Phthalate esters widely exist in the environment. It has been demonstrated that they are harmful to human bodies. Phthalate mono-esters are the first metabolite of phthalate esters. In general,biotransformation or metabolism of xenobiotics most frequently result in detoxification of the chemical and facilitates excretion from the body. However,this may not be the case for phthalates. In the present paper,metabolism of phthalate esters and the toxicity of the phthalate mono-esters were reviewed,not only the effect of their mutagenesis,teratogenesis and carcinogenesis,but also their toxicity on reproduction,development,hormones,nuclear receptor,inflammation and obesity were included.

Objective To study the effect of long-term salmon calcitonin on bone mineral density (BMD), bone metabolism biochemical indicators and subjective score of bone pain in maintenance hemedialysis (MHD) patients with osteopenia. Methods Thirty-four MHD patients diagnosed as osteopenia by dual-energy X-ray absorptiometry (DXEA) were enrolled in this study. All the patients were treated with hypodermic injection of salmon calcitonin (50 U, thrice a week) for 12 months. The detecting parameters were as follows: BMD with DEXA in lumbar spine (L1-L4), femoral neck, troch, inter, and Ward's triangle before and after the study;serum bone metabolism biochemical indicators before and 6 and 12 months after the study;subjective scores of bone pain before and 1, 6, and 12 months after the study. Results Thirty-two patients were followed-up successfully. As compared to BMD parameters before study, the total T-score (-1.98± 2.20 vs 1.26±1.88, P=0.009) and total Z-score (-0.90±2.15 vs 0.08±2.05, P=0.002) of lumbar spine, the total T-score (-1.72±1.53 vs 1.06±1.58, P=0.016) and totle Z-score (-0.66±0.80 vs 0.08±1.08, P=0.029) of hip, the T-score of L3 (-2.02±2.51 vs 1.24±2.02, P=0.033), the Z-score of L2 (-0.44±1.82 vs 0.06±1.63, P=0.016), the Z-score of femoral troch (-0.65±1.11 vs 0.48±1.12, P=0.034) and the Z-score of inter (-0.58±0.94 vs 0.02±1.12, P=0.006) were increased significantly after study. But there were no significant differences in other examined regions and serum biochemical parameters. The subjective scores of bone pain were decreased rapidly for 41.7% after 1 month (P<0.01) and 76.6% after 6 months (P<0.01). The subjective score of bone pain after 12 months was similar to 6 months. The side effects of salmon calcitonin included nausea and vomitting in 5 cases (14.71%, 5/34), dizziness, blushing and flustered in 1 case respectively (3.13%,1/32). Conclusions Long-term hypodermic injection of salmon calcitonin can improve BMD and bone pain for MHD patients with osteopenia but has no significant effect on serum bone metabolism biochemical indicators. Salmon calcitonin is safe for MHD patients with seldom side effects, such as nausea and vomitting.

Objective To explore whether Cdc42 is an independent influence factor in regulating the migration of A549 cells by using quantitative measurement method,and to verify the effectiveness of an automatic measurement and calculation method for in vitro cell migration assay.Methods Cdc42 was overexpressed in human lung adenocarcinoma A549 cell line,and then in vitro scratch assay was applied to evaluate the migration of the cells.Different methods were used to measure the images acquired at different time points for quantitative analysis.Accuracy and repeatability of different measurement methods were analyzed.Results The results showed that overexpression of Cdc42 alone significantly advanced the migration of A549 cells (P＜0.05).The new method is efficient,accurate and reproducible as compared to manual measurement,and has significant advantages in target recognition and noise removal as compared to the existing measurement method in Image Pro Plus 6.0.Conclusions Over expression of Cdc42 significantly increased A549 cell migration ability in vitro.The new method can realize the automatic quantitative analysis of cell migration in vitro.

According to "Cyclic Tutorial System"and the first extra-curricular research competition in School of Public Health and Tropical Medicine in Southern Medical University, the author discussed the significance and the problems during the process,the sense of extracurricular research to the undergraduate students and the problems that both teachers and students should concern,and then put forward some approaches to solve the problems within extracurricular research.

BACKGROUND:In closed fractures, the initial hematoma that is inclined to remove is seldom considered as the important reasons for bone healing. OBJECTIVE:To observe the mechanism and potential role of original fracture hematoma in fracture healing. METHODS:Ninety-six patients with closed fractures of the long bones undergoing open reduction and internal fixation were randomly divided into experimental group (n=48) and control group (n=48). In the experimental group, original fracture hematoma, 1.0-2.0 mL, was first taken out during the internal fixation and placed into a special sterile plastic bag; then, 3-4 pieces of hematomas were filed into the fracture site and sutured layer by layer. On the contrary, original fracture hematomas from the control group were discarded. Blood samples were extracted to detect the biochemical indicators at 1 month after internal fixation. X-ray examination was done at 1, 3, 6 months after internal fixation for observation of fracture healing. RESULTS AND CONCLUSION: X-ray films showed that the healing rate at 3 months after operation was 95% in the experimental group and 78% in the control group, and there was a significant difference between the two groups (P < 0.05). Levels of bone glaprotein, I-type precolagen carboxy terminus peptide and serum bone alkaline phosphatase were significantly higher in the experimental group than the control group (P < 0.01 orP < 0.05). These findings indicate that the original fracture hematoma can accelerate calus formation, promote bone induction, provide nutrition to the fracture site, and participate in revascularization. Therefore, the original fracture hematomas is one of the effectively therapeutic methods for union and nonunion of fractures.

Objective To investigate the expression of bone sialoprotein (BSP) in prostate cancer and its clinical significance. Methods Prostate cancer tissues of different pathological grades (68 cases) and benign prostatic hyperplasia tissues (22 cases) were selected. SP method was used to detect the expression of BSP. Serum total prostate-specific antigen (tPSA) levels of prostate cancer were detected by electrochemiluminescence immunoassay before the operation. Results Compared with no or low expression in the adjacent normal glandular tissues, the detectable levels of BSP were examined in most of the prostate cancer tissues. The expression rate of BSP in prostate cancer tissues was higher than that in benign prostatic hyperplasia tissues [76.47%(52/68) vs 13.64%(3/22),χ2=27.614, P<0.001]. The expression rates of BSP in well differentiated, moderately differentiated and poorly differentiated tissues according to cell differentiating degree (Gleason system) were 75.0 % (12/16), 77.5 % (31/40) and 75.0 % (9/12) respectively. There was no significant difference in various pathological grading (χ2=0.057, P=0.972). The expression rates of BSP in pathological stage pT2, pT3 and pT4 tissues were 62.16%(23/37), 95.24%(20/21) and 90.0%(9/10) respectively. A statistically significant association was found between BSP expression and pathological stage (χ2=9.338, P=0.009). Serum tPSA level of prostate cancer group with BSP expression was higher than that with no BSP expression [(69.06±25.52)μg/L vs (38.00±21.64)μg/L, F=19.355, P<0.001]. Conclusion The high expression of BSP in prostate cancer has a relationship with pathological stage and serum tPSA level, it may play an important role in the biological behaviour of prostate cancer.

ObjectiveTo detect the difference of cytokines and antibodies productions by immunologic system from mice and rabbits vaccinated with the M.RCAg-1 chimeric protein,expressed in E.coli,formulation with different adjuvants,including Freund's adjuvant and three clinically acceptable adjuvants,namely,Al(OH)3,Montanide ISA720 and Montanide ISA51.MethodsSix weeks female BALB/c mice were vaccinated with recombinant protein formulated with different adjuvants through intranasal.Serum were collected to detect specific antibodies of M.RCAg-1 and individual Epitope by ELISA ; natural parasite antigen was recognized by indirect immunofluorescence assay; mouse specific T lymphocyte activation was detected by enzyme-linked immunosorbent spot test (ELISPOT) ; Affinity assay between protein and immune IgG of rabbits with the biosensor,and the growth of Plasmodiumfalciparum in vitro to evaluate by growth inhibition assay(GIA).ResultsDifferent formulation can induce different levels of antibody titers,the effection of ISA51 adjuvant was most closely with Freund's adjuvant,and can induce a higher specific antibody of 11 epitopes within proteins,can effectively stimulate cellular immune response based on the IFN-γ,to avidity Montanide ISA51 adjuvant immune antibodies and M.RCAg-1 protein affinity than the other two adjuvants;and Montanide ISA720 adjuvants and Al (OH)3 adjuvant group in mice can't induce a significant IFN-γresponse(P＞0.05).On avidity assay,the Montanide ISA51 formulation group was better than the other two adjuvants; and Montanide ISA720 and Al (OH)3 adjuvant formulation group can't induce a significant IFN-γresponse in mice(P＞0.05) ; the inhibition rates were 60% and 100% in 3D7 and Dd2 Plasmodium falciparum at a concentration of 2 mg/ml IgG by Montanide ISA51 formulated protein,and IgG of Al( OH)3 formulation could not effectively inhibit the in vitro growth of Plasmodium falciparum( 10% ),while IgG of Montanide ISA720 formulation could not inhibit growth of parasite in vitro.ConclusionBy comparing three clinically acceptable adjuvants and Freund's adjuvant in BALB/c mice and New Zealand rabbit,Montanide ISA51 adjuvants can be acceptable formulated M.RCAg-1 protein induced humoral and cellular immune responses,can be used as one of the candidate adjuvants.

ObjectiveTo explore the effects of vector fusion peptides on the conformation and immune reactivity of recombinant polyepitope antigens,M.RCAg-1.MethodsWe subcloned polyepitope artificial antigen gene,M.RCAg-1,into prokaryotic expression vectors,pDS-ex,that contain different fusion tags at the N-terminus or no any tag by different restriction enzyme cutting site.Three recombinant proteins expressed by these vectors,named P312-1,P312-2,and P312-3,were purified and purity is greater than 95％.Then BALB/c mice were vaccinated with the three proteins formulated with Freund's adjuvant through intranasal.Serum were collected to detect specific antibodies of M.RCAg-1 and individual epitope by ELISA ; natural parasite antigen was recognized by indirect immunofluorescence assay; mouse specific T lymphocyte activation was detected by enzyme-linked immunosorbent spot test (ELISPOT) ; and the growth of Plasmodium falciparum in vitro to evaluate by growth inhibition assay(GIA),and analyze secondary and tertiary structures of recombinant proteins from different expression vectors by bioinformatics and circular dichroism technique.ResultsThe P312-1,P312-2 have almost the same amino acid sequence,and the three proteins have the same immunogenicity in animal models(P＞0.05),however,the different proteins elicited various T-cell responses,the rabbit antibody induced by these proteins showed diverse efficacy in malaria parasite growth inhibition assaysin vitro ( respectively,93.9％,14.7％,54.3％ ).The significant differences of secondary and tertiary structures were shown in recombinant proteins from different expression vectors,analyzed by bioinformatics and circular dichroism technique,which demonstrated the change of protein molecule spaces conformation,and the obviously change of some epitope locations.ConclusionThese results suggest that the expressed polyepitope artificial antigens originating from the different vector fusion peptides indeed affect the protein folding and,subsequently,the epitope exposure.Thus,these proteins are able to induce both distinct humoral and cellular immune responses in animal models,and they affect the efficacy of immune inhibition against the parasite,the enhancement or suppresses.

Objective To investigate the sleep quality and the risk factors for sleep disorders in population at high-risk for stroke:.Methods A cross-sectional survey was conducted in population at highrisk for stroke:in Water Park and Wangdingdi Communities,Nankai District,Tianjin in March 2016.The residents were divided into either a good sleep group or a sleep disorder group according to the Pittsburgh Sleep Quality Index (PSQI).Multivariate logistic regression analysis was used to determine the risk factors affecting sleep quality.They also divided into a stroke history group and a non-stroke history group according to the high-risk population with or without previous history of stroke.The sleep quality was compared between the 2 groups,and the correlation between sleep disorders and stroke outcomes was analyzed.Results A total of 565 residents at high-risk for stroke were enrolled in the study,and 178 01.5％) had sleep disorders.The age in the sleep disorder group was significantly older than that in the good sleep group (66.70 ±8.97 years vs.62.87 ±9.46 years;t =4.540,P＜0.001).The proportions of female (68.0％ vs.49.1％;x2 =16.190,P ＜ 0.001),hypertension (69.7％ vs.57.9％;x2 =7.154,P =0.005),ischemic heart diseases (48.9％ vs.35.4％;x2 =9.253,P =0.002),history of previous stroke or transient ischemic attack (TIA) (30.9％ vs.18.9％;x2 =10.080,P =0.001),and carotid plaques (71.9 vs.53.7％;x2 =16.688,P ＜0.001) in the sleep disorder group were higher than those in the good sleep group.Multivariate logistic regression analysis showed that after adjusting for age and sex,the history of previous stroke or TIA (odds ratio [OR] 1.712,95％ confidence interval [CI] 1.105-2.653;P =0.016),and carotid plaques (OR 1.583,95％ CI 1.003-2.498;P =0.048) were the dependent risk factors for sleep disorders.The total score of PSQI in patients with previous stroke was significantly higher than that in patients without previous stroke (7.25 ±4.71 vs.6.13 ±4.20,t =-2.578,P =0.010).The sleep latency score (1.24 ± 1.06 vs.0.95 ± 1.02;t =-2.868,P =0.004) and sleep disorder score (1.23 ± 0.63 vs.1.07 ± 0.61;t =-2.622,P =0.009) in patients with previous stroke history were significantly higher than those without.According to the modified Rankin Scale scores,the patients with a history of stroke were divided into a good outcome group (0-2) and a poor outcome group (＞2),including 105 (82.0％) and 23 patients (18.0％),resectively.The proportion of patients with sleep disorders (78.3％ vs.35.2％;x2 =14.251,P＜0.001) and the PSQI score (median and four percentile interval:6 [3-8] vs.12 [8-18];Z =-4.392,P ＜0.001) in the poor outcome group were significantly higher than those in the good outcome group.Conclusions The incidence of sleep disorder is high in the high-risk population,the previous stroke or TIA history and carotid plaques are the independent risk factors for sleep disorder in the high-risk population,and sleep disorder is associated with the poor outcomes of strokes.Therefore,attention should be paid to the sleep quality of this stroke high-risk population and control the risk factors of causing sleep disorders,especially for those with a history of stroke.This will help reduce the risk of stroke.

Recently point-of-care testing (POCT) for cardiovascular biomarkers is sharply increasing among subdivision field of POCT,however the relevant quality control is not matched.In this paper,the present situation and problems about selecting biomarkers,managing quality and reporting results during clinical application were expounded.The difference between POCT for cardiovascular biomarkers and routine biomarkers was elaborated.The domestic and international rules and standards about POCT for cardiovascular biomarkers were summarized.Finally,this paper included some rationalization proposal to solve the noticeable problems of quality control in POCT for cardiovascular biomarkers.