Cloning and expression analysis of superoxide dismutase SmMSD2 gene from Salvia miltiorrhiza / 药学学报

Superoxide dismutase (SOD) is a key enzyme that scavenge superoxide anion free radical (O2·-) in vivo, and plays an important role in plant growth and development and stress. In this study, according to the genome and transcriptome data of Salvia miltiorrhizae, 9 SOD genes were identified and the expression patterns of SOD family genes were further analyzed, including 5 Cu/Zn-SOD, 2 Fe-SOD and 2 Mn-SOD. On the basis of proteomic analysis, combined with transcriptome data, one full-length cDNA of Mn-SOD gene, namely SmMSD2 was cloned from Salvia miltiorrhizae. The results of amino acid sequence alignment and phylogenetic analysis showed that SmMSD2 protein belongs to the manganese superoxide dismutase (Mn-SOD) subfamily, and SmMSD2 protein shares high sequence identity with the Mn-SOD proteins of various plants that all contain a C-terminal conserved metal-binding domain "DVWEHAYY". The prokaryotic expression vector pMAL-c2X-SmMSD2 was constructed and transformed into E. coli BL21 expressing strain, and the target recombinant protein was successfully induced and its enzymatic properties were analyzed. Spatiotemporal expression analysis showed that SmMSD2 gene was expressed in all tissues, indicating that SmMSD2 gene was constitutively expressed at a stable level. Real-time quantitative PCR indicated that drought (15% PEG6000), abscisic acid (ABA) and indole-3-acetic acid (IAA) could induce the expression of SmMSD2 gene, suggesting that SmMSD2 may be involved in the response of Salvia miltiorrhizae to abiotic stress such as drought, as well as the signaling pathways of phytohormone ABA and IAA. These results lay the foundation for further elucidating the involvement of superoxide dismutase in the stress response and accumulation of active components of Salvia miltiorrhiza.

Cloning, induction pattern and prokaryotic expression of a small heat shock protein SmHSP21.8 gene from Salvia miltiorrhiza / 药学学报

In order to reveal the molecular mechanism of the small heat shock proteins (sHSPs) involved in stress resistance and active ingredients accumulation in Salvia miltiorrhiza, a small heat shock protein gene was cloned from Salvia miltiorrhiza by reverse transcription PCR according to the transcriptome data of orange root Salvia miltiorrhiza. The gene is named SmHSP21.8 based on the molecular weight of the protein, and it contains an open reading frame of 585 bp, which encodes 194 amino acids. The results of phylogenetic analysis and amino acid sequence alignment showed that SmHSP21.8 protein belongs to the endoplasmic reticulum (ER) subfamily, and contains a conserved endoplasmic reticulum-specific DPFR-I/V-LE-H/Q-x-P motif at N-terminus. The prokaryotic expression vector pMAL-c2X-SmHSP21.8 was constructed and transformed into E. coli BL21 competent cells. The recombinant protein was successfully expressed after inducted. Temporal and spatial expression analysis showed that SmHSP21.8 gene was the highest expressed in flowers and had significant tissue specificity. The relative expression of the gene was significantly increased in seedlings after induction by 38 ℃, PEG6000, abscisic acid(ABA), and indole-3-acetic acid (IAA), indicating that SmHSP21.8 gene may be involved in abiotic stress such as high temperature and drought, as well as the response to exogenous hormones ABA and IAA. These results lay the foundation for further research on the molecular mechanism of small heat shock proteins involved in adversity stress.

Isolation of Growth-Promoting Bacteria and Effect of Compound Bacteria on Yield of Fritillaria przewalskii / 中国实验方剂学杂志

Objective:To explore the effects of anti-microbial compound （T1） from<italic> Bacillus </italic>（Phylum Firmicutes） and anti-microbial compound （T2） from <italic>Pseudomonas</italic> and <italic>Rhizobium</italic>， two growth-promoting agents， on the physiological characteristics and growth of <italic>Fritillaria przewalskii</italic>， in order to lay a foundation for the development of functional microbial agents and the promotion of ecological planting. Method:The endophytic bacteria of <italic>F. przewalskii</italic> were isolated and identified using conventional methods. The leaves of three-year-old <italic>F. przewalskii</italic> were sprayed with T1 and T2， followed by yield determination. The enzyme activities and physiological and biochemical indexes in the plant and microorganisms were measured using the corresponding assay kits， and the contents of related hormones by liquid chromatography-mass spectrometry （LC-MS）. Result:The isolated endophytic bacteria were classified into Firmicutes，Proteobacteria， and Actinomycetes. The activities of superoxide dismutase（SOD） and peroxidase（POD） and auxin content after T2 treatment were significantly higher than those after T1 treatment， while the contents of siderophore，salicylic acid， and gibberellin were lower. Compared with the blank （CK） group， T1 and T2 increased the contents of endogenous gibberellin，cytokinin， and auxin in <italic>F. przewalskii</italic> leaves，but did not significantly change jasmonic acid and abscisic acid. T1 promoted the accumulation of endogenous salicylic acid in <italic>F. przewalskii</italic> leaves， but there was no significant change after T2 treatment. Compared with CK，T1 and T2 enhanced the activities of SOD， POD， and catalase （CAT） and decreased the content of malondialdehyde. T2 promoted the accumulation of hydrogen peroxide in <italic>F. przewalskii</italic> leaves， but no significant difference was observed after T1 treatment. Compared with CK，both T1 and T2 increased chlorophyll，average iron content in rhizosphere soil， and 100-plant weight. Conclusion:T1 and T2 treatments help to increase the yield，and their specific mechanisms differ from each other. T1 exhibits better effect than T2.

Effect of MR Field Strength on the Texture Features of Cerebral T2-FLAIR Images: A Pilot Study / 中国医学科学杂志(英文版)

Objective To investigate effect of MR field strength on texture features of cerebral T2 fluid attenuated inversion recovery (T2-FLAIR) images. Methods We acquired cerebral 3D T2-FLAIR images of thirty patients who were diagnosed with ischemic white matter lesion (WML) with MR-1.5T and MR-3.0T scanners. Histogram texture features which included mean signal intensity (Mean), Skewness and Kurtosis, and gray level co-occurrence matrix (GLCM) texture features which included angular second moment (ASM), Contrast, Correlation, Inverse difference moment (IDM) and Entropy, of regions of interest located in the area of WML and normal white matter (NWM) were measured by ImageJ software. The texture parameters acquired with MR-1.5T scanning were compared with MR-3.0T scanning. Results The Mean of both WML and NWM obtained with MR-1.5T scanning was significantly lower than that acquired with MR-3.0T (0.05). ASM, Correlation and IDM of both WML and NWM acquired with MR-1.5T revealed significantly lower values than those with MR-3.0T (<0.001), while Contrast and Entropy acquired with MR-1.5T showed significantly higher values than those with MR-3.0T (<0.001). Conclusion MR field strength showed no significant effect on histogram textures, while had significant effect on GLCM texture features of cerebral T2-FLAIR images, which indicated that it should be cautious to explain the texture results acquired based on the different MR field strength.

Effects of levosimendan on kidney injury in ventricular fibrillation-induced cardiac arrest rats / 上海交通大学学报（医学版）

Objective • To investigate the effects of levosimendan on kidney injury in the rat model of cardiopulmonary resuscitation. Methods • Twentyfive healthy adult male SD rats were randomly divided into three groups, which were sham group (S group, n=5), levosimendan group (L group, n=10) and control group (C group, n=10). Cardiac arrest and cardiopulmonary resuscitation procedure were created in L group and C group by inducing ventricular fibrillation. L group was treated with levosimendan during and after resuscitation, while C group and S group were given equivalent volume of saline solution. S group was not induced into cardiac arrest and resuscitation. Serum interleukin-6 (IL-6), IL-10, tumor necrosis factor-α (TNF-α), creatinine (SCr), blood urea nitrogen (BUN) and cystatin-C (CysC) levels were compared between L group and C group at 1, 4 and 6 h after resuscitation.Three groups of rats were sacrificed, and the pathological changes of kidney tissues were observed at 6 h after resuscitation. Results • All rats were resuscitated successfully. No differences were found between the three groups about baseline data. After resuscitation, compared with S group, the levels of serum inflammatory cytokines and kidney function indicators increased dramatically (all P<0.05) in the other two groups. In resuscitation after 1, 4 and 6 h, the levels of IL-6 and TNF-α in L group were lower than those in C group, but IL-10 levels were higher in L group (P=0.000, P=0.002, P=0.036) than those in C group, and there were significant differences between the two groups (all P=0.000). In resuscitation after 1, 4 and 6 h, the levels of SCr (P=0.001, P=0.007, P=0.472), BUN (P=0.001, P=0.004, P=0.122) and CysC (P=0.493, P=0.001, P=0.175) were lower in L group than those in C group. Only 1 and 4 hours after resuscitation, the differences in the levels of SCr and BUN were significant, and only 4 hours after resuscitation, the difference in the level of CysC was significant between L group and C group. Both L and C group showed pathological characteristics of severe acute kidney injury, and the pathological injury scores of L group were alleviated compared with those of C group (all P=0.000). Conclusion • Levosimendan can improve kidney injury of cardiac arrest and cardiopulmonary resuscitation model rats.

Comparison between two rat models of cardiac arrest: asphyxiation and ventricular fibrillation / 上海交通大学学报（医学版）

Objective·To compare the differences in cardiac function and neurological function between asphyxia and ventricular fibrillation (VF)induced cardiac arrest rat model.Methods·Twenty healthy adult male SD rats were randomly divided into VF group (n=8),asphyxial group (n=8)and sham group (n=4).Cardiac arrest models were established in VF group and asphyxial group by VF and asphyxia respectively.All animals were observed for 24 h and advanced life support was offered for the first 1 h after resuscitation.During the 24 h,ejection fraction (EF) and cardiac output (CO) were measured with the help of cardiac ultrasonography at 1,3,5 and 6 h post resuscitation.Electrocardiographic changes,24 h survival analysis and neurological deficit score (NDS) were also recorded and analyzed at 6,12,18 and 24 h post resuscitation.Results·Both EF and CO decreased dramatically after resuscitation compared with sham group at the same time point (P=0.000).At 1 h post resuscitation,the CO decreased from (98.84±4.86)mL/min to (59.17±22.99) mL/min in VF group and from (99.86±10.34) mL/min to (46,02±22.32) mL/min in asphyxial group,but there was no difference between the two groups (P=0.792).At 3,5 and 6 h post resuscitation,the CO in VF group was higher than that in asphyxial group (P=0.041,P=0.007,P=0.020).At 1 h post resuscitation,the EF decreased from (82.67±6.21)％ to (70.23±13.24)％ in VF group and from (83.24±3.01)％ to (65.46±13.11)％in asphyxial group,but no difference was observed between the two groups (P=0.877).Then a recovery tendency was observed in both groups,but more obvious in VF group at 3 and 5 h post resuscitation (P=0.031,P=0.024).No difference was found between the two groups in survival rate during 24 h and the NDS after resuscitation,although the neurological function was greatly impaired.Conclusion·VF and asphyxia are most commonly used methods to induce cardiac arrest,but these models may differ in cardiac function post resuscitation.Researchers need to choose appropriate models according to their study objectives.

Exploration on new role of ancient perilous but key acupoint of renying (ST 9) / 中国针灸

The specific functions, stimulating methods and ranges of clinical application of Renying (ST 9) are explored. Because Renying (ST 9) is located in the neck which is a dangerous area, it is perilous if the acupoint is stimulated, so it is mostly forbidden in the past dynasties. In recent years, the main stimulating methods for Renying (ST 9) include acupuncture and press. The keys of acupuncture are to apply correct technique of needle insertion and control the depth and direction of needle. The keys of press are focused on strength and time of press. Renying (ST 9) is effective for vertigo, palpitation, asthma, hemiplegia, aphasia, hysteria, acute attack of pain, hyperplasia of mammary, continuous hiccup, disorder of throat and so on. As long as the acupoint location is accurate, manipulation is careful and stimulating method is correct, not only the safety could be guaranteed, but also the function can be well-played in clinical application.

Geometric active contour model with color and intensity priors for medical image segmentation / 中国医疗器械杂志

A new algorithm using the geometric active contour model with the fusion of color and intensity priors to segment medical images is presented in this paper. The prior knowledge used here are firstly defined in different color spaces and represented as thresholds searched by the genetic algorithm. Then the prior knowledge is merged into active contour model with its contour evolution by the level set technique. The experiments on clinical marrow images and mammograms have successfully demonstrated its superiority of the proposed algorithm over the existing active contour models which deal with image gradient information.