The assay investigated mainly the adhesion phenomenon of heat killed Bifidoba cterium adolescentis DM8504 to human colorectal carcinoma cell line CCL 22 9 Moreover, the assay discussed the adhesion mechanism of Bifidobacterium Resu lts found heat killed bifidobacterium would be the same ability to adhere and c o lonize to human intestinal epithelial cells in vitro as live bifidobacterium, an d their adhesion had to depend on its spent culture supernatant(SCS) The adh esin of Bifidobacterium adolescentis was possibly lipoteichoic acid(LTA)w h ich existed on the cell wall of bacteria and was secreted into the SCS LTA bou nd to the heat resistant proteins of cell surface of bacteria, and extended out from the cell surface Moreover, the bifidobacterial adhesin rece ptor of intestinal epithelial cells was possibly saccharides or glycopolymers

BACKGROUND:The main component of cartilage, type Ⅱ col agen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner.
OBJECTIVE:To observe the variation of SOX9 and type Ⅱ col agen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cel s, and to explore the correlation of SOX9 expression and type Ⅱ col agen.
METHODS:Bone marrow mesenchymal stem cel s were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cel phenotype was identified with flow cytometry. Cel s were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cel s were used as a control group. The total RNA of cel s was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ col agen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cel s were stained by immunofluorescence to observe the differentiation and perform statistical analysis.
RESULTS AND CONCLUSION:Passage 3 bone marrow mesenchymal stem cel s grew wel , and cel phenotype was confirmed as stem cel s by flow cytometry. The staining results showed that, the cel s differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ col agen mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at 3 and 7 days, and then decreased at 14 days. While type Ⅱ col agen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ col agen expression gradual y decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P>0.05), while type Ⅱ col agen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱcol agen, which may alter along with the SOX9 in the early chondrogenic differentiation;SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation.

OBJECTIVEThe study was designed to investigate the changes in CD(69), CD(25) and HLA-DR expressions in peripheral blood T cell in Kawasaki disease (KD).

METHODSThe authors detected CD(69), CD(25) and HLA-DR expressions in peripheral blood T cell by using flow cytometry. The patients who met the diagnostic criteria for KD comprised sixteen boys and fifteen girls (4 - 60 months of age; mean, 26 +/- 18 months). All received intravenous gammaglobulin at a dose of 1 g/(kg.d), for 2 days and oral aspirin at a dose of 30 - 50 mg/(kg.d). In case of persistent fever, a repeated dose of intravenous gammaglobulin or I.V. methylprednisolone at a dose of 20 mg/(kg.d) for three daily doses was attempted. The authors tested blood samples from 17 healthy controls consisting of nine boys and eight girls (3 - 84 months of age; mean, 25 +/- 18 months) and the samples from 31 patients.

RESULTSThe percentage of peripheral blood CD(3)(+) T lymphocyte was (54.4 +/- 9.0)% in acute stage of KD and (65.0 +/- 7.0)% in healthy controls. There was a significant difference between the two groups (P < 0.001). The values of CD(69)(+) [(11.2 +/- 12.6)%, vs. (0.6 +/- 0.4)%], CD(25)(+) [(9.2 +/- 3.5)% vs. (3.9 +/- 1.8)%] and HLA-DR(+) [(8.3 +/- 5.0)% vs. (4.3 +/- 2.3)%] in KD patients were markedly increased compared to those of the healthy controls. After intravenous gammaglobulin treatment, the percentage of CD(3)(+)CD(69)(+) and CD(3)(+)CD(25)(+) significantly decreased [CD(3)(+)CD(69)(+): (14.0 +/- 13.0)% vs. (1.6 +/- 1.2)%, P < 0.05; CD(3)(+)CD(25)(+): (7.8 +/- 4.1)% vs. (2.0 +/- 0.6)%, P < 0.01]. However, the CD(3)(+) T lymphocytes increased [(50.8 +/- 5.0)% vs. (64.9 +/- 5.5)%, P < 0.01]. There was no significant difference in expression of CD(3)(+) T lymphocyte cell activating markers between coronary artery disease group and normal coronary artery group in KD cases (P > 0.05).

CONCLUSIONCD(3)(+) T cell activation in the early and middle stages is involved in the mechanism responsible for cardiovascular injury.

Antigens, CD ; blood ; Antigens, Differentiation, T-Lymphocyte ; blood ; Aspirin ; administration & dosage ; therapeutic use ; Biomarkers ; blood ; Child, Preschool ; Dose-Response Relationship, Drug ; Drug Therapy, Combination ; Female ; Flow Cytometry ; Glucocorticoids ; therapeutic use ; HLA-DR Antigens ; blood ; Humans ; Immunoglobulins, Intravenous ; therapeutic use ; Immunologic Factors ; therapeutic use ; Infant ; Interleukin-2 Receptor alpha Subunit ; blood ; Lectins, C-Type ; blood ; Male ; Methylprednisolone ; therapeutic use ; Mucocutaneous Lymph Node Syndrome ; blood ; diagnosis ; drug therapy ; immunology ; Platelet Aggregation Inhibitors ; therapeutic use ; Prognosis ; T-Lymphocytes ; drug effects ; immunology ; Treatment Outcome

OBJECTIVETo determine the frequency and characteristics of reassortment among Hantaan and Seoul viruses causing hemorrhagic fever with renal syndrome (HFRS).

METHODSMixed infections were initiated in tissue culture, using Hantaan virus strain 76 - 118 and Seoul virus strain SR-11. Potential reassortant virus plaques were picked out by multiplex RT-PCR, using primers specific for individual genome segments (L, M, S) of each strain.

RESULTSMost of the progeny virus plaques (68.19% of 44) had parental genotype of 76 - 118 strain or SR-11 strain while 2 of 44 plaques had mixed genotypes that yielded RT-PCR bands for the same segment of both parental strains. Reassortant viruses were detected in 68.19% of 44 progeny plaques tested, involving the M and S segments. In addition, approximately 4.55% of the progeny virus plaques appeared to contain S or M segments originating from both parental virus strains, showing that they were diploid.

CONCLUSIONGenetic reassortment can occur between Hantaan virus and Seoul virus strains.

Animals ; Cercopithecus aethiops ; Genome, Viral ; Genotype ; Hantaan virus ; genetics ; RNA, Viral ; genetics ; Reassortant Viruses ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Seoul virus ; genetics ; Vero Cells

The inhibition activity of 36 flavonoids against CYP1A2 was determined by our previously developed in vitro method. The Comparative Molecular Similarity Indexes Analysis (CoMSJA) approach was used to probe the quantitative relationships between the flavonoids' molecular structural descriptors and their inhibitory activities. A reliable CoMSIA model with the combined electrostatic and hydrophobic fields was derived with the regression coefficient R2 of 0.948 and the cross-validation regression coefficient q2 of 0.630, separately, which is capable of elucidating the quantitative relationship between the 3D structural descriptors of the flavones and their bioactivities. Comparing with flavone, the larger pi-pi conjugated system of alpha-naphthoflavone significantly improved the biologically inhibitory ability. Based on the core structure of the latter, either electropositive substituents or hydrophobic groups at the 6, 3', and 4' ring positions or electronegative counterparts at the 5 ring position, can enhance the inhibitory potency against CYP1 A2 according to the CoMSIA contour maps.
Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP1A2 Inhibitors ; Flavonoids ; chemistry ; pharmacology ; Humans ; Microsomes, Liver ; metabolism ; Models, Molecular ; Molecular Structure ; Quantitative Structure-Activity Relationship

Objective To explore the correlation between the replacing time of plasma drainage bag and the bacterial growth rate among orthopaedic surgery patients , and to control or reduce the risk of catheter-related infection.Methods The convenient sampling method was used to select 100 patients with postoperative serum drainage tube and disposable drainage bag , from September 2012 to August 2013.Patients were randomly divided into the control group (group A, 33 cases) and the observation group (group B, 32 cases; group C, 35 cases).The amount of drainage were recorded every 24 hours of the three groups of patients , and then the drainage bags were emptied .Group A replaced the drainage bag regularly every day; Group B replaced the drainage bag every 48 hours;Group C did not replace the drainage bag .The outside mouth of the drainage tube and the drainage liquid were taken to bacterial culture . The positive rate of bacterial culture , patients'postoperative body temperature and the rate of incision infection were compared within groups .Results Twenty-four hours after operation, the results of bacterial culture among three groups were negative .However, after 48 hours, six patients (18.1%) in group A, two patients (6.4%) in group B showed positive results of the bacterial culture .None of the patients in group C had the positive results .The comparison among three groups was statistically significant (χ2 =10.32,P<0.05).After 72 hours, eight patients (24.2%) in group A, two patients (6.4%) in group B showed positive results of the bacterial culture .None of the patients in group C had the positive results.The comparison among three groups was statistically significant (χ2 =11.82,P<0.05). The incidence of incision infection of group A , group B and group C were 15.1%(5/33), 6.3%(2/32) and 0.0%(0/35), respectively.There was a significant difference among three groups (χ2 =11.18,P<0.05).The body temperature of the three groups showed no significant difference (F=7.53,P>0.05).Conclusions With the increase of drainage bag replacement frequency , the positive rate of bacterial culture , as well as the drainage bag and drainage tube mouth bacteria culture positive rate increase .Therefore, for orthopaedic patients , the plasma drainage tube and disposable drainage bag should be emptied every 24 hours after the measurement of drainage liquid, but do not have to be replaced every 48 hours or every 24 hours.

Objective To evaluate the localization of transplanted human menstrual-derived stem cells (MenSCs)in the premature ovarian failure (POF)mouse model.Methods Mice were intraperitoneally injected with cisplatin to create the POF mouse model.Ninety mice aged 8 weeks were randomly divided into MenSCs transplantation group,POF model group and blank control group.Green fluorescence protein (GFP)-expressing MenSCs were transplanted to MenSCs transplantation group by the tail vein;meanwhile the same volume of PBS was injected into the mice in POF model group and blank control group.Physiological characteristics of mice,serum concentration of FSH and E2,ovarian histology and distribution of green fluorescence signal in the ovarian tissues were assessed in the three groups at 10 days after transplantation.Results Following anorexia and lethargy,the average weight of mice in POF mouse model decreased compared with that in blank control group. The concentration of FSH increased but E2 level decreased significantly in POF model group.Meanwhile,the number of primordial,growing and mature follicles reduced significantly and interstitial fibrosis was observed in the ovary ofPOF model group.After MenSCs transplantation,the average weight of mice in MenSCs transplantation group increased gradually with the improvement of feeding and activities.Ten days after injection of MenSCs,sex hormone levels and the number of follicles returned to normal values.The level of E2 was significantly higher and the level of FSH was lower,while the number of primordial,growing as well as mature follicles in MenSCs transplantation group was significantly higher than that in POF model group.By observing the location of GFP-MenSCs,we found that GFP-positive cells were located in the interstitium but not in follicles,granulose cells,the liver,the kidney,or lung tissue.Conclusion MenSCs can home to the ovary and be located in the interstitium in the POF mouse model.MenSCs can repair ovarian injury and improve endocrine functions of the ovary.

Objective To obtain comprehensive expression profile of microRNAs(miRNAs)in endometrial cancer stem cells(ECSCs)during differentiation.Methods ECSCs were enriched by spheroid formation and spontaneously differentiated into cancer cells by attached culture.We characterized miRNA expression in ECSCs and its differentiated cells using miRNA microarrays and verified the results by RT-qPCR.Meanwhile,we applied bioinformatic method to predict the target genes of the differentially expressed miRNAs.Furthermore,gene expression profile was detected by genomic microarray.Finally,we summarized the relationship between the predicted target genes of miRNAs and gene expression profile.Results We observed that 10 miRNAs were significantly upregulated in ECSCs,including miR-522,miR-139-3p,miR-520c-5p,miR-518d-5p,miR-146b-5p,miR-34a,miR-526a,miR-193a-3p,miR-221,and miR-4674.Only 1 miRNA,miR-760,was downregulated in ECSCs.A total of 11 differentially expressed miRNAs were identified by RT-qPCR.The results showed these differentially expressed miRNAs(except miR-526a and miR-4674)were in accordance with those obtained by miRNA microarray analysis. By using bioinformatic approach,the target genes of these differentially expressed miRNAs could be found.The results of genomic microarray showed that 207 genes were more highly expressed and 238 genes were more lowly expressed in ECSCs compared with its differentiated cells.Analysis of the gene expression profile and the predicted target genes of miRNAs showed that some common genes except miR-139-3p could be found.Conclusion Our study provides an available information about miRNAs and target genes from different starting points,such as ECSCs differentiation.Further studies are needed to ascertain the role of these miRNAs in ECSCs'differentiation.

Antigens, CD ; genetics ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Tetraspanin 24

Objective:To analyze the effectiveness and safety of the treatment strategies for vascular restenosis after stent implantation in atherosclerosis obliterans (ASO) .Methods:The clinical data of 5 cases who had received drug‐perfused balloon from January to July in 2016 and 15 cases who had received femoral artery retrograde endarterectomy from June 2012 to July 2016 after presenting reocclusion or restenosis atherosclerosis lesions were reviewed retrospectively .The effects of surgery and complications in follow‐up period were analyzed .The effectiveness ,availability and safety of the above two methods were assessed .Results:The 20 patients had acquired an adequate blood perfusion in their distal limbs and got warmer on their feet after underwent drug‐perfused balloon and stent implantation or femoral artery retrograde endarterectomy .In addition ,they could have good palpitation in dorsal artery of feet and have better feelings on account of relieving pain .In the follow‐up ,no restenosis case had been found among them .Conclusions:The application of the stent angioplasty after using drug‐perfused balloon and femoral artery retrograde endarterectomy on vascular restenosis after stent implantation in atherosclerosis lesions proved to be effective and safe ,greatly reducing the restenosis or reocclusion frequency .