SPINK5 inhibits esophageal squamous cell carcinoma metastasis via immune activity.

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is a predominant subtype of esophageal cancer with relatively high mortality worldwide. Serine peptidase inhibitor Kazal-type 5 (SPINK5) is reported to be downregulated in ESCC. However, its explicit role in ESCC remains further investigation. METHODS: The tumor tissues and adjacent non-cancerous tissues were obtained from 196 patients with ESCC for the determination of SPINK5 mRNA levels. Additionally, the relationship between SPINK5 mRNA levels and clinicopathological features of ESCC patients was explored. The effects of SPINK5 on the invasion and migration of ESCC cells were assessed using Transwell assays. Furthermore, SPINK5 mRNA and LEKTI protein were measured in ESCC cell lines after treatment with poly (I:C), lipopolysaccharide (LPS) or unmethylated CpG DNA. Moreover, the correlation between expression of SPINK5 and nuclear factor-kappa B (NF-κB) signaling pathway-related genes was analyzed in the TCGA-ESCC cohort, and the effects of SPINK5 on NF-κB transcription was analyzed using a luciferase reporter gene assay. Finally, the correlations between SPINK5 and infiltration of immune cells, immune scores, stromal scores and ESTIMATE (i.e., Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data) scores were explored. RESULTS: SPINK5 mRNA levels were downregulated in tumor tissues, which was significantly correlated with higher lymph node metastases. Overexpressed SPINK5 inhibited cell invasion and migration in ESCC cell lines. Mechanistically, LPS-induced activation of Toll-like receptor 4 (TLR4) decreased SPINK5 mRNA and LEKTI in KYSE150 and KYSE70 cells. Spearman correlation analysis revealed that SPINK5 mRNA was significantly negatively correlated with a total of seven NF-κB signaling pathway-related genes in TCGA-ESCC patients. Moreover, downregulation of SPINK5 increased and upregulation of SPINK5 decreased the activity of the NF-κB promoter in HEK293T cells. Finally, immune cells infiltration analysis revealed that SPINK5 was significantly correlated with the infiltration of various immune cells, stromal scores, immune scores and ESTIMATE scores. CONCLUSIONS: SPINK5 plays critical roles in the TLR4/NF-κB pathway and immune cells infiltration, which might contribute to the ESCC metastasis. The findings of the present study may provide a promising biomarker for the diagnosis and treatment of esophageal squamous cell carcinoma.

Characterization of urease active calcite-producing strain YX-3 combined with the whole genome.

Urease found in a wide range of microorganisms plays a vital role in ureolytic induced calcite precipitation (UICP). However, the genomic information on urease-producing strains is limited, and there is a need for further in-depth studies on aspects such as the regulation of urease activity by nickel ligand residues. The present study delved into the elucidation of urease activity in a newly isolated strain YX-3 coupled with nickel-ligand residues by employing the genetic architecture of biomineralization-controlled growth, molecular docking, molecular dynamics simulation (MDS), and site-directed mutagenesis. Genome-wide sequencing showed the presence of urease gene clusters, comprising structural genes ureA, ureB, and ureC, alongside auxiliary genes ureD, ureE, ureF, and ureG. RT-qPCR analysis showed that the addition of NiCl2 resulted in a significant up-regulation of ureC expression. His267, His294, and Gly325 in the domain of UreC were further proved to coordinate with nickel ions and urea simultaneously through homology modeling and molecular docking, and molecular dynamics simulations (MDS) showed the urease-urea docking complexes exhibited degressive binding stability by four metrics including root mean square deviations (RMSD) when those residues were mutated into alanine respectively. Western blotting exhibited that mutations of H267A, H294A, and G325A led to a reduction in the relative expression of urease, wherein urease activity was about 62%, 45%, and 20% times that of the wild type (WT), respectively. The overexpression results further confirmed the importance of these residues for urease activity and CaCO3 precipitation. These results would help to deepen the understanding of urease-producing strains at a molecular level and expand the theoretical basis for modulating urease activity.

Targeted and Equally Distributed Delivery of mRNA to Organs with Pentaerythritol-Based One-Component Ionizable Amphiphilic Janus Dendrimers.

Delivery of nucleic acids with viral and synthetic vectors has pioneered genetic nanomedicine. Four-component lipid nanoparticles (LNPs) consisting of ionizable lipids, phospholipids, cholesterol, and PEG-conjugated lipids, assembled by microfluidic or T-tube, are the benchmark synthetic vector for delivery of mRNA. One-component multifunctional sequence-defined ionizable amphiphilic Janus dendrimer (IAJD) delivery systems for mRNA were developed by us to complement LNPs. IAJDs consist of multifunctional hydrophilic low-generation dendrons or minidendrons conjugated to hydrophobic dendrons. They were inspired by amphiphilic Janus dendrimers and glycodendrimers. IAJDs coassemble with mRNA into predictable-size vesicles, named dendrimersome nanoparticles (DNPs), by simple injection in acetate buffer, rather than by the complex technology required by LNPs. Assembly of DNPs by simple injection together with sequence design in the hydrophilic and hydrophobic modules of IAJDs endowed rapid screening to access discovery. Molecular design principles for targeted delivery were elaborated when the branching points of IAJDs were constructed from symmetrically and nonsymmetrically substituted plant phenolic acids interconnected by pentaerythritol (PE). Here, we report the first library containing simplified IAJDs constructed in only three steps from symmetrically trialkylated PE in the hydrophobic domain and four different piperazine-based ionizable amines in the hydrophilic part. Rapid coassembly with mRNA and in vivo screening led to the discovery of the two most active IAJDs targeting the spleen, liver, and lymph nodes, one predominantly to the spleen and liver and six delivering equally to the spleen, liver, lung, and lymph nodes. These IAJDs represent the simplest synthetic vectors and the first viral or synthetic system delivering equally to multiple organs.

The Unexpected Importance of the Primary Structure of the Hydrophobic Part of One-Component Ionizable Amphiphilic Janus Dendrimers in Targeted mRNA Delivery Activity.

Viral and synthetic vectors for delivery of nucleic acids impacted genetic nanomedicine by aiding the rapid development of the extraordinarily efficient Covid-19 vaccines. Access to targeted delivery of nucleic acids is expected to expand the field of nanomedicine beyond most expectations. Both viral and synthetic vectors have advantages and disadvantages. The major advantage of the synthetic vectors is their unlimited synthetic capability. The four-component lipid nanoparticles (LNPs) are the leading nonviral vector for mRNA used by Pfizer and Moderna in Covid-19 vaccines. Their synthetic capacity inspired us to develop a one-component multifunctional sequence-defined ionizable amphiphilic Janus dendrimer (IAJD) delivery system for mRNA. The first experiments on IAJDs provided, through a rational-library design combined with orthogonal-modular accelerated synthesis and sequence control in their hydrophilic part, some of the most active synthetic vectors for the delivery of mRNA to lung. The second experiments employed a similar strategy, generating, by a less complex hydrophilic structure, a library of IAJDs targeting spleen, liver, and lung. Here, we report preliminary studies designing the hydrophobic region of IAJDs by using dissimilar alkyl lengths and demonstrate the unexpectedly important role of the primary structure of the hydrophobic part of IAJDs by increasing up to 90.2-fold the activity of targeted delivery of mRNA to spleen, lymph nodes, liver, and lung. The principles of the design strategy reported here and in previous publications indicate that IAJDs could have a profound impact on the future of genetic nanomedicine.

Targeted Delivery of mRNA with One-Component Ionizable Amphiphilic Janus Dendrimers.

Targeted and efficient delivery of nucleic acids with viral and synthetic vectors is the key step of genetic nanomedicine. The four-component lipid nanoparticle synthetic delivery systems consisting of ionizable lipids, phospholipids, cholesterol, and a PEG-conjugated lipid, assembled by microfluidic or T-tube technology, have been extraordinarily successful for delivery of mRNA to provide Covid-19 vaccines. Recently, we reported a one-component multifunctional sequence-defined ionizable amphiphilic Janus dendrimer (IAJD) synthetic delivery system for mRNA relying on amphiphilic Janus dendrimers and glycodendrimers developed in our laboratory. Amphiphilic Janus dendrimers consist of functional hydrophilic dendrons conjugated to hydrophobic dendrons. Co-assembly of IAJDs with mRNA into dendrimersome nanoparticles (DNPs) occurs by simple injection in acetate buffer, rather than by microfluidic devices, and provides a very efficient system for delivery of mRNA to lung. Here we report the replacement of most of the hydrophilic fragment of the dendron from IAJDs, maintaining only its ionizable amine, while changing its interconnecting group to the hydrophobic dendron from amide to ester. The resulting IAJDs demonstrated that protonated ionizable amines play dual roles of hydrophilic fragment and binding ligand for mRNA, changing delivery from lung to spleen and/or liver. Replacing the interconnecting ester with the amide switched the delivery back to lung. Delivery predominantly to liver is favored by pairs of odd and even alkyl groups in the hydrophobic dendron. This simple structural change transformed the targeted delivery of mRNA mediated with IAJDs, from lung to liver and spleen, and expands the utility of DNPs from therapeutics to vaccines.

Comparison between gravity drainage group and suction drainage group after cervical laminoplasty: a retrospective STROBE-compliant cohort study.

OBJECTIVES: A retrospective study was conducted to compare gravity drainage and suction drainage after cervical laminoplasty. PATIENTS AND METHODS: A total of 375 patients who underwent laminoplasty between January 2011 and December 2015 were engaged in this analysis. We investigated the patients' basic characteristics, drainage characteristics and postoperative complications. RESULTS: During the initial 24 h after laminoplasty, the drainage volume in the suction drainage group was 177.31 ± 92.02 mL, and the drainage volume in the gravity drainage group was 133.33 ± 92.40 mL. The drainage volume showed significant difference (p < 0.01). The total drainage volume was 357.49 ± 195.16 mL and 250.16 ± 27.44 mL in the suction drainage group and gravity drainage group, respectively. The total drainage volume between the two groups was statistically different (p = 0.03). The postoperative Hb was significantly different between the gravity group and suction group on the first day after the operation (108.37 ± 23.92 mL vs. 87.32 ± 21.53 mL, p = 0.02). The number of patients required blood transfusion was significantly different between the two groups as well (p = 0.04). Two cases had symptomatic epidural hematomas (SEH) after laminoplaty. However, the occurrence of SEH among the two groups was not different significantly. Twelve patients had surgical site infection (SSI). Of these 12, nine had applied gravity drainage and three suction drainage. The rate of SSI was similar between the two groups (p = 0.71). CONCLUSION: The initial 24 h' drainage volume and the total drainage volume increased significantly in the suction drainage group. The postoperative Hb was lower in the suction group than the gravity drainage group the first postoperative day. More patients needed blood transfusion if suction drainage was performed. The application of suction drainage cannot decrease the incidence of SSI and SEH after laminoplasty. Gravity drainage is recommended for laminoplasty.

Simultaneous real-time detection of fractional exhaled nitric oxide and end-tidal carbon dioxide by quantum cascade laser absorption spectroscopy.

The simultaneous detection of fractional exhaled nitric oxide (FeNO) and end-tidal carbon dioxide (ETCO2) is of great importance for the distinguishing and diagnosis of asthma and chronic obstructive pulmonary disease (COPD), providing more comprehensive information on respiratory disorders. This work demonstrates a simultaneous ETCO2 and FeNO detection system based on quantum cascade laser absorption spectroscopy (QCLAS) technology was presented. The system employs wavelength modulation spectroscopy (WMS) technology and the Herriott multi-pass cell, achieving a detection limit of 2.82 ppb for nitric oxide (NO) and 0.05 % for carbon dioxide (CO2). Real-time exhalation measurements were performed on volunteers with varying ETCO2 and FeNO levels, and the results of the test can accurately distinguish whether the corresponding volunteer was healthy, had asthma or COPD. The effect of exhalation flow rate on the concentration of the two gases was explored. A range of expiratory flow rates were tested in the flow rate interval from 1 to 4 L/min, and there was always an inverse relationship between expiratory flow rate and FeNO concentration, but flow rate changes did not affect ETCO2 concentration. The results indicate that this detection system can simultaneously and effectively measure ETCO2 and FeNO concentrations in real-time.

Continuous real-time monitoring of carbon dioxide emitted from human skin by quartz-enhanced photoacoustic spectroscopy.

In this study, a skin gas detection system based on quartz enhanced photoacoustic spectroscopy (QEPAS) with a constant temperature collection chamber and an automatic frequency adjustment function was used to collect and monitor carbon dioxide (CO2) emissions from human skin. The detection element of the system is an on-beam structure assembled by a 30.72 kHz quartz tuning fork (QTF). A laser with a wavelength of 4991.26 cm-1 is emitted (with a wavelength adjustment range of 10 cm-1) to excite the QTF. When the integration time is 365 s, the system can achieve a minimum detection limit (MDL) of 2.6 ppmv. The sensitivity of the system is 636.9 ppmv/V. The gas detection system is used to monitor the concentration of CO2 emissions from different parts of the skin and the same part covered by different cosmetics. The CO2 emission rate is defined as the ratio of the skin gas monitoring time of 25 min to the CO2 concentration variable in the gas chamber (volume of 8 mL). The results were collected from three healthy volunteers. Among the six different parts, the cheeks emitted the fastest rate (the average rate was 365.5 ppmv/min) of CO2, and the thighs emitted the slowest rate (the average rate was 56.4 ppmv/min) of CO2. Comparing the experimental results of the six sites at different times, the order of the CO2 emission rate is identical for all six sites. In the experiments with the three cosmetic products (experimental site: forearm), comparing the CO2 emission rate from clean skin with the CO2 emission rate from cosmetic-covered skin shows that sunscreen is the most breathable, followed by barrier cream, and foundation is the least breathable.

Screening Libraries to Discover Molecular Design Principles for the Targeted Delivery of mRNA with One-Component Ionizable Amphiphilic Janus Dendrimers Derived from Plant Phenolic Acids.

Viral and synthetic vectors to deliver nucleic acids were key to the rapid development of extraordinarily efficient COVID-19 vaccines. The four-component lipid nanoparticles (LNPs), containing phospholipids, PEG-conjugated lipids, cholesterol, and ionizable lipids, co-assembled with mRNA via a microfluidic technology, are the leading nonviral delivery vector used by BioNTech/Pfizer and Moderna to access COVID-19 mRNA vaccines. LNPs exhibit a statistical distribution of their four components when delivering mRNA. Here, we report a methodology that involves screening libraries to discover the molecular design principles required to realize organ-targeted mRNA delivery and mediate activity with a one-component ionizable multifunctional amphiphilic Janus dendrimer (IAJD) derived from plant phenolic acids. IAJDs co-assemble with mRNA into monodisperse dendrimersome nanoparticles (DNPs) with predictable dimensions, via the simple injection of their ethanol solution in a buffer. The precise location of the functional groups in one-component IAJDs demonstrated that the targeted organs, including the liver, spleen, lymph nodes, and lung, are selected based on the hydrophilic region, while activity is associated with the hydrophobic domain of IAJDs. These principles, and a mechanistic hypothesis to explain activity, simplify the synthesis of IAJDs, the assembly of DNPs, handling, and storage of vaccines, and reduce price, despite employing renewable plant starting materials. Using simple molecular design principles will lead to increased accessibility to a large diversity of mRNA-based vaccines and nanotherapeutics.

Compact quartz-enhanced photoacoustic sensor for ppb-level ambient NO2 detection by use of a high-power laser diode and a grooved tuning fork.

A compact quartz-enhanced photoacoustic sensor for ppb-level ambient NO2 detection is demonstrated, in which a high-power blue laser diode module with a small divergence angle was employed to take advantages of the directly proportional relationship between sensitivity and power, hence improving the detection sensitivity. In order to extend the stability time, a custom grooved quartz tuning fork with 800-µm prong spacing is employed to avoid complex signal balance and/or optical spatial filter components. The sensor performance is optimized and assessed in terms of optical coupling, power, gas flow rate, pressure, signal linearity and stability. A minimum detectable concentration (1σ) of 7.3 ppb with an averaging time of 1 s is achieved, which can be further improved to be 0.31 ppb with an averaging time of 590 s. Continuous measurements covering a five-day period are performed to demonstrate the stability and robustness of the reported NO2 sensor system.

Integrated analysis of immune-related gene subtype and immune index for immunotherapy in clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is the most common type of renal cell carcinoma (RCC) with high immunogenicity. Research on immune-related gene (IRG) is of great significance in ccRCC in identifying new therapeutic targets and improving patient prognosis. In this study, the IRG patterns of ccRCC were investigated and correlated these patterns with tumor microenvironment infiltrating characteristics in immunotherapy. Moreover, an IRG score was constructed to quantify the pattern of individual tumors through the principal component analysis algorithm. Two distinct molecular subtypes (C1 and C2) were identified based on the IRGs expression profile. Subtype C1 was characterized with significantly high level of immune-checkpoint, immune score, stromal score, showed high drug sensitivity in Sorafenib, Sunitinib, Cisplatin, Vinblastine, Vinorelbine, Vorinostat, and Gemcitabine. Cytokine-cytokine receptor pathway, chemokine signaling pathway, and JAK signaling pathways were found enriched in the subtype C1 account for the poor prognosis. Subtype C2 was linked to a better survival outcome. By using the Connective Map database, subtype specific small molecular drugs identified that could facilitate the treatment of ccRCC patients. In addition, A immune index that used to evaluated the immune modification patterns and further validated in the other types RCC dataset, such as papillary renal cell carcinoma (pRCC) and chromophobe renal cell carcinoma (chRCC). Together, this study identified two distinct molecular subtypes with immune index, aid to the treatment of ccRCC and enhancing our cognition of the tumor microenvironment infiltration characterization in ccRCC immunotherapy.

Modified axial computed tomography classification of cervical ossification of the posterior longitudinal ligament: selecting the optimal operating procedure and enhancing the accuracy of prognosis.

BACKGROUND: Cervical ossification of the posterior longitudinal ligament (OPLL) causes spinal cord compression, which can lead to myelopathy or radiculopathy. Non-surgical treatments have little effect on this condition. Current OPLL classification systems offer little guidance on the selection of an appropriate operating procedure. In this study, we developed a modified axial computed tomography classification (MACTC) scheme. We then examined the usefulness of the MACTC scheme and two other existing classification schemes in guiding OPLL operation choice. METHODS: Following screening in which a defined exclusion criteria was used, a total of 91 patients with OPLL participated in the study. Patients' follow-up data for at least 2 years were obtained. The recovery rate of the Japanese Orthopaedic Association (JOA) scores was compared to two other classification schemes. RESULTS: According to the MACTC, central-sharp-type OPLL had a lower recovery rate of the JOA score than that of central-gentle-type OPLL (36.05±32.38 vs. 83.90±23.52, P≤0.05). The recovery rate of the JOA scores in the ipsilateral open-door OPLL group was significantly lower than that in the contralateral group of the lateral-steep type (36.67±41.5 vs. 88.89±17.21, P=0.04), but not of that in the lateral-gentle type. There was no significant difference in the recovery rates of the JOA scores between groups when using either existing classification scheme (P>0.05). CONCLUSIONS: The MACTC scheme can assist surgeons to choose the most appropriate operating procedure, and provide an accurate prognosis. If operations on central-sharp-type OPLL are not performed using both the posterior and anterior approaches, prognosis will be poor. The contralateral side should be the first choice for door opening in laminoplasty, especially for patients with lateral-steep-type OPLL. Severe OPLL may not be an absolute contraindication for the posterior approach.

Fenton treatment of bio-treated fermentation-based pharmaceutical wastewater: removal and conversion of organic pollutants as well as estimation of operational costs.

The Fenton process is used as a tertiary treatment to remove organic pollutants from the effluent of bio-treated pharmaceutical wastewater (EBPW). The optimal and most appropriate Fenton conditions were determined by an orthogonal array test and single-factor experiments. The removal of chemical oxygen demand (COD) was influenced by the following factors in a descending order: H2O2/Fe(II) molar ratio > H2O2 dosage > reaction time. Under the most appropriate Fenton conditions (H2O2/Fe(II) molar ratio of 1:1, H2O2 dosage of 120 mg L-1 and reaction time of 10 min), the COD and dissolved organic carbon (DOC) were removed with efficiencies of 62 and 53%, respectively, which met the national discharge standard (GB 21903-2008) for the Lake Tai Basin, China. However, the Fenton treatment was inadequate for removal of N compounds, and the removal of organic nitrogen led to an increment in N-NH3 from 3.28 to 19.71 mg L-1. Proteins and polysaccharides were completely removed, and humic acids (HAs) were partly removed with an efficiency of 55%. Three-dimensional excitation/emission matrix spectra (3DEEMs) indicated complete removal of fulvic acid-like substances and 90% reduction in the florescence intensity of humic acid-like substances. Organic pollutants with molecular weights (MW) > 10 kDa were completely removed, MW 5-10 kDa were degraded into smaller MW ones, and some low molecular weight acids (MW 0.1-1 kDa) were mineralized during the Fenton process. Some species, including pharmaceutical intermediates and solvents were detected by gas chromatography-mass spectrometry (GC-MS). The operational costs of the Fenton's treatment were estimated to be 0.58 yuan RMB/m3 EBPW based on reagent usage and iron sludge treatment and disposal.

Developing the Psychological Strain Scales (PSS): Reliability, Validity, and Preliminary Hypothesis Tests.

Since its inception, the Strain Theory of Suicide has been tested and supported in a number of empirical studies. This social psychological theory can be employed as a complementary conceptualization to account for suicidal behaviors as well as mental disorders. However, the lack of consistent measurements of the strains limits the application of the theory in scientific research. Our research team has developed such scales for future testing of the Strain Theory of Suicide in a more systematic approach. For the initial items to measure the four strains (value, aspiration, deprivation, and coping), we solicited approximately 40 items for each strain with high face validity by about 30 fellow researchers. A preliminary examination of about 160 items for consistency and validity, with a sample of about 300 college students, yielded 20 consistent items for each of the four strain scales. Then, a second study was conducted at a different university with approximately 500 students to further streamline each of the four strain scales and test the validity of each with corresponding established scales and variables. As a result, 15 items were selected for each of the four Psychological Strain Scales (PSS). In correlation and multiple regression analyses, we found support for the hypotheses regarding the positive associations between psychological strains measured by the PSS and psychopathology including suicidal ideation. Follow up research with the new scales needs to be carried out in order to test the effects of psychological strains on suicide and mental disorders for various populations.

[Propensity score matching-based therapeutic effect evaluation in patients with hepatocellular carcinoma].

OBJECTIVE: To evaluate the therapeutic effects of transcatheter arterial chemoembolization (TACE) with or without radiofrequency ablation (RFA) in hepatocellular carcinoma (HCC) patients based on propensity score matching. METHODS: A logistic regression model was established with the treatment assignment as the dependent variable and the covariates as the independent variables. For each HCC patient, the propensity score was calculated from the model for caliper matching, and a survival analysis of the matched data were carried out. RESULTS: The covariates between the groups were balanced after caliper matching based on the propensity scores. Before matching, the one-, two-, and three-year survival rates of TACE and TACE+RFA were 52.07% and 59.08%, 32.24% and 36.43%, and 316.54% and 19.39%, with the median survival time of 1.20 and 1.40 years, respectively, showing no significant differences in the overall survival rate between the two groups. After matching, the 1-year, 2-year, and 3-year survival rates of TACE and TACE+RFA groups were 54.39% and 62.28%, 23.15% and 40.08%, and 10.20% and 18.52%, with the median survival time of 1.10 years and 1.50 years, respectively, showing significant differences in the overall survival between the two groups. The survival rate in TACE+RFA group was higher than that of TACE only group. CONCLUSION: Propensity score matching can effectively reduce the confounding bias of non-randomized clinical observational data for a more accurate evaluation of the therapeutic effect in HCC patients.