Red fluorescent proteins (RFPs) produced from a number of Anthozoa species have been subjected to a series of in vitro molecular evolution, resulting in various emission spectra ranging from 570 nm to 655 nm and thus providing powerful tools for cellular imaging or even body imaging. This article briefly reviewed the optical properties, structures and mutagenesis of RFPs and their applications.

Objective:To investigate the optimal extraction and dripping pills preparation of total terpene constituents in platycladi seed. Methods:The extraction processes of ultrasound, reflux, microwave and homogenization were investigated by single factor tests. The effects of extraction time, temperature, solid-liquid ratio and extraction times on the extraction of total terpene constituents in platy-cladi seed were investigated by orthogonal tests. Orthogonal tests were used to investigate the effects of the ratio of polyethylene glycol 6000 to poloxamer 188, the ratio of drug to matrix, the temperature of liquid and the dropping process on the quality of dropping pills. Results:The homogeneous extraction was used, and the optimum extraction conditions were as follows: the extraction time was 10 min, the temperature was 80℃, the solid-liquid ratio was 1 :10, and extraction time was once. The best preparation process of drop-ping pills was as follows:the ratio of polyethylene glycol 6000 to poloxamer 188 was 5 :1, the ratio of drug to matrix was 1 :3, the temperature of liquid was 80℃ and the dropping distance was 10 cm. Conclusion: The method of homogeneous extraction is simple, which can break the materials and extract the active ingredients simultaneously. The preparation process of dropping pills is reasonable and feasible, which is easy for the production and application.

In order to understand metabolic functions essential for methane assimilation, we investigate dribulose monophosphate pathway and adjacent pathways in gammaproteobacterial Methylomicrobium alcaliphilum 20Z by using combined approaches of RNA-seq, LC-MS, and 13C-labeled techniques. The absolute quantification of metabolome showed that the concentrations of intermediates, such as glucose-6-phosphate and 2-dehydro-3-deoxy-phosphogluconate, involved in Entner-Doudoroff (EDD) pathway were (150.95 +/- 28.75) micromol/L and below the limit of detection of mass spectrometry. In contrast, fructose-1, 6-bisphosphate, glyceraldehyde-3-phosphate/dihydroxyacetone and phosphoenolpyruvate in Embden-Meyerhof-Parnas (EMP) pathway had significantly higher concentrations with (1 142.02 +/- 302.88) micromol/L, (1 866.76 +/- 388.55) micromol/L and (3 067.57 +/- 898.13) micromol/L, respectively. 13C-labeling experiment further indicated that the enrichment of [3-13C1]-pyruvate involved in EMP pathway was 4-6 fold higher than [1,13C1]-pyruvate in EDD pathway in a dynamic course. Moreover, gene expression profile showed that the expression levels of genes in EMP pathway (e.g. fbaA, tpiA, gap and pykA) were 2 479.2, 2 493.9, 2 274.6 and 1 846.0, respectively, but gene expressionlevels in EDD pathway (e.g. pgi, eda and edd) were only 263.8, 341.2 and 225.4, respectively. Overall our current results demonstrated that EMP pathway was the main route for methane assimilation in M. alcaliphilum 20Z. This discovery challenged our understanding of methane assimilation pathway in gammaproteobacterial methanotrophic bacteria, and further provided an important insight for efficient methane biocatalysis in the future.
Glycolysis ; Industrial Microbiology ; Methane ; metabolism ; Methylococcaceae ; metabolism ; Pyruvic Acid ; metabolism

Objective To explore the expression level of OTUB1 and its clinical significance in breast cancer.Methods Immunohistochemistry was used to detect the expression level of OTUB1 in 78 cases of breast cancer tissues and 30 cases of normal breast tissue adjacent to carcinoma,and the relationships between OTUB1 and the clinical pathological features of breast cancer were analyzed.Results The positive expression rate of OTUB1 in breast cancer tissues [66.7％ (52/78)] was significantly higher than that in adjacent normal breast tissues [30.0％ (9/30)],with a statistically significant difference (x2 =11.851,P =0.001).OTUB1 expression level was related to the lymph node metastasis (x2 =5.029,P =0.025),postoperative TNM staging (x2 =4.478,P =0.034),expression of human epidermal growth factor receptor-2 (HER-2) (x2 =8.775,P =0.003),expression of P53 (x2 =4.708,P =0.030),expression of estrogen receptor (ER) (x2 =10.364,P =0.001) and molecular subtypes (x2 =10.934,P =0.012).However,OTUB1 expression level in breast cancer was not related to the age (x2 =2.194,P =0.139),menopausal status (x2 =1.843,P =0.175),tumor size (x2 =0.643,P =0.423),histological grade (x2 =3.580,P =0.167),expression of progestin receptor (PR) (x2 =3.371,P =0.066) and expression of Ki-67 (x2 =1.345,P =0.246).Conclusion OTUB1 expression level increases in breast cancer,which is associated with the lymph node metastasis,TNM staging,expressions of HER-2,P53,ER and molecular subtypes of breast cancer.It suggests that the expression of OTUB1 is related to the progression and metastasis of breast cancer.

OBJECTIVETo comparatively determine the genetic variation and differentiation of different breeding strains of Panax notoginseng for providing the basic information for genetic breeding.

METHODThe genetic diversity and genetic structure of the 17 breeding strains of P. notoginseng were assayed by using EST-SSR molecular marker.

RESULTA total of 136 polymorphic loci of EST-SSR were detected in the 17 breeding strains of P. notoginseng, with the PIC (polymorphism information content) being 0.78, H (the gene diversity within population) being 0.139, the I (the Shannon's information index) being 0.208. Gst (coefficient of gene differentiation) was 0.382 among the 17 strains. The cluster analysis of genetic similarity showed that the 17 strains of P. notoginseng and P. stipuleanatus were classified into 4 groups, while the 17 strains of P. notoginseng were classified into three subgroups.

CONCLUSIONThe genetic differentiation was detected among the 17 strains of P. notoginseng from the same cultivation population by bulk selecting. And it was feasible to detect the effect of bulk selection by EST-SSR markers.

Breeding ; Expressed Sequence Tags ; Genetic Variation ; Microsatellite Repeats ; Panax notoginseng ; classification ; genetics ; physiology

OBJECTIVETo investigate the effect of fuzheng quxie granule (FQG) on immune cells and cytokines in populations with respiratory viral infection.

METHODSFifty-nine patients were randomly divided into 3 groups, that is, 19 patients treated with conventional western medicine (WM) plus FQG in the treated group, 19 patients treated with conventional western medicine alone in the WM group, and 21 patients treated with FQG alone in the TCM group. The levels of T lymphocyte subsets, interleukine-2,4,6,10 (IL-2, IL-4, IL-6, IL-10), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (INF-gamma) and Th1/Th2 were determined before treatment, and at the end of 1st and 2nd week of treatment respectively.

RESULTSBefore treatment, levels of TNF-alpha, IL-2, IL-6, IL-10 and INF-gamma in all patients were significantly higher than normal range (P < 0.05). After being treated for 1 week, the levels of serum TNF-alpha, IL-6, and IL-10 were significantly decreased in all groups (P < 0.05), serum IL-2 and INF-gamma decreased to the normal level in the WM group, but in the treated and the FQG group by the end of the 2nd week, the two indexes still remained at the rather higher level (P < 0.05). The ratio of Th1 and Th2 in the treated group and the FQG group increased significantly by the end of 2nd week, reached the level higher than that in the WM group and that before treatment (P < 0.05). No significant difference in, T lymphocytes subsets (CD3+ , CD4+ , CD8+) and percentage of B and NK cells before and after treatment was found in all the 3 groups.

CONCLUSIONFQG can positively regulate the immune function of patients with respiratory tract viral infection in certain degree.

Adult ; Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Killer Cells, Natural ; immunology ; Male ; Middle Aged ; Phytotherapy ; Respiratory Tract Infections ; drug therapy ; immunology ; virology ; T-Lymphocyte Subsets ; immunology ; Tumor Necrosis Factor-alpha ; metabolism ; Virus Diseases ; drug therapy ; immunology

OBJECTIVETo evaluate the effect of Fuzheng Quxie granule (FQG) on immune cells and cytokines of populations susceptible to respiratory viral infection.

METHODSOne thousand four hundred and two subjects selected from 25 hospitals in Shanghai between May and June in 2003, were divided into the FQG group treated with FQG and the control group treated with placebo. Serum levels of interleukin 2 (IL-2), interleukin 4 (IL-4), gamma-interferon (gamma-IFN), blood lymphocyte subsets (CD3+ , CD4+, CD8+), B-cell count and natural killer cell (NK) percent ratio were measured in 130 of the FQG group and 120 of the control group before treatment, by the end of the 2nd week and two weeks after treatment.

RESULTSBy the end of the 2nd week of treatment, as compared with before treatment, the levels of IL-2, gamma-IFN, and NK percent in the FQG group increased significantly (P < 0.05), while IL-4 and CD3+, CD4+, CD8+ and B-cell count were unchanged. Besides, levels of Th1/Th2 ratio markedly increased at the end of the 2nd week and two weeks after treatment, in comparing with that before treatment and in the control group (P<0.05).

CONCLUSIONFQG could improve immune function of population susceptible to respiratory viral infection certain extent.

Adult ; CD4-CD8 Ratio ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interferon-gamma ; Interleukin-2 ; blood ; Interleukin-4 ; blood ; Killer Cells, Natural ; immunology ; Male ; Middle Aged ; Phytotherapy ; Respiratory Tract Infections ; prevention & control ; Virus Diseases ; prevention & control

Objective: To explore the effects of transient receptor potential vanilloid 1 (TRPV1) on autophagy in early hypoxic mouse cardiomyocytes and the mechanism in vitro. Methods: The hearts of 120 C57BL/6 mice aged 1-2 days, no matter male or female, were isolated, and then primary cardiomyocytes were cultured and used for the following experiments, the random number table was used for grouping. (1) The cells were divided into normoxia group and hypoxia 3, 6, and 9 h groups, with one well in each group. The cells in normoxia group were routinely cultured (the same below), the cells in hypoxia 3, 6, and 9 h groups were treated with fetal bovine serum-free and glucose-free Dulbecco′ s modified Eagle medium under low oxygen condition in a volume fraction of 1% oxygen, 5% carbon dioxide, and 94% nitrogen for 3, 6, and 9 h, respectively. The protein expressions of microtubule-associated protein 1 light chain 3 (LC3), Beclin-1, TRPV1 were determined with Western botting. (2) The cells were divided into normoxia group and hypoxia group, with two coverslips in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above. The positive expression of TRPV1 was detected by immunofluorescence assay. (3) The cells were divided into 4 groups, with one well in each group. The cells in simple hypoxia group were treated with hypoxia for 6 h as above, and the cells in hypoxia+ 0.1 μmol/L capsaicin group, hypoxia+ 1.0 μmol/L capsaicin group, and hypoxia+ 10.0 μmol/L capsaicin group were respectively treated with 0.1, 1.0, 10.0 μmol/L capsaicin for 30 min before hypoxia for 6 h. The protein expressions of LC3, Beclin-1, and TRPV1 were detected by Western blotting. (4) The cells were divided into 5 groups, with 5 wells in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ chloroquine group, hypoxia+ capsaicin group, and hypoxia+ capsaicin+ chloroquine group were treated with hypoxia for 6 h after being cultured with 50 μmol/L chloroquine, 10.0 μmol/L capsaicin, and 50 μmol/L chloroquine+ 10.0 μmol/L capsaicin for 30 min, respectively. Viability of cells was detected by cell counting kit 8 assay. (5) The cells were divided into simple hypoxia group and hypoxia+ 10.0 μmol/L capsaicin group, with one well in each group. The cells in hypoxia group were treated with hypoxia for 6 h as above, the cells in hypoxia+ 10.0 μmol/L capsaicin group were treated with 10.0 μmol/L capsaicin for 30 minutes and then with hypoxia for 6 h. The protein expressions of lysosomal associated membrane protein 1 (LAMP-1) and LAMP-2 were detected by Western blotting. Each experiment was repeated for 3 or 5 times. Data were processed with one-way analysis of variance, least significant difference t test, and Bonferroni correction. Results: (1) Compared with those of normoxia group, the protein expressions of LC3, Beclin-1, and TRPV1 were significantly increased in cardiomyocytes of hypoxia 3, 6, and 9 h groups (t_{3 h}=4.891, 5.890, 4.928; t_{6 h}=9.790, 6.750, 10.590; t_{9 h}=6.948, 6.764, 5.049, P<0.05 or P<0.01), which of hypoxia 6 h group were the highest (1.08±0.05, 1.12±0.10, 0.953±0.071, respectively). (2) The density of TRPV1 in cell membrane and inside the cardiomyocytes in hypoxia group was significantly increased with lump-like distribution, and the expression of TRPV1 was higher than that in normoxia group. (3) Compared with those of simple hypoxia group, the protein expression of Beclin-1 in cardiomyocytes of hypoxia+ 0.1 μmol/L capsaicin group was increased (t=10.488, P<0.01), while the protein expressions of LC3 and TRPV1 were increased without statistically significant differences (t=4.372, 3.026, P>0.05); the protein expressions of LC3, TRPV1, and Beclin-1 in cardiomyocytes of hypoxia+ 1.0 μmol/L capsaicin group and hypoxia+ 10.0 μmol/L capsaicin group were significantly increased (t=15.505, 5.773, 13.430; 20.915, 8.054, 16.384; P<0.05 or P<0.01), which of hypoxia+ 10.0 μmol/L capsaicin group were the highest (2.33±0.09, 1.34±0.07, 1.246±0.053, respectively). (4) Compared with 0.585±0.045 in normoxia group, the cardiomyocyte viability in hypoxia group was significantly decreased (0.471±0.037, t=4.365, P<0.05). Compared with that in hypoxia group, the cardiomyocyte viability in hypoxia+ chloroquine group was further decreased (0.350±0.023, t=6.216, P<0.01), while 0.564±0.047 in hypoxia+ capsaicin group was significantly increased (t=3.489, P<0.05). Compared with that in hypoxia+ chloroquine group, the cardiomyocyte viability in hypoxia+ capsaicin+ chloroquine group did not significantly change (0.364±0.050, t=0.545, P>0.05). (5) Compared with 0.99±0.04 and 0.54±0.04 in simple hypoxia group, the protein expressions of LAMP-1 and LAMP-2 in hypoxia+ 10.0 μmol/L capsaicin group were significantly increased (1.49±0.06, 0.81±0.05, t=12.550, 7.442, P<0.01). Conclusions TRPV1 can further promote the expression of autophagy-related proteins in hypoxic cardiomyocytes through autophagy-lysosomal pathway, enhance autophagy activity, and improve autophagic flow for alleviating early hypoxic cardiomyocyte injury.

As the demand for high-performance computing continues to grow, traditional computing models are facing unprecedented challenges. Among the many emerging computing technologies, DNA computing has attracted much attention due to its low energy consumption and parallelism. The DNA circuit, which is the basis for DNA computing, is an important technology for the regulation and processing of the molecular information. This review highlights the basic principles of DNA computing, summarizes the latest research progress, and concludes with a discussion of the challenges of DNA computing. Such integrated molecular computing systems are expected to be widely used in the fields of aerospace, information security and defense system.
DNA/genetics*

【Objective】 To investigate and analyze the polymorphism of RHD gene in RhD-negative population in Jiayuguan using molecular biological technique, so as to accurately identify RhD-negative individuals, and formulate individualized transfusion strategies. 【Methods】 The RhD negative voluntary blood donors and patients (mainly pregnant women) were recruited. After informed consent, history of blood transfusion and pregnancy of them were investigated, and samples were collected for negative D confirmation, gene sequencing as well as antibody screening and identification. 【Results】 Among the 96 samples, 73 cases were RHD gene deletion, 18 RHD^*01EL.01(17 RHD1227A homozygous type and 1 RHD1227A heterozygous type), 2 weak RHD^*15 type (845G/A), 1 partial D type, i. e. RHD-CE(7) -D heterozygous allele, and 2 RHD^*01N.16 variant. Antibody was detected out in 4 cases, among which 2 were positive for anti-D, 1 anti-D plus anti-E, and 1 anti-Di^a. 【Conclusion】 The proportion of DEL gene in RhD negative Chinese Han population in Jiayuguan is slightly lower than that in general Chinese Han population. No anti-D or RHD-HDN was observed in DEL type women due to multiple pregnancy or delivery of D positive newborns.