BACKGROUND:Bone marrow mesenchymal stem cel s are considered to have good proliferation and differentiation potentials. Sox9 is a transcription factor that is essential for chondrogenesis and has been termed as a“master regulator”of the chondrocyte phenotype. OBJECTIVE:To study the therapeutic effects of Sox9-transfected bone marrow mesenchymal stem cel s on knee osteoarthritis. METHODS:The bone marrow mesenchymal stem cel s were transfected with Lenti-Sox9-EGFP in vitro. The model of murine knee osteoarthritis was established by cutting off the anterior cruciate ligament. Thirty model mice were randomly divided into three groups, as normal saline group, bone mesenchymal stem cel group and Sox9-transfected bone mesenchymal stem cel group. 0.1 mL of normal saline, 0.1 mL of normal saline containing non-transfected bone marrow mesenchymal cel s (non-transfected group), or 0.1 mL of normal saline containing Sox9-transfected bone marrow mesenchymal cel s (Sox9-transfected group) was injected into the knee joint cavity of mice in the corresponding group, respectively. After 4, 8, 12 weeks, the repair of articular cartilage lesions was evaluated by toluidine blue and immunohistochemical staining. RESULTS AND CONCLUSION:The lesions of articular cartilage were more serious in the normal saline group, compared with the other two groups, and the difference became more obvious over time. Damaged articular cartilage was improved in the non-transfected group, but the improvement was less than that in the Sox9-transfected group. Immunohistochemistry staining revealed that in the Sox9-transfected group, the positive type II col agen expression was stronger than that in the other two groups, but this positive expression was decreased over time in al the three groups. These results suggest that Sox9-transfected bone marrow mesenchymal stem cel s promote the repair of damaged cartilage in mice with knee osteoarthritis.

OBJECTIVETo discuss the mechanism of scar hypertrophy in adenosine receptor A(2A) (A(2A) R) knockout mice.

METHODSAnimal models of hypertrophic scar were established in 12 A(2A) R knockout mice and 12 wild-type mice as control. The thickness and the size of transverse section of the hypertrophic scar were observed by H-E staining. The hydroxyproline (HYP) in the scar was measured colorimetrically. The TGF-beta expression was tested by Western blotting method.

RESULTSThe hypertrophic scar in wild-type mice was more severe than that in knockout mice. Compared with self-control, the increase of the thickness and the size of transverse section of hypertrophic scar was markedly higher in wild-type group than in the knockout group (P < 0.01). There was significant difference in HYP content between the two groups (P < 0.01). Compared with self-control, the increase of TGF-beta expression in wild-type group was much more than that in knockout group (P < 0.01).

CONCLUSIONSThe TGF-beta expression decreases in the A(2A) R knockout mice. The scar hypertrophy is also much less in the A(2A) R knockout mice.

Animals ; Cicatrix ; metabolism ; pathology ; Disease Models, Animal ; Mice ; Mice, Knockout ; Receptor, Adenosine A2A ; genetics ; Transforming Growth Factor beta ; genetics ; metabolism

OBJECTIVETo research the effects of incorporating tetrapod-like zinc oxide whisker (T-ZnOw) of different proportions on the antibacterial activity of different kinds of soft denture liners.

METHODSThe minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of T-ZnOw against Saccharomyces albicans (S. albicans) were examined by the broth dilution test. Add T-ZnOw 0%, 1%, 2%, 3% to Silagum soft denture liners and soft denture liners respectively. The antimicrobial rate of S. albicans was determined by the membrane covering method.

RESULTSThe MIC of T-ZnOw against S. albicans was 78 microg mL(-1) and the MBC was 156 microg mL(-1). Comparing 1%, 2%, 3% T-ZnOw group with control group, the results showed significant difference (P<0.05). And as the T-ZnOw antibacterial agent added ratio increased, antibacterial increased significantly.

CONCLUSIONThe soft denture liners incorporating with 1%, 2%, 3% of T-ZnOw can improve antibacterial activity. Following the increased proportions of T-ZnOw, antibacterial rate was significantly increasing.

Animals ; Anti-Bacterial Agents ; Denture Liners ; Microbial Sensitivity Tests ; Vibrissae ; Zinc Oxide

BACKGROUNDPneumonia has become the predominant cause of death for the elderly. It is critical to determine the status of oropharyngeal pathogen colonization in the elderly when treating pneumonia. To explore the efficient approaches to treat age-related pneumonia, we determined the status of oropharyngeal pathogenic colonization in the elderly community.

METHODSThroat swab cultures were used to isolate oropharyngeal pathogens from 706 residents older than 65 years living in the community of Shenyang City. Characteristics of bacterial strains were sorted and identified using drug sensitivity tests.

RESULTSResults of bacterial identification showed that 265 out of 706 samples were positive, thereby exhibiting a 37.5% positive rate. There were 290 bacterial strains isolated from the elderly community in total, of which 248 strains were gram-negative bacilli (GNB) and 42 strains were gram-positive cocci (GNC), accounting for 85.5% and 14.5%, respectively. There were 158 Klebsiella pneumoniae strains, representing 54.4% of the all GNB.

CONCLUSIONThe rate of oropharyngeal GNB colonization in the elderly community increases and Klebsiella pneumoniae is the most common strain.

Aged ; Aged, 80 and over ; Bacteria ; classification ; drug effects ; isolation & purification ; Female ; Humans ; Male ; Microbial Sensitivity Tests ; Oropharynx ; microbiology

BACKGROUNDRecent research shows that lasers can inhibit fungal growth and that Nd:YAG 1064-nm lasers can penetrate as deep as the lower nail plate. The aim of this study was to observe the effect of a long-pulse Nd:YAG 1064-nm laser on 154 nails of 33 patients with clinically and mycologically proven onychomycosis.

METHODSThirty-three patients with 154 nails affected by onychomycosis were randomly assigned to two groups, with the 154 nails divided into three sub-groups (II degree, III degree, and IV degree) according to the Scoring Clinical Index of Onychomycosis. The 15 patients (78 nails) in group 1 were given eight sessions with a one-week interval, and the 18 patients (76 nails) in group 2 were given four sessions with a one-week interval.

RESULTSIn group 1, the effective rates at 8 weeks, 16 weeks, and 24 weeks were 63%, 62%, and 51%, respectively, and the effective rates in group 2 were 68%, 67%, and 53% respectively. The treatment effect was not significantly different between any sub-group pair (P > 0.05).

CONCLUSIONSLong pulse Nd:YAG 1064-nm laser was effective for onychomycosis. It is a simple and effective method without significant complications or side effects and is expected to become an alternative or replacement therapy for onychomycosis.

Adolescent ; Adult ; Aged ; Female ; Humans ; Lasers, Solid-State ; therapeutic use ; Male ; Middle Aged ; Onychomycosis ; surgery ; Young Adult

OBJECTIVETo investigate aberrant methylation in the promoter of p16 gene in the sediment cells of pleural effusion and evaluate its clinical significance in the differentiating benign and malignant pleural effusion.

METHODSUsing methylation-specific PCR (MSP), aberrant promoter methylation of p16 gene was detected in the sedimental cells of pleural effusion samples from 66 patients with pleural effusion.

RESULTSOf the 66 patients with pleural effusion, 36 had a definite diagnosis of malignant pleural effusion, and the rest were confirmed to have benign pleural effusion. The positivity rate of p16 gene promoter methylation was 69.4% (25/36) in malignant pleural effusion and 13.3% (4/30) in benign pleural effusion specimens, showing a significant difference between them (χ² = 20.915, P < 0.01). The diagnostic sensitivity, specificity and accuracy of aberrant promoter methylation of p16 gene in the 36 malignant cases were 69.4%, 86.7% and 77.3%, respectively. The positive expression of p16 gene promoter methylation in malignant pleural effusion was not correlated to the histological type or the pathological grade of the tumor (P > 0.05).

CONCLUSIONDetection of aberrant methylation in p16 gene promoter in the sediment cells of pleural effusion specimens by MSP method allows differentiation between benign and malignant pleural effusion.

Adult ; Aged ; Aged, 80 and over ; Base Sequence ; DNA Methylation ; Female ; Genes, p16 ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Pleural Effusion, Malignant ; diagnosis ; genetics ; Promoter Regions, Genetic ; genetics ; Sensitivity and Specificity

OBJECTIVETo investigate the expression of B cell specific MLV integration site-1 (Bmi-1) in colorectal cancer (CRC) and its correlation to the clinicopathological features and prognosis of CRC.

METHODSSixty CRC, 30 adenomas and 20 normal colorectal mucosal tissues were collected to detect the expression of Bmi-1 protein using immunohistochemistry, and the results were analyzed in comparison with the clinicopathological features and survival rate of patients.

RESULTSThe positivity rate of Bmi-1 expression in CRC tissue was 51.7%. In CRC, the rate of Bmi-1 overexpression was 25.0%, significantly higher than that in the adenomas and normal colorectal mucosal tissues (6.67% and 0%, respectively, P<0.05). The overexpression of Bmi-1 protein in CRC was obviously associated with distant metastasis and the TNM stage (P<0.05), but not with gender, age, tumor size, tumor site, histological type, differentiation degree and lymph node metastasis (P>0.05). But logistic regression analysis showed that Bmi-1 protein overexpression in CRC was associated only with distant metastasis (P<0.01,OR>1); Kaplan-Meier survival analysis showed that the survival rate of the patients with high Bmi-1 expression was significantly lower than that in patients with low expression (P<0.05).

CONCLUSIONThe overexpression of Bmi-1 protein was significantly correlated to the tumorigenesis, metastasis and prognosis of CRC, and may serve as an indicator for evaluating the prognosis of CRC.

Adult ; Aged ; Case-Control Studies ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Nuclear Proteins ; genetics ; metabolism ; Polycomb Repressive Complex 1 ; Prognosis ; Proto-Oncogene Proteins ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Young Adult

BACKGROUNDTrichophyton rubrum (T. rubrum) is the most common causative agent of dermatophytosis worldwide. In this study, we examined the effect of laser irradiation on the growth and morphology of T. rubrum.

METHODSColonies of T. rubrum were isolated (one colony per plate), and randomly assigned to 5 treatment groups: Q-switched 694 nm ruby laser treatment, long-pulsed Nd:YAG 1064 nm laser treatment, intense pulsed light (IPL) treatment, 308 nm excimer laser treatment and the blank control group without treatment. Standardized photographs were obtained from grown-up fungal plates prior to treatment. Colonies were then exposed to various wavelengths and fluences of laser light. To compare the growth of colonies, they were re-photographed under identical conditions three and six days post-treatment. To investigate the morphology of T. rubrum, scanning electron microscope (SEM) and transmission electron microscope (TEM) images were obtained from specimens exposed to 24 hours of laser treatment.

RESULTSGrowth of T. rubrum colonies was significantly inhibited following irradiation by 694 nm Q-switched and 1064 nm long-pulsed Nd:YAG lasers. Other treatments exerted little or no effect. Q-switched laser irradiation exerted a stronger growth inhibitory effect than long-pulsed Nd:YAG laser irradiation. Following treatment by the Q-switched ruby laser system, T. rubrum hyphae became shrunken and deflated, and SEM images revealed rough, fractured hyphal surfaces, punctured with small destructive holes. TEM images showed that the hyphae were degenerating, as evidenced by the irregular shape of hyphae, rough and loose cell wall, and obscure cytoplasmic texture. Initially high electron density structure was visible in the cell; later, low-density structure appeared as a result of cytoplasmic dissolution. In contrast, the blank control group showed no obvious changes in morphology.

CONCLUSIONThe Q-switched 694 nm ruby laser treatment significantly inhibits the growth and changes the morphology of T. rubrum.

Lasers, Solid-State ; therapeutic use ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Photochemotherapy ; Trichophyton ; growth & development ; radiation effects ; ultrastructure

AIM: To investigate whether inactivation of extracellular signal-regulated kinase 1/2 ( Erk1/2 ) will affect the function of fibroblast growth factor 21 (FGF21) to regulate glucose and lipid metabolism .METHODS:Male db/db mice (8 weeks old) were treated with U0126 (an inhibitor of Erk1/2 kinase) for 1 week, and then treated with re-combinant human FGF21 protein and adenovirus-mediated FGF21 (Ad-FGF21).The profile changes of blood glucose and blood lipid were evaluated at 120 min or 4 weeks after FGF21 administration.Meanwhile, the molecular mechanism was ex-plored by in vitro study.RESULTS: Treatment of db/db mice with recombinant human FGF21 protein significantly re-duced blood glucose and triglyceride levels at 120 min after FGF21 administration , but these changes were comparable in U0126-treated mice .Furthermore , abnormal glucose and triglyceride levels , and glucose and insulin tolerance were strong-ly improved in db/db mice as accompanied with decreasing body fat content after 4 weeks of ad-FGF21 administration .In-terestingly, treatment with or without U0126 did not influence these effects of FGF21.Mechanically, treatment with Ad-FGF21 significantly upregulated the protein levels of p-Erk1/2 and peroxisome proliferator-activated receptor γ( PPARγ) as well as the expression of adiponectin at mRNA and protein levels in adipose tissues .However , treatment with or without U0126 did not change the profiles .On the other hand , in vitro experiments also indicated that treatment of adipocytes with recombinant human FGF 21 protein significantly activated Erk 1/2 phosphorylation , and upregulated the expression levels of PPARγand adiponectin (P<0.05).However, pre-administration of U0126 did not affect the profiles.CONCLUSION:Pharmaceutical inactivation of Erk 1/2 by U0216 does not affect the biological function of FGF 21 to regulate blood glucose balance and improve abnormal blood lipids in vivo.

OBJECTIVETo clone the recombinant human islet neogenesis-associated protein (rhINGAP) gene for its secretory expression in Pichia pastoris.

METHODSINGAP gene was amplified with PCR and inserted between Xho I and EcoR I downstream sites of the alpha factor of the recombinant plasmid alpha/pUC18. The fusion gene of alpha factor and INGAP was subsequently inserted between BamH I and EcoR I sites of the plasmid pPIC9K of P. pastoris. After confirmation with restriction enzyme digestion and sequencing, the positive recombinant plasmid that integrated INGAP gene was linearized with Sal I digestion and transformed into the yeast host strain GS115 through electroporation. The yeast transformants that harbored the INGAP gene with high copies were selected with the auxotroph medium and G418, followed then by PCR verification of the positive transformants, from which the expression of recombinant human INGAP was induced with methanol as the only carbone source. The antigenic activity of the desired protein was then detected using Western blotting and enzyme-linked immunosorbent assay (ELISA).

RESULTS AND CONCLUSIONThe recombinant expression plasmid INGAP/pPIC9K was successfully constructed, and 3 positive transformants were obtained. The expressed protein showed good antigenic activity as confirmed by Western blotting and ELISA.

Antigens, Neoplasm ; genetics ; metabolism ; Biomarkers, Tumor ; genetics ; metabolism ; Blotting, Western ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; Gene Expression ; Humans ; Islets of Langerhans ; metabolism ; Lectins, C-Type ; genetics ; metabolism ; Pancreatitis-Associated Proteins ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; metabolism