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.

Amyloid fibrils are found in systemic amyloidosis diseases such as Alzheimer's disease, Parkinson's disease, and type II diabetes. Currently, these diseases are diagnosed by observation of fibrils or plaques, which is an ineffective method for early diagnosis and treatment of disease. The goal of this study was to develop a simple and quick method to predict the possibility and speed of fibril formation before its occurrence. Oligomers generated from seven representative peptide segments were first isolated and detected by ion-mobility mass spectrometry (IM-MS). Then, their assemblies were disrupted using formic acid (FA). Interestingly, oligomers that showed small ion intensity changes upon FA addition had rapid fibril formation. By contrast, oligomers that had large ion intensity changes generated fibrils slowly. Two control peptides (aggregation/no fibrils and no aggregation/no fibrils) did not show changes in their ion intensities, which confirmed the ability of this method to predict amyloid formation. In summary, the developed method correlated MS intensity ratio changes of peptide oligomers on FA addition with their amyloid propensities. This method will be useful for monitoring peptide/protein aggregation behavior and essential for their mechanism studies.

Objective To investigate the role of histone lysine methyltransferase G9a in sevoflurane-induced cognitive impairment in the developing brain of neonatal rats.Methods Thirty-six 5-day-old male SD rats were randomly divided into 3 groups (n =12):control group,sevoflurane group and Bix01294 (the inhibitor of histone lysine methyltransferase G9a)group.The rats in the sevoflurane group and the Bix01294 group received 3% sevoflurane anesthesia for 2 hours once a day at postnatal 5-7 days (P5-P7 ).The rats in the Bix01294 group received Bix01294 (10 mg/kg)subcu-taneously at 1 5 min before anesthesia,and the rats in the control group and sevoflurane group received normal saline for injection (0.1 ml)at the same time.The open-field test and fear condition-ing test were performed at P3 5 and P3 9-P41 ,respectively.The tissues of hippocampus were collected at P42 to measure the levels of G9a,histone H3 lysine 9 dimethylation (H3K9me 2 )and synapsin 1. Results Compared with the control group,the percentage of freezing time of sevoflurane group was significantly decreased in the contextual fear condition test,while the levels of G9a and H3K9me 2 were significantly increased and the level of synapsin 1 was significantly decreased (P <0.01).How-ever,the percentage of freezing time of Bix01294 group was significantly increased,while the levels of G9a and H3K9me 2 were significantly decreased and the level of synapsin 1 was significantly in-creased compared with the sevoflurane group (P <0.05).There was no difference in the total distance and residence time in the central grid in the open-field test,and the percentage of freezing time in the cued fear condition test among the three groups.Conclusion Histone lysine methyltransferase G9a is involved in the sevoflurane-induced long-term cognitive impairment in developing brain of neonatal rats,which may be associated with the increase of H3K9me 2 and the down-regulation of synapsin 1 in the hippocampus.

Objective To observe the effects of mitochondrial protectant on hippocampal mito-chondrial function and synaptic plasticity in isoflurane-anesthesia aged mice.Methods Forty-eight fif-teen-month-old male C57BL/6 mice were equally divided into 4 groups (n=12):oxygen+normal sa-line (group CN),oxygen+SS-31 (group CS),isoflurane+normal saline (group IN),and isoflurane+SS-31 (group IS).SS-31(5 mg/kg)or normal saline was intraperitoneally administered with a vol-ume of 0.4 ml/kg 30 min before gas inhalation.After two hours gas inhalation,the hippocampus was removed to determine complex Ⅰ-Ⅳ activities,adenosine triphosphate (ATP)and mitochondrial membrane potential (MMP)levels from isolated mitochondria,and measure the content of synapsin-1,PSD-95,NMDAR2A,NMDAR2B,CaMKⅡα and CaMKⅡβ.Results Compared with the group IN,complex Ⅰ activity,ATP and MMP levels were increased,synapsin-1 and PSD-95 were up-regu-lated,whereas NMDAR2B,CaMK Ⅱα and CaMK Ⅱβ were down-regulated in the group IS (P <0.05).No significant difference was observed in the complex Ⅱ-Ⅳ activities and NMDAR2A expres-sion.Conclusion Mitochondrial protectant SS-31 attenuates isoflurane-induced mitochondrial dysfunc-tion and neuron synaptic plasticity impairments in aged mice.

Various teaching methods were used in occupational health and occupational medicine teaching,including problem based learning,multimedia teaching,bilingual teaching,case based learning and practice teaching methods when being confronted with new situation of occupational health and occupational safety.These methods are mean to encourage students' enthusiasm,cultivate students' comprehensive ability and enhance their sense of social responsibility and mission.Results showed that these methods improved the quality of teaching and achieved good teaching results.

Objective To look for the genes of Schistosoma japonicum related to the Schistosoma-resistance of Microtus fortis.Methods The fresh sera of Microtus fortis were used to screen a T7 phage display cDNA library from worms of Schistosoma japonicum established in our lab.The positive clones were sequenced and functionally analysed through bioinformatics.Results The specific phages binding to the sera of Microtus fortis were enriched 857-fold after three rounds of biopanning,and 58 positive clones picked at random were sequenced and 10 ESTs were obtained.BLASTn results showed that 7 ESTs had 99%-100% similarity to the genes of Shistosoma japonicum reported in GenBank and 1 EST had 82% similarity to a zinc finger protein encoden gene from Pan troglodytes.The results of these ESTs function prediction indicated most of them were involved in the regulation of gene expresion of Schistosoma japonicum.Conclusions Several target genes of Schistosoma japonicum related to the Schistosoma-resistance of Microtus fortis are obtained and those would lay foundation to expatiate the native resistance mechnism of Microtus fortis to Schistosoma japonicum.

Objective To screen protective antigen genes and construct the T7 phage display library from adult worms of Schistosoma japonicum.Methods Total RNA was extracted from adult worms of S.japonicum by Trizol reagent and mRNA was isolated from the total RNA.The ds cDNA was synthesized by reverse transcription using random primer.Directional EcoRⅠ/HindⅢ linkers were ligated into the ends of ds cDNA and the ds cDNA was digested with EcoRⅠand HindⅢ,which resulted in ds cDNA with EcoRⅠand HindⅢ adhering ends.The digested ds cDNA fragments longer than 300 bp in length were fractionated and ligated into T7 Select 10-3b vector.After packaging in vitro,the T7 Select 10-3b vector was transformed into BLT5403 to construct the T7 phage display cDNA library.Plaque assay and PCR were used to evaluate the library.Seven known objective genes of S.japonicum were screened by PCR to detect the representation of the library.Result Primary library capacity was 4.98?106 pfu,and the titer of amplified library was 3.85?1011 pfu/mL.The PCR identification result of 96 clones picked at random showed that recombination rate was 93.8%,in which 95.6% inserted cDNA fragments were longer than 300 bp in length.All the seven known objective genes of S.japonicum were amplified from the library.Conclusion The T7 phage display library from adult worms of Schistosoma japonicum was constructed.

Objective To express the gene encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase (Sj32) and evaluate the potential of the recombinant protein rSj32 in diagnosis of domestic animal schistosomiasis. Methods The DNA fragment encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase was cloned with PCR from pET-28(a)/Sj32, and a recombinant plasmid was previously constructed in the laboratory, which contained the ORF of the gene encoding the pro-enzyme Sj32. The amplified DNA fragment was subcloned into pET-28a( + ) and the recombinant plasmid was transformed into E. coli BL21 (DE3) to express the mature protease of Sj32. Then the recombinant antigen (rSj32) was used in ELISA assay to diagnose schistosomiasis of mice, rabbits and water buffalo artificially infected. The detection effects of soluble Schistosoma japonicum egg antigen (SEA) , rSj32 and the recombinant 23 KDa membrane protein were compared. Results The recombinant antigen rSj32 with a molecular weight 41 KDa was successfully produced in E. coli BL21 ( DE3) and was purified with His Column with a yield of 25 mg/L E. coli culture. By using rSj32 as coating antigen in ELISA assay to detect the specific antibody in artificially infected mice, rabbits and buffalo, the sensitivities were 88.9% , 85.0% and 71.8% , respectively, the specificities were 100% , 96.7% and 96. 9% , respectively. There were no significant differences among the detection results of rSj32, SEA and rSj23. Conclusion rSj32 is a promising antigen for serological diagnosis of domestic animal schistosomiasis.

Objective To establish a simple, feasible and precise quality control method for the determination of contents and related substances of demethyl levophencynonate hydrochloride (L-LPC)tablets.Methods The mobile phase consisted of methanol,acetonitrile and sodium acetate buffer solution(pH 5.0),at a ratio of 4∶3∶3,at a flow rate 1.0 ml/min and a detection wavelength of 220 nm.Samples were injected 100 μl and determined at room temperature.Results The calibration curves showed good linearity (R2 =1) within the test range of 0.1-50μg/ml.The recovery of the method was about (100.15 ±0.73)%, and the stability of working solutions was acceptable in 8 h (RSD=0.36%).Conclusion The results indicated that the developed method can be readily used as a quality control method.