Objective To compare the differences in behavior characteristics among SHR, WKY and SD rat models of attention deficit/hyperactivity disorder (ADHD), and explore an ideal control model of SHR rats.Methods Using open field test to analyze the rat movement distance, speed, wearing numbers and the number of grooming to evaluate the spontaneous movement in SHR, WKY and SD rats.Using the Morris water maze to test the learning and memory ability among the three rat groups.Results The result of open field test showed that the SHR rats had significantly increased (P< 0.01) total amount of exercise, average speed and wearing numbers than WKY and SD rats.Compared with the WKY rats, SD rats had a significantly higher movement distance (P< 0.01), slightly higher movement speed and wearing number (P< 0.05).In the Morris water maze hidden platform period test, the SD rats had a significantly longer latency than the SHR rats (P< 0.05).SD rats showed longer latency distance on the first, third and fourth days of training, as compared with the SHR rats (P< 0.05 or P< 0.01).Compared with the WKY group, SD rats showed a shorter latency distance in each training time (P< 0.05 or P< 0.01).In the probe trial period, the SD rats showed shorter time and distance ratio to the target quadrant than SHR rats (P< 0.05), while significantly longer than the WKY rats (P< 0.05 or P< 0.01).Conclusions There are significant behavioral differences between SHR and WKY rats, showing certain disadvantages in comparison of the two types of rats.To add SD rats as a control group for SHR rats can improve the comparability of behavior characteristics of SHR rats, and to get more objective evaluation of the behavior characteristics of SHR rats.

Objective To discuss the research status of TCM therapy for ADHD in China in recent ten years; To provide references for clinical workers. Methods Clinical research papers about TCM therapy for ADHD in CNKI, VIP, and CBM from 1st January 2005 to 1st April 2015 were retrieved. Metrology analysis was conducted from the aspects of publication time, diagnostic criteria, contrasted medicine, and types and features of TCM therapy. Results By eliminating duplication literature, 670 papers were screened out. After the screening of exclusion criteria, 148 papers were included. DSM-Ⅳ and CCMD-3 were the most commonly used diagnostic criteria in the 148 papers; internal therapy was the main therapy in TCM therapy; decoction was the main dosage form; liver-kidney yin deficiency syndrome and heart-liver fire syndrome were the main patterns of syndrome; Acori Tatarinowii Rhizoma, Rehmanniae Radix Praeparata and other 19 Chinese medicinal herbs were the main Chinese mateia medica; Xiaoer Zhili Syrup was the most commonly used Chinese patent medicine; acupuncture and moxibustion and auricular point therapy were the main external therapy, and external combined with internal therapy had the best efficacy. Conclusion TCM therapy for ADHD is with high effectiveness. External combined with internal therapy has the best efficacy.

Objective To introduce an improved extraction method of prefrontal cortical and striatal synaptosomes from SHR rat. Methods Synaptosomes were prepared from SHR rat brain tissue by Percoll density gradient centrifugation.Transmission electron microscopy was used to assess the morphology and structural integrity of the synaptosomes.Results The obtained synaptosomes showed oval structures surrounded by an intact membrane.Presynaptic components contained one or more mitochondria and a large number of synaptic vesicles.The synaptic clefts were clearly visible, and prominent part of the characteristic compact structure was clear, complete and with higher electron-density. The synaptosome presynaptic membrane, synaptic cleft, and postsynaptic membrane were well preserved, and the synaptosomes were densely distributed, showing typical morphological characteristics of synaptosomes.Conclusions The results of our study improved the traditional preparation method and provide a less time-consuming, highly productive protocol for preparation of structurally typical and intact synaptosomes, suitable for further research on neuroscience and neurological diseases.

BACKGROUND:Some studies have indicated that different genes in tooth germ tissue play a role at different time, contributing to tooth germ development.
OBJECTIVE:To observe the expressions of dentin matrix protein 1, enamel protein, col agen I and homeobox gene 1 at different stage of in vitro culture of tooth germ cells.
METHODS:RNA from tooth germ cells was extracted at days 1, 3, 6 of in vitro culture. After reverse transcription, real-time quantitative PCR detection was adopted to measure relative expression of dentin matrix protein 1, enamel protein, col agen I and homeobox gene 1 mRNA.
RESULTS AND CONCLUSION:Dentin matrix protein 1, enamel protein, and col agen I mRNA expressions increased with culture time, and reached the peak at day 3 (P<0.05), whereas homeobox gene 1 mRNA decreased with culture time (P<0.05).

BACKGROUND:Cobalt chloride (CoCl2) may promote the proliferation of human umbilical cord-derived mesenchymal stem cels (hUC-MSCs) in a time- and concentration-dependent manner, and meanwhile, CoCl2 can regulate the expression of genes and proteins in hUC-MSCs. OBJECTIVE:To explore the effects of CoCl2 induced-hypoxia on the proliferation of hUC-MSCs and gene and protein expressions in hUC-MSCs, thereby establishing an effective method for MSCs culture and amplificationin vitro. METHODS: hUC-MSCs were extracted using tissue explant method. Under hypoxia conditions induced by CoCl2 (0, 100, 150, 200, 250 μmol/L) for different periods (0, 1, 2, 3, 4 days), flow cytometry was used to identify cel surface-associated antigens; cel counting kit-8 was used to detect cel proliferation; RT-PCR was used to determine levels of hypoxia inducible factor-1α, inducible nitric oxide synthase, stromal cel-derived factor-1, interleukin-6, transforming growth factor-β mRNA; western blot assay was used to detect protein expression of hypoxia inducible factor-1α. RESULTS AND CONCLUSION:The cels were positive for CD29, CD73, CD90, CD105, while negative for CD31, CD14, CD34, CD45, CD11b, HLA-DR. Moreover, the antigen expression was not affected by CoCl2 induced-hypoxia. CoCl2 induced-chemical hypoxia could promote the proliferation of hUC-MSCs in a time- and concentration-dependent manner. RT-PCR results showed thatunder hypoxia, hypoxia inducible factor 1α, inducible nitric oxide synthase and stromal cel-derived factor-1 mRNA expressions were significantly up-regulated, but interleukin-6 and transforming growth factor-β mRNA expressions were down-regulated significantly (P < 0.05). Additionaly, the protein expression of hypoxia inducible factor 1α was increased under hypoxia conditions. These findings indicate that CoCl2 induced-hypoxia environment may promote the proliferation of hUC-MSCs and the optimal concentration of CoCl2 is 200μmol/L. However, a higher concentration of CoCl2 (≥ 250μmol/L) inhibits the proliferation of hUC-MSCs, and the mechanism may be related to the increase of hypoxia inducible factor-1α at protein and mRNA levels.

METHODS:Lentiviral vectors carrying bFGF and BMP-2 were constructed to transfect sheep bone marrow mesenchymal stem cells. cells were divided into four groups:bFGF group, BMP-2 group, co-transfection group BACKGROUND:Basic fibroblast growth factor (bFGF) can promote the proliferation of bone marrow stromal cells, and bone morphogenetic protein-2 (BMP-2) has an important significance in the induction of new bone formation.
OBJECTIVE:To analyze the effects of bFGF, BMP-2 and their co-transfection on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and to compare the relative expressions of col agen I, osteocalcin and osteopontin before and after celltransfection, thereby providing theoretical implications for seed cells in the construction of tissue-engineered bone.
and control group. The RNA was extracted using real-time quantitative PCR to detect mRNA levels of col agen I, osteocalcin, and osteopontin.
RESULTS AND CONCLUSION:Significant difference in non-specific osteogenic gene expressions was found among the four groups (P<0.05). bFGF and BMP-2 showed an interaction (P<0.05). Expressions of col agen I and osteocalcin in the co-tranfection group were higher than those in the other three groups (P<0.05), but osteopontin expression exhibited no difference (P>0.05). In vitro experiments showed that the relative expression of col agen I, osteocalcin and osteopontin were higher in the co-transfection group, indicating the cells from the co-transfection group have strongest osteogenic capacity that are suitable for seed cells for bone tissue engineering.

Objective : To develop an analytical method of ibotenic acid (IBA) and muscimol (MUS) in wild mushroom by dansyl chloride (DNSCl) derivatization-liquid chromatography-tandem mass spectrometry (LC-MS/MS), and to provide technical support for etiological identification of mushroom poisoning events. Methods : The sample was extracted with hydrochloric acid solution, derived by bimolecular DNSCl, diluted and inorganic salts precipitated with acetonitrile. The extract was separated by a waters XBridgeTM BEH C18 column and measured by LC-MS/MS. Results : The limits of detection for IBA and MUS in wild mushroom were 0.15 mg/kg and 0.1 mg/kg, respectively. Good linear relationship was obtained for IBA and MUS at the range of 0.5-250 mg/kg with the correlation coefficient of 0.997 and 0.999, respectively. The average recoveries at three spiking levels were 84.5%-102.0% with relative standard deviations (RSDs, n=6) of 4.7%-8.6% for IBA. The average recoveries were 88.6%-95.4% with RSDs (n=6) of 4.9%-7.5% for MUS. Conclusion The optimized sample extraction and bimolecular DNSCl derivatization conditions can achieve rapid and accurate analysis of IBA and MUS in wild mushroom poisoning sample.

To identify SJCHGC01743 gene of Schistosoma japonicum and evaluate the potential of the recombinant protein as a new vaccine candidate for schistosomiasis, polymerase chain reaction (PCR) technique was used to amplify the cDNA of the gene and real-time RT-PCR was used to analyze the transcription profiles of SJCHGC01743 at different development stages. Recombinant plasmid was successfully constructed and transformed into competent Escherichia coli BL21 (DE3). Then the recombinant protein was expressed, purified and emulsified with ISA206 adjuvant to immunize BALB/c mice for three times. The immunogenicity was confirmed by Western blotting and tissue localization was detected by indirect immunofluorescent assay. The specific antibody level was detected by ELISA. The immunoprotection of rSjOST48 was evaluated by the reduction in worm and egg counts in mice. A cDNA with 1 248 nucleotides was isolated from 28-day-old schistosomes cDNAs by PCR. Sequence analysis revealed that SJCHGC01743 was a 48-kDa subunit of the oligosaccharyltransferase complex (OST48) and named as SjOST48. Real-time PCR analysis indicated that this gene was expressed in all investigated stages and had the highest expression level in 28 d worms, the level of gene transcription in female worms was significantly higher than that of male worms. Then recombinant plasmid pET28a(+)-SjOST48 was successfully constructed and expressed in E. coli BL21 (DE3). Western blotting analysis showed that rSjOST48 had good immunogenicity. Indirect immunofluorescent analysis revealed that SjOST48 was mainly distributed on the tegument of the worms. The result of ELISA indicated that the rSjOST48 vaccinated group could induce a significant increase in the level of specific IgG, IgG1 and IgG2a. An immunoprotection experiment showed that the vaccination of rSjOST48 in mice induced 32.62% (P < 0.05) reduction in the numbers of worms and 57.61% (P < 0.01) in eggs in liver, compared with that of the control group. This study provides the foundation for proceeding further research on the biological function of SjOST48 and screening new vaccine candidates for schistosomiasis.
Animals ; Antibodies, Helminth ; blood ; Cloning, Molecular ; DNA, Complementary ; Escherichia coli ; Female ; Genes, Helminth ; Helminth Proteins ; genetics ; immunology ; Immunoglobulin G ; blood ; Male ; Mice ; Mice, Inbred BALB C ; Real-Time Polymerase Chain Reaction ; Recombinant Proteins ; immunology ; Schistosoma japonicum ; genetics ; Schistosomiasis japonica ; prevention & control ; Vaccination

A method for quantitative detection of florfenicol by colloidal gold lateral flow immunoassay was developed.The experimental conditions including pH value, concentrations of antibody in the process of conjugation between the colloidal gold and antibody, amount of gold-labeled antibody, concentration of the antigen sprayed on test lines (T line), and detection time were optimized.With a colloidal gold strip reader, the signal intensity of T lines and control lines (C line) on lateral flow strips was recorded.The T/C ratio of negative control and positive samples was defined as B0 and Bx, and the standard curve was established by plotting the Bx/B0 ratio against the concentration of florfenicol.This assay showed a good linear range from 0.1 to 1.5 ng/mL with the limit of detection of 0.08 ng/mL, while the result could be obtained within 15 min.The result showed that this quantitative method was convenient and rapid, and could be used in screening a large amount of samples on site.

Objective To clone cDNA encoding troponin T of Schistosoma japonicum(SjTnT),and evaluate the protective efficacy induced by recombinant SjTnT in BALB/c mice against S. japonicum challenge infection. Methods The SjTnT gene was amplified from 28-day-schistosome cDNAs by PCR and then subcloned into pET28a(+). The recombinant SjTnT protein (rSjTnT)was expressed in Escherichia coli BL21(DE3)cells. The serum specific to rSjTnT was prepared by immunized BALB/c mice with the recombinant antigen,and the immunogenicity of rSjTnT was detected by Western blotting and ELISA. The immuno-protective efficacy induced by rSjTnT in BALB/c mice was evaluated according to the reduction in worm and egg counts. Results The cDNA encoding SjTnT was successfully cloned and expressed in E. coli. Western blotting showed that rSjTnT had a good immunogenicity. The high level of specific IgG antibodies was detected,and 33.89% worm reduction and 43.94% liver egg reduction were obtained in mice vaccinated with rSjTnT combined with Seppic 206 adjuvant compared with those in the adjuvant control group. Conclusions rSjTnT could induce partial immuno-protection against S. japonicum infec-tion in BALB/c mice. This study provided a basic for understanding the biological function of SjTnT.