[Study on the dynamic changes of peripheral platelet-to-lymphocyte ratio in the prognosis of neoadjuvant chemotherapy patients with gastric cancer].

Objective: To investigate the significance of platelet lymphocyte ratio (PLR) before and after neoadjuvant chemotherapy in advanced gastric cancer (AGC). Methods: The medical records of 247 AGC patients who underwent surgery between May 2015 and October 2016 were retrospectively reviewed. The relationship between PLR value and its changes before and after neoadjuvant therapy and clinicopathological features and prognosis was further analyzed. Results: △PLR was defined according to the different states of PLR before and after neoadjuvant therapy. If negative value was defined as"Reduced Group"(138) and positive value or 0 was defined as "Unreduced group"(109). There were statistical differences between the two groups of△PLR in tumor size, nerve invasion, presence or absence of vascular tumor thrombus, ypT staging, ypN staging, ypTNM staging, and pathological response (all P<0.05), but there was no statistical difference between age, gender, and postoperative adjuvant chemotherapy (all P>0.05). Survival analysis showed that the 5-year disease-free survival rates between the two groups were 39.0% and 54.0%, respectively (P=0.025); the 5-year overall survival rates between the two groups were 41.8% and 58.1%, respectively (P=0.035); the difference were statistically significant. Multivariate analysis showed that ypT3-4 stage, ypN3b stage and △PLR were independent risk factors for 5-year disease-free survival rate (HR=2.731/2.676, 95%CI: 1.026-7.268/1.014-6.985; HR=4.717, 95%CI: 1.922-11.579; HR=2.854, 95%CI: 1.117-4.124; all P<0.05) and 5-year overall survival rate (HR=3.226/2.655, 95%CI: 1.280-9.227/0.945-7.548; HR=4.550, 95%CI: 1.842-11.239; HR=2.897, 95%CI: 1.049-5.251; all P<0. 05). Conclusion: △PLR can better predict the prognosis of AGC patients receiving neoadjuvant chemotherapy.

[Levels of sgRNA as a Major Factor Affecting CRISPRi Knockdown Efficiency in K562 Cells].

To determine how nuclease deactivated Cas9 (dCas9) or single-guide RNA (sgRNA) expression levels affect the knockdown efficiency of CRISPRi, we created K562 cell clones expressing KRAB-dCas9 protein either with the inducible Tet-on system or with the constitutive SFFV promotor. Single clones were selected by fluorescence-activated cell sorting (FACS) for further study. Six genes with various expression levels were targeted using lentiviral sgRNA from two libraries in four cell clones with various KRAB-dCas9 expression levels. The expression level of dCas9 protein/sgRNA levels and the knockdown efficiency were determined by flow cytometry. The cell clone with the highest KRAB-dCas9 expression level achieved effective CRISPRi knockdown. The data describing this clone were statistically different from that on other clones, indicating the strong KRAB-dCas9 expression might be a prerequisite for CRISPRi. By adopting different multiplicity of infection (MOI) in lentiviral transduction of this clone, we modified the expression level of sgRNA and found that the knockdown efficiency was neither affected by the target gene expression level nor correlated with KRAB-dCas9 levels, which remained relatively constant across all knockdown experiments (coefficient of variation = 2.2%). As an example, the following levels of the knockdowns: 74.72, 72.28 and 39.08% for mmadhc, rpia and znf148 genes, respectively, were achieved. These knockdown efficiencies correlated well with the respective sgRNA expression levels. Linear regression models built using this data indicate that the knockdown efficiency may be significantly affected by the levels of both KRAB-dCas9 and sgRNA. Notably, the sgRNA levels have greater impact, being a major factor affecting CRISPRi efficiency.

Chemical mapping of a single molecule by plasmon-enhanced Raman scattering.

Visualizing individual molecules with chemical recognition is a longstanding target in catalysis, molecular nanotechnology and biotechnology. Molecular vibrations provide a valuable 'fingerprint' for such identification. Vibrational spectroscopy based on tip-enhanced Raman scattering allows us to access the spectral signals of molecular species very efficiently via the strong localized plasmonic fields produced at the tip apex. However, the best spatial resolution of the tip-enhanced Raman scattering imaging is still limited to 3-15 nanometres, which is not adequate for resolving a single molecule chemically. Here we demonstrate Raman spectral imaging with spatial resolution below one nanometre, resolving the inner structure and surface configuration of a single molecule. This is achieved by spectrally matching the resonance of the nanocavity plasmon to the molecular vibronic transitions, particularly the downward transition responsible for the emission of Raman photons. This matching is made possible by the extremely precise tuning capability provided by scanning tunnelling microscopy. Experimental evidence suggests that the highly confined and broadband nature of the nanocavity plasmon field in the tunnelling gap is essential for ultrahigh-resolution imaging through the generation of an efficient double-resonance enhancement for both Raman excitation and Raman emission. Our technique not only allows for chemical imaging at the single-molecule level, but also offers a new way to study the optical processes and photochemistry of a single molecule.

[Impact of gender and age on in-hospital major adverse cardiovascular and cerebrovascular events of patients with acute ST-segment elevation myocardial infarction].

Objective: To investigate the impact of gender and age on in-hospital major adverse cardiovascular and cerebrovascular events of patients with acute ST-segment elevation myocardial infarction (STEMI). Methods: This is a retrospective single-center study. A total of consecutive 1 102 patients with acute STEMI admitted to our hospital from January 2001 to December 2010 were recruited and clinical data were analyzed. The primary end point was in-hospital death due to any cause, and the secondary end point was in-hospital composite end point including death, re-infarction and stroke. Multivariate logistic regression analyses were performed to identify the risk factors of in hospital death and composite end point. Results: The study population included 283(25.7%(283/1 102)) female patients and female patients were older than male patients ((68.7±11.2)years vs. (59.2±12.5)years, P<0.001). Compared with male patients, less female patients received primary percutaneous coronary intervention (50.9%(144/283) vs. 70.9%(581/819), P<0.001), had higher rates of in hospital death(10.6%(30/283)vs. 6.0%(36/819), P<0.001) and composite endpoint(14.1%(40/283)vs. 7.0%(57/819), P<0.001). Among STEMI patients aged <60 years, no differences were found in in-hospital mortality (1.7%(1/58)vs. 1.4%(6/437)) and composite endpoint(3.6%(3/58)vs. 3.4%(15/437)) rates between female and male patients (both P>0.05). Among STEMI patients aged ≥60 years, female patients had higher in-hospital mortality (12.9%(29/225)vs. 7.9%(30/382), P<0.001), and there was no difference on composite endpoint between female and male patients (16.4%(37/225)vs. 11.0%(42/382), P=0.054). Multivariate logistic regression analysis showed that female gender was not the independent risk factor of in-hospital death(OR=1.029, 95%CI 0.564-1.877, P=0.926) and composite end point(OR=1.593, 95%CI 0.989-2.566, P=0.055), but age was the independent risk factor of in-hospital death(OR=1.065, 95%CI 1.037-1.094, P<0.001) and composite end point(OR=1.050, 95%CI 1.029-1.071, P<0.001)in STEMI patients. Multivariate logistic regression analysis also showed that female was not the independent risk factor of in-hospital death(OR=1.539, 95%CI 0.572-4.142, P=0.394) and composite end point(OR=1.563, 95%CI 0.689-3.546, P=0.285), but age was the independent risk factor of in-hospital death(OR=1.052, 95%CI 1.011-1.096, P=0.013) and composite end point(OR=1.042, 95%CI 1.008-1.077, P=0.016)in STEMI patients received primary percutaneous coronary intervention. Conclusion: Female patients with STEMI have higher incidence of in-hospital major adverse cardiovascular and cerebrovascular events than male patients, and age is the independent risk factor of in-hospital major adverse cardiovascular and cerebrovascular events of STEMI patients.

A new method to induce molecular low bias negative differential resistance with multi-peaks.

According to a first-principles study of the transport properties of two thiolated anthracene-9,10-diono molecules sandwiching ethyl, a new method to induce molecular low bias negative differential resistance with multi-peaks for strong n- or p-type molecules is proposed. The anthracene-9,10-diono molecule shows strong n-type characteristics when in contact with Au and Ag electrodes via a thiolate. The multiple negative differential resistance effect originated from the molecule-electrode couple is different between Ag and Au electrodes. Our investigations may promise potential for applications in molecular devices with low power dissipation and multifunction in the future.

A brief introduction to chemical proteomics for target deconvolution.

OBJECTIVE: Drug-target relationships provide the basis for network-based polypharmacology, and target deconvolution is a key step in phenotypic-screening based drug discovery. Due to the complexity of the mammalian proteomics and the often-limited affinity of the lead compound, it is challenging to identify the drug targets, especially when the goal is to identify all targets. This paper attempts to provide a brief and comprehensive introduction to the various methods in chemical proteomics for target deconvolution by categorizing them into two groups: the biochemical enrichment and the proteomics-screening methods. Moreover, a brief introduction of related Mass Spectrometry techniques is also provided, together with recent progress. MATERIALS AND METHODS: The data for this review were queried from Web of Science and PubMed, the keywords used were Drug targets, Target deconvolution, and Chemical Proteomics. A total of over 500 relevant articles, with a time limit from 1953 to 2022, were identified according to search strategy. Duplicate records and review articles were excluded by their titles and abstracts. Finally, we found about 120 articles matching our inclusion criteria, which covered representative research and reviews of various target discovery methods. RESULTS: Existing target discovery methods can be grouped into either biochemical enrichment or the proteomics-screening methods, with the recent emergence of a hybrid method combining these two such as lysine reactivity profiling. The advantage of the biochemical enrichment method is the ease of operation and the comprehensive target coverage. However, most biochemical enrichment methods require a high-affinity binding of the drug to the target proteins and cannot differentiate direct/indirect targets. The proteomics-screening methods do not require drug modification but have limited protein coverage, and most of them cannot differentiate direct/indirect targets. CONCLUSIONS: Although existing target discovery methods have greatly facilitated pharmacological research, each of these methods has advantages and disadvantages. New strategies/methods are needed to further improve both the coverage of the proteosome and the specificity.

[A cross-sectional study on psychological needs of family members of critically ill inpatients in semi-closed wards and analysis of influence factors].

Objective: To analyze and discuss the psychological needs of family members of critically ill burn inpatients in semi-closed wards and their related influence factors. Methods: A cross-sectional survey was conducted among the 82 burn patients, who met the inclusion criteria and were admitted to the Department of Burns and Plastic Surgery of Ruijin Hospital of Shanghai Jiao Tong University School of Medicine from October 2018 to April 2019, and their family members. A general condition questionnaire of 15 indexes was applied to investigate the general data of burn patients and their family members including cause of burn, gender, age, total burn area, burn depth, burn severity, burn site, and type of medical insurance of patients, kinship with the patients, gender, age, experience of accompanying in intensive care isolation wards, occupation, education level, and monthly income of their family members. The Critical Care Family Needs Inventory (CCFNI) was used to assess each item, each factor, and total scores of the family members of patients including 5 factors and 45 items such as support scale (SS), comfort scale (CS), information scale (IS), proximity scale (PS), assurance scale (AS). CCFNI was used to assess the total scores of patients and their family members after classification according to the general data, the data were statistically analyzed with one-way analysis of variance or independent sample t test. Indicators with statistical significance in the above analysis were selected for multiple linear regression analysis to screen the independent influence factors of psychological needs of patients' family members. CCFNI was used to assess each dimension scores of patients and their family members after classification according to the aforementioned selected independent influence factors including age and burn area of patients, gender, educational level, and experience of accompanying in intensive care isolation wards of their family members, and the data were statistically analyzed with one-way analysis of variance or independent sample t test. Results: A total of 82 questionnaires were sent out in this study, and 80 were effectively received, with s a recovery rate of 97.6%. In 80 patients, there were 45 males and 35 females; 35 cases (43.75%) were ≤14 years; most of the patients had flame and thermal burns; 33 patients (41.25%), 21 patients (26.25%), and 26 patients (32.50%) had moderate, severe, and extremely severe burns, respectively; and only 3 patients (3.75%) had no health insurance. In 80 family members of patients, 34 (42.50%) were males and 46 (57.50%) were females; 62.50% in the family members of patients were concentrated between 18 and 45 years old; and 12 family members of patients had experience of accompanying in intensive care isolation wards, accounting for 15.00%. The total CCFNI score of the family members was (141±14) points, and the scores of AS, PS, IS, CS, and SS were (3.77±0.23), (3.43±0.37), (3.53±0.34), (2.50±0.59), and (2.69±0.45) points, respectively. The top five items of the psychological needs of family members of patients were concentrated in three factors (AS, PS, and IS), and the item "ensuring that patients receive the best treatment" ranked the first, with a score of (3.99±0.11) points. There were significant differences on the CCFNI total scores of patients with different age, total burns area, and severity of burns and of their family members with different gender, education level, and whether or not experience of accompanying in intensive care isolation unit (F=4.378, 5.481, 5.913, t=4.027, 14.339, 7.265, P<0.05 or P<0.01). Multiple linear regression analysis showed that the patient's age (≤14, 15-45 years) and total burn area (11%-30%, 31%-49%, ≥50% TBSA) and the patient's family members' gender, educational level, experience of accompanying in intensive care isolation unit could affect the psychological needs of family members of patients (t=3.133, 2.260, 2.297, 2.433, 3.015, 2.200, 2.102, 2.463, P<0.05 or P<0.01). There was statistically significant difference in AS score of the patients' family members with different age (F=4.390, P<0.05). There were significant differences in both IS and AS scores of the patients' family members of patients with different burn area (F=5.042, 3.131, P<0.05 or P<0.01). The AS score of the patients' family members with experience of accompanying in intensive care isolation unit was significantly lower than those without (t=-1.040, P<0.05). The SS score of the patients' family members with college education or above was significantly higher than those with high school (technical secondary school) or below (t=0.657, P<0.05). The IS score of the patients' family members with college education or above was significantly lower than those with high school (technical secondary school) or below (t=-1.438, P<0.05). The SS score of male family members of patients was significantly lower than those of female patients (t=4.149, P<0.05). Conclusions: The family members of burn patients in semi-closed ward have the most urgent need to ensure the patients receive the best treatment. Patients' age, total burn area, and their family members' gender, education level, experience of accompanying in intensive care isolation unit are the main factors affecting the psychological needs of family members of patients.

Target inhibition on GSK-3ß by miR-9 to modulate proliferation and apoptosis of bladder cancer cells.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Target inhibition on GSK-3ß by miR-9 to modulate proliferation and apoptosis of bladder cancer cells, by Z.-C. Dong, D. Zhang, S.-B. Wang, Z.-Q. Lin, published in Eur Rev Med Pharmacol Sci 2018; 22 (10): 3018-3026-DOI: 10.26355/eurrev_201805_15059-PMID: 29863246" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/15059.

MiR-155 affects proliferation and apoptosis of bladder cancer cells by regulating GSK-3ß/ß-catenin pathway.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "MiR-155 affects proliferation and apoptosis of bladder cancer cells by regulating GSK-3ß/ß-catenin pathway, by Z.-C. Dong, D. Zhang, X.-X. Zhang, Z.-Q. Yao, H. Wu, C.-H. Chen, J.-Q. Tian, published in Eur Rev Med Pharmacol Sci 2019; 23 (13): 5682-5690-DOI: 10.26355/eurrev_201907_18305-PMID: 31298320" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18305.

Influence of a dielectric layer on photon emission induced by a scanning tunneling microscope.

We investigate theoretically the influence of a dielectric layer on light emission induced by a scanning tunneling microscope through a combined approach of classical electrodynamics and first-principles calculations. The modification of the junction geometry upon the insertion of a dielectric layer is treated first by using the density functional theory to calculate the effective potential along the surface normal and then by solving a one-dimensional Schrodinger equation to obtain the exact distance between the tip and the substrate for a given current and bias voltage. The modified external field with the inclusion of a dielectric layer is evaluated by using the Fresnel formula. The local-field enhancement factor and radiated power are calculated by the boundary element method for two typical systems, W-tip/C(60)/Au(111) and W-tip/Al(2)O(3)/NiAl(110). The calculated results indicate that the insertion of a dielectric layer tends to reduce the light emission intensity considerably but hardly changes the spectral profile with no substantial peak shifts with respect to the layer-free situation, in agreement with experimental observations. The suppression of the radiated power is mainly due to the increase in the tip-metal separation and the resultant reduction in the electromagnetic coupling between the tip and metal substrate.

MiR-155 affects proliferation and apoptosis of bladder cancer cells by regulating GSK-3ß/ß-catenin pathway.

OBJECTIVE: GSK-3ß negatively regulates Wnt/ß-catenin signaling pathway. The abnormal miR-155 expression is associated with bladder cancer. Bioinformatics analysis revealed a complementary binding site between miR-155 and GSK-3ß mRNA. This study investigated the role of miR-155 in the proliferation and apoptosis of bladder cancer cells. PATIENTS AND METHODS: The dual luciferase reporter gene assay validated the targeted regulation between miR-155 and GSK-3ß. Tumor tissues and adjacent tissues were collected from bladder cancer patients and the expression of miR-155 and GSK-3ß mRNA was detected by RT-PCR. Bladder cancer cell line BIU-87 cells were cultured in vitro and divided into miR-NC group and miR-155 inhibitor group. The expressions of miR-155, GSK-3ß and ß-catenin were compared, cell apoptosis was detected by flow cytometry, and cell proliferation was detected by EdU staining. RESULTS: Compared with adjacent tissues, miR-155 expression was significantly increased in bladder cancer tissues, and GSK-3ß mRNA expression was significantly decreased. There was a targeted regulatory relationship between miR-155 and GSK-3ß. Compared with SV-HUC-1 cells, miR-155 expression in bladder cancer BIU-87 and 5637 cells was significantly increased, and GSK-3ß expression was significantly decreased. Transfection of miR-155 inhibitor significantly increased GSK-3ß expression in BIU-87 and 5637 cells, decreased ß-catenin expression, increased cell apoptosis, and decreased cell proliferation. CONCLUSIONS: The increased expression of miR-155 plays a role in reducing the expression of GSK-3ß and in promoting the pathogenesis of bladder cancer. Inhibition of miR-155 can up-regulate the expression of GSK-3ß, inhibit the activity of Wnt/ß-catenin pathway, attenuate proliferation and promote apoptosis of bladder cancer cells.

Target inhibition on GSK-3ß by miR-9 to modulate proliferation and apoptosis of bladder cancer cells.

OBJECTIVE: Glycogen synthase kinase-3ß (GSK-3ß) can negatively regulate Wnt/ß-catenin signaling pathway via degrading ß-catenin protein, and plays suppressing roles in various tumors. Its role in bladder cancer pathogenesis is still unclear. In bladder cancer tissues, expression of microRNA-9 (miR-9) is significantly elevated. This study investigated the effect of miR-9 in modulating GSK-3ß expression, Wnt/ß-catenin pathway activity, and proliferation or apoptosis of bladder cancer cells. PATIENTS AND METHODS: Dual luciferase reporter gene assay confirmed targeted regulation between miR-9 and GSK-3ß. Bladder cancer tissues were collected to measure expression of miR-9, GSK-3ß mRNA using adjacent tissues as the control. Expression of miR-9 and GSK-3ß was also measured in HBEC, RT4 and TCCSUP cells. Cultured RT4 and TCCSUP cells were transfected with miR-9 inhibitor or pSicoR-GSK-3ß. The expression of miR-9, GSK-3ß and ß-catenin was compared, followed by using flow cytometry assay for cell apoptosis and EdU staining for cell proliferation. RESULTS: Comparing to adjacent tissues, bladder cancer tissues illustrated significantly elevated miR-9 expression and lower GSK-3ß mRNA. Bioinformatics analysis revealed complementary binding sites between miR-9 and 3'-UTR of GSK-3ß mRNA, indicating targeted regulation between miR-9 and GSK-3ß. Comparing to HBEC cells, RT4 and TCCSUP cells had significantly elevated miR-9 expression and lower GSK-3ß expression, with enhanced proliferation. Transfection of miR-9 inhibitor or pSicoR-GSK-3ß significantly elevated GSK-3ß expression and suppressed ß-catenin expression, promoted cell apoptosis and inhibited proliferation. CONCLUSIONS: MiR-9 up-regulation plays a role in suppressing GSK-3ß expression and facilitating bladder cancer pathogenesis. Inhibition of miR-9 could potentiate GSK-3ß expression, suppress proliferation of bladder cancer, and facilitate apoptosis.

Recycled cafeteria food waste as a feed for swine: nutrient content digestibility, growth, and meat quality.

This project was undertaken to compare growth, meat quality, and diet digestibility when pigs were fed cafeteria food waste (FW) or a corn/soybean meal (CSM) diet. Cafeteria food waste (36 samples) fed in the growing and finishing experiment averaged 22.4% DM, 21.4% CP, 14.1% ADF, 27.2% ether extract, and 3.2% ash. The first experiment used 50 crossbred pigs randomly assigned to four diets. During the growing phase, pigs fed a CSM diet gained faster (P < .05) than pigs fed FW or FW plus energy supplements. However, the two groups fed FW plus energy supplements (at 25 or 50% of the intake of the CSM diet) gained faster (P < .05) than pigs fed FW alone (.61 and .65 kg/d, respectively, vs .46 kg/d). In the finishing phase, FW plus an energy supplement fed at 50% of the level of CSM intake resulted in gains that did not differ from those of pigs fed the CSM diet (.90 vs .99 kg/d; P > .05). A nutrient digestibility and nitrogen balance trial using eight growing barrows compared FW with the same CSM growing diet fed earlier. Dry matter digestibility was similar for the two diets (P > .05). However, CP digestibility was higher (P < .05) in the FW diet than in the CSM diet (88.2 vs 84.3%). Although the percentage of nitrogen retained was not different between FW and CSM diets (56.0 vs 55.2%; P > .05), the amount of nitrogen retained was greater for pigs fed the CSM diet (29.3 vs. 24.5 g/d; P < .05) because DMI was greater (1.7 vs 1.4 kg/d) for pigs fed CSM compared with FW. At the completion of the finishing experiment, six pigs were selected from both the CSM and FW diets and fed to finishing weight. The pigs were slaughtered, and the pork loins were removed for flavor and texture analysis. A consumer panel rated the meat quality from FW pigs as acceptable and overall flavor comparable to CSM pigs (P > .05). These results indicate that food waste has nutritive value and may be useful in swine diets.

Note: optical optimization for ultrasensitive photon mapping with submolecular resolution by scanning tunneling microscope induced luminescence.

We report the development of a custom scanning tunneling microscope equipped with photon collection and detection systems. The optical optimization includes the comprehensive design of aspherical lens for light collimation and condensing, the sophisticated piezo stages for in situ lens adjustment inside ultrahigh vacuum, and the fiber-free coupling of collected photons directly onto the ultrasensitive single-photon detectors. We also demonstrate submolecular photon mapping for the molecular islands of porphyrin on Ag(111) under small tunneling currents down to 10 pA and short exposure time down to 1.2 ms/pixel. A high quantum efficiency up to 10(-2) was also observed.

Fabrication of silver tips for scanning tunneling microscope induced luminescence.

We describe a reliable fabrication procedure of silver tips for scanning tunneling microscope (STM) induced luminescence experiments. The tip was first etched electrochemically to yield a sharp cone shape using selected electrolyte solutions and then sputter cleaned in ultrahigh vacuum to remove surface oxidation. The tip status, in particular the tip induced plasmon mode and its emission intensity, can be further tuned through field emission and voltage pulse. The quality of silver tips thus fabricated not only offers atomically resolved STM imaging, but more importantly, also allows us to perform challenging "color" photon mapping with emission spectra taken at each pixel simultaneously during the STM scan under relatively small tunnel currents and relatively short exposure time.

Vibrationally resolved fluorescence from organic molecules near metal surfaces in a scanning tunneling microscope.

Intrinsic molecular fluorescence from porphyrin molecules on Au(100) has been realized by using a nanoscale multimonolayer decoupling approach with nanoprobe excitation in the tunneling regime. The molecular origin of luminescence is established by the observed well-defined vibrationally resolved fluorescence spectra. The molecules fluoresce at low "turn-on" voltages for both bias polarities, suggesting an excitation mechanism via hot electron injection from either tip or substrate. The excited molecules decay radiatively through Franck-Condon pi(*)-pi transitions.