BACKGROUND:A preliminary experiment developed a double-chamber stirred bioreactor which can carry out osteogenic and cartilage induction at the same time.
OBJECTIVE:To explore the effects of double-chamber stirred bioreactor on the repair of goat knee cartilage defects with tissue-engineered cartilage.
METHODS:Twelve goats were selected to make bilateral femoral condyle osteochondral defects models and randomized to three groups:experimental group, implanted with the composites ofβ-tricalcium phosphate and bone marrow mesenchymal stem cells that were subjected to 2-week chondrogenic and osteogenic induction simultaneously in the double-chamber stirred bioreactor under mechanical stimulation;control group, implanted with the composites ofβ-tricalcium phosphate and bone marrow mesenchymal stem cells that were subjected to 2-week chondrogenic and osteogenic induction simultaneously in the double-chamber stirred bioreactor;blank control group, without treatment. After 12 and 24 weeks of implantation, general observation, Masson staining, II col agen immunohistochemical staining and histological scoring were performed.
RESULTS AND CONCLUSION:In the experimental and control groups, new cartilage tissue and bone tissue were visible, but the experimental group showed better repair effects than the control group (P<0.05). The blank control group had no cartilage formation. These findings indicate that under the mechanical stimulation by the double-chamber stirred bioreactor in vitro, the repair effect of tissue-engineered osteochondral complex on knee joint cartilage defects can be improved.

Objective To observe the antioxidative dosage-effect relationship of Marigold lutein, and provide experimental data for clinical use. Methods The mice were randomly divided into seven groups:blank control group, model control group, 1 mg/kg lutein group, 5 mg/kg lutein group, 25 mg/kg lutein group, 125 mg/kg lutein group and 625 mg/kg lutein group. The mice in blank control group were dealt with saline solution by intraperitoneal injection, the others were dealt with D-galactose (120 mg/kg) by intraperitoneal injection for seven weeks to make oxidative damage model, meanwhile the mice were given corresponding dose of the drug. Subsequently, the level of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in the serum were measured, and the antioxidative dosage-effect relationship was observed. Results The 1, 5, 25 mg/kg lutein reduced the MDA level and increased SOD activity, and the 125, 625 mg/kg dose of lutein did not show significant antioxidant activity. Conclusion Lutein has significant antioxidant activity in mouse dealt with D-galactose within the dose range of 1-25 mg/kg. The results suggest that the clinical dosage range of lutein should be kept within reasonable limits.

The Escherichia coli (E.coli) strain from intestinal tract of died newborn swine was isolated and cultured.Preliminary identification of the isolated strain was conducted by conventional biochemical tests,and the molecular biology detections of toxicity gene and typing gene were completed by multi-PCR.Stable toxin and heat-labile enterotoxin genes of E.coli were detected from the isolated strain.By amplifying and sequencing bacterial 16s rDNA and fliC gene,the isolated strain was identified as H10 by Blast analysis.The homology of strain H10 was 99% with bacterial 16s rDNA gene and 98% with fliC gene.

Objective To observe the antioxidative dosage-effect relationship of grape seed proanthocyanidin extract (GSPE). Methods The mice were randomly divided into seven groups:blank group, model group, 5 mg/kg GSPE group, 15 mg/kg GSPE group, 45 mg/kg GSPE group, 135 mg/kg GSPE group and 405 mg/kg GSPE group. The mice in blank group were dealt with saline solution by intraperitoneal injection, the others were dealt with D-galactose (120 mg/kg) by intraperitoneal injection for seven weeks to make oxidative damage model. Meanwhile, the mice were given corresponding dose of the drug. Subsequently the level of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) in the serum were measured to observe the antioxidative dosage-effect relationship of GSPE. Results The 45, 135, 405 mg/kg GSPE group reduced the MDA level, and the 15, 45, 135, 405 mg/kg GSPE group increased the SOD activity. Conclusion GSPE has significant antioxidant activity on mice dealt with D-galactose above the dose of 15 mg/kg, suggesting that the clinical use of GSPE should guarantee a certain dose to play a good antioxidant effect.

cuses tumors. Survivin seems to be a promising marker for analyzing clinical stages and predicting the prognosis of TCC.

Objective To investigate the value of HPV E6/E7 mRNA test combining liquid-based cytology test in cervical cancer screening. Methods A total of 377 samples from Wenzhou People's Hospital from June 2014 to September 2015 were collected and screened by HPV E6/E7 mRNA test combining with liquid-based cytology test , and the results was compared with the findings from the gold criteria of histology and pathology. Results The combination of HPV E6/E7 mRNA test and liquid-based cytology test can enhance the testing sensitivity and specificity. The sensitivity of the combination of HPV E6/E7 mRNA test and liquid-based cytology test for the diagnosis of LSIL was 94.41%, and that for the diagnosis of HSIL was 96.36%. Based on the gold criteria of histology and pathology , the sensitivity , specificity , positive-predictive value and negative predictive value of HPV E6/E7 mRNA test for the diagnosis of HSIL was 90%, 60.67%, 48.53% and 92.49%respectively. The sensitivity, specificity, positive-predictive value and negative- predictive value of liquid-based cytology test for the diagnosis of HSIL was 72.73%, 75.28%, 54.79% and 87.01% respectively. Conclusions The sensitivity of HPV E6/E7 mRNA test is superior to that of liquid-based cytology test , while the specificity of HPV E6/E7 mRNA test is inferior to that of liquid-based cytology test. The negative predictive value of HPV E6/E7 mRNA test is more meaningful than that of liquid-based cytology test. The combination of HPV E6/E7 mRNA test and liquid-based cytology test can enhance the testing sensitivity , but it does not increase the specificity.

Purpose To assess the feasibility of dual-energy dual-source virtual noncontrast (VNC) CT in the diagnosis of laryngeal carcinoma,in order to reduce radiation hazard.Materials and Methods Forty-nine patients with clinically identified laryngeal carcinoma underwent conventional non-contrast (CNC) scan as well as arterial and venous dual-phase and dual-energy VNC scan.The VCN images of arterial and venous phase were obtained by using the dual-energy software.The mean CT value,signal-to-noise ratio (SNR),contrast noise ratio (CNR),lesion morphology,image quality and radiation dose were compared between the VNC and CNC images.Results The mean CT value,SNR,CNR and image quality scores of CNC were higher than those of VNC images of arterial and venous phase (P＜0.05).There was no significant difference in the image quality scores of VNC between the arterial and venous phase (P＞0.05).The image quality scores of VNC were all above 3,which could meet the diagnostic requirements.There was no significant difference in the shape,size,boundary and cystic necrosis of the lesions,invasion of the surrounding space,osteolytic destruction and size of the surrounding lymph node between the CNC and VNC image.The radiation dose of dual-phase dual-energy scan was 0.42 mSv,which was 18.3％ lower than that of the conventional three-phase scan.Conclusion For laryngeal carcinoma,the VNC images from the dual-energy CT scan can be used to reduce the radiation dose without affecting image quality and diagnosis accuracy,which has good clinical value.

This study is to report the establishment of an UPLC-MS/MS method for the determination of plasma concentration of UA carried in self-microemulsifying drug delivery system (SMEDDS) and its pharmacokinetics in rats. It was used for determination and analysis when serum with internal standard was extracted from C18 solid-phase column. Acquity UPLC BEH C18 column (100 mm x 2.1 mm, 1.7 microm) was used for separation. The mobile phase was acetonitrile -0.1% ammonia with gradient elution at the flow rate of 0.2 mL x min(-1). The column temperature was 40 degrees C and the detection wave length was 210 nm. It was detected by negative ion using electrospray ionization source (ESI) and scanned by multiple reaction ion monitoring (MRM) mode. The liner relationship of UA was very good in the range of 1.19-3 815.00 ng x mL(-1) (r = 0.999 0). Recovery rate of different concentrations were 87.42%-89.95%. The precision of inter-day and intra-day were less than 11%. The method developed in our study was proved to be sensitive, rapid and simple. It is suitable for the pharmacokinetic study of UA-SMEDDS in rats.
Animals ; Chromatography, High Pressure Liquid ; Drug Delivery Systems ; Emulsions ; chemistry ; Rats ; Tandem Mass Spectrometry ; Triterpenes ; blood ; pharmacokinetics

The main factors which affected the isolation, purification and cultivation of Pinellia cordata protoplasts from leaves were studied. The results indicated that the optimum enzyme solution for P. cordata leaves was 13% CPW + 1.0% Cellulose +0.1% Pectolase, at pH 6.0, temperature (25-28 degrees C ) for 4 h. The sucrose density gradient centrifugation was adopted to purificate the protoplasts collected, when 25% sucrose was used as mediator, centrifugating at 500 rpm for 10 min. When the protoplasts were shallow liquid and liquid-solid double layer cultured on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA + 13% mannitol at the density of 2.5 x 104 protoplasts/mL, or fed and nursed cultured at the density of 100-500 protoplasts/mL, cell division could be observed for 3 days; granular calli appeared for 30 days. Calli was proliferated on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA solidified by 0.55% agar, and differentiated and regenerated after 5-6 months. Plant generation of P. cordata is successfully established.
Cell Separation ; methods ; Culture Media ; Pinellia ; physiology ; Protoplasts ; physiology ; Regeneration

OBJECTIVE: To investigate characteristics of molecular etiology of children with profound sensorineural hearing loss in Hubei province, and to provide reference for deafness treatment and genetic counseling. METHOD: Three hundred and six children with profound sensorineural hearing loss in Hubei province were enrolled, their genomic DNA were extracted from peripheral blood and a deafness gene test chip was used to screen nine hot spot mutation in the GJB2, GJB3, SLC26A4, and mitochondria 12SrRNA gene. All patients with SLC26A4 gene mutation were given temporal bone CT scan. RESULT: One hundred and thirty-two (43.14%) out of 306 children were found carrying at least one pathogenic gene mutation. The mutation rates of GJB2, SLC26A4 and mitochondria DNA 12SrRNA gene were 29.41% (90/306), 13.72% (42/306) and 0.65% (2/306), respectively. None out of 306 children was detected GJB3 gene mutation. Thirty-six patients carrying SLC26A4 gene mutation were detected enlarged vestibular aqueduct by CT scan. CONCLUSION Mutations of GJB2 and SLC26A4 gene are two major pathogenic gene for genetic hearing loss in children. 235delC mutation is the main mutation type, followed by IVS7-2A> G mutation type. The screening of SLC26A4 gene common mutations contribute to the diagnosis of enlarged vestibular aqueduct syndrome.
Adolescent ; Child ; Child, Preschool ; China ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; Deafness ; genetics ; Female ; Genetic Testing ; Humans ; Infant ; Male ; Membrane Transport Proteins ; genetics ; Mutation ; Oligonucleotide Array Sequence Analysis ; Sulfate Transporters