Improvement effect and mechanism of N-butylphthalide on inflammatory injury of bone marrow mesenchymal stem cells in rats / 中国药房

OBJECTIVE To study the improvement effect and possible mechanism of N-butylphthalide on inflammatory injury of bone marrow mesenchymal stem cells （BMSCs） in rats. METHODS BMSCs of rats were divided into control group， model group， N-butylphthalide low-concentration， medium-concentration and high-concentration groups （10， 20， 50 μmol/L）. BMSCs were cultured in vitro and lipopolysaccharide （the final concentration of 10 mg/L） was used to establish the inflammatory injury model. After the intervention of N-butylphthalide， the survival rate， apoptotic rate， the contents of tumor necrosis factor α （TNF- α）， interleukin 1β （IL-1β） and IL-6 in cell culture medium， the mRNA expression of nuclear factor-κB（NF-κB） p65， and the protein expressions of caspase-3， B-cell lymphoma 2 （Bcl-2）， Bcl-2 related X protein （Bax） and NF-κB p65 in cells were detected. RESULTS Compared with control group， the survival rate and protein expression of Bcl-2 were decreased significantly in model group （P＜0.05）； the apoptotic rate， contents of TNF-α， IL-1β and IL-6， the mRNA expression of NF-κB p65， and the protein expressions of caspase-3， Bax and NF-κB p65 were increased significantly （P＜0.05）. Compared with model group， above indexes were significantly reversed in all concentration groups of N-butylphthalide （P＜0.05）， in concentration-dependent manner. CONCLUSIONS N-butylphthalide can ameliorate the inflammatory injury of BMSCs induced by lipopolysaccharide， and its mechanism may be related to the inhibition of NF-κB signaling pathway.

Establishment and preliminary mechanism study of the zebrafish strain of KIAA0196: A candidate pathogenic gene for heart development / 中南大学学报(医学版)

To explore the effects of KIAA0196 gene on cardiac development and the establishment of zebrafish strain.
 Methods: Peripheral blood and gDNA from patients were extracted. Copy number variation analysis and target sequencing were conducted to screen candidate genes. The KIAA0196 knockout zebrafish was generated by CRISPR/Cas9 to detect whether KIAA0196 deficiency could affect cardiac development. Finally, the wild-type and mutant zebrafish were anatomized and histologically stained to observe the phenotype of heart defects.
 Results: The KIAA0196 knockout zebrafish strain was successfully constructed using CRISPR/Cas9 technology. After 60 hours fertilization, microscopic examination of KIAA0196 knockout zebrafish (heterozygote + homozygote) showed pericardial effusion, cardiac compression and severely curly tail. Compared with wild-type zebrafish, the hearts of mutant KIAA0196 zebrafish had cardiac defects including smaller atrium and larger ventricle, and the myocardial cells were looser.
 Conclusion: KIAA0196 gene plays an important regulatory role in the development of heart. It might be a candidate gene for congenital heart disease.

Echocardiography-guided percutaneous closure of patent ductus arteriosus without arterial access: Feasibility and safety for a new strategy / 中南大学学报(医学版)

Objective:To evaluate the feasibility and safety of device closure of patent ductus arteriosus (PDA)using only venous access under echocardiography guidance alone.Methods:A total of 102 consecutive pediatric patients underwent transcatheter PDA closure without arterial access,under the guidance of only echocardiography.The patients were followed up by clinical examination,electrocardiogram,and echocardiogram at 1,3,6 12,and 24 months.Results:Transvenous PDA closure under echocardiographic guidance was successful in 99 (97.1％)patients.There were no acute procedural complications or severe adverse events.The duration ranged from 10 to 65 minutes (median,21 minutes).Immediate complete closure of PDA was achieved in 87 patients (87.9％),and 100％ of the patients were completely closed after 24 h.There were no severe adverse events in the period of 1-24 months (median,12 months) follow up.Conclusion:Transvenous PDA closure without fluoroscopy avoids radiation exposure,contrast agent usage and potential arterial complications.It can be used as an alternative procedure,especially for children.

Effects of leptin on endoplasmic reticulum stress related proteins after focal cerebral ischemia / 中国神经精神疾病杂志

Objective To investigate the effect of leptin on endoplasmic reticulum stress related protein after focal cerebral ischemia in rats. Methods Ischemia was induced by occluding the middle cerebral artery in rats brain using the filament occlusion method. Forty SD rats were randomly divided into sham operation group, cerebral is-chemia group and leptin-preconditioning group. Leptin was injected subcutaneously before occlusion of blood vessel. Longa 5 score neurological function scale, body weight and brain edema changes were measured 6 hours after MCAO, and the brain was removed to detect the endoplasmic reticulum marker protein: glucose-regulated protein 78 (GRP78) and C/EBP-homologous protein (CHOP) by immunohistochemical method. Results There was no difference in the body weight changes between leptin-preconditioning group and ischemic group. In the leptin-preconditioning group, the neurological function score (1.90±0.31 vs. 2.50±0.52, P<0.05) and the degree of brain edema (3.60±0.52 vs. 7.70±0.94, P<0.001) were significantly lower than those in the cerebral ischemia group. Moreover, the expression of GRP78 in leptin-preconditioning group was significantly higher than that in ischemia group (48.69 ±5.06 vs. 35.78± 4.35, P<0.01), and the expression of CHOP was significantly lower than that of ischemia group (60.24 ±4.11 vs. 38.81±5.34, P<0.01). Conclusion Leptin can reduce the neurological deficit and may be associated with the up-reg-ulation of GRP78 protein, and down regulation of CHOP protein to weaken the endoplasmic reticulum stress caused by cerebral ischemia

Comparison of three methods for preparation of bacterial ghosts from avian pathogenic Escherichia coli / 生物工程学报

Bacterial ghosts are bacterial cell envelopes devoid of cytoplasmic contents while maintaining their cellular morphology, which can be used as a new vaccine and delivery vector. In this study, a clinical isolate of avian pathogenic Escherichia coli (APEC) strain DE17 was used to prepare bacterial ghost through three different ways. The results showed that the cleavage efficiency of DE17 bacterial ghost was 99.9% with the lysis plasmid containing the PhiX174 lysis gene E. Scanning electron microscopy showed that transmembrane tunnels were formed in the middle or both ends of the cell envelope of DE17. Furthermore, the DE17 bacterial ghost was prepared with one of cell penetrating peptides (CPPs) named MAP (KLALKLALKALKAALKLA), which will completely inactivate DE17 (OD₆₀₀=0.1) by 10 μmol/L MAP. The cell envelope showed a gully-like structure and obvious transmembrane tunnels were not found through the SEM. However, the DE17 could not be lysed by importing the lysis plasmid (pBV220-MAP), which was used to express MAP. The present study will benefit for research on bacterial ghost preparation methods and provide a reference for biosafety of bacterial ghost vaccines.

Effect of exogenous carbon monoxide donor hematin on seed germination and physiological characteristics of Cassia obtusifolia seedlings under NaCl stress / 中国中药杂志

<p><b>OBJECTIVE</b>In order to get the method to improve the salt resistance of seeds and seedlings for Cassia obtusbifolia under NaCl stress, seed germination and physiological characteristics of C. obtusifolia seedlings were studied.</p><p><b>METHOD</b>Several physiological indexes of C. obtusifolia seeds treated with exogenous carbon monoxide donor hematin under NaCl stress like the germination vigor, germination rate, germination index and vigor index were measured. And other indexes like the relative water content, the contents of photosynthetic pigment, chlorophyll fluorescence parameters, the contents of soluble sugar, protein and proline, malondialdehyde (MDA), the activities of superoxide (SOD), peroxidase (POD) and catalase (CAT) were also measured.</p><p><b>RESULT</b>The germination indexes of C. obtusifolia seeds under NaCl stress had been inhibited obviously. But after the treatment of hematin, every germination indexes were all increased. The result showed that the treatment of exogenous CO donor hematin obviously improved the germination vigor, germination rate, germination index and vigor index, increased the content of chlorophyll a, chlorophyll b, total chlorophyll, improved the photochemical efficiency of photosystem II (Fv/Fm), photochemical efficiency (Fv'/Fm'), PS II actual photochemical efficiency (phiPS II), photochemical quench coefficient (qP), decreased non-photochemical quenching coefficient (NPQ) and the content of malondialdehyde (MDA) , increased the relative water content of leaves and the content of soluble surge, protein and proline. Meanwhile, the results also indicated that CO improved the activities of superoxide (SOD), peroxidase (POD) and catalase (CAT). The effects of CO could be reversed when CO scavenger Hb is added.</p><p><b>CONCLUSION</b>Exogenous CO donor hematin with appropriate concentration could significantly alleviate the damages to the seeds and seedlings of C. obtusifolia under NaCl stress and promote the salt resistance of the seeds and seedlings through improving the germination indexes, the photochemical efficiency and the antioxidase activities of the seedlings.</p>

Effect of zinc sulphate and PEG priming on ageing seed germination and antioxidase activities of Perilla frutescens seedlings / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the method for improving the aging resistance of seeds and seedlings of Perilla frutescens through study on seed germination and physiological characteristics of P. frutescens seedlings.</p><p><b>METHOD</b>Several physiological indexes of P. frutescens seeds treated by different concentrations of ZnSO4 and PEG were measured. And other indexes like the activities of malondialdehyde (MDA), superoxide (SOD), peroxidase (POD) and catalase (CAT) were also determined.</p><p><b>RESULT</b>The germination indexes of P. frutescens aging seeds treated by different concentrations of ZnSO4 and PEG were all increased. And the seeds that treated by ZnSO4 (600 mg x L(-1)) and PEG (20%) showed the most significantly increase in every index. The germination vigor were 64.7% and 66.8%, the germination rate were 78.7% and 79.4%, the germination index were 11.8 and 12.2, the vigor index were 0.091 1 and 0.0939 respectively. The content of MDA was decreased under different treatment. The activities of three enzymes include SOD, POD and CAT were increased by different treatment of ZnSO4 (0.28, 4.71, 3.82 U x mg(-1) respectively) and PEG (0.29, 4.93, 4.18 U x mg(-1) respectively).</p><p><b>CONCLUSION</b>ZnSO4 with concentration of 600 mg x L(-1) and PEG with concentration of 20% could significantly alleviate the damages to the seeds and seedlings of P. frutescens by aging and promote the aging resistance of the seeds and seedlings.</p>

RAPD analysis for genetic diversity of medicinal plant Coptis omeiensis / 中国中药杂志

<p><b>OBJECTIVE</b>To discuss the genetic diversity of Coptis omeiensis.</p><p><b>METHOD</b>The genetic diversity of 110 individuals from 10 populations was analyzed by RAPD.</p><p><b>RESULT</b>14 primers were selected to produce highly reproducible RAPD bands. Among 132 amplified bands, 98 showed polymorphism, the percentage of polymorphic bands reached to 74.24%. Nei's gene diversity index (H) was 0.2863, Shannon's information index (I) was 0.3624, G(st) was 0.2305. The genetic distance coefficient and the similarity were 0.1931-0.5245 and 0.5016-0.8843, respectively.</p><p><b>CONCLUSION</b>There exists a held high genetic diversity in C. omeiensis and the majority of genetic variation occurs in the populations. By cluster analysis, the geographical distribution is very obvious. The RAPD marker can be used for the analysis of the genetic diversity and genetic variation of C. omeiensis.</p>

ISSR analysis of genetic diversity of Coptis deltoidea / 中国中药杂志

<p><b>OBJECTIVE</b>To study the genetic diversity of Coptis deltoidea.</p><p><b>METHOD</b>The genetic diversity of 90 individuals from 8 populations was analyzed by ISSR.</p><p><b>RESULT</b>Twelve primers were selected to produce highly reproducible ISSR bands. Among 128 amplified bands, 94 showed polymorphism, the percentage of polymorphic bands reached 73.44%. Nei's gene diversity index (H) was 0.1925, Shannon's information index (I) was 0.3028, Gst was 0.7212. The genetic distance coefficient and similarity were 0.0858-0.2314 and 0.8046-0.9425, respectively.</p><p><b>CONCLUSION</b>C. deltoidea held a high genetic diversity and the majority of genetic variation occurs among populations. By cluster analysis, the geographical distribution is very obvious. The ISSR marker can be used for the analysis of genetic diversity and genetic variation of C. deltoidea.</p>

RAPD analysis for genetic diversity of Fagopyrum cymosum / 中国中药杂志

<p><b>OBJECTIVE</b>To study the genetic diversity of Fagopyrum cymosum.</p><p><b>METHOD</b>The genetic diversity of 87 individuals from 10 populations was analyzed by random amplified polymorphic DNA (RAPD).</p><p><b>RESULT</b>Twelve primers were selected to produce highly reproducible RAPD bands. Among 85 amplified bands, seventy-five showed polymorphism, the percentage of polymorphic bands reached to 88.24%. Nei's gene diversity index (H) was 0. 3103, Shannon information index (I) was 0.5632, Gst was 0.1924. The genetic similarity coefficient and the genetic distance were 0.6720-0.9678 and 0.0328-0.3975, respectively.</p><p><b>CONCLUSION</b>F. cymosum shows high genetic diversity and the majority of genetic variation occurs in populations. By cluster analysis, the geographical distribution is very obvious. The RAPD marker can be used for the analysis of the genetic diversity and genetic variation of F. cymosum.</p>

Mechanisms on The Regulation of miRNAs Expression / 生物化学与生物物理进展

MicroRNAs(miRNAs),small noncoding RNAs,are essential for posttranscriptional gene regulation and have important roles in a wide range of biological processes,including development,growth and differentiation. Thousands of miRNAs have been identified in animals,plants and microoaganisms. miRNAs regulate their target genes by mRNA degradation or translation suppression. About 30 % of genes in an organism are subject to miRNA regulation. miRNA expression and function are regulated by transcriptional factors,epigenetics,single nucleotide polymorphisms,RNA editing and so on. Additionally,the success in knocking out a specific mouse miRNA gene has provided a valuable model for studying miRNA function.

ISSR Analysis for genetic diversity of Coptis chinensis / 中草药

Objective To discuss the genetic diversity of Coptis chinensis.Methods The genetic diversity of 78 individuals from seven populations was analyzed by inter-simple sequence repeat(ISSR).Results Twelve primers were selected to produce highly reproducible ISSR bands.Among 106 amplified bands,72 showed polymorphism,the percentage of polymorphic bands reached to 67.92%.Nei's gene diversity index(H)was 0.180 3,Shannon information index(I)was 0.283 2,Gst was 0.681 5.The genetic distance coefficient and genetic similarity were 0.089 4—0.184 6 and 0.832 1—0.912 7,respectively.ConclusionC.chinensis holds high genetic diversity and the majority of genetic variation occurs among the populations.By cluster analysis,the geographical distribution is very obvious.The ISSR marker could be used for the analysis of the genetic diversity and genetic variation of C.chinensis.