Application of the teaching model of massive open online course combined with small private online course in the teaching of Fundamentals of Surgery / 中华医学教育探索杂志

Objective:To investigate the application effect of the teaching model of massive open online course (MOOC)+small private online course (SPOC) in the undergraduate clinical teaching of Fundamentals of Surgery.Methods:A total of 30 undergraduate medical trainees in the five-year program were randomly divided into experimental group and control group, with 15 trainees in each group. With the chapter of basic surgical procedures as an example, the trainees in the control group received traditional teaching, and those in the experimental group received MOOC+SPOC teaching. Theoretical assessment and surgery simulation were performed at one week after the course to evaluate the teaching effect. SPSS 17.0 was used for the t-test and the chi-square test. Results:The experimental group had a theoretical assessment score of (92.5±6.4) points and a basic skill operation score of (91.7±2.2) points, while the control group had a theoretical assessment score of (85.3±7.3) points and a basic skill operation score of (83.6±6.4) points. The experimental group had an excellent rate of 60% (9/15) in theoretical assessment and 66.7% (10/15) in basic skill operation test, while the control group had an excellent rate of 40% (6/15) in theoretical assessment and 33.3% (5/15) in basic skill operation test. The experimental group had significantly higher scores and grades than the control group ( P<0.05). Conclusion:MOOC+SPOC teaching can significantly enhance the understanding of theoretical knowledge and the mastery of operational skills among trainees and improve teaching effect, and therefore, it has a good application prospect.

GAO Hong's clinical experience in treatment of tic disorder with acupuncture technique of cultivating the primary and regulating the mind / 中国针灸

The paper introduces the clinical experience of GAO Hong in treatment of tic disorder. GAO Hong believes that tic disorder results from the primary qi deficiency and mind disturbance. Acupuncture for cultivating the primary and regulating the mind is proposed specially for tic disorder. This acupuncture technique focuses on harmonizing and regulating governor vessel and conception vessel. In clinical practice, the conception vessel acupoints on the abdomen and the governor vessel acupoints on the head are selected particularly, e.g. Zhongwan (CV 12), Xiawan (CV 10), Qihai (CV 6) and Guanyuan (CV 4) on the abdomen; Baihui (GV 20), Shenting (GV 24), Benshen (GB 13) and Yintang (GV 24+) on the head. The needling sequence and the insertion depth are emphasized, which affect the curative effect and GV 20 is generally punctured first. Besides, considering to the type of disorder and the affected site, tic disorder is treated in view of both syndrome/pattern differentiation and symptom differentiation.

Generation and phenotypic characterization of S100A9 gene knockout mice by CRISPR/Cas9-mediated gene targeting / 生理学报

S100 calcium binding protein A9 (S100A9) is involved in a variety of biological processes such as inflammation and tumor cell migration and invasion regulation. The purpose of this study was to construct S100A9 gene-edited mice by using CRISPR/Cas9 technology, thereby providing an animal model for exploring the biological functions of this gene. According to the S100A9 gene sequence, the single-stranded small guide RNA (sgRNA) targeting exons 2 and 3 was transcribed in vitro, and a mixture of Cas9 mRNA and candidate sgRNA was injected into mouse fertilized eggs by microinjection. Early embryos were obtained and transferred to surrogate mice, and F

The effect of c( CGRRAGGSC) labeled with 188Re using different methods on targeting the triple?negative breast cancer / 中华核医学与分子影像杂志

Objective To prepare 188 Re-c ( CGRRAGGSC) by different labeling methods, and compare the stability and targeting ability for triple-negative breast cancer (TNBC). Methods 188 Re la-beled c( CGRRAGGSC) was prepared by direct method ( pre-tinning method) and indirect method (the hydrazinonicotinamide (HYNIC) conjugated c(CGRRAGGSC) peptide) respectively. Quality control and sta-bilities of radiolabeled probes were measured. Nude mice bearing TNBC cells (MDA-MB-468) were estab-lished and used for detecting the distribution in vivo. Tumor/ non-tumor (T/ NT) ratios of radiolabeled probes prepared by direct method and indirect method were compared to certify the targeting ability and biological activi-ty. Two-sample t test was used to analyze the data. Results The labeling rate of 188Re-c(CGRRAGGSC) by di-rect method was >87％, but the serum stability was poor (the degradation rate was about 40％ at 2 h), and the T/ NT ratio was 2.82±0.23 (n=3) at 2 h. On the other hand, the labeling rate of 188Re-HYNIC-c(CGRRAGGSC) by indirect method was 61％, while the serum stability was about 10％ at 2 h, and T/ NT ratio was about 6. 27±0.51 (n= 3) at 2 h, which was significantly higher (t = 2.13, P<0.05). Conclusions The direct method may have higher labeling rate than the indirect method, but the radiolabeled compound by the direct method has poor stability and tumor targeting ability. 188 Re-HYNIC-c(CGRRAGGSC) prepared by indirect method may have remarkable advantages on stability and biological activity.

Changes of chemical constituents in Rubiae Radix et Rhizoma before and after carbonized by UPLC-Q-TOF-MS method / 中国中药杂志

In order to explore the effect on chemical constituents after carbonized, the changes of chemical constituents in raw and carbonized Rubiae Radix et Rhizoma were analyzed by UPLC-Q-TOF-MS. The research also used principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) for data statistics to find out the main differences on components before and after carbonized. The accurate m/z values of Q-TOF-MS and Q-TOF-MS-MS fragments were applied to identify the structures. The results showed that 6 more discrepant constituents were existed between raw and carbonized Rubiae Radix et Rhizoma. Three constituents were selected as the main discrepant components according to the peak area (276 nm) and identified, as lucidin, xanthopurpurin and 1,3,6-trihydroxy-2-methylanthraquinone. After carbonized, contents of xanthopurpurin and 1,3,6-trihydroxy-2-methylanthraquinone were observably increasing, while lucidin was obviously decreasing. They could be used as the chemical markers for the differentiation between raw and carbonized Rubiae Radix et Rhizoma. The results of this experiment played an important role in the study of processing principle of carbonized Rubiae Radix et Rhizoma. It also provided important evidences for the interpretation of effective material based on carbonized Rubiae Radix et Rhizoma.

Protein Kinase C Mediates the Corticosterone-induced Sensitization of Dorsal Root Ganglion Neurons Innervating the Rat Stomach

BACKGROUND/AIMS: Gastric hypersensitivity contributes to abdominal pain in patients with functional dyspepsia. Recent studies showed that hormones induced by stress are correlated with visceral hypersensitivity. However, the precise mechanisms underlying gastric hypersensitivity remain largely unknown. The aim of the present study was designed to investigate the roles of corticosterone (CORT) on excitability of dorsal root ganglion (DRG) neurons innervating the stomach. METHODS: DRG neurons innervating the stomach were labeled by DiI injection into the stomach wall. Patch clamp recordings were employed to examine neural excitability and voltage-gated sodium channel currents. Electromyograph technique was used to determine the responses of neck muscles to gastric distension. RESULTS: Incubation of acutely isolated DRG neurons with CORT significantly depolarized action potential threshold and enhanced the number of action potentials induced by current stimulation of the neuron. Under voltage-clamp mode, incubation of CORT enhanced voltage-gated sodium current density of the recorded neurons. Pre-incubation of GF109203X, an inhibitor of protein kinase C, blocked the CORT-induced hyperexcitability and potentiation of sodium currents. However, pre-incubation of H-89, an inhibitor of protein kinase A, did not alter the sodium current density. More importantly, intraperitoneal injection of CORT produced gastric hypersensitivity of healthy rats, which was blocked by pre-administration of GF109203X but not H-89. CONCLUSIONS: Our data strongly suggest that CORT rapidly enhanced neuronal excitability and sodium channel functions, which is most likely mediated by protein kinase C but not protein kinase A signaling pathway in DRG neurons innervating the stomach, thus underlying the gastric hypersensitivity induced by CORT injection.

In vitro expression and analysis of the 826 human G protein-coupled receptors

G protein-coupled receptors (GPCRs) are involved in all human physiological systems where they are responsible for transducing extracellular signals into cells. GPCRs signal in response to a diverse array of stimuli including light, hormones, and lipids, where these signals affect downstream cascades to impact both health and disease states. Yet, despite their importance as therapeutic targets, detailed molecular structures of only 30 GPCRs have been determined to date. A key challenge to their structure determination is adequate protein expression. Here we report the quantification of protein expression in an insect cell expression system for all 826 human GPCRs using two different fusion constructs. Expression characteristics are analyzed in aggregate and among each of the five distinct subfamilies. These data can be used to identify trends related to GPCR expression between different fusion constructs and between different GPCR families, and to prioritize lead candidates for future structure determination feasibility.

Metabolism study of asperosaponin VI by using zebrafish / 药学学报

Model organism zebrafish was used to study metabolism of asperosaponin VI from Dipsacus asper Wall. ex Henry for the first time. Metabolic components of asperosaponin VI after exposing to zebrafish for 24 h were identified by high performance liquid chromatography--electrospray mass spectrometry (HPLC-ESI-MS), the separation was performed with a Zorbax C18 column using a binary gradient elution of 0.05% formic acetonitrile--0.05% formic acid water. The quasi-molecular ions of compounds in both negative and positive mode were observed for molecule mass information, and the potential structures were identified by attentive study on the deglycosylation metabolites and one hydroxylation metabolite of asperosaponin VI. The results were highly in consistent with metabolism of asperosaponin VI in rat. It can be concluded that zebrafish model can wonderfully imitate current metabolic model with advantages of small amount of lower cost, far less amount compound, higher efficiency and more simple, and can reflect integrated metabolism results of in vivo method. Zebrafish metabolic model may become a novel organism model for quick predication on metabolism of even mircoamount compound, which can enrich the available models greatly.

Dendritic cell subsets and function in newborns from mothers of different HBV infection status / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>The aim of this study was to explore the frequency of mDC and pDC and expression of surface markers of the neonates and to discuss the effect of different status of HBV infection of mother on biological characteristics of DC.</p><p><b>METHODS</b>Umbilicus cord blood in neonates of HBeAg positive HBV infected mother, HBeAg negative HBV infected mother, and normal mother were collected respectively; peripheral blood of healthy adults were selected as control group. Flow cytometry was employed to detect frequency of the mDC and its expression of CD86, frequency of pDC and its expression of CD80, CD83, CD86, and FlowJo software was used to compare these indicators among the groups.</p><p><b>RESULTS</b>Compared with control group, the frequency of mDC of cord blood (0.29 +/- 0.16 vs 0.81 +/- 0.17), CD86 positive rate of mDC (10.72 +/- 10.01 vs 32.13 +/- 7.46), the frequency of pDC (0.15 +/- 0.07 vs 0.30 +/- 0.07), and CD86/CD83 positive rate of pDC (31.61 +/- 12.81 vs 74.96 +/- 9.78; 42.66 +/- 20.83 vs 82.00 +/- 6.94) were lower (t = -7.86, P = 0.00; t = -5.36, P = 0.00; t = -5.43, P = 0.00; t = -8.49. P = 0.00; t = -4.90, P = 0.00).</p><p><b>CONCLUSIONS</b>The frequency of mDC and pDC in umbilical cord blood was lower than the peripheral blood of healthy adult, which was the possible mechanism of newborns easier to chronicity after the infection of hepatitis B virus. A significant correlation was found between different status of HBV infection and costimulatory molecule CD86 positive rate of mDC, but not for the frequency of mDC and pDC, and the expression of pDC molecules.</p>

The correlation study of HBV serological index in neonate's venus, cord blood and mother's venus blood / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>In this study, we discussed the consistency and correlation of HBV serological indexes between neonates' venous blood and cord blood whose mothers had chronical HBV infection, as well as the correlation of thoses indexes with the mothers'.</p><p><b>METHOD</b>Chronically HBV infected mothers who were postive of both HBsAg and HBeAg and also had a HBV DNA virus load above 10(5) copies/ ml and their infants were enrolled. The mothers' venous blood were collected before delivery. The neonates' cord blood were collected at birth after removal of contaminants and disinfected with alcohol on the cord's surface, and the venous blood were collected before hepatitis B virus immune globin(HBIG) and hepatitis B vaccine were given. The levels of HBsAg, anti-HBs, HBeAg and anti-HBeAg were tested with Abbott microparticle chemiluminescence method (Abbott Laboratories, Abbott Architac i2000). HBV DNA quantification were tested by COBAS TagMan real-time PCR Assay.</p><p><b>RESULTS</b>383 mothers and their infants were enrolled. The positive rates of HBsAg in cord blood and venous blood were 61.2% and 63.9%. The positive rates of HBeAg level in cord blood and venous blood were 83.2% and 83.5%. The positive rates of HBV DNA level in cord blood and venous blood were 56.0% and 59.4%. The state of HBsAg, HBeAg and HBV DNA in cord blood and venous blood were consistency, and significant correlation was observed in their levels with correlation coefficients of 0.766, 0.857, and 0.692, respectively (P < 0.000). Significant correlation of the HBeAg levels were observed between mothers' venous blood and neonates' venous blood, as well as neonates' cord blood with correlation coefficients of 0.362 and 0.352 (P < 0.000). However, there was no significant correlation of HBsAg levels between them (r = 0.023, P = 0.785; r = 0.04, P = 0.604).</p><p><b>CONCLUSIONS</b>The HBV serological index of neonate's cord blood could reflect the HBV serological indexes in venous blood because of the good correlation and consistency between them.</p>

Molecular cloning, expression pattern, and putative cis-acting elements of a 4-coumarate: CoA ligase gene in bamboo (Neosinocalamus affinis)

Background: 4-coumarate:CoA ligase (4CL) plays an important role at the divergence point from general phenylpropanoid metabolism to several branch pathways. Although 4CL sin higher plants have been extensively studied, little has known about the 4CL gene of bamboo. Results: In current study, a Na4CL gene putative encoding 4-coumarate:CoA ligase (4CL) and its 5’-flanking region were isolated from bamboo (Neosinocalamus affinis) by RACE-PCR and genomic DNA walker, respectively. Na4CL encodes a predicted protein of 557 amino acids, with conserved motifs of adenylate-forming enzymes. Phylogenetic analysis showed that Na4CL shared 62~85 percent identity with other known plant 4CLs, and cluster closely with some known 4CLs in monocots. Sequence analysis revealed conserved cis-acting elements (Box A and AC-II element) present in the Na4CL promoter. Additionally, a Na4CL RNAi construct was transformed into tobacco. Transgenic tobaccos displayed significant down-expression of endogenesis 4CL and reduced lignin contents. Conclusion: These results contribute to the knowledge of the presence of Na4CL gene and its possible role in phenylpropanoid metabolism.

Gene optimization, protein expression, purification and characterization of the HDV antigen for diagnosis / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To prepare HDAg with biological activities as a candidate of diagnostic reagent.</p><p><b>METHODS</b>To synthesize HDV gene fragment after codon optimization. To construct a thio-fused recombinant plasmid based on M48 expression vector. To express in E. coli induced by IPTG. To purify the protein by affinity chromatography followed by characterization in ELISA:</p><p><b>RESULTS</b>Plasmid construction was verified by enzyme digestion. SDS-PAGE indicated the molecular weight of the protein was the same as we expectation. ELISA proved its affinity with HDV antibodies.</p><p><b>CONCLUSION</b>HDAg was obtained successfully and it will pave the road to the research of HDV diagnostic reagent.</p>

The exploration of the antibacterial mechanism of fe3+ against bacteria

This study demonstrated that the bacteria could adsorb Fe3+ and reduce Fe3+ to Fe2+. Iron had significant bacteriostatic effects, which were directly proportional to the iron concentration and under the influence of pH and chelator. It presumed that the inhibition of Fe3+ acts through the formation of hydroxyl free radicals.

Screening and enzymatic study of a composite microbial system FH3

Five strains of fungi and 18 strains of bacteria were isolated from the soil and horse manure with cellulose as the sole carbon source. Among them, two fungal, F9 and F13, and two bacterial, B16 and B21, showed the highest filter paper activities, which were 7.79 U g-1, 9.84 U g-1, 7.34 U g-1 and 9.68 U g-1, respectively. Four microbial systems, designed as FH1 (F13+B21), FH2 (F13+B16+B21), FH3 (F9+B16+B21) and FH4 (F13+F9+B16+B21) were developed. The fermentation studies showed that the filter paper activity of the composite microbial system FH3 was higher than the others, which was 21.34 U g-1. The medium with bran and filter paper as the carbon source and peptone as nitrogen source was optimal and the maximum cellulase activity was reached at 30~35ºC and pH 6.0~6.5 when FH3 was incubated for 48 h. The enzymatic reaction conditions were estimated at 45~55 ºC, pH 4.5~5.5 and the thermal stability temperature was up to 60 ºC.

Chemical constituents of Geranium eristemon / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Geranium eristemon.</p><p><b>METHOD</b>Chromatography and spectral analysis were used to isolate the constituents and elucidate their structures.</p><p><b>RESULT</b>Five compounds were isolated from acetone extract of the whole grass of G. eristemon, and identified as beta-sitosterol, protocatechuic acid, myricetin, kaempferol-7-O-alpha-L-arabifuranoside and kaempferol-3-O-alpha-L-arabifuranoside.</p><p><b>CONCLUSION</b>kaempferol-7-O-alpha-L-arabifuranoside and kaempferol-3-O-alpha-L-arabifuranoside were isolated from G. genus for the first time.</p>