Three types of enzymes complete the furanocoumarins core skeleton biosynthesis in Angelica sinensis.

Furanocoumarins (FCs) are widely distributed secondary metabolites found in higher plants, including Apiaceae, Rutaceae, Moraceae, and Fabaceae. They play a crucial role in the physiological functions of plants and are well-known for their diverse pharmacological activities. As a representative plant of the Apiaceae family, Angelica sinensis is highly valued for its medicinal properties and FCs are one of the main ingredients of A. sinensis. However, the biosynthetic mechanism of FCs in A. sinensis remains poorly understood. In this study, we successfully cloned and verified three types of enzymes using genome analysis and in vitro functional verification, which complete the biosynthesis of the FCs core skeleton in A. sinensis. It includes a p-coumaroyl CoA 2'-hydroxylase (AsC2'H) responsible for umbelliferone formation, two UbiA prenyltransferases (AsPT1 and AsPT2) that convert umbelliferone to demethylsuberosin (DMS) and osthenol, respectively, and two CYP736 subfamily cyclases (AsDC and AsOD) that catalyze the formation of FCs core skeleton. Interestingly, AsOD was demonstrated to be a bifunctional cyclase and could catalyze both DMS and osthenol, but had a higher affinity to osthenol. The characterization of these enzymes elucidates the molecular mechanism of FCs biosynthesis, providing new insights and technologies for understanding the diverse origins of FCs biosynthesis.

Identification of transcription factor genes responsive to MeJA and characterization of a LaMYC2 transcription factor positively regulates lycorine biosynthesis in Lycoris aurea.

Jasmonates (JAs) are among the main phytohormones, regulating plant growth and development, stress responses, and secondary metabolism. As the major regulator of the JA signaling pathway, MYC2 also plays an important role in plant secondary metabolite synthesis and accumulation. In this study, we performed a comparative transcriptome analysis of Lycoris aurea seedlings subjected to methyl jasmonate (MeJA) at different treatment times. A total of 31,193 differentially expressed genes (DEGs) were identified by RNA sequencing. Among them, 732 differentially expressed transcription factors (TFs) comprising 51 TF families were characterized. The most abundant TF family was WRKY proteins (80), followed by AP2/ERF-EFR (67), MYB (59), bHLH (52), and NAC protein (49) families. Subsequently, by calculating the Pearson's correlation coefficient (PCC) between the expression level of TF DEGs and the lycorine contents, 41 potential TF genes (|PCC| >0.8) involved in lycorine accumulation were identified, including 36 positive regulators and 5 negative regulators. Moreover, a MeJA-inducible MYC2 gene (namely LaMYC2) was cloned on the basis of transcriptome sequencing. Bioinformatic analyses revealed that LaMYC2 proteins contain the bHLH-MYC_N domain and bHLH-AtAIB_like motif. LaMYC2 protein is localized in the cell nucleus, and can partly rescue the MYC2 mutant in Arabidopsis thaliana. LaMYC2 protein could interact with most LaJAZs (especially LaJAZ3 and LaJAZ4) identified previously. Transient overexpression of LaMYC2 increased lycorine contents in L. aurea petals, which might be associated with the activation of the transcript levels of tyrosine decarboxylase (TYDC) and phenylalanine ammonia lyase (PAL) genes. By isolating the 887-bp-length promoter fragment upstream of the start codon (ATG) of LaTYDC, we found several different types of E-box motifs (CANNTG) in the promoter of LaTYDC. Further study demonstrated that LaMYC2 was indeed able to bind the E-box (CACATG) present in the LaTYDC promoter, verifying that the pathway genes involved in lycorine biosynthesis could be regulated by LaMYC2, and that LaMYC2 has positive roles in the regulation of lycorine biosynthesis. These findings demonstrate that LaMYC2 is a positive regulator of lycorine biosynthesis and may facilitate further functional research of the LaMYC2 gene, especially its potential regulatory roles in Amaryllidaceae alkaloid accumulation in L. aurea.

Simulation of seasonal transport of microplastics and influencing factors in the China Seas based on the ROMS model.

Elucidating the mechanisms governing microplastic transport and spatial distribution in offshore waters is essential to microplastic control. However, current research on microplastic transport in the China Seas is largely restricted to small-scale investigations, which do not provide a comprehensive result. Therefore, in this study, we used the Regional Ocean Modeling System (ROMS) combined with the Lagrangian Transport (LTRANS v.2) model to investigate how microplastics are transported around the China Seas during different seasons and under climatological river discharge. Our findings showed that the microplastic pathways and spatial distributions exhibit marked seasonal variations controlled by circulation patterns in the China Seas, river discharge values, and the characteristics of the microplastic materials. Floating microplastics exhibited the longest transport distance in summer, when microplastics from the Pearl River could be transported up to 1375.8 km through the Tokara and Tsushima straits. The heavy pollution areas in summer were located in the South Yellow Sea and East China Sea, mainly resulting from the contribution of the Yangtze River (>66%). In autumn and winter, more than three-quarters of the microplastics beached off the south-central Chinese coast. In addition, simulating the vertical velocity of the water prolonged the time required for microplastics to reach the open ocean, thereby reducing the amount of microplastics entering the Pacific Ocean by 6% compared to the simulation without the vertical velocity of the water in summer. Microplastics with higher densities were generally transported shorter distances. The transmission distances of PET and PS were two orders of magnitude smaller than that of PE. This study enhances knowledge of the sources and fates of offshore microplastics and provides scientific support for offshore microplastic control.

Silencing Rac1 and Prex1 Inhibit Epithelial-Mesenchymal Transition in Human Gastric Cancer Cells Induced by Transforming Growth Factor-ß1.

BACKGROUND/AIMS: Transforming growth factor-beta can influence tumor cells, causing epithelial-mesenchymal transition and enhancing their invasion and metastasis ability. Rac1 protein could be used as an independent tumor diagnostic marker and survival predictor. Prex1 is closely related to cell metastasis. In this study, the impact of silencing Rac1 and Prex1 on transforming growth factor-beta 1-induced epithelial-mesenchymal transition and apoptosis of human gastric cancer cells MGC-803 and MKN45 was investigated. MATERIALS AND METHODS: MGC-803 and MKN45 cells received recombinant transforming growth factor-beta 1 (rTGF-ß1) treatments at various concentrations. Cell Counting Kit-8 kit was used to determine cell viability. Rac1 and Prex1 interference vectors were transfected into the rTGF-ß1-treated MGC-803 and MKN45 cells. Cell apoptosis and migration were detected by flow cytometry and scratch test, respectively. Western blot was used to detect the epithelial-mesenchymal transition-related markers E-cadherin, N-cadherin, vimentin, and PDLIM2 expression levels. RESULTS: The rTGF-ß1 (10 ng/mL) could promote MGC-803 and MKN45 cell viability. Silencing Rac1 and Prex1 could increase E-cadherin and PDLIM2 expression, decrease N-cadherin and vimentin expression, inhibit cell viability and migration, and promote apoptosis in rTGF-ß1-treated MGC-803 and MKN45 cells. CONCLUSIONS: Silencing Rac1 and Prex1 could inhibit epithelial-mesenchymal transition, reduce cell viability and migration, and promote apoptosis in human gastric cancer cells.

Retina-inspired narrowband perovskite sensor array for panchromatic imaging.

The retina is the essential part of the human visual system that receives light, converts it to neural signal, and transmits to brain for visual recognition. The red, green, and blue (R/G/B) cone retina cells are natural narrowband photodetectors (PDs) sensitive to R/G/B lights. Connecting with these cone cells, a multilayer neuro-network in the retina provides neuromorphic preprocessing before transmitting to brain. Inspired by this sophistication, we develop the narrowband (NB) imaging sensor combining R/G/B perovskite NB sensor array (mimicking the R/G/B photoreceptors) with a neuromorphic algorithm (mimicking the intermediate neural network) for high-fidelity panchromatic imaging. Compared to commercial sensors, we use perovskite "intrinsic" NB PD to exempt the complex optical filter array. In addition, we use an asymmetric device configuration to collect photocurrent without external bias, enabling a power-free photodetection feature. These results display a promising design for efficient and intelligent panchromatic imaging.

Analysis of Clinical and Pathological Features of Malignant Pulmonary Nodules.

Context: Early detection of pulmonary nodules in lung cancer and timely intervention can improve the number of diagnoses at early stages of lung cancer and can reduce mortality. At present, it's not possible to accurately determine the degree of pathological invasion of ground-glass nodules and the probability of regional lymph node metastasis using an imaging examination before surgery. Objective: The study intended to analyze the clinical, imaging, and pathological characteristics of malignant pulmonary nodules and to explore the high-risk factors for lymph node metastasis, using logistic regression multivariate analysis. Design: The research team retrospectively analyzed lung-cancer patients' demographic and clinical data. Setting: The study took place in the Department of Thoracic Surgery at Zhangzhou Municipal Hospital, affiliated with Fujian Medical University, in Zhangzhou, China. Participants: Participants were 1168 patients with malignant pulmonary nodules at the hospital between January 2018 and December 2020. Outcome Measures: The research team: (1) collected participant's pulmonary nodules after surgical resection, which the hospital had confirmed were primary lung cancer and (2) analyzed the clinical characteristics of the malignant pulmonary nodules using the World Health Organization's (WHO's) 2021 classification standard for lung-cancer tissue. The research team also collected participants' data, including gender, age, smoking status, nodular size, imaging characteristics, pathological type, degree of invasion, and lymph node metastasis, and analyzed the clinical characteristics of the malignant pulmonary nodules and explored the risk factors for lymph node metastasis. Results: Participants' average age was 56.79 ± 11.53 years, and the study included 675 females (57.79%) and 493 males (42.21%), 932 of whom didn't smoke (79.8%). Imaging indicated that most participants had nodules in the upper lobes of the lungs, 424 participants in the right lung (36.30%) and 303 in the left (25.94%). Imaging also showed that 400 participants had pure ground-glass nodules (34.25%) and 371 had solid nodules(31.76%), 355 had partial solid nodules (30.39%), the other 42 had cavitary nodules (3.60%) , and 1098 participants had adenocarcinoma (94.00%). Regarding the incidence of lymph node metastasis, 67 participants had N1 metastasis (5.74%) and 34 had N2 metastasis (2.91%). The multivariate logistic regression analysis showed that an increase in the nodular size (P < .001); the presence of lower-lobe pulmonary nodules, the nodular site (P = .025); and the amount of solid components in the nodule, the nodule's features (P < .001), were significant adverse factors for N1 lymph node metastasis, while gender, age, and smoking status didn't affect that outcome. Conclusions: Adenocarcinoma was the most common pathological type, and the probability of lymph node metastasis was low. N1 lymph node metastasis was associated with increased nodular size and solid components and the presence of lower lobe nodules.

Colorimetric and Fluorescence Dual-Mode Biosensors Based on Peroxidase-Like Activity of the Co3O4 Nanosheets.

The mimic enzyme has become a research hotspot in recent years because of its advantages of high stability, convenient preparation, and low price. In this article, Co3O4 nanosheets synthesized by a simple hydrothermal method possess the characteristics of a peroxidase-like activity. The results demonstrated that 3,3',5,5'-Tetramethylbenzidine (TMB) could be oxidized by H2O2 to produce a typical blue product (oxTMB) which has a strong absorption at 650 nm wavelength with the help of the Co3O4 nanosheets. Thus, a simple and sensitive colorimetric detection method for H2O2 was established with a good linear relationship (2-200 µM) and a low limit of detection (0.4 µM). Meanwhile, the colorimetric product can effectively quench the fluorescence emitted by Ru(bpy)3 2+. Therefore, a colorimetric and fluorescence dual detection mode photochemical sensor for H2O2 detection is constructed based on the principle of the inner filter effect (IFE) between the colorimetric product (oxTMB) and Ru(bpy)3 2+. It can effectively avoid the false positive problem of a single detection mode. In the presence of glucose oxidase, glucose can be catalyzed to produce gluconic acid and H2O2; therefore, the sensor can also be used for the determination of glucose with a good linear relationship (0.02-2 µM) and a low limit of detection (5 nM). Experimental results showed that the sensor has a high sensitivity and strong anti-interference ability which can be used for the detection of actual samples.

Efficacy and Safety of Temozolomide Combined with Radiotherapy in the Treatment of Malignant Glioma.

MG is a clinical common intracranial tumor, with the characteristics of strong invasion. In our study, we aim to explore the efficacy and safety of temozolomide combined with radiotherapy in the treatment of malignant glioma (MG) and its influence on postoperative complications and survival rate of patients. 120 MG patients admitted to our hospital (January 2019-January 2020) were chosen as the research objects and were randomly divided into group A (n = 60) and group B (n = 60). All patients were treated with radiotherapy, and patients in group A were additionally treated with temozolomide. The clinical efficacy, quality of life, incidence of adverse reactions, incidence of postoperative complications, survival rates, and average survival time of the two groups were compared. The objective remission rate (ORR), disease control rate (DCR), survival rates after one year and two years of follow-up, and the number of patients with improved quality of life in group A were markedly higher compared with group B (P < 0.05). The incidence of postoperative complications in group A was remarkably lower compared with group B (P < 0.05). The average survival time of group A was dramatically longer compared with group B (P < 0.001). There was no significant difference in the incidence of adverse reactions between the two groups (P > 0.05), and no new adverse reactions occurred in the patients. Temozolomide combined with radiotherapy can effectively improve the quality of life, treatment effect, and survival rate of MG patients, with a lower incidence of postoperative complications and better tolerance. Our finding indicates that temozolomide combined with radiotherapy has a high clinical application value. In addition, it indicates that this treatment method should be promoted in practice.

Genome-Wide Identification, Expansion Mechanism and Expression Profiling Analysis of GLABROUS1 Enhancer-Binding Protein (GeBP) Gene Family in Gramineae Crops.

The GLABROUS1 enhancer-binding protein (GeBP) gene family encodes a typical transcription factor containing a noncanonical Leucine (Leu-)-zipper motif that plays an essential role in regulating plant growth and development, as well as responding to various stresses. However, limited information on the GeBP gene family is available in the case of the Gramineae crops. Here, 125 GeBP genes from nine Gramineae crops species were phylogenetically classified into four clades using bioinformatics analysis. Evolutionary analyses showed that whole genome duplication (WGD) and segmental duplication play important roles in the expansion of the GeBP gene family. The various gene structures and protein motifs revealed that the GeBP genes play diverse functions in plants. In addition, the expression profile analysis of the GeBP genes showed that 13 genes expressed in all tested organs and stages of development in rice, with especially high levels of expression in the leaf, palea, and lemma. Furthermore, the hormone- and metal-induced expression patterns showed that the expression levels of most genes were affected by various biotic stresses, implying that the GeBP genes had an important function in response to various biotic stresses. Furthermore, we confirmed that OsGeBP11 and OsGeBP12 were localized to the nucleus through transient expression in the rice protoplast, indicating that GeBPs function as transcription factors to regulate the expression of downstream genes. This study provides a comprehensive understanding of the origin and evolutionary history of the GeBP genes family in Gramineae, and will be helpful in a further functional characterization of the GeBP genes.

Engineering Burkholderia xenovorans LB400 BphA through Site-Directed Mutagenesis at Position 283.

Biphenyl dioxygenase (BPDO), which is a Rieske-type oxygenase (RO), catalyzes the initial dioxygenation of biphenyl and some polychlorinated biphenyls (PCBs). In order to enhance the degradation ability of BPDO in terms of a broader substrate range, the BphAES283M, BphAEp4-S283M, and BphAERR41-S283M variants were created from the parent enzymes BphAELB400, BphAEp4, and BphAERR41, respectively, by a substitution at one residue, Ser283Met. The results of steady-state kinetic parameters show that for biphenyl, the kcat/Km values of BphAES283M, BphAEp4-S283M, and BphAERR41-S283M were significantly increased compared to those of their parent enzymes. Meanwhile, we determined the steady-state kinetics of BphAEs toward highly chlorinated biphenyls. The results suggested that the Ser283Met substitution enhanced the catalytic activity of BphAEs toward 2,3',4,4'-tetrachlorobiphenyl (2,3',4,4'-CB), 2,2',6,6'-tetrachlorobiphenyl (2,2',6,6'-CB), and 2,3',4,4',5-pentachlorobiphenyl (2,3',4,4',5-CB). We compared the catalytic reactions of BphAELB400 and its variants toward 2,2'-dichlorobiphenyl (2,2'-CB), 2,5-dichlorobiphenyl (2,5-CB), and 2,6-dichlorobiphenyl (2,6-CB). The biochemical data indicate that the Ser283Met substitution alters the orientation of the substrate inside the catalytic site and, thereby, its site of hydroxylation, and this was confirmed by docking experiments. We also assessed the substrate ranges of BphAELB400 and its variants with degradation activity. BphAES283M and BphAEp4-S283M were clearly improved in oxidizing some of the 3-6-chlorinated biphenyls, which are generally very poorly oxidized by most dioxygenases. Collectively, the present work showed a significant effect of mutation Ser283Met on substrate specificity/regiospecificity in BPDO. These will certainly be meaningful elements for understanding the effect of the residue corresponding to position 283 in other Rieske oxygenase enzymes.IMPORTANCE The segment from positions 280 to 283 in BphAEs is located at the entrance of the catalytic pocket, and it shows variation in conformation. In previous works, results have suggested but never proved that residue Ser283 of BphAELB400 might play a role in substrate specificity. In the present paper, we found that the Ser283Met substitution significantly increased the specificity of the reaction of BphAE toward biphenyl, 2,3',4,4'-CB, 2,2',6,6'-CB, and 2,3',4,4',5-CB. Meanwhile, the Ser283Met substitution altered the regiospecificity of BphAE toward 2,2'-dichlorobiphenyl and 2,6-dichlorobiphenyl. Additionally, this substitution extended the range of PCBs metabolized by the mutated BphAE. BphAES283M and BphAEp4-S283M were clearly improved in oxidizing some of the more highly chlorinated biphenyls (3 to 6 chlorines), which are generally very poorly oxidized by most dioxygenases. We used modeled and docked enzymes to identify some of the structural features that explain the new properties of the mutant enzymes. Altogether, the results of this study provide better insights into the mechanisms by which BPDO evolves to change and/or expand its substrate range and its regiospecificity.

[DNA fingerprinting identification of bile power(bile)medicines].

The pig bile powder, bovine bile powder, snake bile, sheep bile, goose bile powder, and bear bile powder were contained by the Chinese Pharmacopoeia. The bile power medicine has a long history in traditional Chinese medicine and definite effect. However, the medicine of bile powder(bile) are similar in morphology. Besides, many medicine lack specific microscopic identification characteristics and chemical characteristics. There is a risk of adulteration, especially when the fake medicine were mixed in authentic medicine, it is difficult to detection. The key to control the quality and ensures the clinical efficacy is the good or bad, true or false of the bile power medicine. The STR typing technology is a method that according to differential typing of PCR amplified lengths to compare and identify individual organisms. Based on the principle of STR typing, the easily, rapid DNA fingerprinting method to identify the bile power and adulteration was established.The original animal or bile powder of pigs, cattle, sheep, chickens, ducks, geese, snakes, bears, fish were collected, the 12 S-L1091/12 S-H1478 and 16 S-L3428/16 S-H3667 was obtained by sifted, the DNA fingerprinting of the bile power and adulteration was obtained by STR typing. Every species has different STR fingerprints, so different species can be identified. Besides, the fingerprints have both the authentic and fake's information, the adulteration of authentic and fake can be identified. Therefore, the method to identify the bile power and adulteration was achieved through the combination of two primers. The DNA fingerprinting method established in this study can also be used for other animal medicine.

Simultaneous detection of five flavoring agents in chewing gum by ultrasound-microwave synergistic extraction coupled with gas chromatography.

So far, the identification and determination of flavor additives in food has gained extensive attention in the area of food safety. However, it remains a big challenge for simultaneous detection of diverse flavor agents. In this work, a novel gas chromatography method coupled with ultrasound-microwave synergistic extraction was developed for simultaneous detection of five flavor compounds, including butyl butyrate, menthol, methyl salicylate, eugenol and vanilline. In this strategy, ultrasound-microwave synergistic extraction was used to extract the five flavoring agents from chewing gum. The effects of extractants, solid-liquid ratio, extraction time and microwave power on extraction yield were researched by using orthogonal test. After the optimization of programme temperature and splitless injection, the five flavoring agents were well separated and simultaneously detected with wide linear ranges, low limits of detection, high accuracy and good repeatability. Therefore, this proposed method would hold great promise for assay application on the food safety.

Health monitoring through wearable technologies for older adults: Smart wearables acceptance model.

In the context of a fast aging population, ubiquitous usage of smart wearable systems can alleviate the social burden caused by the increasing need of older adults for healthcare and assistance. To facilitate and encourage the use of smart wearable systems among older adults, this study investigated the factors that contribute to the acceptance of such systems, and smart wearables acceptance model for older adults was developed using structural equation modeling. The model was validated using 146 survey samples collected from adults aged 60 years and above. The results indicated that perceived usefulness, compatibility, facilitating conditions, and self-reported health status significantly and positively affect older adults' intention to use such technologies. Useful implications and insights were provided to future researchers and practitioners to enhance older adults' acceptance of smart wearable systems.

Custom-built tools for the study of deer antler biology.

Deer antlers can be developed into multiple novel models to study growth and development of tissues for biomedical research. To facilitate this process, we have invented and further refined five custom-built tools through three decades of antler research. These are: 1. Pedicle growth detector to pinpoint the timing when pedicle growth is initiated, thus stimuli for pedicle and first antler formation can be investigated and identified. 2. Thin periosteum slice cutter to thinly slice (0.2mm or 0.7 mm thick) a whole piece of antlerogenic periosteum (AP) or pedicle periosteum (PP), which facilitates gene delivery into cells resident in these tissues, thus making transgenic antlers possible. 3. The porous periosteum multi-needle punch to effectively loosen the dense AP or PP tissue. This allows most cells of the periosteum to come into direct contact with treating solutions, thus making artificial manipulation of antler development possible. 4. The intra-dermal pocket maker to cut the thin dermal tissue (less than 2 mm in thickness) of a male deer calf horizontally into two layers to make an intra-dermal pocket. This allows loading of AP tissue intra-dermally to test the theory of "antler stem cell niche" in vivo. 5. The sterile periosteum sampling system to allow aseptic collection of the AP, PP or the antler growth centre tissue on farm, thus allowing antler generation, regeneration or rapid growth to be investigated in vitro. Overall, we believe the application of contemporary cellular and molecular biological techniques coupled with these custom-built tools would greatly promote the establishment of this unique and novel model for the benefits of biomedical research, and hence human health.

[Design and implementation of real-time control of changes to national Chinese medicine resources fill system based on GIS].

The dynamic monitoring data of traditional Chinese medicine resources is one of the important tasks of the dynamic monitoring system of Chinese medicine resources,the system has formed a periodic monitoring data reporting mechanism. Data authenticity and accuracy are the basis for the sustainable and healthy development of Chinese medicine resources dynamic monitoring,information technology is an effective means to improve the efficiency of data reporting, reporting quality. Data production based on dynamic monitoring is of great significance for grasp the trend of change and development of Chinese medicine resources. In order to achieve the real-time control of changes to the national Chinese medicine resources, we build the Chinese medicine resources dynamic monitoring system. In order to solve the problems in practice, we have upgraded the fill system by using the data of GIS. In order to achieve the multidimensional, improve safety, practicality and standardization of the data, which laid a foundation for subsequent processing of data. The system can collect the information of the cultivation of Chinese herbal medicines,production and sales of daily reporting data, provide the Chinese herbal medicine market,fast growing industry environment such as positioning center. In this paper, the design and implementation of the system are expounded.According to the business requirements, we designed 12 forms, 98 collection indicators to meet the needs of dynamic monitoring of traditional Chinese medicine resources. This paper will introduce the development content, design and implementation, main function characteristics and application effect of the national Chinese medicine resources fill System. To explain the role that GIS technology plays in the system and how to realize the cultivation of Chinese herbal medicines, production and sales of daily reporting data, provide the Chinese herbal medicine market,fast growing industry environment such as positioning center,and information collecting.

A new dipole index of the salinity anomalies of the tropical Indian Ocean.

With the increased interest in studying the sea surface salinity anomaly (SSSA) of the tropical Indian Ocean during the Indian Ocean Dipole (IOD), an index describing the dipole variability of the SSSA has been pursued recently. In this study, we first use a regional ocean model with a high spatial resolution to produce a high-quality salinity simulation during the period from 1982 to 2014, from which the SSSA dipole structure is identified for boreal autumn. On this basis, by further analysing the observed data, we define a dipole index of the SSSA between the central equatorial Indian Ocean (CEIO: 70°E-90°E, 5°S-5°N) and the region off the Sumatra-Java coast (SJC: 100°E-110°E, 13°S-3°S). Compared with previous SSSA dipole indices, this index has advantages in detecting the dipole signals and in characterizing their relationship to the sea surface temperature anomaly (SSTA) dipole variability. Finally, the mechanism of the SSSA dipole is investigated by dynamical diagnosis. It is found that anomalous zonal advection dominates the SSSA in the CEIO region, whereas the SSSA in the SJC region are mainly influenced by the anomalous surface freshwater flux. This SSSA dipole provides a positive feedback to the formation of the IOD events.

[Effects of different stocking density and diet on development of Bufo gargarizans tadpoles].

The effects of four different stocking densities and five different diets on the growth of Bufo gargarizans tadpoles were studied to determine the optimum stocking density and diet. For stocking density experiment, the tadpoles were fed respectively at different density of 200, 500, 1 000 and 2 000 tadpoles per square meter. For diet experiment, the tadpoles were divided into five groups fed respectively with five different diets. The body weight, snout-vent length and tail length were measured every seven days, and mortality was recorded. The results showed that: the survival rates of tadpole before metamorphosis and after metamorphosis were from 68.7% to 96.3% and from 5.7% to 36.0%, respectively; the optimum stocking density is 1 000 tadpoles per square meter for the stocking density had no effect on the survival rate of tadpole before metamorphosis, and the tadpoles had the relative large body weight and survival numbers in 1000 tadpoles per square meter; the diet Ⅱ(37.9% crude protein and 5.7% crude fat), Ⅳ (25.1% crude protein and 4.0% crude fat), and Ⅴ (egg yolk) were all the optimum diets for the diet had no effect on the survival rate of tadpole before metamorphosis and the tadpoles fed with three kinds of diet above had relatively large body weight, and one of these three diets based on their availability and cost should be adopted during breeding period.

[Research progress on breeding standard of medicinal animals and discussion on several key problems].

Medicinal animal breeding standards is regarded as the law to normalize relevant production that can guarantee the quality of traditional Chinese medicine of animal category. The article summarized the medicinal animal resources in our country and the present condition of medicinal animal breeding standards. It considered the current animal breeding standards system was in adequate, not only the quantity of breeding standards, the standard content and index were also uncomprehensive, which is not conducive to the scientific and orderly development and utilization of medicinal animal resources. The article pointed out that the development of the basic standards, environmental control, feed quality, raising management, inspection and quarantine should be included into the medicinal animal breeding standards, and the medicinal animal breeding standards content framework was introduced. Meanwhile, animal welfare, biological safety and file management should be concerned during the process of research. Hope the article has good reference value to medicinal animal breeding standards establishment and production management.

[Identification of original species of Mantidis Oötheca (Sangpiaoxiao) based on DNA barcoding].

Both market research and literature reports both found that the ootheca of mantodea was all used as medicine. However, Chinese Pharmacopoeia only records the ootheca of three mantis species. The clinical use of ootheca unrecorded in Chinese Pharmacopoeia, will pose potential risks to drug safety. It's urgent to identify the origin of Mantidis Oötheca. The current researches about original animal in Mantidis Oötheca are based on morphology and unanimous. DNA barcoding fill gaps of the traditional morphological identification, which is widely used in animal classification studies. This study first use DNA barcoding to analyze genetic distance among different Mantidis Oötheca types, align COI sequences between mantis and Mantidis Oötheca and construct the phylogeny tree. The result confirmed that Tenodera sinensis and Hierodula patellifera were the origin insects of Tuanpiaoxiao and Heipiaoxiao, respectively, and Statilia maculate and Mantis religiosa were the origin insects of Changpiaoxiao.

Cytochrome C oxidase subunit I barcodes provide an efficient tool for Jinqian Baihua She (Bungarus parvus) authentication.

OBJECTIVE: To test the feasibility of DNA barcoding for accurate identification of Jinqian Baihua She and its adulterants. MATERIALS AND METHODS: Standard cytochrome C oxidase subunit I (COI) gene fragments were sequenced for DNA barcoding of 39 samples from 9 snake species, including Bungarus multicinctus, the officially recognized origin animal by Chinese Pharmacopoeia, and other 8 adulterate species. The aligned sequences, 658 base pairs in length, were analyzed for divergence using the Kimura-2-parameter (K2P) distance model with MEGA5.0. RESULTS: The mean intraspecific K2P distance was 0.0103 and the average interspecific genetic distance was 0.2178 in B. multicinctus, far greater than the minimal interspecific genetic distance of 0.027 recommended for species identification. A neighbor-joining (NJ) tree was constructed, in which each species formed a monophyletic clade with bootstrap supports of 100%. All the data were submitted to Barcode of Life Data system version 3.0 (BOLD, http://www.barcodinglife.org) under the project title "DNA barcoding Bungarus multicinctus and its adulterants". Ten samples of commercially available crude drugs of JBS were identified using the identification engine provided by BOLD. All the samples were clearly identified at the species level, among which five were found to be the adulterants and identified as Dinodon rufozonatum. CONCLUSION: DNA barcoding using the standard COI gene fragments provides an effective and accurate means for JBS identification and authentication.