Temperature affects the tolerance of Liriomyza trifolii to insecticide abamectin.

The leafminer fly, Liriomyza trifolii, is an invasive pest of horticultural and vegetable crops that possesses a robust competitive ability when compared to congeneric species, especially with respect to temperature and insecticide tolerance. Abamectin, which is commonly used to control L. trifolii in the field, was selected as the target insecticide in this study. Our objective was to study the effect of abamectin and high temperature stress on L. trifolii mortality and the expression of genes encoding cytochrome P450 (CYP450s) and heat shock proteins (Hsps) by quantitative real-time reverse transcriptase PCR (qRT-PCR). When L. trifolii was exposed to abamectin followed by exposure to 40 °C (LC50 +HT40), mortality showed a significant increase, whereas exposure to 40 â„ƒ followed by abamectin (HT40+LC50) reduced mortality relative to abamectin or HT40 alone. Expression of three CYP450s in the CYP4 family was highest in the HT40+LC50 treatment, followed by the LC50+HT40 treatment. The expression levels of CYP18A1 (CYP18 family) were not significantly different among treatments, and CYP301A1 (CYP301 family) was only sensitive to temperature (HT40). The expression of five sHsps showed similar expression patterns and were highly responsive to the LC50+HT40 treatment, followed by the HT40 and HT40+LC50 treatments. Based on CYP450s and Hsps expression levels, our findings that suggest that L. trifolii exhibits adaptive cross-tolerance to high temperature and abamectin. This study provides a framework for selecting the most effective application time for abamectin with respect to controlling L. trifolii, which will ultimately reduce the overuse of pesticides.

High temperature stress induces expression of CYP450 genes and contributes to insecticide tolerance in Liriomyza trifolii.

Liriomyza trifolii is an invasive leafminer fly that inflicts damage on many horticultural and vegetable crops. In this study, the effects of elevated temperatures on L. trifolii tolerance to insecticides abamectin (AB), monosultap (MO) and a mixture of abamectin and monosultap (AM) were firstly investigated, then five CYP450 genes (LtCYPs) were cloned, and expression patterns and NADPH cytochrome C reductase (NCR) activity in L. trifolii were compared in response to high temperature stress and insecticide exposure. Results showed elevated temperatures induced expression of LtCYP450s, the expression level of LtCYP4g1, LtCYP4g15 and LtCYP301A1 after exposed to different high temperature were significantly up-regulated compared with the control (25 °C), while there was no significant difference in LtCYP4E21 and LtCYP18A1. Under the joint high temperature and insecticide stress, the expression of LtCYP4g15, LtCYP18A1 and LtCYP301A1 was significantly higher under elevated temperatures than that of only under AB exposure. For MO and AM exposure, only 40 °C could induce the expression of LtCYP4g15, LtCYP18A1 and LtCYP301A1. In general, the LtCYPs expression pattern was correlated with increased NCR activity and decreased mortality in response to insecticide exposure under elevated temperatures. These all demonstrated that insecticide tolerance in L. trifolii could be mediated by high temperature. This study improves our understanding of L. trifolii physiology and offers a theoretical context for improved control that ultimately reduces the abuse of insecticides and decreases exposure to non-target organisms.

A genetic study of the regeneration capacity of embryonic callus from the maize immature embryo culture.

We carried out a study of regeneration capacities of embryonic callus from maize immature embryo culture with 144 different inbred lines of natural groups from different countries, and found that the regeneration capacity was affected by three factors: environment, genotype and the interaction between the environment and genotype. We found that green embryonic callus rate (GCR), embryonic callus differentiating rate (CDR) and the plantlet number of embryonic callus regeneration (CPN) have significant positive correlations with each other, and they all have significant negative correlations with embryonic callus browning rate (CBR). Moreover, embryonic callus cloning index for the first subculture (CCI1) and embryonic callus cloning index for the second subculture (CCI2) have a significant positive correlation with each other, and CCI2 is positively correlated with green GCR, and is negatively correlated with CBR. Embryonic callus rooting rate (CRR) is positively correlated with GCR, CDR and CPN to some degree. Furthermore, we calculated Broad-Sense Heritability of each trait, and uncovered that the heritability index of CCI1, CCI2 and CRR was lower, and the heritability index of others was higher. In addition, by using the Ward method for two-way cluster analysis, we found eleven inbred lines with high regenerating abilities, and the rooting situation of regenerating plantlet was excellent by rooting culture, which could be used as the elite inbred lines of the maize transgenic receptor.

Fine mapping of the qLOP2 and qPSR2-1 loci associated with chilling stress tolerance of wild rice seedlings.

KEY MESSAGE: Using leaf osmotic potential and plant survival rate as chilling-tolerant trait indices, we identified two major quantitative trait loci qLOP2 and qPSR2 - 1 (39.3-kb region) and Os02g0677300 as the cold-inducible gene for these loci. Chilling stress tolerance (CST) at the seedling stage is an important trait affecting rice production in temperate climate and high-altitude areas. To identify quantitative trait loci (QTLs) associated with CST, a mapping population consisting of 151 BC(2)F(1) plants was constructed by using chilling-tolerant Dongxiang wild rice (Oryza rufipogon Griff.) as a donor parent and chilling-sensitive indica as a recurrent parent. With leaf osmotic potential (LOP) and plant survival rate (PSR) as chilling-tolerant trait indexes, two major QTLs, qLOP2 (LOD = 3.8) and qPSR2-1 (LOD = 3.3), were detected on the long arm of chromosome 2 by composite interval mapping method in QTL Cartographer software, which explained 10.1 and 12.3% of the phenotypic variation, respectively. In R/QTL analyzed result, their major effects were also confirmed. Using molecular marker RM318 and RM106, qLOP2 and qPSR2-1 have been introgressed into chilling-sensitive varieties (93-11 and Yuefeng) by marker-assisted selection procedure (MAS), which resulted in 16 BC(5)F(3) BILs that chilling tolerance have significantly enhanced compare with wild-type parents (P < 0.01). Therefore, two large segregating populations of 11,326 BC(4)F(2) and 8,642 BC(4)F(3) were developed to fine mapping of qLOP2 and qPSR2-1. Lastly, they were dissected to a 39.3-kb candidate region between marker RM221 and RS8. Expression and sequence analysis results indicated that Os02g0677300 was a cold-inducible gene for these loci. Our study provides novel alleles for improving rice CST by MAS and contributes to the understanding of its molecular mechanisms.

pH-sensitive fluorescent dyes: are they really pH-sensitive in cells?

Chemically synthesized near-infrared aza-BODIPY dyes displayed off-on fluorescence at acidic pH (pKa = 6.2-6.6) through the suppression of the photoinduced electron transfer and/or internal charge transfer process. The apparent pKas of the dyes were shifted well above physiological pH in a hydrophobic microenvironment, which led to "turned-on" fluorescence in micelles and liposomes at neutral and basic pH. Bovine serum albumin also activated the fluorescence, though to a much lesser extent. When these small molecular dyes entered cells, instead of being fluorescent only in acidic organelles, the whole cytoplasm exhibited fluorescence, with a signal/background ratio as high as ∼10 in no-wash live-cell imaging. The dye 1-labeled cells remained highly fluorescent even after 3 days. Moreover, slight variations of the dye structure resulted in significantly different intracellular fluorescence behaviors, possibly because of their different cellular uptake and intracellular activation capabilities. After the separation of cellular components, the fraction of plasma membrane and endoplasmic reticulum showed the highest fluorescence, further confirming the fluorescence activation by membrane structures. The fluorescence intensity of these dyes at different intracellular pHs (6.80 and 8.00) did not differ significantly, indicating that intracellular pH did not play a critical role. Altogether, we showed here for the first time that the fluorescence of pH-sensitive aza-BODIPY dyes was switched intracellularly not by acidic pH, but by intracellular membranes (and proteins as well). The excellent membrane permeability, ultrahigh fluorescence contrast ratio, persistent fluorescent signal, and minimal biological interference of dye 1 make it an ideal choice for live-cell imaging and in vivo cell tracking. These findings also imply that the intracellular fluorescence properties of pH-sensitive dyes should be carefully examined before they are used as pH indicators.

Lipid-mediated DNA and siRNA Transfection Efficiency Depends on Peptide Headgroup.

A series of amphiphiles with differing cationic tri- and di- peptide headgroups, designed and synthesized based on lysine (K), ornithine (O), arginine (R), and glycine (G), have been characterized and evaluated for DNA and siRNA delivery. DNA-lipoplexes formed from the tri- and di- lipopeptides possessed lipid:nucleic acid charge ratios of 7:1 to 10:1, diameters of ~200 nm to 375 nm, zeta potentials of 23 mV to 41 mV, melting temperatures of 12 °C to 46 °C, and lamellar repeat periods of 6 nm to 8 nm. These lipid-DNA complexes formed supramolecular structures in which DNA is entrapped at the surface between multilamellar liposomal vesicles. Compared to their DNA counterparts, siRNA-lipoplexes formed slightly larger complexes (348 nm to 424 nm) and required higher charge ratios to form stable structures. Additionally, it was observed that lipids with multivalent, tripeptide headgroups (i.e., KGG, OGG, and RGG) were successful at transfecting DNA in vitro, whereas DNA transfection with the dipeptide lipids proved ineffective. Cellular uptake of DNA was more effective with the KGG compared to the KG lipopeptide. In siRNA knockdown experiments, both tri- and di- peptide lipids (i.e., RGG, GGG, KG, OG, RG, GG) showed some efficacy, but total cellular uptake of siRNA complexes was not indicative of knockdown outcomes and suggested that the intracellular fate of lipoplexes may be a factor. Overall, this lipopeptide study expands the library of efficient DNA transfection vectors available for use, introduces new vectors for siRNA delivery, and begins to address the structure-activity relationships which influence delivery and transfection efficacy.

Prevalence and risk factors of stroke in China: a national serial cross-sectional study from 2003 to 2018.

Stroke imposes a substantial burden worldwide. With the rapid economic and lifestyle transition in China, trends of the prevalence of stroke across different geographic regions in China remain largely unknown. Capitalizing on the data in the National Health Services Surveys (NHSS), we assessed the prevalence and risk factors of stroke in China from 2003 to 2018. In this study, data from 2003, 2008, 2013, and 2018 NHSS were collected. Stroke cases were based on participants' self-report of a previous diagnosis by clinicians. We estimated the trends of stroke prevalence for the overall population and subgroups by age, sex, and socioeconomic factors, then compared across different geographic regions. We applied multivariable logistic regression to assess associations between stroke and risk factors. The number of participants aged 15 years or older were 154,077, 146,231, 230,067, and 212,318 in 2003, 2008, 2013, and 2018, respectively, among whom, 1435, 1996, 3781, and 6069 were stroke patients. The age and sex standardized prevalence per 100,000 individuals was 879 in 2003, 1100 in 2008, 1098 in 2013, and 1613 in 2018. Prevalence per 100,000 individuals in rural areas increased from 669 in 2003 to 1898 in 2018, while urban areas had a stable trend from 1261 in 2003 to 1365 in 2018. Across geographic regions, the central region consistently had the highest prevalence, but the western region has an alarmingly increasing trend from 623/100,000 in 2003 to 1898/100,000 in 2018 (P trend<0.001), surpassing the eastern region in 2013. Advanced age, male sex, rural area, central region, hypertension, diabetes, depression, low education and income level, retirement or unemployment, excessive physical activity, and unimproved sanitation facilities were significantly associated with stroke. In conclusion, the increasing prevalence of stroke in China was primarily driven by economically underdeveloped regions. It is important to develop targeted prevention programs in underdeveloped regions. Besides traditional risk factors, more attention should be paid to nontraditional risk factors to improve the prevention of stroke.

[Study on therapeutical effect of Canghuopingwei granules on chronic gastritis in rats].

OBJECTIVE: To investigate the therapeutical effect of Canghuopingwei Granules on chronic gastritis in rats. METHODS: Rat models of chronic gastritis and bile reflux gastritis were used. After rat models were established, the rats were divided into 6 groups and were treated with different drugs. The tissue samples were obtained after one week. The volume of gastric juice, acidity of gastric juice and pepsase activity were determined, and changes of the gastric mucosa were studied by microscopy. RESULTS: The acidity of gastric juice was reversed with Canghuopingwei granules treatment. Gastric pathologic examination suggested that Canghuopingwei granules could markedly attenuate the pathological changes of gastric mucosa in rats. CONCLUSION: Canghuopingwei granules has remarkably therapeutical effect on chronic gastritis and bile reflux gastritis in rats.

Comparative transcriptome analysis of Liriomyza trifolii (Burgess) and Liriomyza sativae (Blanchard) (Diptera: Agromyzidae) in response to rapid cold hardening.

The ability of insets to react efficiently to fluctuation in temperature is crucial for them to survive in variable surroundings. Rapid cold hardening (RCH) is a process that increase cold tolerance in most insect species. The molecular mechanisms of RCH remain largely unknown, and whether it is associated with transcriptional changes is unclear. In this study, we compared the transcriptomes of Liriomyza trifolii and L. sativae exposed to RCH to investigate the transcript abundance due to RCH in both species. RNA-seq revealed 93,166 assembled unigenes, and 34,303 of these were annotated in the L. trifolii and L. sativae transcriptome libraries. After a 4-h treatment at 1°C (RCH) compared with control, 268 and 606 unigenes were differentially expressed in L. trifolii and L. sativae, respectively. When comparing pupae exposed to 2h cold shock directly with pupae went through 4h acclimation prior to 2h cold shock, 60 and 399 unigenes were differentially expressed in L trifolii and L sativae, respectively. Genes that were commonly expressed in both L. trifolii and L. sativae, included cytochrome P450, cuticular protein, glucose dehydrogenase, solute carrier family 22 and cationic amino acid transporter. Additionally, several pathways including galactose metabolism and peroxisome were significantly enriched during RCH. Our results show that the transcriptional response is correlated with RCH in the pupal stage of the two Liriomyza species, but more transcriptional changes were identified in L sativae than in L. trifolii.

Synthesis, characterization, and in vitro transfection activity of charge-reversal amphiphiles for DNA delivery.

A series of charge-reversal lipids were synthesized that possess varying chain lengths and end functionalities. These lipids were designed to bind and then release DNA based on a change in electrostatic interaction with DNA. Specifically, a cleavable ester linkage is located at the ends of the hydrocarbon chains. The DNA release from the amphiphile was tuned by altering the length and position of the ester linkage in the hydrophobic chains of the lipids through the preparation of five new amphiphiles. The amphiphiles and corresponding lipoplexes were characterized by DSC, TEM, and X-ray, as well as evaluated for DNA binding and DNA transfection. For one specific charge-reversal lipid, stable lipoplexes of approximately 550 nm were formed, and with this amphiphile, effective in vitro DNA transfection activities was observed.

Macropinocytosis is the major pathway responsible for DNA transfection in CHO cells by a charge-reversal amphiphile.

The cellular uptake of a functional charge-reversal amphiphile:DNA lipoplex is described. First, pharmacological inhibitors were applied to block different endocytosis pathways. By examining the resulting transfection activities, it was found that endocytosis was the pathway leading to transfection in Chinese hamster ovary (CHO) cells. When the specific pathway of macropinocytosis was inhibited, ß-galactosidase expression was significantly depleted (90%); meanwhile the inhibition of clathrin-mediated pathway only brought a 30% decrease in expression; and the inhibition of caveolae-mediated pathway did not affect expression. Furthermore, a transfection kinetics study revealed that the cellular uptake responsible for gene expression was a slower process compared to clathrin-mediated endocytosis, consistent with fluid-phase uptake compared to receptor-mediated uptake. Next, a fluorescence colocalization study was used to visualize the DNA lipoplex uptake pathways. The colocalization of the DNA lipoplex and Cascade Blue, a fluid-phase uptake marker, was observed. Meanwhile, the colocalization of the DNA lipoplex and transferrin, a clathrin-mediated endocytosis marker, was also seen. However, no colocalization was observed with the endosome/lysosome marker Lysotracker. Our results indicate that macropinocytosis, not the commonly seen clathrin-mediated endocytosis for cationic lipids, is the major pathway leading to gene transfection in CHO cells for this charge-reversal amphiphile.

Differential Response of Leafminer Flies Liriomyza trifolii (Burgess) and Liriomyza sativae (Blanchard) to Rapid Cold Hardening.

Rapid cold hardening (RCH) is a rapid and critical adaption of insects to sudden temperature changes but is often overlooked or underestimated as a component of survival. Thus, interspecific comparisons of RCH are needed to predict how phenotypes will adapt to temperature variability. RCH not only enhances cold survival but also protects against non-lethal cold injury by preserving essential functions such as locomotion, reproduction, and energy balance. This study investigated the difference in basal cold tolerance and RCH capacity of L. trifolii and L. sativae. In both species, the cold tolerance of pupae was significantly enhanced after short-term exposure to moderately cold temperatures. The effect of RCH last for 4 h in L. sativae but only 2 h in L. trifolii. Interestingly, L. trifolii adults had a RCH response but L. sativae adults failed to acclimate. Short-term acclimation also lowered the supercooling point significantly in the pupae of both species. Based on these results, we propose a hypothesis that these differences will eventually affect their competition in the context of climate change. This study also provides the basis for future metabolomic and transcriptomic studies that may ultimately uncover the underlying mechanisms of RCH and interspecific competition between L. trifolii and L. sativae.

RNA Interference of Genes Encoding the Vacuolar-ATPase in Liriomyza trifolii.

The leafminer fly, Liriomyza trifolii, is an invasive pest of vegetable and horticultural crops in China. In this study, a microinjection method based on dsRNA was developed for RNA interference (RNAi) in L. trifolii using genes encoding vacuolar-ATPase (V-ATPase). Expression analysis indicated that V-ATPase B and V-ATPase D were more highly expressed in L. trifolii adults than in larvae or pupae. Microinjection experiments with dsV-ATPase B and dsV-ATPase D were conducted to evaluate the efficacy of RNAi in L. trifolii adults. Expression analysis indicated that microinjection with 100 ng dsV-ATPase B or dsV-ATPase led to a significant reduction in V-ATPase transcripts as compared to that of the dsGFP control (dsRNA specific to green fluorescent protein). Furthermore, lower dsRNA concentrations were also effective in reducing the expression of target genes when delivered by microinjection. Mortality was significantly higher in dsV-ATPase B- and dsV-ATPase D-treated insects than in controls injected with dsGFP. The successful deployment of RNAi in L. trifolii will facilitate functional analyses of vital genes in this economically-important pest and may ultimately result in new control strategies.

The Novel Application of Three-Dimensional Printing Assisted Patient-Specific Instrument Osteotomy Guide in the Precise Osteotomy of Adult Talipes Equinovarus.

OBJECTIVE: This current research is aimed at assessing clinical efficacy and prognosis of three-dimensional (3D) printing assisted patient-specific instrument (PSI) osteotomy guide in precise osteotomy of adult talipes equinovarus (ATE). METHODS: We included a total of 27 patients of ATE malformation (including 12 males and 15 females) from June 2014 to June 2018 in the current research. The patients were divided into the routine group (n = 12) and 3D printing group (n = 15) based on different operative methods. The parameters, including the operative time, intraoperative blood loss, complications, time to obtain bony fusion, functional outcomes based on American Orthopedic Foot and Ankle Society (AOFAS), and International Congenital Clubfoot Study group (ICFSG) scoring systems between the two groups were observed and recorded regularly. RESULTS: The 3D printing group exhibits superiorities in shorter operative time, less intraoperative blood loss, higher rate of excellent, and good outcomes presented by ICFSG score at last follow-up (P < 0.001, P < 0.001, P = 0.019) than the routine group. However, there was no significant difference exhibited in the AOFAS score at the last follow-up and total rate of complications between the two groups (P = 0.136, P = 0.291). CONCLUSION: Operation assisted by 3D printing PSI osteotomy guide for correcting the ATE malformation is novel and feasible, which might be an effective method to polish up the precise osteotomy of ATE malformation and enhance the clinical efficacy.

Transcriptional regulation of small heat shock protein genes by heat shock factor 1 (HSF1) in Liriomyza trifolii under heat stress.

Small heat shock proteins (sHSPs) function as molecular chaperones in multiple physiological processes and are active during thermal stress. sHSP expression is controlled by heat shock transcription factor (HSF); however, few studies have been conducted on HSF in agricultural pests. Liriomyza trifolii is an introduced insect pest of horticultural and vegetable crops in China. In this study, the master regulator, HSF1, was cloned and characterized from L. trifolii, and the expression levels of HSF1 and five sHSPs were studied during heat stress. HSF1 expression in L. trifolii generally decreased with rising temperatures, whereas expression of the five sHSPs showed an increasing trend that correlated with elevated temperatures. All five sHSPs and HSF1 showed an upward trend in expression with exposure to 40 ℃ without a recovery period. When a recovery period was incorporated after thermal stress, the expression patterns of HSF1 and sHSPs in L. trifolii exposed to 40 °C was significantly lower than expression with no recovery period. To elucidate potential interactions between HSF1 and sHSPs, double-stranded RNA was synthesized to knock down HSF1 in L. trifolii by RNA interference. The knockdown of HSF1 by RNAi decreased the survival rate and expression of HSP19.5, HSP20.8, and HSP21.3 during high-temperature stress. This study expands our understanding of HSF1-regulated gene expression in L. trifolii exposed to heat stress.

Stress analysis of the thoracolumbar junction in the process of backward fall: An experimental study and finite element analysis.

The aim of the present study was to evaluate the biomechanical mechanism of injuries of the thoracolumbar junction by the methods of a backward fall simulation experiment and finite element (FE) analysis (FEA). In the backward fall simulation experiment, one volunteer was selected to obtain the contact force data of the sacrococcygeal region during a fall. Utilizing the fall data, the FEA simulation of the backward fall process was given to the trunk FE model to obtain the stress status of local bone structures of the thoracolumbar junction during the fall process. In the fall simulation test, the sacrococcygeal region of the volunteer landed first; the total impact time was 1.14±0.58 sec, and the impact force was up to 4,056±263 N. The stress of thoracic (T)11 was as high as 42 MPa, that of the posterior margin and the junction of T11 was as high as 70.67 MPa, and that of the inferior articular process and the superior articular process was as high as 128 MPa. The average stress of T12 and the anterior margin of lumbar 1 was 25 MPa, and that of the endplate was as high as 21.7 MPa, which was mostly distributed in the back of the endplate and the surrounding cortex. According to the data obtained from the fall experiment as the loading condition of the FE model, the backward fall process can be simulated to improve the accuracy of FEA results. In the process of backward fall, the front edge of the vertebral body and the root of vertebral arch in the thoracolumbar junction are stress concentration areas, which have a greater risk of injury.

The effect of charge-reversal amphiphile spacer composition on DNA and siRNA delivery.

A series of charge-reversal amphiphiles with different spacers separating the headgroup from the hydrophobic chains are described for delivery of DNA and siRNA. Among them, the amphiphiles possessing a glycine spacer (e.g., B-GlyGly) showed effective DNA transfection in CHO and NIH 3T3 cells, as well as siRNA gene knockdown in HepG2 and UASMC cells. Ethidium bromide quenching assays revealed that DNA was released the fastest from the lipoplex of B-GlyGly in the presence of esterase. Also, X-ray diffraction results indicated that the DNA was located between the adjacent lipid bilayers in the lipoplex of B-GlyGly. These distinct features appear to be required for high transfection activity.

Cationic nucleoside lipids derived from universal bases: A rational approach for siRNA transfection.

Cationic nucleoside lipids (CNLs) derived from 5-nitroindole and 4-nitroimidazole bases were prepared from d-ribose by using a straightforward chemical synthesis. TEM experiments indicate that these amphiphilic molecules self-assemble to form supramolecular organizations in aqueous solutions. Electrophoresis and standard ethidium bromide (EB) fluorescence displacement assay shows that CNLs are able to bind siRNA. We demonstrated that both the nature of the universal bases and the stereochemistry of the anomeric position (alpha, beta) have an impact on the CNLs-siRNA complex formation. Correlations among chemical structure, stereochemistry, siRNA knockdown effect, and binding affinities for all the compounds were shown and analyzed with a simple molecular modeling study. The best binding affinities for siRNA were found for the beta anomer of the 5-nitroindole CNL which exhibits protein knockdown activity similar to the standard siPORT NeoFX positive control. It is noteworthy that no significant cytotoxicity at the tested concentration was observed for the novel CNLs.

[Study on the interaction of ofloxacin and levofloxacin with DNA].

The interaction of ofloxacin (OFLX) and levofloxacin (L-OFLX) with calf thymus DNA(ctDNA)was studied by absorption spectra, fluorescence spectra, fluorescence polarization, and K3Fe(CN)6 quenching tests to show the interaction differences and binding modes. The experimental results of absorption spectra indicated that when ctDNA was added to the solution of OFLX or L-OFLX and its concentration was increased, the hypochromic effects in the absorption spectra of OFLX or L-OFLX were observed, and no changes in its maximum absorption wavelengths were found. Fluorescence spectroscopy was an appropriate method to study the interactions between small molecule ligands and biomacromolecule. From the measurements of emission peaks, transfer efficiency of energy, lifetime and so on, a vast amount of information about the interaction will be given. The experimental results indicated that the fluorescence quenching effects of OFLX and L-OFLX were not initiated by the dynamic collision according to the Stern-Volmer equation, and caused by the static quenching of compounds formation. From the Scatchard equation, its association constants were obtained for OFLX and L-OFLX to be 1.15 x 10(5) and 3.75 x 10(5) L x mol(-1), respectively. On the basis of the absorption spectra and the association constants, it was showed that the interaction between L-OFLX and ctDNA was stronger than that of OFLX and ctDNA. There are three modes for the binding of small molecules to DNA double helix: electrostatic binding, groove binding and intercalative binding. The interaction modes between OFLX, L-OFLX and ctDNA were studied by fluorescence polarization, the K3Fe (CN)6 quenching tests and so on. The results showed that the interaction modes of OFLX and L-OFLX with ctDNA were both groove binding.

[Effect of different coating materials on hygroscopicity of compound Luhui granules].

OBJECTIVE: To study the effect of different coating materials (hydroxy-propyl methylcellulose, ethylcellulose, polyacrylic resin) on hygroscopicity of granules of traditional Chinese medicine. METHODS: The methods coated with the single coating material, the complex membrane (coated with two or three kinds of coating materials in turn), the mixed membrane (coated with two or three kinds of coating materials mixed) and the combination of mixed membrane with complex membrane were used, respectively. Taking decreasing percentage of moisture absorption of coated granules as index, the optimum condition of coating technology (the optimum coating method, coating material and its dosage) was optimized by orthogonal test. RESULTS: The damp-proof effect of coated with the combination of mixed membrane with complex membrane (decreasing percentage of moisture absorption of coated granules: 35.39%) was better than that of coated with the single coating material (decreasing percentage of moisture absorptions < or = 21.82%), the complex membrane (decreasing percentage of moisture absorption: 30.28%), or the mixed membrane (decreasing percentage of moisture absorption: 31.37%) (P < 0.05). CONCLUSION: The method coated with the combination of mixed membrane (mixed coating materials composed of ethylcellulose and polyacrylic resin) and with complex membrane (coating material composed of HPMC; The granules were coated firstly with the mixed coating materials, followed by HPMC) , obviously reduces the hygroscopicty of compound Luhui granules, and raises the stability of compound Luhui capsules. It is advantageous to the preparation production, storage and application.