Objective:To understand the roles of PTHrP in the pathogenesis of temporomandibular joint after over mechanical loading. Methods:Fifteen adult New Zealand rabbits were subjected to traction between the mandibular ramus and zygomatic arch in the postero-superior direction unilaterally using elastic force. The rabbits were killed respectively at 2, 4 and 6 weeks and the histologic changes were observed by Hematoxylin & Eosin staining. The expression of PTHrP in the condyle of TMJ was observed by immunohistochemistry. Results:Stronger expression of PTHrP could be found in proliferation cell layers and upper hypertrophy cell layers in the early stage after operation, and weaker expression in mid stage, but stronger near the chondrocyte clusters. Conclusion:It is suggested that PTHrP might relate to the regeneration of condyle cartilage.

Objective To study the occurrence of BRCA1 MSI in endometrial cancer and its relationship with clinic pathologic features; to explore the correlation between MSI and protein expression in BRCA1 gene. Methods Application of PCR-SSCP and DNA sequence analysis method was used to study D17S579 and D17S1349 in 49 sporadic endometrial cancer tissues, 20 cases with endometrial atypical hyperplasia and 28 cases with normal endometrial tissues. Results In the total samples of D17S579 and D17S1349, the three groups were significantly different: 34.69%(17/49) in the endometrial cancer group, 10%(2/20) in the endometrial atypical hyperplasia group and 7.14%(2/28) in the normal endometrium group (χ2= 11.208, P = 0.004). BRCA1 MSI positive rate related to the pathology grade and clinical stage, but no relationship was found in muscular infiltration depth, lymph node metastasis and histopathology types. In the endometrial cancer group, BRCA1 MSI positive rate and BRCA1 protein expression were in moderate correlated negatively (r = -0.779, P = 0.000). Conclusion BRCA1 MSI might play a role in the development of endometrial cancer, and low expression of BRCA1 protein. BRCA1 MSI might be associated with pathology grade and clinical stages in EC.

Objective To investigate the short term effect of insulin on proinsulin gene expression of HIT-T15 insu-linnoma cells(pancreatic isletβ-cell). Methods The HIT-T15 cells were randomly divided into four groups.Blank Con-trol Group (LG):complete medium contain 1.4 mmol/L glucose. Control group (LGC):co-cultured nifedipine with medium in order to restain endogenous insulin release. Experimental group (LINS or HINS) add 0.5 U/L insulin or 5 U/L insulin on top of LGC. After being stimulated for 0, 30, 60, 90, 120 mins, proinsulin (PI) mRNA level were assessed by semi-quantitative RT-PCR. Insulin receptor substrate1 (IRS1) tyrosine phosphorylation was detected by immunocytochemistry. Results (1) Expression of PI was up regulated by both LINS and HINS, and peak at 60 mins. (2) After stimulation for 30 mins, the level of IRS1 tyrosine phosphorylation in the experimental group was significantly higher than control group, and the peak time be-tween LINS and HINS was different. (3) Between group of LG and LGC, the expression of PI mRNA and IRS1 tyrosine phos-phorylation show no difference. Conclusion Short term exogenous insulin stimulation can promote expression of proinsulin genes,which is concentration dependent. The expression and regulation of PI were related with IRS1 signal transduction.

OBJECTIVE: To analyze the rate of 235delC mutation in GJB2 gene in patients with idiopathic sudden hearing loss, and to explore its possible correlation with pathogenesis of idiopathic sudden hearing loss. METHOD: Two hundred and thirty-four patients with diagnosis of idiopathic sudden hearing loss in otolaryngology department were recruited as experimental group. Eighty people with normal hearing level were enrolled as control group. Their peripheral blood samples were obtained and genomic DNA was extracted. Using polymerase chain reaction, the coding region of GJB2 gene was amplified, and 235delC mutation is screened for in GJB2 gene by restriction endonuclease. At same time the clinical data of 234 patients was collected to analyze. RESULT: In 234 cases of idiopathic sudden hearing loss, 5 cases were found to have heterozygous 235delC mutation, none of them harbored homozygous 235delC mutation, the 235delC mutation rate was 2.1% (5/234). No 235delC mutation was found in control group. The rate of 235delC mutation in two group showed no statistically significant difference (P > 0.05). CONCLUSION This research shows that the rate of 235delC mutation in GJB2 is low in patients with idiopathic sudden hearing loss, and suggest that 235delC mutation possible has no correlation with idiopathic sudden hearing loss.
Adolescent ; Adult ; Aged ; Child ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; Female ; Hearing Loss, Sudden ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Young Adult

Objective To analyze the characteristics of laboratory test resuits of dengue fever(DF)patients in Guangzhou area.Methods Routine tests were performed in the patients admission to hospital. Serology examination was performed in the patients in acute phase or recovery phase.The clinieal symptoms and teatures were analyzed and positive numbers and positivity ratios were calculated.Results The clinical symptoms of the dengue fever were typieal,with the features of fever,headache,myalgia and rash.The leukopenia rate was 76.0%,and the thromboeytopenia rate was 62.6%.The levels of ALT increased in 56.7%patients,and the levels of AST increased in 84.0%patients.Hypopotassemia was found in 46.1%patients.Dengue virus antibody IgM(DF-IgM)was detected positive from the first day to the 16th day of the onset,and the positive rate was 85.9% on the 8th day.Virus loads were positive by fluorescence real-time PCR in seven acute serum samples(within 3 days of the onset)of 51 cases whose DE-IgM were negative all the time, and the results was 105 -106 copies/ml(＜103 copies/ml means negative).Conclusions Clinical manifestations of this DF epidemic were typical including fever,headache,myalgia and skin rash.Most of the patients had decreased leukocyte and thrombocyte obviously.Liver damage was common but kidney damage was seldom.Halt of the patients got hypopotassemia.DF-IgM appeared in very early and persisted for a long time.The detection of DF-IgM within 7 days of the onset was helpful for diagnosis as early as possible.Viral load detected by real-time PCR could be another indicator of early pathogen diagnosis which provides complementation for antibody detection.

Objective:This work aims to investigate diallyl disulfide (DADS) inhibition of cell migration and invasion in human colon cancer SW480 cells through the Rac1-ADF/cofilin1 pathway. Methods:The potential of cell migration and invasion was examined by scratch healing assay and transwell membrane assay. The expression of Rac1-ADF/cofilin1 pathway was detected by RT-PCR and Western blot. Results:After the SW480 cells were treated with 40 and 50 mg·L-1 of DADS for 24 h, the number of transmembrane cells through the Matrigel obviously decreased by 57.12%and 64.59%, respectively (P<0.05). After cell treatment for 48 h, the cell migration rates were 23.23%and 12.87%, which were significantly lower compared with the control group (75.86%;P<0.05). After the cells were treated with 45 mg·L-1 of DADS for 24 and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and destrin mRNA respectively decreased compared with the control group (P<0.05). However, no significant difference was observed in the expression of cofilin1 mRNA (P>0.05). After the treatment with 45 mg·L-1 of DADS for 6, 12, 24, and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and Destrin proteins respectively decreased in a time-dependent manner compared with the control group (P<0.05). However, no significant differences were observed in the expression of the cofilin1 protein (P>0.05). Moreover, the expression of p-LIMK1 and p-cofilin1 notably decreased in a time-dependent manner (P<0.05). Conclusion:DADS inhibits cell migration and invasion, which is related to the down-regulation of Rac1, Rock1, PAK1, LIMK1, p-LIMK1, p-cofilin1, and destrin through the Rac1-ADF/cofilin1 pathway.

Objective:To investigate whether chitosan-polyelectrolyte complex (CS-PEC) can be used as scaffold for chondrocyte culture and for cartilage regeneration in vivo.Methods:Condylar chondrocytes of fetal mouse were seeded onto the three-dimension gel scaffolds of CS-PEC and cultured.The cultured chondrocytes/CS-PEC complex samples were transplanted subcutaneously into nude mice and the CS-PEC scaffold without chondrocyte was used as the control.The animals were sacrificed 4 and 8 weeks after operation respectively.Cartilage formulation was observed by histological and immunohistochemical methods.Results:In the in vitro culture the majority of cells attached to the CS-PEC surface and expanded rapidly. 4 weeks after transplantation,in the chondrocytes/CS-PEC complex the scaffold maintained mostly the original structure. Hypertrophic chondrocytes appeared in scaffold materials. CollagenⅡwas positive in the new cartilage. 8 weeks after transplantation the scaffold degraded almost completely and new cartilage could be observed. CollagenⅡ and cartilage matrix was positive in the new cartilage and the collagen I was positive in the surrounding fibroblast-like cells. In control transplants,8 week after transplantation some fibre-like tissue formed in the circumference, but there was no new cartilage formation and the collagen II and the cartilage matrix was negative.Conclusions:CS-PEC may be used as scaffold for fibre-cartilage regeneration.

Objective To lay the foundation for further exploration on parasitifer's defence reaction to pneumolysin through constructing ply gene-deletion strain of Streptococcus pneumoniae and researching on its virulence change. Methods A linker fragment with erm gene in middle and homologous upstream and downstream fragment of ply gene at both sides was prepared by long flanking homology-polymerase chain reaction(LFH-PCR). The linker fragment was transformed into Streptococcus pneumoniae. ply-deficient strain was then screened out from blood plate which contains erythromycin and identified by PCR. ply-deficient strain growth in vitro was observed and virulence change was observed through infecting mouse model. Results PCR results showed that ply gene was replaced completely by erm gene. The ply deficient strain was successfully constructed. The growth of single strain culture medium showed that ply genetic defect made no influence on bacterial's external growth. While in the mice nasal cavity infecting experiment, deficient strain enter into blood after 6 h from infecting which obviously slower than that did wild-type(2 h). And the number of bacteria at each point was much smaller than that of wild-type(P ＜0. 01 ). The mice peritonaeum infecting experiment showed that median lethal time of wild-type was 3 d, while that of deficient strain was 18 d(P＜0. 01). Conclusion It is a good way to completely substitute ply gene using LFH-PCR. ply deletion made no influence on baterial's growth in vitro, but it resulting in reduction of bacterial virulence in vivo.

Objective To observe changes of cell cycle after stable silencing of SNCG gene in human endometrial carcinoma HEC-1A cells.Methods Flow cytometry was used to observe the changes of SNCG stable silencing HEC-1A cell growth cycle.The percentage of G2/M phase of HCE-1A cells after acridine orange staining was observed by laser scanning confocal microscopy.Results Flow cytometry results showed that the group of SNCG stable silencing in G1 and G2/M phase of the cells increased,but the percentage of cells in the S phase were decreased,the difference were both statistically significant (P ＜ 0.05).The results of cells after confocal microscopy observation of acridine orange staining,the group of SNCG stable silencing were observed in G2/M phase high percentage were statistically significant (P ＜ 0.05).Conclusion Two experimental methods commonly validated that the cell cycle of SNCG stable silencing on HEC-1A cells was arrested in G2/M phase,suggesting that SNCG gene is closely related to the cell growth cycle of endometrial carcinoma.

Objective: To study the effect of different proportions of piper chinaroot and rheum palmatum on the dissolution rate of five effective components (aloe emodin, emodin acid, emodin, emodin, emodin methyl ether). Methods: The high performance liquid chromatography (HPLC) method was used to analyz the contents of five effective components of rheum palmatum in the extracts of different combination of piper chinaroot and rheum palmatum. The tests were carried out by Thermo C18 (250 mm×4.6 mm, 5 μm) by gradient elution with methanol and 0.1% phosphoric acid water solution as mobile phase at a flow rate of 1ml/min, the column temperature was 30 ℃ and the detection wavelength was 254 nm. Results: The linear ranges of aloe emodin, emodin acid, emodin, emodin, emodin methyl ether were 0.018 5-0.741 8, 0.017 9-0.717 8, 0.015 9-0.635 5, 0.054 2-2.167 2, 0.016 2-0.646 4 μg, respectively. The average recoveries of aloe emodin, emodin acid, emodin, emodin, emodin methyl ether were 94.35%, 95.50%, 100.61%, 96.27%, 97.39%, and the RSDs were 1.81%, 1.99%, 2.84%, 2.71%, and 1.86%, respectively. Compared with rheum palmatumby single extract, the content of aloe emodin and emodin were increased by 5 proportions (3:1, 2:1, 1:1, 1:2, 1:3), while the content of emodin and emodin methyl ether were decreased. Conclusions The optimal compatibility proportion of piper chinaroot and rheum palmatum is 1:1.