Objective To study the intervention effect of the fluorosis with the high aluminum plus soybean, selenium powder, spiral algae. Methods Two week-long SD rats were randomly allocated into 5 groups bases on body weight with 8 in each group, respectively being control, the high fluoride aluminum,the high fluoride aluminum plus soybean, the high fluoride aluminum plus selenium and the high fluoride aluminum plus spiral algae group. The control group was fed with corn produced in non-endemic area containing 5.20 mg/L of fluoride and 6.80 mg/L of aluminum and tap water of fluoride 0.70 mg/L and aluminum 0.20 mg/L. Other groups were fed primarily with corn produced in the endemic area with the content of fluoride of 110.00 mg/L and aluminum of 19.70 mg/L and tap water with high aluminum also with fluoride of 0.70 mg/L and aluminum of 90.00 mg/L. The high fluoride aluminum plus soybean group was added soybean in the beginning of the experiment, approximately 30% of the total amount. Selenium (3.00 mg/kg) was given to the high fluoride aluminum plus selenium groups and spiral algae (1000.00 mg/kg) to the high fluoride aluminum plus spiral algae groups after dental fluorosis was formed. The experiment lasted for 22 weeks. In the end of the experiment 24-hour urine and bones of the limbs were collected to test the content of fluorine and aluminum. The ultrastructure of femur spongiosa was observed under electron microscope. Results ① Bone fluoride in high fluoride aluminum group [(204.07 ± 63.78)mg/kg] was higher than that in the control group, high fluoride aluminum plus soybean group, high fluoride aluminum plus selenium group and high fluoride aluminum plus spiral algae group[(30.06 ± 6.11), (54.12 ± 14.56), (30.44 ± 5.02), (105.08 ± 21.07)mg/kg, t = 0.62,0.53,0.62,0.35, all P < 0.01], indicating that it was lower in high fluoride aluminum plus selenium group than high fluoride aluminum plus spiral algae group(t = 0.27, P < 0.01). ② The urinary fluoride levels in high fluoride aluminum group [(4.52 ± 3.09)mg/L] was higher than that in the control group [(0.89± 0.37)mg/L, t = 0.23, P < 0.01] and lower than that in the fluoride aluminum plus selenium group[(10.38 ± 1.58) mg/L, t = 0.34, P < 0.01], it was higher in high fluoride sluminum with selenium group than that in the high fluoride aluminum plus soybean group and high fluoride aluminum plus spiral algae group[(5.56 ± 1.69), (4.38 ± 3.36)mg/L, t = 0.28,0.35, all P < 0.05]. ③ The bone aluminum level in the control group[(3.32 ± 3.02)mg/kg] was lower than that in the high fluoride aluminum group, high fluoride aluminum plus soybean group, high fluoride aluminum plus selenium group and high fluoride aluminum plus spiral algae group [(374.21 ± 56.11), (118.20 ± 23.59), (114.01 ± 22.84), (67.11 ± 11.53)mg/kg, t = 1.42,0.44,0.42,0.24, all P < 0.05], it was higher in the high fluoride aluminum group than high fluoride aluminum plus the soybean and selenium and spiral algae groups (t = 0.56,0.57,0.68, all P < 0.01)lower in high fluoride aluminum plus spiral algae group than the soybean plus selenium group(t = 0.11,0.10, all P < 0.05). ④The urine aluminum level in high fluoride aluminum group [(1.14 ± 0.32)mg/L] was higher than that in the control group and the high fluoride aluminum plus the soybean group [(0.66 ± 0.10) mg/L, t = 1.92,2.24, all P< 0.05] and that in high fluoride aluminum plus the soybean group was lower than aluminum plus the selenium[(1.03 ± 0.22)mg/L] and aluminum plus spiral algae group[(1.10 ± 0.28) mg/L, t = 1.73,2.06, all P < 0.05]. ⑤ Under electron microscope, the collagenous fiber was arranged in disorder in high aluminum fluorine group, the trabeculum of bone merged into patches or mostly vanished, This phenomenon was alleviated in the intervened group compared to the high aluminum fluorine group, in an attenuating order of the selenium group, the spiral algae and the soybean group. Conclusion In this experiment the intervention measure alleviates fluorosis, the selenium is the best, spiral algae the second and soybean the last.

AIMTo evaluate the effects of surfactants on the pharmacokinetics and distribution in rats after intravenous administration of SOD liposomes.

METHODSThe liposomes were prepared by reverse phase evaporation method. The activity of SOD was assayed by method of xanthine oxidase.

RESULTSThe T1/2 of SOD solution, common SOD liposome, SOD liposomes modified by DSPE-PEG2000 and Tween 80 were 0.25, 0.34, 0.66 and 0.41 h, respectively; AUC were 12.48, 24.66, 41.16 and 33.02 microg x h x mL(-1), respectively. Compared with the common liposome, the liposomes modified by DSPE-PEG and Tween 80 decreased the content of SOD in liver and spleen, but increased in brain.

CONCLUSIONThe three kinds of liposomes could increase T1/2 and AUC in some extent, especially in PEG-L group. Tween-L could increase the SOD content in brain, and PEG-L could decrease the SOD content in the liver and spleen compared with the common liposome.

Animals ; Area Under Curve ; Brain ; enzymology ; Injections, Intravenous ; Liposomes ; Liver ; enzymology ; Male ; Polysorbates ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spleen ; enzymology ; Superoxide Dismutase ; administration & dosage ; pharmacokinetics ; Surface-Active Agents ; pharmacology ; Tissue Distribution

BACKGROUND: Peripheral nerve injury can lead to extensive changes in central nervous system, and exercise training can promote the recovery of locomotor function following central nervous system injury. OBJECTIVE: To observe the changes of locomotor function and the expression levels of vesicular glutamate transporter VGLUT1 in the spinal cord in a rat model of dorsal root ganglion resection after treadmill exercise and to explore the effect of treadmill training on the locomotor function after peripheral nerve injury. METHODS: Thirty-nine 10-week-old male Wistar rats were randomized into experimental (n=15), control (n=15) and sham operation (n=9) groups. The rats in the experimental and control groups received the dorsal root ganglion resection at L3and L4segments to establish the model of peripheral nerve injury under local anesthesia, while the rats in the sham operation group were only subjected to dorsal root ganglion exposure. The rats in the experimental group underwent 15 m/minute treadmill training at postoperative 7 days, while rats in the other two groups were in free movement. Gait analysis was performed at preoperative 3 days, postoperative 7, 14, 21, and 28 days, respectively, and the behavioral changes of rats were observed. The tissue sections were obtained from L3segment at postoperative 7, 14, and 28 days to detect the expression levels of VGLUT1 in the spinal cord by immunohistochemistry. RESULTS AND CONCLUSION: The peroneal nerve function index in the experimental and control groups was lower than that before surgery and that in the sham operation group at postoperative different time points (all P ＜ 0.05). The index in the experimental and control groups was the lowest on day 7 postoperatively (P ＜ 0.05), then the index gradually increased, but was still lower than the preoperative level (P ＜ 0.05). The index in the experimental group was significantly higher than that in the control group at postoperative 21 and 28 days (P ＜ 0.05). The expression levels of VGLUT1 in the lamina IX in the experimental and control groups were significantly lower than those in the sham operation group at different time points after surgery (P ＜ 0.01). The levels in the experimental group were significantly higher than those in the control group at postoperative 14 and 28 days (P ＜ 0.05). The levels in both groups on a decline after surgery, especially the control group (P ＜ 0.05). These results suggest that treadmill can promote the recovery of locomotor function post peripheral nerve injury.

Objective: To observe the changes of PD-1 expression, mRNA level and cytotoxic activity of CD19 CAR-T cells during the culture process of CAR-T cells. Methods: The peripheral blood T cells of 6 lymphoma patients with high expression of PD-1 and 6 healthy volunteers were the source of CAR-T cells. The expression of PD-1 was analyzed by flow cytometry. The mRNA level of PD-1 was analyzed by PCR. The cell proliferation was analyzed by CCK-8 assay. The cytotoxicity was analyzed by LDH assay. Results: ①The transfection efficiency of high PD-1 expression T cells and healthy volunteer T cells were as the same (P>0.05) . ②The cell proliferation capacity of CD19 CAR-T cells from high PD-1 expression T cells or healthy volunteer T cells, with or without PD-1 inhibitor were as the same (P>0.05) . ③The cytotoxicity to lymphoma cells of high PD-1 expression T cells and CAR-T cells were lower than that of these two T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer T cells (P<0.001) . There was no difference of the cytotoxicity between the CAR-T cells from high PD-1 expression T cells combined with PD-1 inhibitor and the CAR-T cells from healthy volunteer (P>0.05) . ④There was no difference of the expression of PD-1 in all CAR-T cell groups during the culture process (P>0.05) . There was no difference of mRNA level of PD-1 in all groups during the culture process (P>0.05) . ⑤The PD-1 expression of CAR-T cells increased by the time of culture after contacting with lymphoma cells (P<0.001) . The PD-1 inhibitors could antagonize this effect. There was no difference of mRNA level of PD-1 in all groups after contacting with lymphoma cells (P>0.05) . Conclusion: The PD-1 expression of CAR-T cells from high PD-1 expression T cells increased by the time of culture after contacting with lymphoma cells. However, the mRNA level of PD-1 of all groups did not change, even if PD-1 inhibitor was applied.
Antigens, CD19 ; Humans ; Programmed Cell Death 1 Receptor/genetics* ; RNA, Messenger ; Receptors, Antigen, T-Cell ; T-Lymphocytes

Objective:To explore the influence of high altitude hypoxia on the phagocytosis and killing functions of peritoneal macrophages in mice by establishing mouse model in high altitude hypoxic environment.Methods:①After exposure of mice to an altitude of 4 200 m,2 200 m and 400 m for 30 d respectively,flow cytometry was used to detect the phagocytosis and killing functions of peritoneal macrophages on staphylococcus aureus labeled with FITC.②The respiratory burst level of the cultured macrophage in mice was detected in 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe method. ③The concentration of NO2- as a stable oxidative metabolite of NO in the supernatant of the cultured macrophages was measured with ELISA kits;④The release level of IL-6 and TNF-α in the cultured mice macrophage supernatant was also determined with ELISA kits.Results:After exposed under high altitude hypoxia for 30 d,compared with the control group (400 m),the phagocytosis,respiratory burst level and NO release in high altitude groups (4 200 m and 2 200 m) were all than those in the control group(400 m) (P<0.05).While the concentration of IL-6 and TNF-α in the Mφ cultured supernatant showed an obvious increase (P<0.05).Conclusion:Exposure under high altitude hypoxia (at altitude of 4 200 m and 2 200 m) for 30 d compromised the phagocytosis and oxygen dependent cytolyticactivity functions,and also raised the cytokines secretion level of IL-6 and TNF-α in Mφ,thereby affecting the innate immune response ability of Mφ in body.

Objective To explore factors influencing the incidence of carotid atherosclerosis in different genders so as to provide reference for the specific prevention of the disease. Methods A nested case-control study was conducted to analyze factors influencing the incidence of carotid atherosclerosis in Jinchang cohort population who were randomly selected through stratified sampling by age and followed up. A risk prediction model was established and the goodness of fit was evaluated by the area under the receiver operator characteristic curve (ROC). Results The standardized incidence of carotid atherosclerosis in this follow-up population was 12.32%, and the incidence rate of males (13.65%) was greater than that of females (11.29%). The difference was statistically significant ( 2=4.267, P<0.001). Age, education, elevated systolic blood pressure, and elevated low-density lipoprotein cholesterol were common risk factors for carotid atherosclerosis in both men and women. Elevated fasting plasma glucose (OR=2.556, 95% CI: 1.618-4.038) and elevated triglyceride (OR=1.535, 95% CI: 1.058-2.227) were only associated with men. Abdominal obesity (OR=1.414, 95% CI: 1.013-1.974) was only associated with women. The area under ROC of male and female prediction models was 0.835 (95% CI: 0.815-0.856) and 0.809 (95% CI: 0.788-0.831), respectively. The sensitivity was 78.0% and 78.9%, the specificity was 78.8% and 73.1%, and the diagnostic coincidence rate was 91.3% and 82.4%, respectively. Conclusions There are different risk factors for carotid atherosclerosis in males and females, and targeted prevention and control measures should be taken according to gender. The risk prediction model established by Logistic regression had certain guiding value.

OBJECTIVETo investigate the impact of college students' evening exercise on their sleep quality, so as to provide a scientific basis for college students to choose an appropriate method of exercise and improve their sleep quality.

METHODSFrom September to October in 2012, Multi-stage cluster random sampling method was used to select the 5997 college students in Anhui province. The status of college students' exercise and their sleep quality were investigated by the general situation questionnaire, Physical activity rating scale-3(PARS-3), Rating of perceived exertion(RPE) and Pittsburgh sleep quality index(PSQI). Kruskal-Wallis test was used to analyze the impact of evening exercise on sleep quality and multivariate unconditional logistic regression was used to analyze the factors of sleep quality in evening excise students.

RESULTSThe median of PSQI total score among 5806 college students was 5 and 1030(17.7%) students had poor sleep quality. The median of the PSQI scores was the same (5 points) for evening exercise group, daytime exercise group,daytime and evening exercise group and non-exercise group (1406, 1514, 1244, 1642 respectively). The difference was not statistically significant (χ(2) = 2.80, P = 0.42). Compared to non-exercise population, the OR (95%CI) value of evening exercise' impact on sleep quality was 0.90(0.73-1.10). Compared to very light evening exercise, the OR (95%CI) value of moderate and large amount of evening exercise' impact on sleep quality was 0.58 (0.44-0.75) and 0.67 (0.48-0.93) respectively; Compared to other sports, the OR (95%CI) value of badminton, rope skipping and jogging' impact on sleep quality was 0.72 (0.55-0.93), 0.38 (0.21-0.70) and 0.76 (0.60-0.95) respectively and they were all protective factors of sleep quality. Compared to small exercise intensity, the OR (95%CI) value of moderate, vigorous and very vigorous exercise intensity' impact on sleep quality was 1.68 (1.13-2.52), 2.38 (1.48-3.83) and 3.18 (1.72-5.90) respectively and they were harmful factors of sleep quality.

CONCLUSIONThere was no impact of evening exercise on sleep quality for college students. Type of sports should be adequately chosen for evening exercise. College students can take moderate and large amount of evening exercise but should avoid activities of vigorous intensity.

Exercise ; Female ; Humans ; Male ; Sleep ; Students ; Surveys and Questionnaires ; Time Factors ; Young Adult

OBJECTIVETo investigate the expression of indoleamine 2, 3-dioxygenase (IDO) in breast cancer and its correlation with clinicopathologic factors and prognosis.

METHODSThe expression of IDO, CD31, CD105 proteins in 40 specimens of breast cancer were assessed by immunohistochemistry.

RESULTSThe overexpression rate of IDO in breast cancer was 67.5% (27/40), and expression of IDO was closely associated with clinical stage and lymph nodes metastasis. The disease-free survival rate in patients with IDO overexpression was not significantly lower than that in patients with negative or low expression of IDO (P > 0.05). Moreover, the expression of IDO was positively correlated with CD105-labeled microvessel density (r = 0.659, P < 0.05).

CONCLUSIONSExpression of IDO is associated with clinical stage and lymph nodes metastasis, and microvessel densitty. IDO expression may promote the growth and metastasis of breast cancer, probably via the increased agiogenesis. A larger sample study is needed to verify whether the prognosis of beast cancer is significantly correlated with IDO expression.

Adenocarcinoma ; enzymology ; immunology ; pathology ; Adult ; Aged ; Antigens, CD ; metabolism ; Breast Neoplasms ; enzymology ; immunology ; pathology ; Carcinoma, Ductal, Breast ; enzymology ; immunology ; pathology ; Carcinoma, Medullary ; enzymology ; immunology ; pathology ; Disease-Free Survival ; Endoglin ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; metabolism ; Lymphatic Metastasis ; Microvessels ; enzymology ; immunology ; Middle Aged ; Neoplasm Staging ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Receptors, Cell Surface ; metabolism ; Survival Rate

OBJECTIVESTo facilitate gene therapy research using recombinant adeno-associated virus type 2 (rAAV2) vector as gene transfer vehicle, and to construct a rAAV2 based vector carrying bone morphogenetic protein-7 (BMP7) and observe its expression in bone mesenchymal stem cells.

METHODSThe coding sequence (1.3 kb) of BMP7 was amplified by polymerase chain reaction (PCR) from the pcDNA1.1(+) plasmid containing the human BMP-7 cDNA. After purified, the gene fragment was cloned into a plasmid pUC18 and termed plasmid pUC18-hBMP7. The recombinant pUC18-hBMP7 was digested by Kpn I and Sal I and further ligated to the pSNAV by T4DNA ligase. The resultant plasmid PSNAV-hBMP7 was transformed into DH5a Escherichia coli, and positive colonies were screened by PCR and digest with restriction enzyme to identify the correct recombinant clones. BHK-21 cells were transfected with the purified pSNAV-BMP7 plasmid according to a standard calcium phosphate precipitation method. The cells were then cultured in selection media containing 800 micro g/ml G418 (Gibco/BRL). G418-resistant BHK-21 cell clones were isolated and the integrity of hBMP7 gene was determined by PCR using the above PCR primers. To package the virus, stably transfected BHK-21 cells were subsequently infected with recombinant herpes simplex virus type 1 (rHSV-1). The collected cells were processed by chloroform treatment, PEG8000/NaCl precipitation and chloroform extraction for purification. The titer was determined using quantitative DNA dot blots and the purity was examined by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Following infection with rAAV2-BMP7 at multiplicities of infection of 1 x 10(5) vector genomes per cell and subsequent culture, MSCs were assessed qualitatively for BMP7 production.

RESULTSTransient transfection showed an efficiency of 98.8% in MSCs. RT-PCR showed that MSCs had transcription of BMP7 that was enhanced by the gene transfer. BMP-7 expression in MSCs was identified by Western-blot.

CONCLUSIONSThe hBMP7 recombinant adeno-associated virus vector is successfully constructed. The present in vitro study demonstrates that rAAV2-BMP7 could infect MSCs.

Animals ; Blotting, Western ; Bone Marrow Cells ; cytology ; metabolism ; Bone Morphogenetic Protein 7 ; Bone Morphogenetic Proteins ; genetics ; metabolism ; Cell Line ; Cells, Cultured ; Dependovirus ; genetics ; Gene Expression ; Genetic Vectors ; genetics ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Rabbits ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Transforming Growth Factor beta ; genetics ; metabolism

OBJECTIVETo explore the relationship between gene polymorphism of GABAA receptors and childhood autism by detecting rs140682, rs2081648 and rs140679 site of single nucleotide polymorphism (SNP) in GABAA receptors gene.

METHODSA total of 94 children with autism and 124 normal children were enrolled in a hospital from November 2010 to May 2011. Childhood autism rating scale (CARS) and autism behavior checklist (ABC) were used to evaluate or investigate the case group. After collecting venous blood and extracting the genome DNA, the allele and genotype of SNP rs140682, rs2081648 and rs140679 site in GABAA receptors gene were detected by PCR-RFLP. The allele and genotype of case group and control group were analyzed by χ(2) test, while the score of scales was analyzed by Spearman rank correlation analysis.

RESULTSThe age of the case group was 5.12 ± 0.32, and it was 5.25 ± 0.27 in the control group (P < 0.05). In case group, the frequency of genotype CC, CT and TT of rs140682 site was 44, 41 and 9, while it was 48, 65, and 11 in control group (P > 0.05), respectively. The frequency of genotype AA, AG and GG of rs2081648 site was 8, 58 and 28 in case group, while it was 12, 49 and 63 in control group (P < 0.05), respectively. In case group, the frequency of genotype CC, CT and TT of rs140679 site was 15, 36 and 43, while it was 18, 59 and 47 in control group (P > 0.05), respectively. It was revealed by Spearman rank correlation analysis that of rs2081648 site, there was a positive correlation between genotype AG and sensation factor (S), social intercourse factor (R), and language factor (L) of autism behavior checklist (ABC) (r values were 0.149, 0.165 and 0.155, all P values < 0.05). A negative correlation between genotype GG and S, R, L and self-help factor (V) was proved (r values were -0.140, -0.173, -0.158 and -0.135, all P values < 0.05). There was a positive correlation between allele A and R and L factors (r values were 0.153 and 0.137, all P values < 0.05), while a negative correlation between allele G and R and L factors (r values were -0.153 and -0.137, all P values < 0.05). In case group, 42 children were diagnosed with mild-to-moderate autism, while 52 children were severe autism. There was no statistically significant correlation between allele or genotype of SNP rs140682 and rs140679 site and the degree of autism (P > 0.05). There was a positive correlation between allele A and genotype AG and the degree of autism (r values were 0.147 and 0.616, all P values < 0.05), while a negative correlation between allele G and genotype GG and the degree of autism (r values were -0.159 and -0.616, all P values < 0.05).

CONCLUSIONThe SNP rs2081648 site which located in GABAA receptors gene may be related to autism. No evidence for significant association between rs140682 and rs140679 site and autism was found.

Alleles ; Autistic Disorder ; genetics ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Polymorphism, Single Nucleotide ; Receptors, GABA-A ; genetics