BACKGROUND: Whether choosing a suitable biological scaffold compounding with mesenchymal stem cells (MSCs) can construct an ideal tissue-engineering cartilage or not should be researched further.OBJ ECTIVE: To investigate the feasibility of constructing artificial cartilage by using amplified rabbit MSCs which were inoculated on poly-glycolic acid (PGA).DESIGN: Single sample observation.SETTING: Department of Otolaryngology-Head & Neck Surgery, Affiliated Hospital of Sichuan Luzhou Medical College.MATERIALS: The experiment was carried out in the Luzhou Medical College from October 2004 to October 2005. A total of 8 Japanese large ear rabbits, of both genders, clean grade, aged from 2 to 3 months, weighing 1.5-2.0 kg, were fed in normal temperature and humidity. Poly-glycolic acid was provided by Albany Company, USA.METHODS: Rabbit MSCs were separated, obtained and amplified. In addition, poly-glycolic acid was sheared into pieces with the size of 1 cm × 1 cm × 1 cm and embedded with poly-L-lysine. Amplified MSCs were inoculated on the surface of poly-glycolic acid, and then, they were averagely grown on pre-wet scaffold of poly-glycolic acid according to 4 mL/cm3multi-points spreading style and cultured in vitro for 3 weeks. After the operations mentioned above, samples were regarded as the experimental group. Scaffolds of poly-glycolic acid without MSCs were considered as the control group.Samples in both experimental group and control group were transplanted into abdominal cavity of 4 rabbits, respectively,cultured in vivo for 6-12 weeks, taken out and observed generally. Meanwhile, the samples were fixed with 100 g/L neutrality formaldehyde, cut into sections with the thickness of 5 μm, stained with haematine-eosin (HE) and observed their histomorphological characteristics. Moreover, the samples were stained with alcian blue for observation of glycosaminoglycans formation, with toluidine blue for observation of metachromasia matrix formation, and with immunohistochemical staining for detection of type- Ⅱ collagen expression.MAIN OUTCOME MEASURES: Generally observational results of two kinds ofscaffolds at 6 and 12 weeks after transplanting into experimental rabbits, various staining results and expression of type- Ⅱ collagen.RESULTS: A total of 8 experimental rabbits were involved in the final analysis. ① Generally observational results of two kinds of scaffolds at 6 and 12 weeks after transplanting into experimental rabbits: At 6 weeks after transplanting scaffold into abdominal cavity of rabbits, samples in the experimental group were still coated with greater omentum of abdominal cavity. After clearing greater omentum, three samples were light yellow, smooth and moderate quality; meanwhile, their appearances were coincidence with those before transplantation. However, one sample was gray black and soft quality;meanwhile, it was not able to take shape. Twelve weeks later, appearances of samples in the experimental group were still coincidence with those before transplantation. They were gray white, smooth but hard quality. However, samples in the control group were mostly absorbed at 6 and 12 weeks after transplantation, especially, samples were remarkably absorbed at 12 weeks after transplantation. Any tissue like cartilage did not form in both two durations. ② Various staining results and expression of type- Ⅱ collagen at 6 and 12 weeks after transplanting scaffolds into experimental rabbits: Results of HE staining showed that, at 6 weeks after transplantation, structure of cartilage lacuna-like was started form, and PGA scaffold began to be degraded. In contrast, at 12 weeks postoperatively, some cartilage-like tissue were observed in compound, cartilage lacuna-like structure formed obviously; cells arrayed regularly and geminately existed In cartilage lacuna-like atructure; the smaller cell sizes observed at borders and the bigger ones observed at the center;PGA degraded completely. Results of alcian blue staining showed that, at 6 weeks after transplantation, a partial of regions in tissue were light blue; meanwhile, at 12 weeks after transplantation, matrixes in tissue were mostly blue. This suggested that a lot of glycosaminoglycans were formed. Results of toluidine blue staining suggested that, at 6 weeks after transplantation, blue metachromasia matrixes were observed in tissue; meanwhile, at 12 weeks after transplantation, blue metachromasia matrixes were stronger and stronger in tissue. Results of immunohistochemical staining indicated that, at 6 weeks after transplantation, buffy positive granules were observed in plasma and a few of type-Ⅱ collagen expressed in matrix; meanwhile, at 12 weeks after transplantation, powerfully positive expressions were observed in both plasma and matrix. Expression of type- Ⅱ collage showed that, at 6 weeks after transplantation, dark buffy positive granules were observed in plasma and a few of type- Ⅱ collagen mRNA expressed in matrix; meanwhile, at 12 weeks after transplantation, powerfully positive expressions were observed in both plasma and matrix. Samples in the control group were completely absorbed and any tissue-engineering samples did not form.CONCLUSION: Affecting by osteogenic inducer, rabbit MSCs can generate tissue-engineering cartilage after culture in vitro and in vivo.

Aged, 80 and over ; Chordoma ; complications ; Humans ; Male ; Pharyngeal Neoplasms ; complications ; Respiration Disorders ; etiology

Objective To establish the in vitro biofilm model of Laribacter hongkongensis(LH),to analyze the type Ⅰ integron related genes carried by LH and to investigate the mechanism of LH resistance to quinolones.Methods The biofilm forming abilities of LH clinical isolates were determined by Giemsa staining qualitative method and by crystal violet staining semi-quantitative method.The sensitivity of LH to norfloxacin,ofloxacin,levofloxacin,ciprofloxacin and lomefloxacin in both planktonic and biofilm conditions were dectermined by broth microdilution susceptibility tests.Type I integron related genes carried in 18 LH strains resistant to quinolone were detected by PCR amplification method.Results The detection results by Giemsa staining demonstrated that 36 strains in 55 LH clinical isolates formed visible biofilm,and the biofilm formation rate was 65.4%(36/55).In the biofilm forming ability detected by crystal violet staining semi-quantitative method,OD560≤0.15 was in 8 strains of LH,0.150.20 in 7 strains respectively.The levels of minimal biofilm inhibitory concentration in norfloxacin,ofloxacin,levofloxacin,ciprofloxacin and lomefloxacin to LH were respectively higher than the minimal inhibitory concentration(MIC) in the corresponding floating bacteria(P<0.05).Among 55 strains of LH,18 strains were resistant to quinolones,the resistance rate was 32.7%,and the type I integron in 18 strains of LH carried the drug resistant genes,these drug resistant genes played the drug resistance role in corresponding bacterial strains.Conclusion Drug resistance gene formation and widespread of LH may be associated with type I integron.

BACKGROUND: Both aperture and porosity are mainly evaluating markers for three-dimensional poly materials. The higher the porosity is, the easier the growth and proliferation of cartilage cells are. However, with the successive increasing of porosity, compressive strength of scaffolds decreases and utility of aperture also decreases. Therefore, it is extremely significant for tracheal cartilage tissue engineering to establish three-dimensional poly scaffolds which have suitable aperture and porosity.OBJECTIVE: To establish tube foam scaffolds by using solvent casting/particulate leaching method so as to find out practical and ideal scaffolds for tracheal cartilage tissue engineering.DESIGN: Observational study.SETTING: Department of Otolaryngology, Taihe Hospital, Yunyang Medical College; Department of Otolaryngology, West China Hospital, Sichuan University.MATERIALS: The experiment was carried out in Chemical Institute, Chengdu Sub-college of Chinese Academy of Sciences from March to May 2002. Poly-D, L-lactic acid (PDLLA, Mr= 4.23×104) and sodium chloride granules (50-200 μm in diameter) were used as porogenic agent.METHODS: PDLLA was dissolved in chloroform in spherical-shape glass container to dispend 100 g/L solution and then add with sodium chloride granules (50-200 μm) based on various mass fractions of 800, 850, 900, 920, 940 and 960 g/L. Sodium chloride granules were regarded as porogenic agent (scaffolds numbered from 1 to 6) to stir and make paste suspension. Continuously, suspension was cast into tube models, heated at 90 ℃, compressed,and maintained in ventilation cabinet for 48 hours for solvent volatilization. And the resting solvent was drawn out.Form-fitting drying tube foam scaffolds were taken out and dipped in double distilled water for 48 hours so as to remove sodium chloride. The double distilled water was changed every 8 hours. Then, all tube materials were dried in vacuum drying oven for 24-48 hours. While, three-dimensional PDLLA scaffolds were successfully established.Form and intensity of scaffolds were observed with gross and scanning electron microscope; meanwhile, pore parameter was measured and analyzed.MAIN OUTCOME MEASURES: ①Gross observation of tube foam scaffolds;② measurement of pore parameters.RESULTS: ①Scaffolds were appeared as white tube foam with 8 mm in internal diameter and 12 mm in outside diameter.Scaffolds with 80-250 μm in bore and 90.6% in porosity had defined strength and ductility. ②Scanning electron microscope demonstrated that there were many holes distributed in PDLLA scaffolds in various sizes. Otherwise, hole of scaffolds was connected to each other, while big hole also contained numerous small holes. ③Porosity of scaffolds increased with the increasing mass fraction of sodium chloride; but effective porosity did not increase with the increasing mass fraction of sodium chloride. There were different effective porosities of bore (80-250 μm). Effective bore of number 4 sample was 76% and relative porosity was 90.6%. Therefore, number 4 sample was an ideal scaffold for tracheal cartilage tissue engineering.CONCLUSION: Tube foam scaffold fabricated by solvent casting/particulate leaching method is suitable for tracheal cartilage tissue engineering.

Objective To investigate the expression of YAP,LPA and TAZ in head and neck squamous cell carcinoma and its clinical significance.Methods Selective collection of Ya'an People's Hospital pathology department from January 2010 to December 2016,the surgical resection of the pathological tissue paraffin specimens amounted to 136 cases,including the normal tissue of head and neck squamous cell carcinoma 37 cases,head and neck benign tumor-like tissue 35 cases,head and neck squamous cell carcinoma 64 cases,all specimens without chemotherapy history,The relationship between the expression of LPA,YAP and TAZ in various tissues by immunohistochemistry and real-time quantitative PCR assay and its clinical correlative parameters.Results The expression of LPA mRNA in normal tissues,benign neoplasia tissues and squamous carcinoma tissues of the head and neck squamous cell carcinoma was similar (P ＞ 0.05).The expression of YAP mRNA in squamous cell carcinoma tissues was higher than that in the adjacent tissues and tumor tissues (P ＜ 0.05).TAZ mRNA was low expression in the three tissues of the head and neck,but the expression of squamous cell carcinoma was significantly higher than that of benign tumor tissues and adjacent normal tissues (P ＜ 0.05).The expression of YAP and TAZ protein was significantly higher than that in the tissues of normal tissues and benign tumor tissues (P ＜ 0.05).The expression of YAP in squamous cell carcinoma was significantly higher than the control group (P ＜ 0.05).The expression of TAZ was low differentiation in tumor,tumor ＞3.0 cm,lymph node positive and Ⅲ-Ⅳ period were higher than the control group (P ＜ 0.05).Condusions The expression of YAP and TAZ in head and neck squamous cell carcinoma was closely related to tumor staging,differentiation degree and lymph node metastasis,while the expression of LPA showed no significant difference and correlation.

Objective To investigate the colonization of Gram-negative bacilli carrying mcr-1 gene in intestinal tracts of inpatients and people having physical examination for further elucidating the molecular and epidemiological features of mcr-1 gene. Methods A total of 1263 and 750 fecal specimens were col-lected from inpatients in the First Affiliated Hospital of Guangzhou Medical University and people having physical examination in the Kingmed Physical Examination Centre, respectively. Drug-resistant bacteria were isolated using Maconkey agar supplemented with colistin. PCR was performed to detect the bacteria carrying mcr-1 gene. Multilocus sequence typing ( MLST) and enterobacterial repetitive intergenic consensus-PCR ( ERIC-PCR) were used for homology analysis. The transferability of mcr-1 gene was verified by plasmid transfer assays. Plasmids of mcr-1-carrying strains were typed by PCR-based replicon typing techniques. Twelve virulence-related genes were also detected by PCR. Results Ninety-two colistin-resistant strains were isolated from the 1263 samples from inpatients(7. 3％, 92/1263) and two of them were positive for mcr-1 gene ( one strain also carried the blaNDM-5 gene) . Thirty-six colistin-resistant strains were isolated from the 750 samples of physical examination group (4. 8％, 36/750) and one of them carried the mcr-1 gene. MLST analysis showed that three mcr-1-carrying Escherichia coli strains ( minimum inhibitory concentration of colistin:8 μg/ml) belonged to three different sequence types. Moreover, they exhibited different banding patterns in ERIC-PCR analysis. All of the mcr-1-carrying isolates could transfer mcr-1 gene to the recipient strains successfully. Six types of incompatibility plasmids were detected in the mcr-1-carrying isolates ( IncFⅡ, IncX2, IncHI2, IncFIB, IncX4 and IncX1). Virulence-related genes fimH, iutA and fyuA were detec-ted in all mcr-1-carrying Escherichia coli strains. Conclusions Colistin-resistant strains and mcr-1 gene are prevalent in inpatients and people having physical examination, which brings potential risk for the control of clinical infections.

OBJECTIVE: To investigate the features of nasal septum cellule in computed tomographic (CT) images and its clinical significance. METHOD: CT scans data of nasal septum in 173 patients were randomly obtained from January 2001 to June 2005. Prevalence and clinical features were summarized in the data of 19 patients with nasal septum cellule retrospectively. RESULT: (1) Nineteen cases with nasal septum cellule were found in 173 patients. (2) All nasal septum cellule of 19 cases located in perpendicular plate of the ethmoid bone, in which 8 cases located in upper part of nasal septum and 11 located in middle. (3) There were totally seven patients with nasal diseases related to nasal septum cellule, in which 3 cases with inflammation, 2 cases with bone fracture, 1 case with cholesterol granuloma, 1 case with mucocele. CONCLUSION Nasal septum cellule is an anatomic variation of nasal septum bone, and its features can provide further understanding of some diseases related to nasal septum cellule.
Adolescent ; Adult ; Aged ; Ethmoid Bone ; diagnostic imaging ; Female ; Humans ; Male ; Middle Aged ; Nasal Bone ; diagnostic imaging ; Nasal Septum ; diagnostic imaging ; Retrospective Studies ; Tomography, X-Ray Computed ; Young Adult

Rabbit bone marrow-derived mesenchymal stem cells(MSCs) are multipotent. We studied the adipogenic and osteogenic differentiation potent using adipogenic supplement (AS) or osteogenic supplement (OS) in vitro. Specific markers of this induced adipogenic and osteogenic lineage were identified. The findings showed that the rabbit MSCs are capable of differentiating into adipogenic and osteogenic lineages spontaneously. On the 21st day, approximately 75% rabbit MSCs were induced to adipogenic or osteogenic cells in medium containing AS or OS, respectively. These results demonstrated that the differentiation of MSCs could be regulated in vitro. The underlying molecular mechanisms of adipogenic or osteogenic differentiation await elucidation.
Adipose Tissue ; cytology ; Animals ; Bone and Bones ; cytology ; Cell Differentiation ; Cell Lineage ; In Vitro Techniques ; Mesoderm ; cytology ; Rabbits ; Stem Cells ; cytology