Different methods establish a rat model of morphine drug discrimination with a fixed ratio 10 / 药学学报

In this study, a rat morphine drug discrimination model with a fixed ratio (FR) of 10 (FR10) was established using different methods to explore which methods can shorten the modeling time and test the dose-response relationship and median effective dose (ED50) value. Animal welfare and experimental procedures are in accordance with the provision of the Animal Ethics Committee of Shanghai InnoStar Bio-tech Co., Ltd. Forty rats were initially shaped to press lever under a fixed-ratio schedule of food reinforcement. The animals that were successfully trained under a FR10 schedule of food reinforcement were divided into two groups, namely the single-lever + double-lever training group 1 and the double-lever training group 2. In each group, rats were trained to discriminate morphine at 5.6 mg·kg-1 from saline by the intraperitoneal route. After training, different doses of morphine were used to substitute for training dose of morphine, the dose-response curve for morphine were identified in rats, and the ED50 value was calculated. The results showed that, in food training phase: 34 rats successfully entered the discrimination training during food training; in discrimination training phase: 14 animals in group 1 met the discrimination training standard for the first time, which took about (40.71 ± 2.93) days, and there were 13 animals in group 2 that met the discrimination training criteria for the first time, and it took about (51.15 ± 2.55) days. It can be seen that the method of single-lever + double-lever training is better than single-lever training, and the difference is significant compared with group 1 (P ˂ 0.05); in generalization test phase: there are 17 rats completed morphine generalization test, and the percentages of morphine-lever responses produced by the generalization test of different doses of morphine (0, 0.1, 0.5, 1, 3, 5.6, and 10 mg·kg-1) were (9.56 ± 3.13) %, (9.01 ± 5.83) %, (13.82 ± 7.95) %, (29.04 ± 10.13) %, (41.70 ± 10.65) %, (85.36 ± 7.16) %, (94.56 ± 2.76) %, respectively. The results showed that the discriminative stimulative effect induced by morphine dose between 0-10 mg·kg-1 increased in a dose-dependent manner, producing a good dose-response curve, and the ED50 value of morphine was 4.74 mg·kg-1 by linear fitting. The above results showed that, the FR10 morphine drug discrimination model has been successfully established using different methods; the single-lever + double-lever training method is better than the single-lever training, and can relatively shorten the discrimination training cycle.

Comparison study of whole exome sequencing and targeted panel sequencing in molecular diagnosis of inherited retinal dystrophies / 北京大学学报(医学版)

OBJECTIVE@#To evaluate and compare whole exome sequencing (WES) and targeted panel sequencing in the clinical molecular diagnosis of the Chinese families affected with inherited retinal dystrophies (IRDs).@*METHODS@#The clinical information of 182 probands affected with IRDs was collected, including their family history and the ophthalmic examination results. Blood samples of all probands and their relatives were collected and genomic DNA was extracted by standard protocols. The first 91 cases were subjected to the WES and the other 91 cases were subjected to a specific hereditary eye disease enrichment panel (HEDEP) designed by us. All likely pathogenic and pathogenic variants in the candidate genes were determined by Sanger sequencing and co-segregation analyses were performed in available family members. Copy number variations (CNVs) detected by HEDEP were further validated by multiplex ligation-dependent probe amplification (MLPA). As PRGR ORF15 was difficult to capture by next generation sequencing (NGS), all the samples were subjected to Sanger sequencing for this region. All sequence changes identified by NGS were classified according to the American College of Medical Gene-tics and Genomics and the Association for Molecular Pathology (ACMG/AMP) variant interpretation guidelines. In this study, only variants identified as pathogenic or likely pathogenic were included, while those variants of uncertain significance, likely benign or benign were not included.@*RESULTS@#In 91 cases with WES, pathogenic or likely pathogenic variants were determined in 30 cases, obtaining a detection rate of 33.00% (30/91); While in 91 cases with HEDEP sequencing, pathogenic or likely pathogenic variants were determined in 51 cases, achieving the diagnostic rate of 56.04% (51/91), and totally, the diagnostic rate was 44.51%. HEDEP had better sequencing coverage and read depth than WES, therefore HEDEP had higher detection rate. In addition, HEDEP could detect CNVs. In this study, we detected disease-causing variants in 29 distinct IRD-associated genes, USH2A, ABCA4 and RPGR were the three most common disease-causing genes, and the frequency of these genes in Chinese IRDs population was 11.54% (21/182), 6.59% (12/182) and 3.85% (7/182), respectively. We found 43 novel variants and 6 cases carried variants in RPGR ORF15.@*CONCLUSION@#NGS in conjunction with Sanger sequencing offers a reliable and effective approach for the genetic diagnosis of IRDs, and after evaluating the pros and cons of the two sequencing methods, we conclude that HEDEP should be used as a first-tier test for IRDs patients, WES can be used as a supplementary molecular diagnostic method due to its merit of detecting novel IRD-associated genes if HEDEP or other methods could not detect disease-causing va-riants in reported genes. In addition, our results enriched the mutational spectra of IRDs genes, and our methods paves the way of genetic counselling, family planning and up-coming gene-based therapies for these families.

Association of folate metabolism genes MTRR and MTHFR with complex congenital abnormalities among Chinese population in Shanxi Province, China / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To explore the association of polymorphisms in folate metabolism genes, methionine synthase reductase (MTRR) gene and 5,10-methylenetetrahydrofolate reductase (MTHFR) gene, with complex congenital abnormalities and to further investigate its association with complex congenital abnormalities derived from three germ layers.</p><p><b>METHODS</b>A total of 250 cases of birth defects (with complex congenital abnormalities including congenital heart disease, neural tube defects, and craniofacial anomalies) in Shanxi Province, China were included in the study. MTRR single nucleotide polymorphism (SNP) (rs1801394) and MTHFR SNP (rs1801133) were genotyped by the SNaPshot method, and the genotyping results were compared with those of controls (n=420).</p><p><b>RESULTS</b>SNPs rs1801394 and rs1801133 were associated with multiple birth defects. For the recessive model, individuals with GG genotype at rs1801394 and CC genotype at rs1801133 had a relatively low risk of developing birth defects, so the two genotypes were protective factors against birth defects. The homozygous recessive genotype at rs1801133, which served as a protective factor, was associated with ectoderm- or endoderm-derived complex congenital abnormalities, while the homozygous recessive genotype at rs1801394, which served as a protective factor, was associated with ectoderm-, mesoderm- or endoderm-derived complex congenital abnormalities.</p><p><b>CONCLUSIONS</b>Among the Chinese population in Shanxi Province, the SNPs in folate metabolism genes (MTRR and MTHFR) are associated with complex congenital abnormalities and related to ectoderm, mesoderm or endoderm development.</p>

Optimization of extraction process for tannins from Geranium orientali-tibeticum by supercritical CO2 method / 中国中药杂志

In order to optimize extraction process conditions of tannins from Geranium orientali-tibeticum by supercritical CO2, the content of tannins was determined by phosphomolybdium tungsten acid-casein reaction, with extraction pressure, extraction temper- ature and extraction time as factors, the content of tannins from extract of G. orientali-tibeticum as index, technology conditions were optimized by orthogonal test. Optimum technology conditions were as follows: extraction pressure was 25 MPa, extraction temperature was 50 °C, extracted 1.5 h. The content of tannins in extract was 12.91 mg x g(-1), extract rate was 3.67%. The method established could be used for assay the contents of tannin in G. orientali-tibeticum. The circulated extraction was an effective extraction process that was stable and feasible, and that provides a way of the extraction process conditions of tannin from G. orientali-tibeticum.

Effect of two kinds of endophytic fungi on salt resistance of Chrysanthemum morifolium / 中草药

Objective: The NaCl stress conditions were simulated to study the effect of the endophytic fungi C1, C4 on antisalty characteristic of Chrysanthemum morifolium in the adverse circumstance. Methods: Endophytic Botrytis sp. (C1) and Chaetomium globosum (C4) were inoculated to the C. morifolium plantlets which were planted in the pots in order to research the effects of salt stress on physiological indicators of C. morifolium. Results: With the increase of NaCl concentration, the water content of root and leaf decreased in every group. The loss of root and leaf's water in fungi-treated group was smaller than that in the control group. SOD activities in every group increased with the increase of NaCl concentration, and achieved the peak value at 20 g/L NaCl. The SOD activity in fungi-treated group was higher than that in the control group. Soluble protein of fungi-treated group was higher than that in the control group, and C4 group surpassed C1 group. POD activity increased firstly and then decreased, and compared to the control group, the POD activities in C4 and C1 groups increased by 25.50% and 1.35%, respectively at 15 g/L NaCl. PAL activity of C4 treated group was seven folds compared to the control group at 15 g/L NaCl. Conclusion: Endophytic fungi could enhance the salt-tolerant ability of C. morifolium, and the effect in C4 group was better than C1 group.

Effects of trajectory wind direction on ion concentration of PM10 / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the characterization apportionment of main ion concentrations of PM10 under the influence of trajectory wind direction in London.</p><p><b>METHODS</b>PM10 samples from 1 May 1995 to 30 October 1995 of Oxford Street of Central London were collected, the metals and anions of which were measured using atomic absorption spectrometry (AAS) and ion chromatography (IC). Composite trajectories representative of the air mass arriving in London at the same period were calculated based on basic routine back trajectories from the British Atmospheric Data Centre (BADC).</p><p><b>RESULTS</b>Concentration apportionments of main ions were similar when the trajectory was plotted back at 6 h, 12 h, and 24 h, some were obviously different. Mg, Ba, Pb, and Cu had similar peak apportionments at the area 180 degrees-320 degrees, but Zn and Ni at the area of 90 degrees-270 degrees, NO3 and SO4(2-) at the area of 100 degrees-220 degrees. Cl- concentration peak apportionment was at the area of 220 degrees-300 degrees, which showed that Cl- mainly came from the North Sea.</p><p><b>CONCLUSION</b>Trajectory wind direction has important effect on ion concentration apportionment of PM10 in London. The ions have similar concentration peak apportionments or their correlation coefficients are statistically significant.</p>