Effect of Hei Xiaoyaosan on Neuroinflammation in AD Model Rats： Based on Wnt/β-catenin Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore whether Hei Xiaoyaosan can inhibit the inflammatory response in the hippocampi of Alzheimer's disease （AD） rats by regulating and activating the Wnt/β-catenin signaling pathway to improve the cognitive and memory dysfunction. MethodAmong the 90 male Wistar rats， 12 were randomly selected as the blank group （normal saline） and 12 as the sham operation group （normal saline）. For the remainder， amyloid β-protein42 （Aβ42） was injected in the left and right hippocampus to induce AD， and then the AD rats were randomized into model group， low-， medium-， and high-dose Hei Xiaoyaosan groups （corresponding doses of Hei Xiaoyaosan， ig）， and donepezil group （donepezil hydrochloride，ig）， with 12 in each group. The administration lasted 42 days. The pathological changes of hippocampal CA1 region was observed based on Nissl staining. The escape latency on the 1st to 5th day in Morris water maze was recorded and the spatial memory on the 6th day was tested. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the expression of interleukin （IL）-6， IL-10， and tumor necrosis factor-α （TNF-α） in rat hippocampus and serum， Western blotting to examine the protein expression of glycogen synthase kinase-3β （GSK-3β）， β-catenin， and peroxisome proliferator-activated receptor gamma （PPARγ）， and real-time polymerase chain reaction （Real-time PCR） to determine the mRNA expression of rat GSK-3β， β-catenin， and PPARγ. ResultCompared with the blank group， the number of neurons in the hippocampal CA1 area of model group was significantly reduced， the arrangement was uneven， the cell body was damaged more obviously， and the Neisser body was unclear. The treatment group was significantly prolonged （P<0.01）， and the number of crossing stations was significantly reduced （P<0.01）， the levels of IL-10 in serum and hippocampus of rats in the model group were significantly decreased， while the levels of IL-6 and TNF-α were significantly increased （P<0.01）， the GSK-3β protein and mRNA in the model group were significantly increased， and the protein expressions of β-catenin and PPARγ were significantly decreased （P<0.01）， and the difference was more obvious. The number of neurons in the donepezil group was more distributed， neatly arranged， the structure was intact， and the Nissl bodies were clear and definite， the escape latency on the 3rd to 5th days in middle and high dose groups of Hei Xiaoyaosan and the donepezil group was significantly shortened （P<0.01）， the number of crossing platforms increased significantly （P<0.01）， the expression levels of IL-10 in the rat hippocampus and serum were significantly increased， while IL-6 and TNF-α were significantly decreased （P<0.01）， GSK-3β in the rat hippocampus was significantly increased. The expressions of GSK-3β protein and mRNA were significantly decreased， while the expression levels of β-catenin and PPARγ protein and mRNA were significantly increased （P<0.01）. There was no significant difference in each index between the donepezil hydrochloride group and the high-dose Hei Xiaoyaosan group. ConclusionHei Xiaoyaosan can inhibit the inflammatory response in the hippocampus of AD rats by regulating the Wnt/β-catenin signaling pathway， thereby alleviating the cognitive and memory impairment of AD rats.

Analysis of influencing factors and predictive parameter of acute kidney injury in sepsis shock patients / 中国实用内科杂志

OBJECTIVE: To investigate the influencing factors in patients with septic shock and the predictive value of related parameters for AKI.METHODS: Totally 256 patients with septic shock treated in our hospital from January 2015 to December 2018 were collected.The patients were divided into acute kidney injury(AKI)group and non-AKI group.The miRNA-125,IL-6,neutrophil gelatinase associated lipocalin(NGAL)and tumor necrosis factor-a(TNF-a)were compared between two groups.Multi-factor Logistic regression analysis was used to assess the variables in predicting the incidence rate.The patients in AKI group were divided into mild-AKI group(AKI stage 1-2)and severe AKI group(AKI stage 3).The factors were statistically compared between two groups.RESULTS: The levels of miRNA-125 and NGAL in AKI group were significantly higher than those in non-AKI group.The creatinine(OR 1.03,95%CI 0.88-1.36),glomerular filtration rate(OR1.23,95% CI 0.75-2.01),miRNA-125(OR 1.56,95% CI 1.02-2.10)and NGAL(OR 1.32,95%CI 0.83-1.67)were associated with AKI(P<0.05).The levels of miRNA-125,NGAL and TNF-a in severe AKI group were significantly higher than those in mild and moderate AKI group(P<0.05).The area under curve of miRNA-125 was 0.80(95%CI 0.75-0.83),the best cut-off value was 32.1,and the sensitivity and specificity were 81.5% and 76.2%.CONCLUSION: The creatinine,glomerular filtration rate and the level of miRNA-125 and NGAL were independently associated with AKI.The level of miRNA-125 can predict the incidence of AKI.

Trans-differentiation from osteoblasts to adipocytes in indirect co-culture / 中国组织工程研究

BACKGROUND:Co-culture technique makes different kinds of ceils cultured in the same system.The trans-differentiation from osteoblasts to adipocytes is usually analyzed under the action of adipogenesis inducers in vitro,but the cellular interactions in vivo are neglected.OBJECTIVE:To investigate the trans-differentiation from osteoblasts to adipocytes in indirect co-culture using Transwell system.METHODS:Mouse preadipocytes 3t3-l1 were induced to adipocytes.There were three groups:group A:mature adipocytes in the lower chamber of Transwell system;group B:mouse osteoblasts MC3T3-E1 in the upper chamber of Transwell system according to a ratio of 1:4 (MC3T3-E1:3t3-l1);group C:mouse osteoblasts MC3T3-E1 alone.At 7,14,and 21 days the cell morphology was observed under inverted phase contrast microscope,the relative level of triglyceride,expression of peroxisome proliferator-activated recoptor y2 in each group were detected,and red oil O staining and alizarin red staining were performed.The cell proliferation inhibition rate in the groups B and C were detected at 0,24 and 36 hours.RESULTS AND CONCLUSION:At 2 weeks after culture,spindle-shaped 3t3-l1 coils changed into round,the light and round lipid droplets in the cytoplasm were increased and were reserved after identified by oil red O staining.In the group B,the cells presented with spindle shape with no transparent lipid droplets after 7-day co-culture until black granules and small round lipid droplets appeared on day 14;and the cells changed from spindle shape to oval or round,and larger lipid droplets were found on day 21.Alizarin red staining results:the staining region in the group B was on a decline with time,while the group C showed no significant changes at each time point and all appeared with large staining region.Oil red O staining results:the staining region in the group B increased gradually in a time-dependant manner,while the group C was negative for oil red O staining and showed no significant changes at different time points.The relative level of triacylglycerol in the group B was increased with time,and there was significant difference between groups A and B (P ＜0.05),and the group C showed no significant differences (P ＞ 0.05).The coll proliferation inhibiting rate in the group B was increased with time,which showed significant difference from the group C (P ＜ 0.05).The expression level of peroxisome proliferator-activated recoptor y2 in the group B was on a rise with time,which had significant difference compared with the groups A and C (P ＜ 0.05).These results indicate that the trans-differentiation from osteoblasts to adipocytes appears in the Transwell system,and metabolic products and cytokines of adipocytes obviously inhibit the proliferation of osteoblasts,but all above conclusions need to be studied in depth.

Differentially expressed genes in heads and tails of Angelica sinensis diels: Focusing on ferulic acid metabolism / 中国结合医学杂志

<p><b>OBJECTIVE</b>To explore the scientific connotation of the discrepant pharmaceutical activities between the head and tail of Angelica sinensis diels (AS), an important herb extensively utilized in Chinese medicine, by the approach of transcriptome sequencing.</p><p><b>METHODS</b>Ten samples of AS were randomly collected in Min County, Gansu Province of China. Transcriptome sequencing of AS was accomplished in a commercial ILLumina HiSeq-2000 platform. The transcriptome of each head and tail of AS were fixed in a gene chip, and detected under the procedure of Illumina HiSeq-2000. Differentially expressed unigenes between the heads and tails of AS were selected by Shanghai Biotechnology Corporation (SBC) online analysis system, based on Gene Ontology, Kyoto Encyclopedia of Genes and Genomes and relevant bioinformatic database.</p><p><b>RESULTS</b>Totally 63,585 unigenes were obtained from AS by high-throughput sequencing platform. Among which 3359 unigenes were identified as differentially expressed unigenes between the heads and tails of AS by SBC analysis system scanning. Of which 15 differentially expressed unigenes participate in the metabolic regulation of phenylpropanoid biosynthesis (PB) pathway and ferulic acid metabolites, in response to the distinguished pharmaceutical actions of the heads and tails of AS.</p><p><b>CONCLUSION</b>Different content of ferulic acid in the heads and tails of AS is related to the differentially expressed genes, particularly involved in the PB pathway.</p>

Online Two Dimensional Liquid Chromatography and Its Application on TCM Quality Control Analysis / 世界科学技术-中医药现代化

Separation of complex mixtures needs more resolving power. In two dimensional liquid chromatography (2D LC), the sample is subjected to two different separation conditions or mechanisms, which significantly increases the peak capacity,resolving power and reduces peak overlap. Meanwhile, 2D separation is more reliable for peak i-dentification from run to run than single-dimension chromatography, because two independent elution times are ob-tained for each peak. Among different 2D liquid chromatographic combinations/configurations, online comprehensive 2D-LC has appeared to be very attractive due to its high peak capacity, minimal sample loss, good reproducibility, and easy to automation, especially for separation of very complex samples. The present review discusses the theoreti-cal aspects and hardware configuration of online 2D-LC, particularly online comprehensive 2D-LC. It highlights its concept, features, advantages, applications and prospective in TCM quality Control analysis.

Screening and Identification of a New Elastase-producing Strain / 微生物学通报

The study provide a theoretical basis for the industrial production of a elastase-high-yield strain which was isolated from the straw,and the selected strain was identified. The screening strategy included casein(skim milk) plate selecting and elastin(beef tendon) re-screening. And then,the morphological,physiological and biochemical characteristics as well as 16S rRNA sequence homology of the selected strain were studied. Finally,the strain LSF-97 which had a excellent decomposing ability to beef tendon was obtained. The results showed that the strain LSF-97 is relative to the Bacillus pumilus with 100 % similar in sequence under the phylogenetic tree,the morphological and physiological and biochemical characteristics are also consistent with pattern of bacteria. So it was identified as Bacillus pumilus.

Mutational Screening of Zuelacmycin-producing Strain RL-2 / 微生物学通报

In this research, the zuelacmycin-producing strain Streptomyces venezuelaevar. qinlingensis RL-2 was used as original strain, the spore suspension of which was induced by UV, LiCl and the compound treatmeat (UV+LiCl) respectively. The zuelacmycin-high-yield strain UVL-108 was obtained by the treatment that the exposure time under UV irradition was 45 s and the concentration of LiCl was 0.4%. The heredity characters of mutant UVL-108 were stable in succesive six generations. The antibacterial activity and the fermentation titer of mutant UVL-108 were determinded by bidirectioned culture and mycelial linear growth respectively. The results demonstrated the antibaterial activity of mutant UVL-108 was improved by 77%, and the relative toxicity of fermentation to Phyricularia grisea was improved by two times compared with the original strain.

Transgenic mice produced by intracytoplasmic sperm injection / 生物工程学报

In our previous study, normal and fertile mice were successful produced from oocytes following intracytoplasmic sperm injection (ICSI). In the present study, the possibility of producing transgenic embryos and offspring with this procedure was evaluated. After freezing-thawed once using HEPES-CZB medium without cryoprotectants, the cauda sperm from KM fertile male were exposed to the circular or linear pEGFP-N1 DNA for 1 min and then co-injected into metaphase II oocytes of B6D2F1 strain. When the zygotes with two pronuclei were cultured in CZB medium to day 3.5, 39.1% (9/23) of them, derived from oocytes co-injected with sperm head and pEGFP-N1 plasmid DNA, were expressed GFP protein. After transfer of the ICSI embryos with two pronuclei from co-injection of sperm head and foreign DNA, seven recipients delivered 30 pups (23.8%, 30/126). Southern blot results revealed that three of sixteen offspring integrated with GFP and neomycin genes together (18.8 %). Interestingly, all of them were produced from oocytes co-injected sperm head and linear DNA (33.3%, 3/9), while none of seven ICSI offspring integrated either GFP or neomycin gene in the group of co-injection of sperm head and circular plasmid DNA. These results indicated that the high efficiency of transgenic mouse could be produced by ICSI. It may be shown that linear DNA is more easily to integrate into host genome than circular DNA when ICSI was used to produce transgenic animals.

Normal mice derived from oocytes following intracytoplasmic sperm injection (ICSI) / 生物工程学报

This paper describes the use of piezo-driven micropipette for intracytoplasmic sperm injection of mice eggs. The head of fresh spermatozoa from KM (Kunming) fertile mice was individually injected into mature oocytes of hybrid mice B6D2F1. Approximately eighty three percent of sperm-injected oocytes survived, and 84.0% of them fertilized normally (extrusion of the second polar body and formation of male and female pronuclei). The eggs fertilized by sperm injection could develop in vitro to 2-cell (98% vs 94.7%), 4-cell (89.5% vs 92.1%) stages, no significantly (P > 0.05) different from embryos fertilized in vivo but there were significantly (P < 0.01) few morulae (63.8% vs 84.2%) and blastocysts (25.7% vs 68.4%) developed in vitro after further culture in vitro in the group of ICSI. When 120 embryos at the pronuclear stage were transferred to seven pseudopregnant KM female, 23.3% of the embryos (0 - 50%, depending on the host) reached the full term. Except for three that were cannibalized soon after birth, all of the young (25 pups) developed into normal and fertile adult. Here we report the first birth of mouse offspring following ICSI in China. These studies may increase understanding of the fertilization process and of how ICSI works.

Temperament characteristics of children with cerebral palsy / 中国康复理论与实践

@#ObjectiveTo explore temperament characteristics of children with cerebral palsy.Methods364 children with cerebral palsy, ages from four months to seven years, were tested with modified Carey\'s Children Temperament Questionnaires. Temperament characteristics of children with different types of cerebral palsy, able and unable walk independently were compared.ResultsChildren with cerebral palsy had negative temperament including less active, low persistence, high distractibility and negative mood. Approach was different between children with diplegia and athtosis. There were no different temperaments among children with different lever of IQ. Children they could not walk independently had an obvious negative temperament.ConclusionChildren with cerebral palsy have significantly negative temperament.

The effects of semiconductor laser irradiation and a high voltage static electric field on small vessel impairment in diabetic rats / 中华物理医学与康复杂志

Objective To study the effects of semiconductor laser irradiation and high voltage static electric fields on small vessel damage in diabetic rats.Methods Fifty healthy male Wistar rats were randomly divided into two groups:8 rats in a normal group and 42 in a diabetic model group.The diabetic models were created by intrape- ritoneal injection of streptozocin.The diabetic model rats were randomly divided into four subgroups:a diabetes group,a semiconductor laser treatment group,a high voltage static electric field treatment group and a comprehensive treatment group receiving combined semiconductor laser and high voltage static electric field exposure.The rats in each treatment group were subjected to the corresponding intervention.After 20 days of treatment,the venous blood, kidney tissue and myocardium tissue of the rats were collected,and the concentrations of blood glucose,insulin,en- dothelin and tissue were detected.Results Compared with the normal group,a significant increase in blood glu- cose and endothelin was observed in the diabetic model group,along with significantly decreased blood insulin and significant small vessel endothelium proliferation in the kidney tissue.Compared with the diabetes group,endothelin levels were significantly lower in all 3 treatment groups,and blood insulin was also higher in the comprehensive treat- ment group.Conclusion There were severe abnormalities in blood glucose,insulin and endothelin as well as mild impairment of small vessel endothelium proliferation in the diabetic rats.Semiconductor laser and high voltage static electric field exposure have a role in treating and preventing these conditions in diabetic rats.

Molecular cloning, expression mutation of myostatin and study on biochemical activity of its C-terminal peptide / 生物工程学报

Myostatin, a member of the TGF-beta family, negatively regulates skeletal muscle development. Mutation of myostatin activity leads to increases muscle growth and carcass lean yield. The bovine myostatin mutation cDNA was amplified by polymerase chain reaction, and then sub-cloned into the expression vector pET-30a( + ) to form the expression plasmid pET30a (+)-action/ Myostatin. The recombinant plasmid was transformed into E. coli BL21. The overexpression product of pET30a (+)-action/ Myostatin was been showed in vitro. Sheep skeletal muscle cell were cultured with the purified myostatin mutation C-terminal peptide. The results of this study suggest that had a powerful activity to stimulate the hyperplasia and proliferation of sheep muscle cells and shows high biochemical activity.

Bioavailability and pharmacokinetics of anastrozole in healthy male volunteers / 中国药理学通报

AIM: To evaluate bioavailability and pharmacokinetics of domestic and imported anastrozole tablets. METHODS: Twenty Chinese healthy male volunteers were enrolled in a randomized crossover study with a single oral dose of 2 mg of the two formulations respectively. The anastrozole in plasma was measured by gas chromatography with electron-capture detection. The linear range was from 1. 325 to 106 μg·L-1 plasma. The extraction recovery rates for plasma concentration of 5.3, 21.2 and 53.0 μg·L-1 were 76.8%, 87.0% and 78.7% respectively. Inter-day and intra-day precisions of the method were < 9%. Area under concentration-time curve (AUC0-t), maximum plasma concentration (Cmax) and reach peak time (Tmax) were evaluated by variance analysis and two one-side t-test. RESULTS: A two-compartment model was adopted in anasreozole plasma concentration-time data analysis. The main pharmacokinetic parameters of domestic and imported anastrozole tablets such as Cmax, Tmax, AUC0-t and T1/2β were (36.5 ± 6.9) and (35.6 ± 9.4) μg·L-1, (1.56±0.41) and (1.53±0.49) h, (1403.6 ± 321.2) and (1371.6±329.4) μg·h·L-1, (42.57±10.15) and (43.41±8.59) h, respectively, and there were no significant differences between the two formulations. CONCLUSION: Domestic and imported anastrozole tablets are of bioequivalence. The relative bioavailability of the domestic tablet is (102.7±5.6) %.

Transgenic animals bioreactors / 生物工程学报

The production of human recombinant proteins in milk of transgenic farm animals offers a safe, very cost-effective source of commercially important proteins that cannot be produced as efficiently in adequate quantities by other methods. This review has summarized the current status of gene selection, vector construct, transgenic methods, economics, and obvious potential in transgenic animals bioreactors. Recently, a more powerful approach was adopted in the transgenic animals founded on the application of nuclear transfer. As we will illustrate, this strategy presents a breakthrough in the overall efficiency of generating transgenic farm animals, product consistency, and time of product development. The successful adaptation of Cre-/lox P-mediated site-specific DNA recombination systems in farm animals will offer unprecedented possibilities for generating transgenic animals.