BACKGROUND:Calcitonin gene-related peptide has been confirmed to induce osteogenic differentiation, but whether it can induce the osteogenic differentiation of adipose-derived stem cells under three-dimensional culture to construct tissue-engineered bone is rarely reported.
OBJECTIVE:To investigate the feasibility of exogenous calcitonin gene-related peptide to induce osteogenic differentiation of adipose-derived stem cells combined with calcium alginate gel in three-dimensional condition.
METHODS:Adipose-derived stem cells were gained by col agenase I digestion of the subcutaneous adipose tissue of both inguinal regions of New Zealand rabbits. Passage 3 cells were mixed with sodium alginate to prepare calcium alginate gel, and then the cells were assigned into two groups and cultured in 24-wel plates. Adipose-derived stem cells in the control group were cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 supplemented with 10-2 mol/Lβ-glycerophosphate sodium, 10-7 mol/L dexamethasone, 50 mg/L ascorbic acid, 10%fetal bovine serum. While, adipose-derived stem cells in the experimental group were incubated with the same medium as above, but 1.5 μg/L calcitonin gene-related peptide was added. The cells proliferation and the mRNA expressions of col agen I and osteocalcin were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and reverse transcription-PCR respectively, and alkaline phosphatase and calcium concentration were detected at different induction time.
RESULTS AND CONCLUSION:The cellproliferation curves were S shaped. The absorbance values of the experimental group were higher than those of the control group at1, 3, 5, 7, 14, 21 days after osteogenic induction (P<0.05). Alkaline phosphatase and alizarin red staining of adipose-derived stem cells was al positive, but golden round nodes became more and bigger in the experimental group compared with the control group after 2 weeks. At 7 and 14 days, col agen I and osteocalcin mRNA expressions were higher in the experimental group than in the control group. Alkaline phosphatase activity and calcium concentration of the experimental group were higher than those of the control group at 1, 2, 3, 4 weeks after osteogenic induction (P<0.05). Results showed that the calcitonin gene-related peptide can induce the osteogenic differentiation of adipose-derived stem cells combined with calcium alginate gel.

Objective To investigate the prevalence of Toxoplasma gondii infection and related knowledge and behavior among special population in Changzhou City,so as to provide the evidence for formulating effective measures of toxoplasmosis prevention. Methods The pregnant women and patients with neoplasia in Changzhou hospitals,and livestock and poultry breeding or processing workers were selected as the subjects of the study. Venous blood samples were collected from each partici-pant for detecting IgG and IgM antibodies against T. gondii by ELISA. A questionnaire investigation on knowledge and behavior about T. gondii infection was conducted. Results Among the total 300 respondents investigated from March to May,2015,the prevalence of T. gondii infection was 16.3％(49/300). Totally 52 respondents knew the knowledge about the prevention and con-trol of T. gondii infection with an awareness rate of 17.3％(52/300). The proportion of participants who frequently contacted with cats/dogs(25.0％,13/52)in the group who knew the knowledge about the prevention and control of T. gondii infection was lower than that of participants(50.8％,126/248)in the group who did not know the knowledge about the prevention and control of T. gondii infection,and the difference was statistically significant(X2=11.51,P<0.05). The proportion of participants sepa-rating chopping boards for raw and cooked food(61.5％,32/52)in the group who knew the knowledge about the prevention and control of T. gondii infection was statistically higher than that of the participants(9.3％,23/248)in the group who did not know the knowledge about the prevention and control of T. gondii infection,and the difference was statistically significant (X2=78.43,P<0.001). There was a negative correlation between the awareness rate of the knowledge about the prevention and con-trol of T. gondii infection and the infection rate of T. gondii. The infection rate of T. gondii(5.8％,3/52)in the group who knew the knowledge about the prevention and control of T. gondii infection was lower than that(18.5％,46/248)of the group who did not know the knowledge about the prevention and control of T. gondii infection,and the difference was statistically significant (X2=5.14,P<0.05). Conclusions The awareness rate of the knowledge about the prevention and control of T. gondii infec-tion among special population in Changzhou City is low. The health education on the knowledge of the prevention and control of toxoplasmosis should be strengthened,in order to improve the awareness of personal hygiene and change the unhealthy lifestyles and dietary habits.

Objective To understand the prevalence of Toxoplasma gondii infection and related factors among special popu?lation in Changzhou City,so as to provide the evidence for formulating toxoplasmosis prevention measures. Methods The ve?nous blood was collected from participants in Changzhou City for detecting IgG and IgM antibodies against T. gondii by ELISA;and an epidemiological questionnaire investigation was also conducted. Results Among the total 400 respondents detected,the prevalence of T. gondii infection was 15.0%and the positive rates of IgG and IgM were 15.0%and 0.8%,respectively. The T. gondii infection rates of pregnant women,HIV/AIDS patients,patients with neoplasia,and livestock and poultry breeding and processing workers were 11.0%,11.0%,24.0%and 14.0%,respectively. As the age increased,the infection rate of T. gondii showed an upward trend,with the highest infection rate(21.6%)among the group of respondents aged above 50 years. The in?fection rates of groups with different education levels were statistically different(χ2=11.443,P<0.05),and there was a trend that the infection rate decreased with the increase of education level. The prevalence of T. gondii infection in pregnant women was not significantly associated with the number of pregnancies and gestational age. The infection rate of T. gondii in the live?stock and poultry breeding and processing workers increased with the length of their working years. Conclusions The preva?lence of T. gondii infection among the special population in Changzhou City is high. Therefore,it is necessary to enhance the health education on the knowledge and information of prevention and control of toxoplasmosis,and to improve the awareness of personal protection,the hygiene practices and diet habits. In addition,more attention should be paid to T. gondii monitoring in the special population.

Objective As SELDI-TOF-MS (Surface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) has been broadly used to screen biomarkers for a variety of diseases, the identification and validation of the revealed biomarkers requires more focused attention.Method In this paper, the serum samples from 60 cholangiocarcinoma, 146 lung cancer, 65 LGC and 58 LPC, 49 benign diseases of hepatobiliary and 53 normal individuals were analyzed by SELDI-TOF-MS. Results Among a set of proteins automatically selected as specific biomarkers by Biomarker Wizard software, three protein peaks, with molecular weights of 13. 71 × 103 , 13.83 × 103 and 13. 99 ×103 , were found significantly decreased in cholangiocarcinoma samples. The candidate biomarkers obtained from Tricine-SDS-PAGE gel bands by matching the molecular weight with peaks on CM10 chips were identified by Mass spectrometry as the native transthyretin(native TTR),cysTTR and glutTTR.These preliminary results were further proven by immunoprecipitation using commercial TTR antibodies. This allowed us to re-measure the TTR levels in all the groups more simply by ELISA assay. It showed a firm consistency between ELISA and SELDI analysis. In addition, while TTR levels in cholangiocarcinoma were found to be lower than those in normal healthy controls, TTR levels in benign diseases of the hepatobiliary system were found to be higher than those in healthy controls.Conclusion TTR could be a biomarker that better discriminates cholangiocarcinoma patients from the benign diseases compared to other biomarkers presently available.

Objective: To evaluate the efficacy of Tuina (Chinese therapeutic massage) manipulation plus horse-riding squat exercise in treating knee osteoarthritis (KOA) and optimize the combining protocol. Methods: Based on a 2×2 factorial design, 120 eligible KOA patients were randomized into a manipulation group (group A1B2), a manipulation plus horse-riding squat group (group A1B1), a sitting knee-adjustment group (group A2B2 group), and a sitting knee-adjustment plus horse-riding squat group (group A2B1), with 30 cases in each group. The intervention was conducted three times a week, lasting for four weeks. The Western Ontario and McMaster Universities osteoarthritis index (WOMAC) was taken as the major measure for efficacy evaluation (including three component scores, pain, stiffness, and daily function, and total score). Results: The three component scores (pain, stiffness, and daily function) and the total score of WOMAC showed significant differences after the intervention in the four groups (P<0.05). There were significant inter-group differences in the WOMAC stiffness score amongst the four groups after the intervention (P<0.05). In group A1B1, the step length, stride, walking speed, and knee joint flexion angle changed significantly after treatment (P<0.05). After the intervention, the step length changed significantly in group A1B2 (P<0.05), and the walking speed changed significantly in group A2B1 (P<0.05). There were no significant differences in the step length, stride, walking speed, or knee joint flexion angle among the four groups (P>0.05). The extensor peak torque at 180 °/s changed significantly in group A1B2 after treatment (P<0.05). Neither the intra-group nor the inter-group comparisons of the four groups revealed significant differences in the other isokinetic muscle strength parameters (P>0.05). The main effect of manipulation showed significant in affecting the WOMAC pain and total scores (P<0.05). The main effect of horse-riding squat exercise showed significant in affecting the WOMAC pain and stiffness scores (P<0.05). Conclusion: The four treatment protocols all can improve the symptoms of KOA, for instance, relieving pain and stiffness, and enhancing daily function. Group A2B1 produces the most eminent effect in relieving joint stiffness. The main effects of both manipulation and horse-riding squat exercise are significant in reducing pain. Besides, the main effect of horse-riding squat exercise is significant in relieving joint stiffness.