OBJECTIVEThis study is performed to investigate the cell topographies and biomechanical properties of two different types of temporomandibular joint (TMJ) discs from goats by using JPK Nano Wizard 3 biological atomic force microscopy (AFM). This process provides a guideline for selecting seed cells for TMJ disc tissue engineering.

METHODSTMJ disc cells from primary goats were cultured by monolayer culture method. AFM was used to contact scan the topographies of the two types of TMJ disc cells under physiological environment. Approximately 20 chondrocyte-like and fibroblast-like cells were selected randomly to plot the force-versus-distance curves of the cytoplasm and nucleus. Young's modulus and adhesion were analyzed by JPK Data Processing.

RESULTSThe triangle-shapednucleus of the chondrocyte-like cell occupied a large portion of the cell. Cytoskeleton was arranged dendritically on the surface. Pseudopodia were extended from cell edges. The spindle-shaped nucleus of the fibroblast-like cell occupied a significantly larger region compared with the cytoplasmic region. Cytoskeleton was arranged regularly. Cell edges were smooth with less pseudopodia extended. No difference was found in the surface roughness between the two types of cells. According to the force-versus-distance curves, the Young's moduli of the two types of cells were not statistically different (P>0.05), but differences were found in the cytoplasmic regions (P=0.047). No statistical difference was found in the adhesions between the two types of cells (P>0.05).

CONCLUSIONThe AFM topography and curves were compared and analyzed. The two types of TMJ disc cells exhibited significantly different topographies, but only slight difference in their mechanical abilities.

Objective:To investigate the effect of the intact parathyroid hormone (iPTH) on residual renal function (RRF). Methods: The relationship between iPTH and calcium, phosphorum, product Ca?P, hypertension, triglycende, cholesterol,left ventricular mass index(LVMI) and RRF in 120 hemodialysis patients with chronic renal failure. Results: The results showed that 95. 9% of the hemodialysis patients with chronic renal failure had secondary parathyroldism. It was found that iPTH was positively correlated with SBP,DBP, product Ca?P, triglycende and LVMI, and negatively correlated with endogenous creatinine clearance rate and KT/V. RRF had positive correlation with KT/V and SBP,DBP, calcium, product Ca?P, triglyceride, cholesterol and LVMI. Conclusion: iPTH level is elevated in hemodialysis patients, which may lead to RRF loss.

Objective:To investigate the inhibitory effect of RNA interference (RNAi) on the expression of Aurora-A in SKOV3 cells and on proliferation of SKOV3 cells.Methods:Two pairs of oligo small interference RNA (Oligo siRNA) specific for Aurora-A were designed for RNAi and were transferred into SKOV3 cells.The expression of Aurora-A were de- tected by RT-PCR and Western blot.Furthermore,the cell proliferation and apoptosis were observed by MTT and FCM af- ter transfection.Results:After transfection with Oligo siRNA,mRNA and protein level of Aurora-A gene in SKOV3 cells were obviously reduced,while the inhibitory rate of proliferation and apoptosis rate in SKOV3 cells were increased signifi- cantly.Conclusion:The Oligo siRNA specific for Aurora-A can reduce the expression of Aurora-A gene and induce apop- tosis of SKOV3 cells.

Objective To study whether the organs besides digestive system of musca domestica Ⅲ instar larvae have the capability of produceing musca β-glucosidase.Methods Tissues of malpighian tubules,trachea,epiploon and body wall of musca domestica Ⅲ instar lar-vae were dissected under anatomic microscope,and the expression of β-glucosidase gene in these dissected tissues were detected by reverse transcription PCR.And the tissue localization of β-glucosidase mRNA was further identified by in situ hybridization.Moreover,anti-cellulase was used to determinate the tissue distribution with immunohistochemical staining.The relative mRNA expression levels of musca domesticaβ-glucosidase gene in these organs were tested by real-time quantitative PCR.Results The reverse transcription PCR showed that the ampli-fication products of β-glucosidase gene were observed in tissues of malpighian tubules,trachea and body wall.β-glucosidase mRNA was shown in the epithelium cells of malpighian tubules,trachea and body wall by in situ hybridization,and it was almost the same in the results of im-munohistochemical staining.The real-time quantitative PCR showed that the relative expression quantity of β-glucosidase gene in malpighian tubules and body wall were higher than that in foregut,while it was lower in itrachea than that in foregut.And it was of statistical difference in gene expression level of β-glucosidase among these organs (P <0.05).Conclusion Malpighian tubules,trachea and body wall of musca domestica Ⅲ instar larvae have the function of secreting β-glucosidase.Combining with the characteristics of secreting β-glucosidase in most organs of digestive system,it may provide a new biological method for the prevention and treatment of human diseases transmitted by musca domestica with the use of taget gene β-glucosidase.

Objective To investigate the relationship between the expression of MMP-7 in peripher- al blood and esophageal cancer.Methods By using Real time RT-PCR, blood samples from 82 cases with postoperative esophageal cancer were detected.Results 37.8 %(32/82)patients were positive for MMP-7 mRNA expression.MMP-7 mRNA positivity significantly correlated with lymph node metastasis,stage,and hematogeneous metastasis.Recurrent disease was found in 28 of 82 cases. Of 28 patients experiencing re- lapse,15 patients were serially follow-up in every three months.The number positive of MMP-7 mRNA be- fore detection by imaging,at the same time of detection by imaging,and after detection by imaging was 5 cas- es,8 cases and 2 cases respectively.Conclusion Examination of MMP-7 mRNA in peripheral blood during follow-up is useful for early detection of occult recurrence.

Objective To investigate the clinical value of low-dose decitabine (DAC) in elderly patients with acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS) patients with intermediate-or high-risk. Methods Low-dose DAC (10 mg/d, 7 days) combined with CAG regimen were given to 19 elderly patients with AML and intermediate- or high-risk MDS patients. The efficacy and adverse reactions were evaluated after a course of treatment, and the patients were followed up for survival. Results After a course of treatment, 8 patients achieved complete remission (CR), 7 patients achieved partial remission (PR). After 4 courses of treatment, 68.4 % (13/19) of patients achieved CR, the overall response rate reached 78.9% (15/19). Fewer side effects were seen associated with chemotherapy. After 42 months of follow-up, there were 12 survival cases, the median survival time was 13.5 months (3-42 months). Conclusion Low-dose DAC combined with CAG regimen have a better efficacy, higher safety, and lower economic burden for elderly AML patients and intermediate- or high-risk MDS patients, which is beneficial to greatly improve patients' compliance.

OBJECTIVE:To explore the regularity of relative correction factor (RCF) in multi-components by single marker (QAMS). METHODS:With the source of CNKI,literature retrieval was used to collect the relevant documents of multi-compo-nents by single marker (QAMS) and extract the calculated values by quantitative analysis of multi-components by single marker QAMS and measured values by external standard method (ESM),SPSS 19.0 software was used to calculate the percentage value (RSD,%) of relative standard deviation between calculated values and measured values,and the correlation between RSD and RCF was analyzed by drawing a scatter plot. RESULTS:Most of RCF values distributed in the range of >0.62-1.53,accounting for 76.7%to all data. Calculated values and measured values showed high similarity in this range and the RSD<3.0%data account-ed for 88.8% to all data,which had good credibility. Meanwhile,the similarity of calculated values and measured values was influ-enced by the differences of structure and physicochemical property of parent nucleus between other components under measured and internal references. The high degree of similarity between calculated values and measured values were decisive by very similar struc-tures of parent nucleus;the low degree was decisive by discrepant structures;and the low degree was also decisive by very similar structures and discrepant physicochemical property of parent nucleus. CONCLUSIONS:RCF will affect the credibility of content de-termination results for components under measured in QAMS,it is necessary to establish database of RCF,RSD and related param-eters.

BACKGROUND:Studies have found that combination of two of chitosan (CS), nano-hydroxyapatite (nHA) and poly(lactide-co-glycolide) (PLGA) can improve the mechanical properties and biocompatibility of the composite stent in certain extent as wel as improve osteogenic differentiation of the cels, but there is a certain distance from the ideal bone tissue engineering scaffolds.
OBJECTIVE:To study biocompatibility and osteoinductive activity of nHA/CS/PLGA scaffolds with different proportions in vitro.
METHODS: nHA/CS/PLGA scaffolds were prepared at mass ratio of 10:10:80, 10:20:70, 20:10:70 respectively by particle leaching method. And human bone marrow stem cels (hBMSCs) were co-cultured with these scaffolds in vitro. Adhesion, proliferation, and osteoinductive activity of these scaffolds were examined qualitatively and quantitatively by growth curve of hBMSCs on scaffolds. Gene expression of alkaline phosphatase activity and osteocalcin was detected by RT-PCR.
RESULTS AND CONCLUSION: hBMSCs could be attached, proliferated, and osteoinduced better on the nHA/CS/PLGA scaffold with the mass ratio of 20:10:70, compared to the other two groups of scaffolds. The differences were significant statisticaly (P< 0.05). Alkaline phosphatase and osteocalcin expressions were respectively higher in the scaffold with the mass ratio of 20:10:70 after 9-27 days of co-culture and 15-27 days of co-culture, in comparison with the other two groups of scaffolds. These findings indicate that the nHA/CS/PLGA scaffolds with the mass ratio of 20:10:70 demonstrated preferable biocompatibility and osteogenic inductivity, which is expected to be a promising scaffold material for bone tissue engineering.

Objective To discuss the prevalence of non-EV71,non-CA16 virus strains of hand,foot and mouth disease(HFMD) in Guangdong province between 2008 and 2009,and analyze the genetic evolution of these non-EV71,non-CA16 virus strains.Methods Isolated viruses from stool samples collected from outpatient and in-patient cases of HFMD between 2008 and 2009 by human rhabdomyosarcoma(RD) cell and HEp-2 cell,cultures that exhibited a characteristic enterovirus cytopathic effect were evaluated by RT-PCR.Those strains which identified non-EV71,non-CA16 were analyzed by VP1 sequencing and then were identified by BLAST program.A phylogenetic tree was constructed using the Neighor-Joinning method in the MEGA 4.0 software.Results Twenty-two virus strains of non-EV71,non-CA16 were obtained,and nine of the twenty-two virus strains in 2008 were classified into CA2,CA4,and CB3 by BLAST; thirteen of the twenty-two virus strains in 2009 were classified into EV80,Echo13,Echo30,CBS,Echo24,CA10,CA6,and poliovirus 1 by BLAST.The honology of all strains was low,and all the strains belonged to CA,CB,Echoviruses,Enterovirus and poliovirus subgroup.Conclusion Except for EV71 and CA16 was a major causative agent in prevail of HFMD in Guangdong province between 2008 and 2009,there also existed other subgroup Enterovirus.The other twenty-two strains respectively belonged to CA,CB,Echoviruses,Enterovirus and poliovirus subgroup,and none of those strains was predominant.Muti-species Enterovirus occurred concomitantly.

Objective: To investigate the long-term effect of submandibular salivary gland transfer on xerostomia induced by radiation in patients with nasopharyngeal carcinoma (NPC). Methods: A total of 70 eligible patients with NPC were divided into the test group (36 cases) and the control group (34 cases). In the test group, the submandibular salivary glands were transferred to the submental space before conventional radiotherapy (XRT) and shielded during XRT. Submandibular gland function and salivary fluid before and after radiotherapy, questionnaire of xerostomia at 60 months after XRT, and 5-year survival rate were compared between the two groups. Results: At 5 years after XRT, the trapping and excretion function of submandibular glands were significantly better in the test group (P=0.000 and P=0.000, respectively). The mean weight of saliva after XRT was greater in the test group than in the control group (1.65gvs.0.73g, P=0.000). Incidence of moderate to severe degree of xerostomia was significantly lower in the test group than in the control group (12.9%vs.78.6%, P=0.000). No significant difference was found in 5 year survival rate between the two groups (86.1%vs.82.4%, P＞0.05). Conclusion: Submandibular gland transfer procedure is safe for NPC patients. It can prevent XRT induced xerostomia and improve the quality of life of NPC patients.