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The results of ellipse fitting can be considerably distorted by outliers in the fitted point set.To tackle this problem, three improved ellipse fitting algorithms, one of which is based on least trimmed square, and the other two on dual point removal, are proposed.The least trimmed square algorithm starts from a random sample of the original complete fitted set, and then in each iteration, new fitted set is formed by points with the least residual errors, till the process converges to an ellipse fitting a subset whose members are mostly non-outliers.Dual point removal algorithms, on the other hand, starts from the whole fitted set, removes the two points respectively with the maximal positive and the minimal negative residual errors, and halts when the number of points in the remaining set does not exceeds a user-defined threshold.The two proposed algorithms and existing methods are compared on an image base of actual accessories.Experimental results show that when the number of reserved ellipse points is relatively small, the dual removal-based algorithms present the best fitting accuracies, but are slower than the least trimmed square fitting algorithm.When the best performance with parameter tuning is concerned, however, the least trimmed square algorithm achieves a shape-location matching accuracy of 0.62 pixels and an orientation matching accuracy of 0.6°, at an average execution time of 6.5ms, outperforming other algorithms.Other advantages of the proposed algorithms include the small number of algorithm parameters, the intuitiveness of the parameters, and the insensitivity of the algorithm performance to the parameters.These experimental results provide solid evidences for the effectiveness of the proposed algorithms, especially the least trimmed square algorithm.

[Objectives] To estimate the risk of Monochamus saltuarius Gebler spreading in China. [Methods] Three species distribution models within the BIOMOD2 modeling platform were used to predict the future distribution of M. saltuarius. In addition, we analyzed five potential consequences of the spread of this pest: its future distribution, its potential economic impact, the economic importance of its host plants, the possibility of transmission and the difficulty of control and management. [Results] The synthetic risk index, R, is 2.05 indicating that M. saltuarius has the potential to become widely distributed. [Conclusion] M. saltuarius has the potential to become a widespread, economically significant, pest in China. Quarantine management and removal measures need to be formulated to reduce the risk posed by this pest. [ABSTRACT FROM AUTHOR]

[Objectives] The study aims to clarify the potential distribution of Dendrolimus tabulaeformis in China to reduce the economic and ecological losses caused by the spread of the pest. [Mehtods] In this study, we used CLIMEX software, combined with the known geographical distribution and biological data of Dendrolimus tabulaeformi and Pinus tabulaeformis, to predict the global potential distribution of the pest and its host plant under historical climatic condition (1987-2016) and future climatic condition (2071-2100). Besides, we analyzed the impact of climate change on the climatic suitability of the pest and its host plant. We also identified the impacts of host plant on the climatic suitability of the pest. [Results] The results show that the pest had a suitable distribution area in all continents (except Antarctica) under historical and future climatic conditions. The marginal region was widely distributed, while the very favorable region was only distributed in some areas, such as the United States and China. Furthermore, the potential habitat of the pest tended to move northward, and the total suitable area increased in the future. The host plant only had slight impacts on the potential distribution of the pest. [Conclution] We clarified the impact of climate change on the potential distribution of D. tabulaeformi and provided a theoretical basis for the future monitoring and control of the pest. [ABSTRACT FROM AUTHOR]

[Objective]The breeding system and promotion model of improved crop variety in Burundi were studied to provide references to enhance the cooperation between China and the Republic of Burundi (Burundi for short). [Method] Aimed to agricultural improved varities of Burundi, the basic utilization situation, administrative management and promotion systems, breeding models and main achievements were introduced and analysed. As for existing problems, some targeted suggestions for development were put forward. [Result]In recent years, the amounts of main food crop seeds used in Burundi remained relatively stable. Among which, there was an increasing trend of amount for seeds used of pea bean, peanut, potato, rice and so on. Burundi had mainly established systems for seed breeding and promotion and had achieved mainly self-sufficiency of improved seeds. There were two breeding systems viz., the folk breeding system and national breeding system. While due to the backwardness of breeding technology, poor quality and high breeding cost, the outputting efficiency of improved varities was relatively lower, especially the commercial seeds, which was not consistent with utilization situation of improved seeds. While the organization for agricultural technology promotion system was relatively perfect in Burundi, the operating efficiency for promotion was relatively lower, owing to the single promoting subject, lack of promotion expense, low-quality technicians to spread agricultural technology and backwardness of promotion means and conditions. [Suggestion] Some suggestions of development were put forward to establish breeding system and promotion model dominated by government, put science ahead to play a role of scientific support in development, and open market depending on enterprise innovation. [ABSTRACT FROM AUTHOR]

For multiple linear regression model which coefficients are fuzzy numbers, this paper uses least- squares approach based on fuzzy structured element theory to study analytical expression of the model. First, a new distance formula of fuzzy numbers is defined using structured element method, and also proved equivalent with one given by the paper [1] under certain conditions. But it can avoid the inconvenience of the latter caused by interval arithmetic. Fuzzy parameters are expressed by the structured element and analytical expression of fuzzy linear regression model is provided. Then we give a parameter estimation method for the model whose fuzzy parameters are LR-type fuzzy numbers. At last, an application example is presented in order to illustrate the simplification of the proposed method. [ABSTRACT FROM AUTHOR]

The main goal of this paper is to solve fuzzy linear differential systems with fuzzy initial valued problem by fuzzy structured element method. Firstly, we introduce the basic principle of fuzzy structured element, define the fuzzy linear differential systems which are linear formed by fuzzy structured element method. Secondly, we investigate the fuzzy linear differential systems by fuzzy-valued function and the derivative of fuzzy-valued function which are linear formed by fuzzy structured element method, and give the necessary and sufficient condition of a solution of the fuzzy linear differential systems. Finally, we put forward an example to illustrate effectivity. [ABSTRACT FROM AUTHOR]

Extreme of fuzzy-valued function was not thoroughly researched until we resolved the analytical expression of fuzzy-valued functions, in this paper, based on that the fuzzy-valued function could be expressed by structured element, we present adjoint function of fuzzy-valued function by defining a type of structured sequence of fuzzy numbers, and also prove that the research of extreme of fuzzy-valued function can be transformed into general extreme of its adjoint function under structured sequence. Meanwhile, we not only present the generalized extreme of fuzzy-valued function, but also give the methods for solving the generalized extreme value and generalized extreme points which are formulated by structured element, the research results enrich the research content of fuzzy optimization theory and methods, as well as provide a reasonable and useful analysis tool for studying optimization problem of fuzzy operations research. [ABSTRACT FROM AUTHOR]

In this paper, a class of new fuzzy linear programming problem with intuitionistie fuzzy elastic constraints is constructed and discussed based on the fuzzy structured element method. By introducing the basic principle of the weighted characteristic number (WCN), the authors define an order relation and expand the Verdegay's method of the fuzzy linear programming. Then, the authors transform the new fuzzy linear programming problem into two equivalent and clear linear programming models with parameter constrains. The compared optimal feasible solution of the two equivalent and clear linear programming models is given. Finally, the authors give a numerical example to illustrate the general method for solving the proposed fuzzy linear programming problem. [ABSTRACT FROM AUTHOR]

The textbooks of psychological history published in China three decades ago abandoned humanism and adopted scientism, leading Chinese psychological studies to an unprecedented paradox: on the one hand, we were offered ancient Chinese psychological thoughts based on the ethical culture, which is totally different from the western culture; on the other hand, the texts adhered to the scientific research model imported from the western world. In order to address this problem, Professor Pan Shu advanced "the consistency principle". Moreover, the western psychology, which emphasized cognition research and paid little attention to emotion research, is now beginning to turn to the spiritual nature, which takes emotion as its core. This spiritual turn in the western psychology provides an important chance to solve the problem about how to combine Chinese psychology and western psychology. [ABSTRACT FROM AUTHOR]

In this paper, we define the multiplication for fuzzy number and fuzzy-valued function and investigates the full fuzzy linear differential systems which are linear formed by symmetrical fuzzy structured element, we also give the necessary and sufficient condition of a solution of the full fuzzy linear differential system and obtain the structured element method of solving homogeneous and non-homogeneous full fuzzy linear differential systems. At the same time, some examples in economic and military are considered. [ABSTRACT FROM AUTHOR]

Increasing concentrations of greenhouse gases in the atmosphere caused by human activities are the main cause of climate warming. Global warming is a severe challenge confronted by human society today. Reducing greenhouse gas emissions and increasing carbon sinks are the keys to addressing climate warming. Biochar addition is considered to be a promising way to reduce greenhouse gas emissions and increase carbon sinks, due to its unique physical, chemical, and biological properties. Therefore, it is of great significance to study the effects of biochar on soil greenhouse gas emissions to mitigate the greenhouse effect and achieve "carbon neutrality." The long-term and short-term effects of biochar on soil greenhouse gas emissions and their influencing mechanism were reviewed. It was found that the effects of biochar on soil greenhouse gas emissions varied with the types of biochar feedstock, pyrolysis temperature, application ratio, and soil and vegetable types. In addition, due to the different aging times and modes and cultivation methods, the mitigation effect of aged biochar on soil greenhouse gas could be enhanced or weakened or even disappeared. Further, based on the deficiencies of the previous research, the direction and focus of future research on the effects of biochar on soil greenhouse gas emissions were analyzed and prospected. It was proposed to strengthen simultaneous research on the effects of biochar on CO 2 , N 2 O, and CH 4 emissions; reducing greenhouse gas emissions and carbon sequestration; different aging modes and cultivation methods of biochar; and revealing the influencing mechanism at the process level, through exploring the effects of biochar on soil carbon and nitrogen dynamics and tracing the source of greenhouse gases using 13 C and 15 N tracer technology.

In this study, the authors cloned a glycosyltransferase gene PpUGT2 from Paris polyphylla var. yunnanensis with the ORF length of 1 773 bp and encoding 590 amino acids. The phylogenetic tree revealed that PpUGT2 belonged to the UGT80A subfamily and was named as UGT80A49 by the UDP-glycosyltransferase(UGT) Nomenclature Committee. The expression vector pET28a-PpUGT2 was constructed, and enzyme catalytic reaction in vitro was conducted via inducing protein expression and extraction. With UDP-glucose as sugar donor and diosgenin and pennogenin as substrates, the protein was found with the ability to catalyze the C-3 hydroxyl β-glycosylation of diosgenin and pennogenin. To further explore its catalytic characteristic, 15 substrates including steroids and triterpenes were selected and PpUGT2 showed its activity towards the C-17 position of sterol testosterone with UDP-glucose as sugar donor. Homology modelling and molecule docking of PpUGT2 with substrates predicted the key residues interacting with ligands. The re-levant residues of PpUGT2-ligand binding model were scanned to calculate the corresponding mutants, and the optimized mutants were obtained according to the changes in binding affinity of the ligand with protein and the surrounding residues within 5.0 Å of ligands, which had reference value for design of the mutants. This study laid a foundation for further exploring the biosynthetic pathway of polyphyllin as well as the structure of sterol glycosyltransferases.

This study used the zebrafish model to explore the hepatotoxicity of Rhododendri Mollis Flos(RMF). The mortality was calculated according to the number of the survival of zebrafish larvae 4 days after fertilization under different concentration of RMF, and the dose-toxicity curve was fitted to preliminarily evaluate the toxicity of RMF. The liver phenotypes under the sublethal concentration of RMF in the treatment group and the blank control group were observed by hematoxylin-eosin(HE) staining and acridine orange(AO) staining. Meanwhile, the activities of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were determined to confirm the hepatotoxicity of RMF. Real-time quantitative polymerase chain reaction(real-time PCR) and Western blot were used to determine the expressions of genes and proteins in zebrafish larvae. Gas chromatography time-of-flight mass spectrometry(GC-TOF-MS) was used to conduct untargeted metabolomics testing to explore the mechanism. The results showed that the toxicity of RMF to zebrafish larvae was dose-dependent, with 1 100 μg·mL~(-1) of the absolute lethal concentration and 448 μg·mL~(-1) of sublethal concentration. The hepatocyte apoptosis and degeneration appeared in the zebrafish larvae under the sublethal concentration of RMF. The content of ALT and AST in zebrafish larvae at the end of the experiment was significantly increased in a dose-dependent manner. Under the sublethal concentration, the expressions of genes and proteins related to apoptosis in zebrafish larvae were significantly increased as compared with the blank control group. The results of untargeted metabolomics showed that the important metabolites related to the he-patotoxicity of RMF were mainly enriched in alanine, aspartic acid, glutamic acid, and other pathways. In conclusion, it is inferred that RMF has certain hepatotoxicity to zebrafish larvae, and its mechanism may be related to apoptosis.

The transcription factor X-box binding protein-1 (XBP1) plays a key role in unfolded protein reaction. This study was aimed to investigate the expression pattern and regulation of XBP1 in the mouse uterus during early pregnancy. The methods of immunohistochemistry (IHC) and real time quantitative RT-PCR were used to test XBP1 expression in early pregnancy, artificial decidualization, oestrous cycle and hormone-regulated mouse models. The results showed that XBP1 was spatiotemporally expressed in mouse uterus during early pregnancy. The XBP1 protein was mainly detected in the luminal and glandular epithelia on days 1-4 of pregnancy, and was strongly detected in the decidual area on days 5-8 of pregnancy. Similarly, XBP1 expression was also mainly expressed in decidual cells following artificial decidualization. During the oestrous cycle, Xbp1, Xbp1u, and Xbp1s mRNA was predominantly present in proestrus. In the ovariectomized uterus, the expression of XBP1 in luminal and glandular epithelia was up-regulated after estrogen treatment. These results suggest that XBP1 is associated with embryo implantation and decidualization during early pregnancy in mice, and the expression of XBP1 in luminal and glandular epithelia may be regulated by estrogen.

To reveal the toxic mechanism of Kansui stir-baked with vinegar(VEK), conducta comparative study on the metabolites of fecal samples of rats before and after being treated with chemical constituents group B and C(VEKB/VEKC) extracted from VEK by metabolomics approach. The fecal samples of each group were analyzed using ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UFLC-Q-TOF-MS). Then the data was processed by principal component analysis(PCA) and partial least square discriminant analysis(OPLS-DA) to screen and identify biomarkers relating to the toxicity of VEK. Besides, t-test was adopted for univariate statistical analysis, so as to study the changes of these biomarkers in drug groups before and after being treated with VEKB/VEKC and explore the effect of VEKB/VEKC on the metabolism of rat feces. Furthermore, the toxic mechanism of VEKB/VEKC was explored based on the results of the metabolic pathway analysis. The results displayed that compared with control group, the metabolism of fecal samples of VEKB and VEKC treated groups show obvious changes, and the VEKB treated group show more significant changes. A total of 16 potential biomarkers and 5 metabolic pathways relating to the toxicity of VEK were found and identified. And the toxicity of VEK might be associated with the disorder of such metabolic pathways as tryptophan metabolism, primary bile acid biosynthesis, amino sugar and nucleotide sugar metabolism, purine metabolism, and degradation of valine, leucine and isoleucine. This study provides a scientific basis for the clinical safety application of VEK.

To prepare Helix aspersa muller-paeonol nanogel( PAE-HAM-Gels) with anti-proliferative scar effect,evaluate its skin penetration,retention and irritation,and to investigate its prevention and treatment effect for hypertrophic scar in rabbit ears. The dermal retention,transdermal rate and cumulative permeability of paeonol were investigated in vitro by using the modified Franz diffusion cell and the abdominal skin of suckling pigs,SD rats and KM mice,respectively,and the in vitro permeation curves were drawn. The normal skin of the back of New Zealand rabbits was continuously treated with PAE-HAM-Gels for 7 days,and the physiological state of the skin was observed under light microscope after HE staining by using homologous left and right contrast method. The hypertrophic scar model in rabbit ears was established,and the New Zealand rabbits were randomly divided into blank group,model group,positive drug group,PAE-Gels group and PAE-HAM-Gels group. After 28 days of administration,the scar hyperplasia rate and scar elevation index( SEI) of each group were calculated; the scar tissues were taken and stained with Masson for observation of collagen fibers and muscle fibers hyperplasia under light microscope,and the expression level of TGF-β1 in each group was detected. The Qnof PAE-HAM-Gels in aqueous solution was in line with the Higuchi equation,and its transdermal rate,cumulative permeation and dermal retention in different animal skins were all higher than those of PAE-Gels. The skin of the drug-administered group was intact,without erythema,edema or other phenomena; under light microscope,the subcutaneous tissue and the epidermal cells were neatly arranged with uniform thickness,which showed no difference from the blank group. The scar hyperplasia rate of the PAE-HAM-Gels group was 62. 50%; SEI was 2. 17±0. 33 and TGF-β1 was( 815. 4±34. 69) ng·L~(-1),significantly different from those in model group( P<0. 01). Masson staining showed that as compared with the model group,the number of collagen fibers and muscle fibers was small and the arrangement was loose and tidy in the PAE-HAM-Gels group,with regular arrangement of chondrocytes and a small number of inflammatory cells and microvessels.PAE-HAM-Gels have good transdermal properties and dermal retention without skin irritation,offering a promising therapeutic strategy for transdermal delivery during the prevention and treatment of hypertrophic scar in rabbit ears.

Light is a key factor affecting seed germination and seedling growth. In this study, seed germination and seedling growth of Cunninghamia lanceolata and Schima superba were compared under controlled conditions with five light treatments (100%, 60%, 40%, 15% and 5% of full sunlight). The results showed that light intensity significantly impacted seed germination and seedling growth of both species. With decreasing light intensity, the germination rate and germination index of C. lanceolata increased, while those of S. superba showed a trend which increased first and then decreased, with the maximum at 40% light intensity. The seedling survival rate of both species was 0 under full sunlight, while significantly decreased with decreasing light intensity from 60% to 5%. Root length, basal stem diameter and height showed a consistent trend with the change of light availability in both species. Root length significantly decreased, basal stem diameter and height increased first and then decreased with decreasing light intensity, with the minimum at 5% light intensity. With decreasing light intensity, root biomass, stem biomass, leaf biomass and total biomass of C. lanceolata seedlings declined, while high biomass accumulation of S. superba seedlings were observed in 15%-60% light intensities, and lowest at 5% light intensity. Biomass accumulation in each organ of S. superba seedlings was greater than that of C. lanceolata seedlings under the same light intensity. High stem biomass and leaf biomass, low root biomass and root to shoot ratio were a phenotypic response to low light intensity in C. lanceolata and S. superba seedlings grown under poor light condition. The growth of C. lanceolata is better under relatively high light intensity than S. superba. Whereas S. superba is moderately shade-tolerant at the seedling stage, thus is more suitable for planting under closed canopy.

Tripterygium wilfordii is a medicinal plant commonly used in the treatment of rheumatoid arthritis,and with pharmacological activities in anti-tumor and obesity treatment. The known active ingredients in T. wilfordii are mainly terpenoids,but with very low content. Therefore,the analysis of the biosynthesis pathway of terpenoids in T. wilfordii has become a research hotspot to solve the problem of its resources. Terpenoid synthase( TPS) is a key enzyme that catalyzes the formation of a wide variety of terpenoid skeletons. In this study,a gene fragment with an ORF of 1 785 bp was cloned from T. wilfordii. Bioinformatics analysis was performed using NCBI's BLASTP,ProtParam and Interpro online tools and MEGA 6.0 software. The response of this gene to methyl jasmonate was also detected by real-time fluorescent quantitative PCR,and its catalytic function was verified by prokaryotic expression and in vitro enzymatic assay. Bioinformatics analysis indicated that the amino acid sequence encoded by this gene had both N-terminal domain and C-terminal domain of TPS,as well as the DDxx D conserved domain of the class I of TPS family. And Tw MTS gathered together with TPS-b subfamily in the Neighbor-Joining Tree constructed with known homologous TPSs. The results of RT-PCR showed that 50 μmol·L-1 MeJA 12 h could increase the expression of Tw MTS to 735 times in the control group at 12 h,and 1 644 times at 24 h. In addition,in vitro enzymatic reaction results showed that Tw MTS can catalyze the production of β-citronellol with GPP as substrate,indicating that Tw MTS was a monoterpene synthase. The above results provided a new element for the synthetic biology database of T. wilfordii terpenoids,and laid the foundation for future biosynthesis research.

Hypertrophic scar( HS) is a very common skin fibrosis disorder after human skin injury and wound healing. The objective of this study was to investigate the efficacy of cell penetrating peptide TAT-modified liposomes loaded with salvianolic acid B( SAB-TAT-LIP) on proliferation,migration and cell cycle of human skin fibroblasts( HSF),and preliminarily evaluate its effect on prevention and treatment of HS. HSF were cultured in vitro,and MTT assay was used to detect the inhibitory effect of SAB-TAT-LIP on cell proliferation. Cell migration was assessed by Transwell chamber method and scratch method; and cell cycle change was detected by flow cytometry. In vitro cell studies showed that blank liposome basically had no toxic effect on HSF. Different concentrations of SABTAT-LIP inhibited proliferation on HSF in varying degrees after intervention for different periods in a dose and time dependent manner;meanwhile,SAB-TAT-LIP significantly inhibited the migration and invasion of HSF. At the same time,SAB-TAT-LIP could block the cell cycle at G0/G1 phase after intervention for 48 h,P<0.01 as compared with the blank control group. Conclusively,our experimental data quantitatively demonstrate that SAB-TAT-LIP has significant inhibitory effect on cells proliferation,invasion and migration,with blocking effect on G0/G1 phase. This may offer a promising therapeutic strategy for transdermal delivery in prevention and treatment of HS.

The increasing antibiotic resistance is now threatening to take us back to a pre-antibiotic era. Bacteria have evolved diverse resistance mechanisms, on which in-depth research could help the development of new strategies to control antibiotic-resistant infections. Epigenetic alterations and protein post-translational modifications (PTMs) play important roles in multiple cellular processes such as metabolism, signal transduction, protein degradation, DNA replication regulation and stress response. Recent studies demonstrated that epigenetics and PTMs also play vital roles in bacterial antibiotic resistance. In this review, we summarize the regulatory roles of epigenetic factors including DNA methylation and regulatory RNAs as well as PTMs such as phosphorylation and succinylation in bacterial antibiotic resistance, which may provide innovative perspectives on selecting antibacterial targets and developing antibiotics.

Objective: To investigate the role of TXNDC5 in serum starvation-induced proliferation inhibition of HeLa cell., Methods: TXNDC5 was either over-expressed or knocked down by small interfering RNA (siRNA) in HeLa cells which were then cultured in conventional medium or serum starvation medium. The protein level of TXNDC5 was evaluated by Western blot analysis. The mRNA level of TXNDC5 was measured by quantitative real-time PCR. Cell growth rate was determined by cell proliferation assay kit (MTS method). Cell cycle distribution and apoptosis were detected by flow cytometry., Results: Serum starvation mildly reduced the mRNA level of TXNDC5 (P<0.05), but dramatically increased the protein level of TXNDC5 in HeLa cells. The stability of TXNDC5 mRNA remained unchanged. Cycloheximide abolished the serum starvation-induced up-regulation of TXNDC5 protein. Over-expression of TXNDC5 had no effect on cell proliferation. However, suppression of TXNDC5 attenuated the proliferation inhibition of HeLa cell induced by serum starvation (P<0.05), increased the proportion of cells in S phase (P<0.05), but had no effect on cell apoptosis., Conclusion: TXNDC5 mediates serum starvation-induced proliferation inhibition of HeLa cell.

Arundo donax linn, as a perennial energy crop, has promising application prospect. In the present study, Fourier transform Raman (FT Raman) spectroscopy was applied to determine the structural information of materials, milled wood lignin (MWL), and alkaline lignins (AL, under different treated time) from A. donax stem nondestructively. The results indicated that, extractable compounds in A. donax had negative contribution to the Raman spectra without rising new Raman peaks. FT Raman spectrum of MWL indicated that MWL from A. donax was HGS type lignins. Compared with the spectra of MWL from wood materials, the peak at 1173 cm(-1) was much higher in intensity for the MWL from A. donax stem, which may be assigned to hydroxycinnamic acid by analyzing the standard. With respect to FT Raman spectra of ALs, the relatively highest intensity of 1173 cm(-1) was found in alkaline lignin (AL2), which was treated for 40 min by alkaline. Moreover, the peak of coniferaldehyde/sinapaldehyde (1630 cm(-1)) was lowest in intensity while the band attributed to coniferyl alcohol/sinapyl alcohol (1660 cm(-1)) was almost disappeared in AL2. It could be inferred that AL2 demonstrated a highest content of phenolic acid, which may improve its potential application, such as for antioxidant activity. Furthermore, the results obtained by FT Raman spectra were verified by two dimensional heteronuclear singlequantum coherence nuclear magnetic resonance analyses. Above all, FT Raman spectroscopy provided alternative safe, rapid, accurate, and nondestructive technology for lignin structure determination.

Objective: To observe the primary prevention role of Wuling Capsule (WC) on poststroke depression (PSD) patients., Methods: Acute stroke patients were recruited and randomized into 2 groups by stratification, 55 in each group. All patients received same routine treatment of cardiovascular diseases. Patients in the experimental group additionally took WC (0.33 g each pill), 3 pills per day, three times per day; while those in the control group additionally took placebos, 3 pills per day, three times per day. Two weeks consisted of one therapeutic course. The diagnosis of PSD was performed once every other week. Those in accordance with PSD diagnosis discontinued any drug therapy. Those not in accordance with PSD diagnosis continued the drug therapy for 1-12 therapeutic course(s) (in total of 6 months). If they were still not in accordance with PSD diagnosis, then they discontinued the drug therapy. The morbidity of PSD, the average time of depression occurrence, Hamilton depression rating scale (HAMD) score, and adverse reactions were observed., Results: The 1-, 3-, and 6-month morbidity of PSD was 8%, 16%, and 34% in the experimental group, while they were 19.6%, 29.4%, and 54.9% in the control group. The occurrence rate was lower in the experimental group than in the control group. Besides, there was statistical difference in the 6-month occurrence rate between the two groups (chi2 = 4.465, P < 0.05). The average time of PSD occurrence was longer in the experimental group than in the control group (14.96 +/- 8.31 weeks vs. 9.36 +/- 6.06 weeks; t=6.762, P < 0.05). The HAMD score at the PSD occurrence was 11.96 +/- 2.14 in the experimental group, lower than that of the control group (14.57 +/- 4.24), showing statistical difference (t=5.641, P < 0.05)., Conclusion: WC was superior to the placebos in lowering the incidence of PSD, delaying the occurrence time of PSD, attenuating the depression degree of PSD, and had certain preventive effect on the incidence of PSD.

Aim: To construct, express and identify a recombinant prokaryotic expression vector pET-3872 carrying Rv3872 of Mycobacterium tuberculosis H37Rv strain, and to construct the recombinant Bacillus Calmette-Guerin (rBCG) strain expressing Rv3872 (PE35)of Mycobacterium tuberculosis H37Rv strain., Methods: The Rv3872 gene was amplified by PCR from Mycobacterium tuberculosis H37Rv strain and cloned into prokaryotic expression vector pET32a(+). The recombinant plasmid pET-3872 was sequenced and transformed into E.coli BL21(DE3), and was induced with IPTG to express a 35 kD fusion protein, which was confirmed as His-Rv3872 by Western blot. The expression product was purified and the new Zealand rabbits were immunized. Rv3872 was amplified by PCR and cloned into E.coli-Mycobacteria shuttle vector pMV361. The recombinant vector was named as pMV-3872, and then pMV-3872 was transformed the to BCG via electroporation, the recombinant BCG-3872 strain. The PE35 protein expression of BCG-3872 was induced with heat shock reaction. Then BCG-3872 culture supernatant and bacterial precipitation were collected respectively and analyzed by Western blot., Results: A recombinant fused expression vector pET-3872 was constructed and His-3872 protein was confirmed by Western blot. BCG-3872 strain was constructed and Western blot confirmed the presenle of the PE35 protein in the supernatant of BCG-3872 strain culture., Conclusion: The prokaryotic expression vector pET-3872 was constructed, and the 35 kD fusion protein His-3872 was expressed and purified successfully, which provides a tool for further functional study of the Rv3872 . The recombinant BCG strain expressing PE35 protein, BCG-3872 strain, was constructed successfully, which facilitates further study on BCG-3872 immune function.

Objective: To investigate the correlation between viral factors and liver histological changes of HBeAg-negative chronic hepatitis B patients with persistently normal serum ALT levels (PNAL)., Methods: HBV DNA level, HBV genotype, basal core promoter (BCP) and precore mutation were examined in 52 HBeAg-negative chronic hepatitis B patients with PNAL (defined as normal ALT measured on at least 3 occasions in the intervals of about two months over a period of 12 months or more prior to the biopsy)., Results: Subjects with both BCP and precore mutations had significantly higher HBV DNA levels than those without mutations [(4.9+/-1.4) vs (4.1+/-1.1) log(10)copies/ml, t = 2.308, P < 0.05]. A higher proportion of patients with histological activity index (HAI) > or = to 4 was found in patients with both mutations (32.1% vs 16.7%) than in patients without mutation, however, the proportion of patients with histological activity index (HAI) > or = to 3 in patients with mutations was not significantly different from that in patients without mutations (14.3% vs. 12.5%, x(2)=0.000, P > 0.05). In patients without precore or BCP mutations, there was a strong positive correlation between viral load and liver inflammation as well as fibrosis (precore: r=0.626, 0.592, P < 0.01; BCP: r=0.730, 0.641, P < 0.01). In patients without both mutations, HBV DNA has shown a high accuracy for predecting fibrosis (F > or = 3) (AUC = 0.905, 95% CI: 0.771-1.039, P < 0.05) with the cutoff value of 4.5 log(10) copies/ml (sensitivity = 1.000, specificity = 0.778, PPV = 42.9%, NPV = 100.0%). Results of both genotypes and mutations were successfully obtained in 40 samples with HBV DNA is > or = to 10(4) copies/ml. The higher viral load was observed in the patients with genotype B than genotype C (5.1 vs 4.3 log(10)copies/ml, t = 2.059, P < 0.05), but no difference was seen of liver pathologic changes between these two genotypes., Conclusions: Virus harboring both BCP and precore mutants has the higher replication level than wild type virus. 32.1% and 14.3% of the patients with both mutations have moderate or severe inflammation and fibrosis. There was a strong positive correlation between viral load and liver histological changes in patients without precore or BCP mutations, and viral load shows a high accuracy for predecting significant fibrosis (F > or = 3).

Objective: By summarize the characteristics of death cases caused by road traffic accident, to provide information and data for prevention of traffic accident., Methods: To retrospectively analyze 4148 death cases caused by road traffic accident in Shenzhen. The characteristics studied include the age and sex of the dead, the cause of death, time and place of the accidents and vehicle types, etc., Results: The death was mainly male and the proportion of male to female is 2.45:1; the accidents mainly occured in 6:00-8:00 and 18:00-2:00; 72% of the accidents took place on the main suburban roads. The proportions for each traffic modes of the death were: 44% of walking, 19% of bike, 15% of motorbike. Most traffic accidents were induced by truck, 83.2% of the death causations were severe head injury and 13.3% were complex injuries., Conclusion: The death cases of road-traffic accident in Shenzhen has obvious characteristic and maybe is preventible.

The interaction of two kinds of sulfated chitosans with DNA was investigated using fluorescence spectra, absorption spectra, alkaline denaturation curves, ionic strength, fluorescence quenching etc. The results showed that there were two modes of the interaction between sulfated chitosans and DNA/ ethidium bromide (EB): intercalative and electrostatic binding. Hyperchromic and bathochromic effects in the absorption spectra were observed when sulfated chitosans with a low degree of substitution (CT-L) or a high degree of substitution (CT-L) were bound to herring sperm DNA. The stability of DNA/EB was increased with the increase in alkaline denaturation pH in the presence of CT-H and CT-L. Meanwhile, a weak interaction of cationic Mg2+ and phosphate groups of DNA by electrostatic force binding was observed. The fluorescence was efficiently quenched in the presence of a micro-amounts of sulfated chitosans, which confirmed that there was a strong competitive binding interaction among EB, CT-H or CT-L and DNA. All of these demonstrated that different sulfated chitosans can be used as a type of hopeful target-pointed molecule of gene therapy.

In order to investigate the expression of heavy chain of HLA-B * 2705 in prokaryotic system and identify its activity, the extra-membrane gene fragment of HLA-B * 2705 was amplified from full-length HLA-B*2705 cDNA by PCR and cloned into pGEM-T vector. After identification by sequencing, the prokaryotic expressing vector pET32a (+)-B * 2705 was constructed. The antigenic activity of expressed protein was identified by Western blot and antibody blocking reaction. The results indicated that the fused HLA-B * 2705 protein expression with high efficiency was obtained. The expressed product was more than 50% of the total bacteria protein. The antigenic activity of expressed protein was confirmed by Western blot and antibody blocking reaction. It is concluded that HLA-B * 2705 fusion protein are obtained as basis for the further studies.

Objective: To elucidate the roles of Toll-like receptor 3 (TLR3) on dendritic cells (DCs) in HBV infection., Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from 48 healthy volunteers (HV) and 50 chronically HBV-infected patients (CH). DCs were induced and proliferated in a culture medium with rhGM-CSF and rhIL-4. We stimulated DCs with poly I:C and then TLR3, HLA-DR, and CD86, and CD1a expressions were examined by flow cytometry at 0 h, 12 h, 24 h and 48 h. The mRNA expressions of TLR3 were quantified by real-time PCR., Results: TLR3 expression on DCs before the poly I:C stimulation and afterwards on the 12 h, 24 h, and 48 h were 69.2%+/-20.4%, 76.0%+/-18.6%, 78.2%+/-19.5% and 85.5%+/-6.9% respectively in the CH group, and 70.8%+/-11.2%, 67.5%+/-20.9%, 86.3%+/-14.7%, 68.6%+/-16.9% in the HV group. The expressions of TLR3 were up-regulated significantly at 24 h and 48 h after stimulation with poly I:C in the HV group, and in the CH group they were not significantly increased at 24 h but obviously increased at 48 h. The mRNA expressions of TLR3 increased significantly at 12 h in the HV groups, and at 48 h in CH group. The rate of CD86 expressions increased after poly I:C stimulation, and the increased rates were 12.6%+/-9.8%, 23.8%+/-20.0%, 20.7%+/-14.3% in the CH group, and 31.0%+/-25.0%, 43.4%+/-24.7%, 44.6%+/-25.5% in the HV group at 12 h, 24 h and 48 h after poly I:C stimulation. There was a marked increase of the expression level of CD86 in the HV group. In contrast, the level was only slightly increased in the CH group (31.0% vs 12.6%). The differences between the two groups were significant at 24 h and 48 h. No significant differences were detected in HLA-DR and CD1a between the two groups., Conclusions: The increase of expression level of TLR3 is slower in the CH group than that in the HV group. A marked increase of the expression level of CD86 is observed in the HV group. Our results suggest that abnormal expression of TLR3 and CD86 may relate to the persistence of HBV infection.

Effects of environmental factors and denaturants on the variation rules of the specific absorption spectrum and fluorescence emission intensity of B-phycoerythrin were systematically investigated in the present paper. The results showed that B-phycoerythrin was relatively stable under certain conditions: the pH range from 4. 5 to 9. 5, temperature range from 25 to 60 degrees C, and illumination intensity at 2 800 lx in 24 h. The specific absorption spectrum and fluorescence emission intensity of B-phycoerythrin changed slightly. The conformation of B-phycoerythrin changed remarkably with 0. 5 %-3% SDS or 0. 25 %-1% beta-mercaptoethanol, the fluorescence emission intensity declined greatly, and the specific absorption peaks disappeared. The fluorescence emission intensity declined, while the specific absorption spectrum did not change, but the peak intensity reduced slightly with 10%-70% acetonitrile or 2-8 mol x L(-1) urea. The possible reasons for the conformation changes of B-phycoerythrin with various environmental factors and different denaturants were briefly discussed.

The interaction of chitosan (CTS), carboxymethyl chitosans (CMCTS), and carboxymethyl cellulose (CMCL) with DNA was investigated through ethidium bromide (EB) as a sensitive fluorescence probe, and ultraviolet absorption spectra. The constant D can directly denote the degree of interaction of polysaccharide with DNA. The results indicated that the interraction sequence of biological polysaccharides with DNA is as follows: CTS>CMCTS>CMCL. The polysaccharides were mainly bound to DNA by intercalated binding, which resulted in fluorescence quenching effect of EB-DNA.

Objective: To investigate the expression of oncoprotein c-erbB2 in primary breast cancer and to analyze its relation to its prognosis., Methods: Immunohistochemical staining for c-erbB2 was performed on paraffin-embedded specimens of primary breast cancer from 284 patients, and the relation to its prognosis was statistically analyzed., Results: Positive expression rate of c-erbB2 was 26.8% (76/284) in 284 primary breast cancer patients. Expression of c-erbB2 was positively correlated with the status of lymph node metastasis (P = 0.003). Univariate analysis indicated that c-erbB2 expression is a significant prognostic factor for the disease-free survival (DFS) (P = 0.024) and overall survival (OS) (P = 0.002), while multivariate analysis demonstrated that c-erbB2 is an independent prognostic factor for OS (P = 0.023). Moreover, tumors with c-erbB2 positive expression are more tend to metastasis to other viscera than those with c-erbB2 negative. c-erbB2 expression has different prognostic values for patients with different status of estrogen receptor (ER) and lymph node metastasis., Conclusion: c-erbB2 expression is an independent prognostic factor for total survival time in primary breast cancer patients, and its prognostic values are different according to the different ER status and lymph node metastasis.

This study was aimed to establish downstream purification procedure by which the protein of interest can be purified to higher purity rapidly and efficiently. The different combinations of various purification strategies, methods and conditions were compared, including reversed phase chromatography, metal chelating chromatography, anion exchange chromatography, blue dye affinity chromatography, filtration chromatography and so on. The results showed that in reversed phase chromatography, isolated protein of interest was denatured and precipitated immediately after chromatography because methanol or acetonitrile were adopted as the organic phase. In blue dye affinity chromatography expecting to purify the protein of interest in one step, protein of interest was difficultly differentiated from mixed protein as much proteins bound to the chromatography media by non-specific affinity. While there is a translation-enhancing sequence T7-g10 in the PRSETA-B7-2-PE40KDEL expression vector, so it adds 6 histidines to the N terminus of the protein of interest, this allows to purify the protein of interest by metal chelating chromatography. Based on this characteristic, a three-step chromatography line including metal chelating chromatography, anion exchange chromatography and filtration chromatography was finally established after repeated experiments. By this way the purity of protein of interest reached 95% and the total recovery rate was 8%. The result of Western blot indicated that the expressed and purified recombinant B7-2-PE40KDEL could specifically bind with mAb against human B7-2 and multiple antibody against PEA. The cytotoxicity of the recombinant toxin tested by MTT method showed that the B7-2-PE40KDEL could selectively kill Jurkat cell line expressing CD28 receptor well and had no killing effect on the Raji cell line unexpressing CD28 receptor. It is concluded that a high efficient and speedy three-step purification procedure for the purifying recombinant protein B7-2-PE40KDEL was established, and this procedure possess selective killing activity on CD28 positive T lymphocytes.

Objective: To study the expression of recombinant human SCF-TPO fusion protein and its biological function., Methods: Four primers were designed according to known sequences of TPO and SCF. The functional amino acid domains of TPO and SCF were amplified by RT-PCR from fetus hepatocytes, respectively. The expression plasmid pET32a/SCF-TPO was constructed by VOE gene fusion technique and expressed in E. coli BL21(DE3) plysS as inclusion body after isopropyl-beta-D-1-thiogalactopyranoside (IPTG) induction. The fusion protein was tested by SDS-PAGE and Western blot. The biological functions of SCF-TPO fusion protein in MO7e cells was investigated by MTT method after purification with metal chelating chromatography., Results: The high expression SCF-TPO fusion protein was obtained, reaching up to 30% of the total cellular protein. Western blot verified the correct fusion expression and MTT results showed the growth promoting effect of the SCF-TPO fusion protein on MO7e cells, with a higher promoting activity at 100 ng/ml., Conclusions: Expressed SCF-TPO fusion protein after renaturation has biological activity in promoting the proliferation of MO7e cells.

The objective was to identify some biochemical and physical properties for fusion protein IL6D24-PE40KDEL. Edman degradation, SDS-PAGE, peptide mass fingerprinting, Western blot and MTT were used for identification of the protein. The results showed that the sequence of N-terminus is Met-Ile-Asp-Lys-Gln-Ile, Met was added because of prokaryotic expression system; Western blot revealed that the purified protein could react with IL6 and PEA antibody. The purified protein IL6D24-PE40KDEL could kill the multiple myeloma cell lines U266 expressing high affinity IL6R, but it could not kill the cell lines CEM which not expressed IL6R; The molecular weight was 58.7 kD measuring by SDS-PAGE; peptide mass fingerprinting (PMF) confirmed that the construction of IL6D24-PE40KDEL was correct. A novel protein by Peptident database in EXPASY web site was identified. In conclusion, IL6D24-PE40KDEL is a new targeting protein with bioactivity of specific killing effect.

Aim: Design, construction, expression in E coli and protein characteristics prediction of bimolecular thrombopoietin (T-T) with more stability, efficiency, and lower toxicity., Methods: The expression vectors of TPO and T-T, pET32 a(+)/TPO and pET32 a (+)/T-T, had been constructed by molecular cloning methods. Then, they were expressed in host bacterium. Their products were identified by Western blot. The protein characteristics, such as second structure, antigenicity, hydrophilicity, flexibility and isoelectric point, were predicted by DS Gene and Protscale software., Results: The expressing vectors pET32a(+)/TPO and T-T were constituted correctly and expressed in origami (DE3), and their expression efficiency were more than 40 percent of total protein. T-T was identified correctly by Western blot. DS Gene and Protscale software predict the protein characteristics of TPO sequences in T-T molecule were no change, there was high flexibility in the linker domain. But two amino acids in T-T molecule have been mutated, and an insert fragment with 34 amino acids following the linker had antigenicity, hydrophilicity, and beta-sheet structure., Conclusion: We have constructed correctly and expressed T-T with high level in E Coli. Protein characteristics prediction of T-T accords with our design.

Objective: A novel recombinant interleukin6-Pseudomonas exotoxin fusion protein, having targeted cytotoxicity for leukemic cells highly expressing IL6R has been molecularly designed and constructed in this study., Methods: Firstly, REDLK at C-terminus of Pseudomonas exotoxin PE40 was replaced with KDEL using point mutagenesis technology. Secondly, a cDNA encoding interleukin-6 devoid of N-terminal 24 amino acids [IL6D24] was fused to 5' terminus of PE40KDEL DNA by the method of gene splicing by overlap extension, which could generate recombinant IL6D24-PE40KDEL and IL6D24-Linker-PE40KDEL fusion genes. Thirdly, recombinant fusion genes IL6D24-PE40KDEL and IL6D24-Linker-PE40KDEL were ligated into the EcoR I and Sma I cloning sites in the pBV220 plasmids respectively and then transformed into E.coli HB101 cells. The expressed recombinant exotoxin fusion proteins were purified to electrophoresis purity by Mono Q column chromatography. Its selectively killing was tested by the MTS colorimetric method using both U937 and CEM cells lines., Results: Recombinant exotoxin fusion proteins IL6D24-PE40KDEL was expressed as a form of inclusion bodies at higher level of 40% approximately of total proteins in bacterial cells. Western blot showed that the purified products could react specifically with IL6 monoclonal antibody and PEA antiserum, respectively. IL6D24-PE40KDEL was selectively cytotoxic to U937 cells expressing IL6R-positive with ID50 of 250 ng/ml, and but not CEM cells expressing IL6R-negative., Conclusions: Two novel recombinant interleukin6-Pseudomonas exotoxin fusion proteins IL6D24-PE40KDEL and IL6D24-Linker-PE40KDEL have been successfully constructed and they can selectively kill the leukemic cells expressing highly IL6R.

After search at hematopoietic stem cell banks for transplant donors, there may be several donors matched with given standards. To determine the most appropriate donor for a specific patient, the potential donors were analyzed and compared by three methods. The first is cross-reactive group (CREG) antigens, which defined as the public antigens that shared specific serological reaction patterns. The second is residue match theory, which concerned the three residues oriented upward toward the T-cell receptor. The third is comparing HLA three-dimensional structure models. The results of the three methods were not completely accorded in our case. However, some less matched donors could be excluded from the candidates and the range of selection was further reduced. It is concluded that combined application of three methods would contribute in selecting donor for hematopoietic stem cell transplantation in clinics.

To probe into the expression of alpha subunit for IL-6R at both mRNA and protein levels in human leukemic cells and to discuss its implication in targeted treatment for leukemia with recombinant IL-6-PE40 exotoxin fusion protein mediated by IL-6/IL-6R system, semi-quantitative RT-PCR, sequencing and FCM were used to analyze the gene and protein expression levels of alpha subunit for IL-6R in leukemic cells. Our results showed that not only mRNA but also protein of alpha subunit for IL-6R are highly expressed in the myelogenous leukemic cell lines, the relative expression levels of mRNA were KG-1(1.22) > (1.02) > U266(1.00) > U937(0.99) > HL-60(0.76). Among lymphoblastic leukemic cell lines, Raji expressed a certain amount of alpha subunit mRNA (0.77), whereas its alpha subunit protein was not detected. Expression of alpha subunit mRNA and protein were negative in lymphoblastic leukemic cell lines, HuT28 and CEM, and chronic myelocytic leukemic cell line K562. These results correlate with those of FCM highly. Noteworthily, normal human peripheral blood mononuclear cells expressed hardly protein of IL-6R alpha subunit. So this study provides sufficient experimental evidence that the targeted treatment by recombinant IL-6-PE40 can specifically kill leukemic cells highly expressing IL-6R without toxicity to normal hematopoietic cells.

The purpose of the research is to provide a new standard for matching of HLA three-dimensional structure, and summarize the major permissible mismatch and immunogenic mismatch antigens. The molecular modeling method was used to create HLA molecular structures by Swiss Model Server, and the comparison of the differences among the alleles was done by SPDV software with the function of iterative magic fit. The results were recorded by relative mean square deviation (RMSD, nm). The differences among alleles were scattered below 0.06 nm for HLA-A and -B molecules, and below 0.03 nm for HLA-DRB1 molecules. On the basis of the statistical analysis, when RMSD is greater than 0.04 nm for -A and -B molecules and 0.02 nm for -DRB1 molecules, the difference is meaningful and can be related with graft versus host disease. When RMSD is lower than 0.02 nm for -A and -B molecules and 0.01 nm for -DRB1 molecules, the difference is decided unmeaningful. From the data, the permissible mismatch and immunogenic mismatch alleles within HLA-A, HLA-B and HLA-DRB1 molecules were summarized. Three-dimensional structure matching is a new area in the transplantation field, much research should be done in the future.

Objective: To analyze the hereditary susceptibility for HLA-DQA1 alleles according to trait of multiple genes leading to disease in atherothrombotic brain infarction (ABI)., Methods: HLA-DQA1 alleles in ABI patients (n=81) and healthy controls (n=99) were detected by PCR-SSP techniques., Results: (1)The frequency of HLA-DQA1*0301 was significantly higher in ABI patients. (2)The frequency of HLA-DQA1*0301 was significantly higher in patients with family history of essential hypertension (EH). (3)The frequency of HLA-DQA1*0103 was significantly lower in ABI patients., Conclusion: HLA-DQA1*0301 allele might be a correlative gene with hereditary susceptibility of ABI and HLA-DQA1*0301 allele might be a correlative gene in ABI patients with the family history of essential hypertension, whereas HLA-DQA1*0103 allele might be a protective gene in the patients with ABI.