biosynthesis ; genetics ; Transfection ; Vascular Endothelial Growth Factor A ; biosynthesis

Objective To study the routine examination of Aconitum sinomontanum Nakai and determine the contents of lappacontine and ranaconitine; To provide basis for establishing the quality standard of Aconitum sinomontanum Nakai.Methods Aconitum sinomontanum Nakai were collected from different areas.A method of TLC was used for qualitative discrimination. The methods in the Chinese Pharmacopoeia were adopted for the determination of moisture content, ash content and extractives. Determination of lappacontine and ranaconitine were performed by HPLC. Results The TLC showed that the spots were clear and the separation was good. Individual provisional standards:the moisture,total ash and acid-insoluble ash content of Aconitum sinomontanum Nakai were not more than 11.0%, 12.0%, and 7.0%, respectively; water soluble and alcohol soluble extractives were not less than 18.2% and 10.6%,respectively.The content of ranaconitine and lappacontine in Aconitum sinomontanum Nakai were not less than 0.125% and 0.815%, respectively. Conclusion The method established by the study is accurate and reliable,and can be used for quality evaluation of Aconitum sinomontanum Nakai.

[Objective] To observe the dynamic changes of intestinal IL-17,occludin,and ZO-1 in mice with postinfectious irritable bowel syndrome (PI-IBS).[Methods] Forty C57BL/6 mice were randomly divided into 5 groups:control group and infection groups (2 weeks,4 weeks,6 weeks,and 8 weeks after trichinella infection).Infection groups were given by gavaging of 400～500 Trichinella spiralis in 0.2 mL of normal saline.The body weight of mice were recorded at week 2,4,6,and 8 after infection.The visceral sensitivity of mice was measured by the abdominal withdrawal reflex (AWR).The stool was collected continuously for 8 hours to calculate the percentage of fecal water content.Pathological changes of gut were observed by HE staining.The expressions of IL-17,occludin,and ZO-1 in ileocecus and colon were detected by immunohistochemistry and Western blotting.[Results] At week 2 after infection,the acute inflammation of the intestinal tract was observed and the body weight of mice were significantly decreased (P=0.000).Until week 8 after infection,the intestinal inflammation and body weight of mice recovered to normal.When the colorectal dilatation capacity was 0.35 and 0.5 mL,the AWR scores in the infection groups were significantly higher than those in the control group (P＜0.01).The percentages of fecal water content in the infection groups were significantly higher than those in the control group (P＜0.05).Compared with the control group,the expressions of IL-17 were significantly decreased in week 2 group (P＜0.05) and increased in week 8 group (P＜0.05).The expressions of occludin and ZO-1 in the infection groups were significantly lower than those in the control group (P＜0.05).[Conclusion] The dynamic changes of IL-17 and the decrease of Tight junction proteins may be one of the mechanisms of visceral hypersensitivity and increased percentages of fecal water content.They may be involved in the development of PI-IBS.

BACKGROUNDIn bone marrow transplant patients, the microenvironment in bone marrow is damaged after chemotherapy or radiotherapy. Subsequent to allogenic hematopoietic stem cell transplantation in patients with clinically successful engraftments, the source of mesenchymal stem cells (MSCs) remains controversial. To further verify the stimulatory effect of the simultaneous transplantation of cells from second donors on engraftment success for hematopoietic stem cell transplantation in support of donor MSCs engraftments, the aim of this study is to monitor the dynamics of the engraftment of bone marrow-derived MSCs in patients after transplantation with mismatched-sex hematopoietic stem and third-party cells.

METHODSIn this study, the hematopoietic stem cells from 32 clinical donors of different sexes that resulted in successful engraftments were selected for transplantation and were classified into three groups for research purposes: group A consisted of 14 cases of transplantation with bone marrow and recruited peripheral hematopoietic stem cell transplantation, group B contained 8 cases of simultaneous re-transfusion of MSCs from the second donor, and group C contained 10 cases of simultaneous re-transfusion of umbilical blood from the second donor. The bone marrow from 32 patients with successful engraftments of hematopoietic transplantation were selected and sub-cultured with MSCs. Flow cytometry (FCM) was used to measure the expression of surface antigens on MSCs. Denaturing high-performance liquid chromatography (DHPLC) in combination with polymerase chain reaction amplification of short tandem repeats (STRPCR) was used to measure the engraftment status of fifth-generation MSCs in patients. Fluorescence in situ hybridization (FISH) revealed the sex origin of the fifth-generation MSCs in 32 patients. Dynamic examinations were performed on patients receiving donor transplantations.

RESULTSThe progenies of fifth-generation MSCs were successfully cultured in 32 cases. The results of FCM demonstrated that the expression levels of CD14+ and CD45+ cells were lower than 0.04% in the fifth-generation MSCs. The analysis using DHPLC and FISH showed similar results. One patient from group B also received a temporary transplantation of MSCs from the donor. The MSCs in the remaining 31 patients all originated from the patients themselves.

CONCLUSIONSAfter transplantation, the MSCs present in patients originated from the host. In patients transplanted with MSCs from a second donor, the phenomenon of temporary chimerization of MSCs was observed.

Adolescent ; Adult ; Cells, Cultured ; Chromatography, High Pressure Liquid ; Female ; Flow Cytometry ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukocyte Common Antigens ; metabolism ; Lipopolysaccharide Receptors ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Middle Aged ; Young Adult