Synergistic effects and competitive relationships between DOC and DOX as acting on DNA molecules: Studied with confocal Raman spectroscopy and molecular docking technology.

Docetaxel (DOC) is one of the second-generation antineoplastic drugs of the taxanes family with excellent antitumor activity. However, the mechanism of DOC inducing tumor cell apoptosis and treating cancer diseases, especially its interaction with DNA in the nucleus, and its adjuvant or combined Doxorubicin (DOX) acting on DNA molecules are unclear. In this study, the interaction mechanism between DOC and DNA, as well as the synergistic effects and competitive relationships among DOC and DOX when they simultaneously interact with DNA molecules were studied by laser confocal Raman spectroscopy combined with UV-visible absorption spectroscopy and molecular docking technology. The spectroscopic results showed that the binding constant of DOC to DNA is 5.25 × 103 M-1, the binding modes of DOC and DNA are non-classical intercalation and electrostatic binding, and the DNA-DOC complex has good stability. When DOC or DOX interacts with DNA alone, both of them can bind with bases and phosphate backbone of DNA, and also lead to DNA conformation changes; when DOC and DOX interact with DNA at the same time, the orders of interaction not only affect their binding sites with DNA, but also cause changes in the surrounding environment of the binding sites. In addition, the molecular docking results further verified that DOC and DOX have synergy and competition when they interact with DNA molecules simultaneously. The docking energies of DNA-DOC and DNA-DOX indicate the important role of van der Waals forces and hydrogen bonds. This study has practical significance for the design and development of antitumor drugs with less toxic based on the taxanes family and the combination with other drugs for the treatment of cancer.

Effects of soaking glutinous sorghum grains on physicochemical properties of starch.

Glutinous sorghum grains were soaked (60-80 °C, 2-8 h) to explore the effects of soaking, an essential step in industrial processing of brewing, on starch. As the soaking temperature increased, the peak viscosity and crystallinity of starch gradually decreased, while the enzymatic hydrolysis rate and storage modulus first increased and then decreased. At 70 °C, the content of amylose, the enzymatic hydrolysis rate of starch, and the final viscosity first increase and then decrease with the increase of soaking time, reaching their maximum at 6 h, increased by 53.1 %, 11.0 %, and 10.4 %, respectively, as compared with the non-soaked sample. At 80 °C (4 h), the laser confocal microscopy images showed a network structure formed between the denatured protein chains and the leached-out amylose chains. The molecular weights of starch before and after soaking were all in the range of 3.82-8.98 × 107 g/mol. Since 70 °C is lower than that of starch gelatinization and protein denaturation, when soaking for 6 h, the enzymatic hydrolysis rate of starch is the highest, and the growth of miscellaneous bacteria is inhibited, which is beneficial for subsequent processing technology. The result provides a theoretical basis for the intelligent control of glutinous sorghum brewing.

Application of confocal microscopy and flow cytometry to identify physiological responses of Prorocentrum micans to the herbicide glyphosate.

The physiological response of the dinoflagellate P. micans to the effect of the herbicide glyphosate at a concentration of 25-200 µg L-1 was evaluated. It has been shown that P. micans is able to grow due to the consumption of dissolved organic phosphorus formed as a result of the mineralization of glyphosate by bacteria. The addition of glyphosate to the medium inhibits the photosynthetic activity of cells; there is a pronounced inhibition of the relative electron transfer rate along the electron transport chain and the maximum quantum efficiency of the use of light energy. Morphological and ultrastructural changes in P. micans cells were evaluated at sublethal (150 µg L-1) and lethal (200 µg L-1) glyphosate concentrations. It has been shown that at a herbicide concentration of 150 µg L-1, the first signs of apoptosis appear in most P. micans cells: a decrease in lateral light scattering, cytoplasmic retraction, partial destruction of cytoplasmic organelles, a change in the morphology of nuclei, mitochondria, a change in the potential of mitochondrial membranes, and a decrease in the autofluorescence of chlorophyll in cells. At a glyphosate concentration of 200 µg L-1, P. micans showed signs of a late stage of apoptosis: violation of the integrity of intracellular organelles and chromatin organization, fragmentation of nuclei, condensation of cytoplasm, disorganization of chloroplasts in the cells, and the release of cell contents beyond the cell membrane. The effectiveness of using flow cytometry and laser scanning confocal microscopy methods for identifying signs and stages of cell apoptosis when exposed to glyphosate is discussed.

In Vivo Ultrafast Doppler Imaging Combined with Confocal Microscopy and Behavioral Approaches to Gain Insight into the Central Expression of Peripheral Neuropathy in Trembler-J Mice.

The main human hereditary peripheral neuropathy (Charcot-Marie-Tooth, CMT), manifests in progressive sensory and motor deficits. Mutations in the compact myelin protein gene pmp22 cause more than 50% of all CMTs. CMT1E is a subtype of CMT1 myelinopathy carrying micro-mutations in pmp22. The Trembler-J mice have a spontaneous mutation in pmp22 identical to that present in CMT1E human patients. PMP22 is mainly (but not exclusively) expressed in Schwann cells. Some studies have found the presence of pmp22 together with some anomalies in the CNS of CMT patients. Recently, we identified the presence of higher hippocampal pmp22 expression and elevated levels of anxious behavior in TrJ/+ compared to those observed in wt. In the present paper, we delve deeper into the central expression of the neuropathy modeled in Trembler-J analyzing in vivo the cerebrovascular component by Ultrafast Doppler, exploring the vascular structure by scanning laser confocal microscopy, and analyzing the behavioral profile by anxiety and motor difficulty tests. We have found that TrJ/+ hippocampi have increased blood flow and a higher vessel volume compared with the wild type. Together with this, we found an anxiety-like profile in TrJ/+ and the motor difficulties described earlier. We demonstrate that there are specific cerebrovascular hemodynamics associated with a vascular structure and anxious behavior associated with the TrJ/+ clinical phenotype, a model of the human CMT1E disease.

Effect of different obturation techniques on sealer penetration into dentinal tubules after final irrigation with XP-Endo finisher file

The first objective of this research was to evaluate the effectiveness of XP-Endo Finisher on dentinal tubule penetration of irrigation solution using confocal laser scanning microscopy. The main purpose of this research was to compare the effect of cold lateral condensation, continuous wave obturation and core-carrier based techniques on sealer penetration. Sixty mandibular premolars were prepared and allocated into two experimental groups (n=30) as the final irrigation technique and obturation technique experiment. In the final irrigation technique experiment, final irrigation was performed with XP-Endo Finisher, passive ultrasonic irrigation (PUI) and conventional needle irrigation (CNI) (n=10). The roots in the obturation technique experiment were also assigned into 3 groups and obturated with cold lateral condensation, continuous-wave obturation and core-carrier techniques (n=10). The most effective activation method, which emerged as a result of the first part of this study, was used as the final irrigation method in the obturation technique experiment. Then, all roots were sectioned in 1-mm-thick slices at 3mm from the apex for scanning. In terms of depth and percentage of material penetration, CNI exhibited significantly the lowest values and no significant difference was found between others. Also, there was no significant difference among obturation methods. In conclusion, XP-Endo Finisher and PUI are more effective than CNI on irrigant penetration. Sealer penetration into dentinal tubules is independent of obturation techniques.

El objetivo principal de esta investigación fue evaluar la eficacia de XP- Endo Finisher en la penetración de la solución de irrigación en los túbulos dentinarios mediante microscopía de láser confocal. Se prepararon sesenta premolares mandibulares y se distribuyeron en dos grupos experimentales (n=30) según el tipo de método de evaluación utilizado. En el experimento de la técnica de irrigación final, la irrigación final se realizó con XP-Endo Finisher, irrigación ultrasónica pasiva (PUI) e irrigación con aguja convencional (CNI) (n=10). Las raíces en el experimento de la técnica de obturación también se asignaron en 3 grupos y se obturaron con técnicas de condensación lateral fría, obturación de onda continua y portador de núcleo (n=10). El método de activación más eficaz, que surgió como resultado de la primera parte de este estudio, se utilizó como método de irrigación final en el experimento de la técnica de obturación. Luego, todas las raíces se seccionaron en muestras de 1mm de espesor. En términos de profundidad y porcentaje de penetración del material, CNI exhibió significativamente los valores más bajos y no se encontraron diferencias significativas entre los demás. Además, no hubo diferencias significativas entre los métodos de obturación. En conclusión, XP-Endo Finisher y PUI son más efectivos que CNI en la penetración del irrigante. La penetración del sellador en los túbulos dentinarios es independiente de las técnicas de obturación.

[Treatment of rheumatoid arthritis by injection of sinomenine solid lipid nanoparticles under a fluorescence endoscopic laser confocal microscope].

A fluorescence endoscopic laser confocal microscope(FELCM) was used to direct the injection of sinomenine solid lipid nanoparticles(Sin-SLN) into the joint, and the in vitro effectiveness of Sin-SLN in the treatment of rheumatoid arthritis(RA) was evaluated. Sin-SLN was prepared with the emulsion evaporation-low temperature curing method. The Sin-SLN prepared under the optimal conditions showed the encapsulation efficiency of 64.79%±3.12%, the drug loading of 3.84%±0.28%, the average particle size of(215.27±4.21) nm, and the Zeta potential of(-32.67±0.84) mV. Moreover, the Sin-SLN demonstrated good stability after sto-rage for 30 days. The rabbit model of RA was established by the subcutaneous injection of ovalbumin and complete Freund's adjuvant. Five groups were designed, including a control group, a model group, a Sin(1.5 mg·kg~(-1)) group, a Sin-SLN(1.5 mg·kg~(-1)) group, and a dexamethasone(positive drug, 1.0 mg·kg~(-1), ig) group. The control group and the model group only received puncture treatment without drug injection. After drug administration, the local skin temperature and knee joint diameter were monitored every day. The knee joint diameter and the local skin temperature were lower in the drug administration groups than in the model group(P<0.05, P<0.01). FELCM recorded the morphological alterations of the cartilage of knee joint. The Sin-SLN group showed compact tissue structure and smooth surface of the cartilage. Enzyme-linked immunosorbent assay(ELISA) was employed to determine the serum le-vels of interleukin-1(IL-1) and tumor necrosis factor-α(TNF-α). The findings revealed that the Sin-SLN group had lower IL-1 and TNF-α levels than the model group(P<0.05, P<0.01). Hematoxylin-eosin(HE) staining was employed to reveal the pathological changes of the synovial tissue, which were significantly mitigated in the Sin-SLN group. The prepared Sin-SLN had uniform particle size and high stability. Through joint injection administration, a drug reservoir was formed. Sin-SLN effectively alleviate joint swelling and cartilage damage of rabbit, down-regulated the expression of inflammatory cytokines, and inhibited the epithelial proliferation and inflammatory cell infiltration of the synovial tissue, demonstrating the efficacy in treating RA.

Performance Evaluation and Oil Displacement Effect of Amphiphilic Polymer Heavy Oil Activator.

A heavy oil activator is an amphiphilic polymer solution that contains hydrophilic and oleophobic groups. It can enhance heavy oil recovery efficiency. This paper studied the changes in the distribution of the remaining oil after activator flooding and the performance of heavy oil's active agent. Nuclear magnetic resonance spectroscopy, laser confocal microscopy, microscopic visualization, and CT scanning techniques were used to analyze crude oil utilization, and the distribution characteristics of the remaining oil during activator flooding of heavy oil. The results showed that the heavy oil activator solution presented a dense spatial network and good viscosification ability. The activator could reduce the interfacial tension of oil and water, disassemble the heavy components of dispersed heavy oil and reduce the viscosity of heavy oil. The utilization degree of the remaining oil in small and middle pores increased significantly after activator flooding, the remaining oil associated with membranous-like and clusterlike structures was utilized to a high degree, and the decline of light/heavy fraction in heavy oil slowed down. Heavy oil activator improved the swept volume and displacement efficiency of heavy oil, playing a significant role in improving the extent of recovery of heavy oil reservoirs.

Super-resolution reconstruction, recognition, and evaluation of laser confocal images of hyperaccumulator Solanum nigrum endocytosis vesicles based on deep learning: Comparative study of SRGAN and SRResNet.

It is difficult for laser scanning confocal microscopy to obtain high- or ultra-high-resolution laser confocal images directly, which affects the deep mining and use of the embedded information in laser confocal images and forms a technical bottleneck in the in-depth exploration of the microscopic physiological and biochemical processes of plants. The super-resolution reconstruction model (SRGAN), which is based on a generative adversarial network and super-resolution reconstruction model (SRResNet), which is based on a residual network, was used to obtain single and secondary super-resolution reconstruction images of laser confocal images of the root cells of the hyperaccumulator Solanum nigrum. Using the peak signal-to-noise ratio (PSNR), structural similarity (SSIM) and mean opinion score (MOS), the models were evaluated by the image effects after reconstruction and were applied to the recognition of endocytic vesicles in Solanum nigrum root cells. The results showed that the single reconstruction and the secondary reconstruction of SRGAN and SRResNet improved the resolution of laser confocal images. PSNR, SSIM, and MOS were clearly improved, with a maximum PSNR of 47.690. The maximum increment of PSNR and SSIM of the secondary reconstruction images reached 21.7% and 2.8%, respectively, and the objective evaluation of the image quality was good. However, overall MOS was less than that of the single reconstruction, the perceptual quality was weakened, and the time cost was more than 130 times greater. The reconstruction effect of SRResNet was better than that of SRGAN. When SRGAN and SRResNet were used for the recognition of endocytic vesicles in Solanum nigrum root cells, the clarity of the reconstructed images was obviously improved, the boundary of the endocytic vesicles was clearer, and the number of identified endocytic vesicles increased from 6 to 9 and 10, respectively, and the mean fluorescence intensity was enhanced by 14.4% and 7.8%, respectively. Relevant research and achievements are of great significance for promoting the application of deep learning methods and image super-resolution reconstruction technology in laser confocal image studies.

Characteristics of corneal stromal demarcation line after different cross-linking methods in keratoconus and its influence on the cross-linking effect / 中华实验眼科杂志

Objective:To compare the characteristics of corneal stromal demarcation line after different surgical methods of riboflavin/ultraviolet A corneal collagen cross-linking (CXL) in early keratoconus, and analyze the influence of the demarcation line on the cross-linking effect.Methods:A non-randomized controlled clinical study was conducted.Sixty-nine eyes of 69 patients treated with riboflavin/ultraviolet A CXL in the Eye Hospital of Shandong First Medical University from May 2019 to February 2021 were included.According to the cross-linking methods, the patients were divided into epithelium-on treatment group (21 eyes) and epithelium-off treatment group (48 eyes). There were 25 eyes in 5.4 J energy group and 44 eyes in 7.2 J energy group.The morphology and changes of corneal stromal cross-linking reaction (corneal stromal demarcation line) were observed at 2 weeks, 1, 3 and 4 months after operation.Changes in the thinnest corneal thickness (TCT), uncorrected visual acuity (UCVA, LogMAR), best corrected visual acuity (BCVA, LogMAR) and corneal maximum curvature (Kmax) were recorded.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Eye Hospital of Shandong First Medical University (No.2019.05). Written informed consent was obtained from each subject.Results:Of the 69 eyes after operation, 44 eyes (63.77%) had demarcation lines, and 25 eyes (36.23%) had no demarcation lines.The occurrence rate of demarcation lines in the epithelium-on treatment group was 79.17%(38/48), which was significantly higher than 28.57%(6/21) in the epithelium-off treatment group ( χ2=16.186, P<0.01). The occurrence rate of demarcation line in 5.4 J energy group was 72.00%(18/25), and the 7.2 J energy group was 56.80%(25/44), with no significant difference ( χ2=1.565, P=0.302). Slit lamp microscopy and anterior segment-optical coherence tomography showed that the demarcation line appeared at 1-2 weeks after operation, gradually converged and strengthened after 1 month, turned diffuse, blurred and faded by degrees after 2-3 months, and basically disappeared after 4 months.The depth of the demarcation line reached 141-423 μm, with an average depth of (263.44±84.22)μm.Scanning laser confocal microscopy showed that corneal stromal cells were activated and light reflection was enhanced after CXL.Collagen fibers extended vertically and horizontally, crisscrossed, and were in a reticular arrangement.The TCT decreased from preoperative (458.69±38.28)μm to (443.86±36.54)μm at 4 months after operation, showing a statistically significant difference ( t=6.705, P<0.001). There was no significant difference in the TCT reduction between groups with and without demarcation lines ( t=1.684, P=0.100). At 4 months postoperatively, the UCVA of all eyes increased from preoperative 0.74±0.37 to 0.69±0.38, and the difference was statistically significant ( t=2.109, P=0.039). There was no significant difference in BCVA between before and after operation ( t=1.006, P=0.319). There was no significant difference in change of UCVA and BCVA between groups with and without demarcation lines ( t=0.065, P=0.949; t=0.346, P=0.730). There was no significant difference in Kmax in all patients between before and after operation ( t=0.050, P=0.950). There was no significant difference in the Kmax change between groups with and without demarcation lines ( t=-0.739, P=0.464). The change in TCT in the epithelium-off treatment group was significantly greater than that in the epithelium-on treatment group ( t=2.815, P=0.008). There was no significant difference in UCVA, BCVA and Kmax changes between epithelium-on and epithelium-off treatment groups (all at P>0.05). There was no obvious corneal scarring, infectious keratitis, corneal endothelial decompensation or other complications. Conclusions:The demarcation line after CXL may be a sign of the depth of cross-linking reaction, which is more prone to occur after the epithelium-off operation method.Both the epithelium-on and epithelium-off operation methods have similar therapeutic effects.Demarcation line after different cross-linking methods has no significant influence on the cross-linking effect in keratoconus.

Treatment of rheumatoid arthritis by injection of sinomenine solid lipid nanoparticles under a fluorescence endoscopic laser confocal microscope / 中国中药杂志

A fluorescence endoscopic laser confocal microscope(FELCM) was used to direct the injection of sinomenine solid lipid nanoparticles(Sin-SLN) into the joint, and the in vitro effectiveness of Sin-SLN in the treatment of rheumatoid arthritis(RA) was evaluated. Sin-SLN was prepared with the emulsion evaporation-low temperature curing method. The Sin-SLN prepared under the optimal conditions showed the encapsulation efficiency of 64.79%±3.12%, the drug loading of 3.84%±0.28%, the average particle size of(215.27±4.21) nm, and the Zeta potential of(-32.67±0.84) mV. Moreover, the Sin-SLN demonstrated good stability after sto-rage for 30 days. The rabbit model of RA was established by the subcutaneous injection of ovalbumin and complete Freund's adjuvant. Five groups were designed, including a control group, a model group, a Sin(1.5 mg·kg~(-1)) group, a Sin-SLN(1.5 mg·kg~(-1)) group, and a dexamethasone(positive drug, 1.0 mg·kg~(-1), ig) group. The control group and the model group only received puncture treatment without drug injection. After drug administration, the local skin temperature and knee joint diameter were monitored every day. The knee joint diameter and the local skin temperature were lower in the drug administration groups than in the model group(P<0.05, P<0.01). FELCM recorded the morphological alterations of the cartilage of knee joint. The Sin-SLN group showed compact tissue structure and smooth surface of the cartilage. Enzyme-linked immunosorbent assay(ELISA) was employed to determine the serum le-vels of interleukin-1(IL-1) and tumor necrosis factor-α(TNF-α). The findings revealed that the Sin-SLN group had lower IL-1 and TNF-α levels than the model group(P<0.05, P<0.01). Hematoxylin-eosin(HE) staining was employed to reveal the pathological changes of the synovial tissue, which were significantly mitigated in the Sin-SLN group. The prepared Sin-SLN had uniform particle size and high stability. Through joint injection administration, a drug reservoir was formed. Sin-SLN effectively alleviate joint swelling and cartilage damage of rabbit, down-regulated the expression of inflammatory cytokines, and inhibited the epithelial proliferation and inflammatory cell infiltration of the synovial tissue, demonstrating the efficacy in treating RA.

Effect of cavity disinfection protocols on microtensile bond strength of universal adhesive to dentin / Efecto de los protocolos de desinfección de cavidades en la fuerza de adhesión microtensil del adhesivo universal a la dentina

Abstract The purpose of this study was to compare the effect of different disinfection protocols of dentin on bond strength of an MDP-containing universal adhesive. Twelve extracted mandibular third molars were separated horizontally at the mid-coronal of crown to get smooth and sound dentin surfaces using low-speed diamond saw. The teeth were randomly fallen into four groups: chlorhexidine (CHX), ozone, Er,Cr:YSGG laser irradiation (LASER) and no treatment (control). After cavity disinfection application, a universal adhesive (G-Premio Bond) was applied to the surface of dentin according to self-etch mode as instructed by the manufacturer. After incremental built-up of composite resin (Charisma Smart), the specimens were immersed in distilled water at 37°C for 24h. Dentin/composite beams with 1 mm² cross sectional area were produced and micro-tensile bond strength (µTBS) was applied on these beams (n=20). Failure mods were determined under a stereomicroscope at ×40. The resin penetration of samples stained with Rhodamine B fluorochrome dye was examined with a confocal laser scanning microscope. Statistical analysis was performed with SPSS-22. Test results were analyzed using One-way Anova and Tukey HSD Post-Hoc tests (p0.5). All applications of cavity disinfection procedures decreased the µTBS of the resin-dentin interface.

Resumen El propósito de este estudio fue comparar el efecto de diferentes protocolos de desinfección de la dentina sobre la fuerza de unión de un adhesivo universal que contiene MDP. Doce terceros molares mandibulares extraídos se quebraron horizontalmente en la mitad de la corona para obtener superficies de dentina lisas y sólidas utilizando una sierra de diamante de baja velocidad. Los dientes se dividieron aleatoriamente en cuatro grupos: clorhexidina (CHX), ozono, irradiación con láser Er,Cr:YSGG (LASER) y ningún tratamiento (control). Después de la aplicación de la desinfección de la cavidad, se aplicó un adhesivo universal (G-Premio Bond) a la superficie de la dentina según el modo de autograbado indicado por el fabricante. Después de la obturación con resina compuesta (Charisma Smart), las muestras se sumergieron en agua destilada a 37°C durante 24h. Se produjeron porciones de dentina/resina con un área de sección transversal de 1 mm² y se aplicó una fuerza de adhesión microtensile (µTBS) (n=20). Los modos de falla se determinaron bajo un microscopio estereoscópico a ×40. La penetración de la resina de las muestras teñidas con colorante fluorocromo rodamina B se examinó con un microscopio de barrido láser confocal. El análisis estadístico se realizó con SPSS-22. Los resultados de las pruebas se analizaron utilizando las pruebas post-hoc Anova unidireccional y Tukey HSD (p0.5). Todas las aplicaciones de procedimientos de desinfección de cavidades redujeron el µTBS de la interfaz resina-dentina.

Ripening induced degradation of pectin and cellulose affects the far infrared drying kinetics of mangoes.

The quality parameters of mangoes change during ripening, which has a vital impact on processing characteristics. Effects of ripening stage (four stages from the lowest to highest degree-RS-1, RS-2, RS-3, RS-4) on cell wall polysaccharides and far infrared drying kinetics of mangoes were investigated. As ripening progressed, the water-soluble pectin contents increased by 213.5%; while the chelate-, sodium carbonate-soluble pectin and hemicellulose contents decreased by 44.0%, 59.5% and 65.8%, respectively. Moreover, the molecular weight reduction confirmed the degradation of pectin. These further caused the alteration of cell wall structure and changes in water distribution. Meanwhile, the drying time of mangos with different ripeness were in the order: RS-3 > RS-4 > RS-2 > RS-1. It correlated with the degradation of cell wall polysaccharides, the destruction of cell wall and the increases in free water during ripening. The ripeness classification could effectively improve the uniformity and efficiency of fruits drying processing.

Evaluation of Laser Confocal Raman Spectroscopy as a Non-Invasive Method for Detecting Sperm DNA Contents.

RESEARCH QUESTION: Is Raman spectroscopy an efficient and accurate method to detect sperm chromosome balance state by DNA content differences? DESIGN: Semen samples were provided by diploid healthy men, and the analysis parameters met the current World Health Organization standards. The DNA content was assessed by analysis of the corresponding spectra obtained from a laser confocal Raman spectroscope. The sperm sex chromosome information was obtained by fluorescence in situ hybridization (FISH). Comparative analysis was performed between FISH results and Raman spectral analysis results. RESULTS: Different parts of the sperm head showed different spectral signal intensities, which indicated that there were different chemical components. Standard principal component analysis (PCA) can preliminarily classify sperm with different DNA contents into two groups. Further analysis showed that there were significant differences in the 785 DNA backbone peaks and 714-1,162 cm-1 DNA skeleton regions among sperm with different DNA contents. The peak and regional peak of the DNA skeleton of X sperm were significantly higher than those of Y sperm (X vs. Y, p < 0.05). The above sperm types were confirmed by FISH. ROC curve analysis shows that there is a correlation between the Raman spectrum data and FISH results. CONCLUSION: Raman spectroscopy can identify X and Y sperms by analyzing the DNA content difference. However, the accuracy of the detection still needs to be improved. Nevertheless, Raman spectroscopy has a potential application value in the field of sperm aneuploidy detection and may even be used as a non-invasive predictor of sperm aneuploid state in preimplantation genetic testing (PGT-A).

In Situ Micro-Observation of Surface Roughness and Fracture Mechanism in Metal Microforming of Thin Copper Sheets with Newly Developed Compact Testing Apparatus.

A better understanding of material deformation behaviours with changes in size is crucial to the design and operation of metal microforming processes. In order to facilitate the investigation of size effects, material deformation behaviours needed to be determined directly from material characterizations. This study was aimed at the design and manufacture of a compact universal testing machine (UTM) compatible with a 3D laser-confocal microscope to observe the deformation behaviour of materials in real-time. In this study, uniaxial micro tensile testing was conducted on three different thin (0.05 mm, 0.1 mm, and 0.3 mm) copper specimens with characteristic dimensions at micro scales. Micro tensile experimental runs were carried out on copper specimens with varying grain sizes on the newly developed apparatus under a 3D laser-confocal microscope. Microscale experiments under 3D laser-confocal microscope provided not only a method to observe the microstructure of materials, but also a novel way to observe the early stages of fracture mechanisms. From real-time examination using the newly developed compact testing apparatus, we discovered that fracture behaviour was mostly brought about by the concave surface formed by free surface roughening. Findings with high stability were discovered while moving with the sample grasped along the drive screw in the graphical plot of a crosshead's displacement against time. Our results also showed very low mechanical noise (detected during the displacement of the crosshead), which indicated that there were no additional effects on the machine, such as vibrations or shifts in speed that could influence performance. The engineering stress-strain plots of the pure copper-tests with various thicknesses or samples depicted a level of stress necessary to initiate plastic flowing inside the material. From these results, we observed that strength and ductility declined with decreasing thickness. The influence of thickness on fracture-strain, observed during tensile testing, made it clear that the elongation-at-break of the pure-copper foils intensely decreased with decreases in thickness. The relative average surface-roughness Ra was evaluated, which showed us that the surface-roughness escalated with the increasing trend of plasticity deformation (plastic strain) ε. For better understanding of the effects of plastic strain on surface roughness prior to material fractures, micro tensile tests were performed on the newly developed machine under a 3D laser-confocal-microscope. We observed that homogeneous surface roughness was caused by plastic strain, which further formed the concave surface that led to the fracture points. Finally, we concluded that surface roughness was one of the crucial factors influencing the fracture behaviour of metallic sheet-strips in metal microforming. We found that this type of testing apparatus could be designed and manufactured within a manageable budget.

A Novel qPCR Method for the Detection of Lactic Acid Bacteria in Fermented Milk.

The number of live lactic acid bacteria (LAB) is an important quality indicator for yogurt, the quantitative testing of LAB has become an important task in the evaluation of product quality and function. By analyzing and comparing the performance of 16S rRNA gene and tuf gene used in absolute quantification, the tuf gene with copy number 1 was selected as the target gene of six LAB. By drawing a standard curve to achieve qualitative and quantitative detection of six strains of LAB, the detection range was found to be 1 × 103-1 × 108 copies/µL. The traditional plate colony count and Flow Cytometry (FCM) were compared with the method of qPCR, which was used in this experiment. Meanwhile, the confocal laser microscope combined with STYO 9 and propidium iodide dyes was used to determine that the content of viable bacteria in the yogurt was more than 90%, which proved that the detection result using qPCR method was closer to the true level of LAB in yogurt. Compared with the existing methods, the method in this study allowed the qualitative and quantitative detection of the six kinds of LAB in yogurt, and the distribution of live and dead bacteria in yogurt could be calculated.

Wheat Varietal Response to Tilletia controversa J. G. Kühn Using qRT-PCR and Laser Confocal Microscopy.

Tilletia controversa J. G. Kühn is a causal organism of dwarf bunt in wheat. Understanding the interaction of wheat and T. controversa is of practical and scientific importance for disease control. In this study, the relative expression of TaLHY and TaPR-4 and TaPR-5 genes was higher in a resistant (Yinong 18) and moderately resistant (Pin 9928) cultivars rather than susceptible (Dongxuan 3) cultivar at 72 h post inoculation (hpi) with T. controversa. Similarly, the expression of defensin, TaPR-2 and TaPR-10 genes was observed higher in resistant and moderately resistant cultivars after exogenous application of phytohormones, including methyl jasmonate, salicylic acid, and abscisic acid. Laser confocal microscopy was used to track the fungal hyphae in the roots, leaves, and tapetum cells, which of susceptible cultivar were infected harshly by T. controversa than moderately resistant and resistant cultivars. There were no fungal hyphae in tapetum cells in susceptible cultivar after methyl jasmonate, salicylic acid and abscisic acid treatments. Moreover, after T. controversa infection, the pollen germination was of 80.06, 58.73, and 0.67% in resistant, moderately resistant and susceptible cultivars, respectively. The above results suggested that the use using of resistant cultivar is a good option against the dwarf bunt disease.

Synthesis, biological evaluation, and molecular docking analysis of phenstatin based indole linked chalcones as anticancer agents and tubulin polymerization inhibitors.

A library of new phenstatin based indole linked chalcone compounds (9a-z and 9aa-ad) were designed and synthesized. Of these, compound 9a with 1-methyl, 2- and 3-methoxy substituents in the aromatic ring was efficacious against the human oral cancer cell line SCC-29B, spheroids, and in a mouse xenograft model of oral cancer AW13516. Compound 9a exhibited anti-cancer activity through disrupting cellular integrity and affecting glucose metabolism-which is a hallmark of cancer. The cellular architecture was affected by inhibition of tubulin polymerization as observed by an immunofluorescence assay on 9a-treated SCC-29B cells. An in vitro tubulin polymerization kinetics assay provided evidence of direct interaction of 9a with tubulin. This physical interaction between tubulin and compound 9a was further confirmed by Surface Plasmon Resonance (SPR) analysis. Molecular docking experiments and validations revealed that compound 9a interacts and binds at the colchicine binding site of tubulin and at active sites of key enzymes in the glucose metabolism pathway. Based on in silico modeling, biophysical interactions, and pre-clinical observations, 9a consisting of phenstatin based indole-chalcone scaffolds, can be considered as an attractive tubulin polymerization inhibitor candidate for developing anti-cancer therapeutics.

[Early diagnosis of diabetic polyneuropathy based on the results of corneal nerve fibers examination]. / Vozmozhnosti rannei diagnostiki diabeticheskoi polineiropatii na osnove issledovaniya nervnykh volokon rogovitsy.

Laser corneal confocal microscopy (CCM) is a method of objective visualization of thin corneal nerve fibers (CNF), the structure of which changes in patients with diabetes mellitus (DM). PURPOSE: To conduct comparative analysis of the results of CNF assessment using CCM and other known neurological instrumental techniques as well as evaluate their applicability to the early diagnosis of diabetic polyneuropathy (DPN). MATERIAL AND METHODS: We examined a total of 46 patients (85 eyes) with type 1 DM and either subclinical (24 patients), or clinical-stage DPN (22 patients) and 50 patients (87 eyes) with type 2 DM (subclinical DPN in 27 patients and clinical-stage DPN in 23 patients). The control group consisted of 34 healthy volunteers (68 eyes). All patients underwent standard ophthalmological examination, CCM with nerve tortuosity assessment (including calculation of coefficients of CNF orientation anisotropy, KΔL, and symmetry, Ksym) and interocular asymmetry, electroneuromyography (ENMG), and quantitative sensory testing (QST). RESULTS: Analysis of the CCM results revealed a reliable decrease in the average KΔL values in patients with type 1 and type 2 DM compared with the control group. In the group of patients with type 1 DM and subclinical DPN, correlations were revealed between the CNF tortuosity coefficients and a number of ENMG parameters, such as the M-response amplitude of the peroneal nerve (r=0.73, p≤0.02), M-response amplitude of the tibial nerve (r=0.58, p≤0.01), residual latency (r= -0.62, p≤0.05), and peroneal nerve conduction velocity (r=0.57, p≤0.01). Ksym values correlated with the warm sensitivity threshold (r=0.6, p≤0.008). Among patients with type 2 DM and subclinical DPN, the KΔL coefficient correlated with the peroneal nerve conduction velocity (r=0.46, p≤0.02), M-response amplitude of the tibial nerve (r=0.6, p≤0.04), and residual latency of the peroneal nerve (r=-0.56, p≤0.05). CONCLUSION: The state of thin corneal nerves correlates with functional changes in the peripheral nerves. Pathological changes in CNF in patients with DM can be detected at an early (subclinical) stage of DPN using laser CCM and a program for corneal nerve tortuosity analysis.

Spectroscopic Evidence of Energy Transfer in BODIPY-Incorporated Nano-Porphyrinic Metal-Organic Frameworks.

Metal-organic frameworks (MOFs) represent a class of solid-state hybrid compounds consisting of multitopic organic struts and metal-based nodes that are interconnected by coordination bonds, and they are ideal for light harvesting due to their highly ordered structure. These structures can be constructed with chromophore organic ligands structures for the purpose of efficient light harvesting. Here, we prepared porphyrin-based nano-scaled MOFs (nPCN-222) with BODIPY and I2BODIPY photosensitizers by incorporating BODIPY/I2BODIPY into nPCN-222 (nPCN-BDP/nPCN-I2BDP) and demonstrated resonance energy transfer from the donor (BODIPY/I2BODIPY) to the acceptor (nPCN-222) resulting in greatly enhanced fluorescence of nPCN-222, as visually manifested by time-resolved and space-resolved fluorescence imaging of the nano-scaled MOFs.

Synthesis, crystal structure, polymorphism and microscopic luminescence properties of anthracene derivative compounds.

Anthracene derivative compounds are currently investigated because of their unique physical properties (e.g. bright luminescence and emission tunability), which make them ideal candidates for advanced optoelectronic devices. Intermolecular interactions are the basis of the tunability of the optical and electronic properties of these compounds, whose prediction and exploitation benefit from knowledge of the crystal structure and the packing architecture. Polymorphism can occur due to the weak intermolecular interactions, requiring detailed structural analysis to clarify the origin of observed material property modifications. Here, two silylethyne-substituted anthracene compounds are characterized by single-crystal synchrotron X-ray diffraction, identifying a new polymorph in the process. Additionally, laser confocal microscopy and fluorescence lifetime imaging microscopy confirm the results obtained by the X-ray diffraction characterization, i.e. shifting the substituents towards the external benzene rings of the anthracene unit favours π-π interactions, impacting on both the morphology and the microscopic optical properties of the crystals. The compounds with more isolated anthracene units feature shorter lifetime and emission spectra, more similar to those of isolated molecules. The crystallographic study, supported by the optical investigation, sheds light on the influence of non-covalent interactions on the crystal packing and luminescence properties of anthracene derivatives, providing a further step towards their efficient use as building blocks in active components of light sources and photonic networks.