Characterization and Genome Analysis of the Delftia lacustris Strain LzhVag01 Isolated from Vaginal Discharge.

Delftia has been separated from freshwater, sludge, and soil and has emerged as a novel opportunistic pathogen in the female vagina. However, the genomic characteristics, pathogenicity, and biotechnological properties still need to be comprehensively investigated. In this study, a Delftia strain was isolated from the vaginal discharge of a 43-year-old female with histologically confirmed cervical intraepithelial neoplasm (CIN III), followed by whole-genome sequencing. Phylogenetic analysis and average nucleotide identity (ANI) analysis demonstrated that it belongs to Delftia lacustris, named D. lacustris strain LzhVag01. LzhVag01 was sensitive to ß-lactams, macrolides, and tetracyclines but exhibited resistance to lincoamines, nitroimidazoles, aminoglycosides, and fluoroquinolones. Its genome is a single, circular chromosome of 6,740,460 bp with an average GC content of 66.59%. Whole-genome analysis identified 16 antibiotic resistance-related genes, which match the antimicrobial susceptibility profile of this strain, and 11 potential virulence genes. These pathogenic factors may contribute to its colonization in the vaginal environment and its adaptation and accelerate the progression of cervical cancer. This study sequenced and characterized the whole-genome of Delftia lacustris isolated from vaginal discharge, which provides investigators and clinicians with valuable insights into this uncommon species.

In vitro biofilm formation of Gardnerella vaginalis and Escherichia coli associated with bacterial vaginosis and aerobic vaginitis.

Objective: To determine the optimum biofilm formation ratio of Gardnerella vaginalis (G. vaginalis) in a mixed culture with Escherichia coli (E. coli). Methods: G. vaginalis ATCC14018, E. coli ATCC25922, as well as five strains of G. vaginalis were selected from the vaginal sources of patients whose biofilm forming capacity was determined by the Crystal Violet method. The biofilm forming capacity of E. coli in anaerobic and non-anaerobic environments were compared using the identical assay. The Crystal Violet method was also used to determine the biofilm forming capacity of a co-culture of G. vaginalis and E. coli in different ratios. After Live/Dead staining, biofilm thickness was measured using confocal laser scanning microscopy, and biofilm morphology was observed by scanning electron microscopy. Results: The biofilm forming capacity of E. coli under anaerobic environment was similar to that in a 5% CO2 environment. The biofilm forming capacity of G. vaginalis and E. coli was stronger at 106:105 CFU/mL than at other ratios (P<0.05). Their thicknesses were greater at 106:105 CFU/mL than at the other ratios, with the exception of 106:102 CFU/mL (P<0.05), under laser scanning microscopy. Scanning electron microscopy revealed increased biofilm formation at 106:105 CFU/mL and 106:102 CFU/mL, but no discernible E. coli was observed at 106:102 CFU/mL. Conclusion: G. vaginalis and E. coli showed the greatest biofilm forming capacity at a concentration of 106:105 CFU/mL at 48 hours and could be used to simulate a mixed infection of bacterial vaginosis and aerobic vaginitis in vitro.

Physalin A Induces Apoptosis and Autophagy in Hepatocellular Carcinoma via Inhibition of PI3K/Akt Signaling Pathway.

Physalin A (PA) is a bioactive withanolide with multiple pharmacological properties and has been indicated to be cytotoxic to hepatocellular carcinoma (HCC) cell line HepG2. This study aims to explore the mechanisms underlying PA antitumor activity in HCC. HepG2 cells were exposed to various concentrations of PA. Cell counting kit-8 assay and flow cytometry were implemented for evaluating cell viability and apoptosis, respectively. Immunofluorescence staining was utilized for detecting autophagic protein LC3. Western blotting was employed for measuring levels of autophagy-, apoptosis- and phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) signaling-related proteins. A xenograft mouse model was established to verify the antitumor activity of PA in vivo. PA impaired HepG2 cell viability, and triggered apoptosis as well as autophagy. Inhibiting autophagy augmented PA-evoked HepG2 cell apoptosis. PA repressed PI3K/Akt signaling in HCC cells and activating PI3K/Akt reversed PA-triggered apoptosis and autophagy. PA treatment inhibited tumor growth in tumor-bearing mice. PA triggers HCC cell apoptosis and autophagy by inactivating PI3K/Akt signaling.

Bioinspired Fast Room-Temperature Self-Healing, Robust, Adhesive, and AIE Fluorescent Waterborne Polyurethane via Hierarchical Hydrogen Bonds and Use as a Strain Sensor.

Developing a new generation of ecofriendly water-based polymeric materials that integrate mechanical robustness, fast room-temperature self-healing, adhesive, and fluorescence remains a formidable challenge. Herein, inspired by titin protein, a series of novel waterborne polyurethanes (WPU-CHZ-NAGA) containing irregular 6-fold and diamide hydrogen bonds are synthesized by introducing carbohydrazide (CHZ) and N,N-bis(2-hydroxyethyl)-3-amino propionyl glycinamide (HO-NAGA-OH) groups. The representative WPU-CHZ2-NAGA3 exhibits outstanding mechanical properties (tensile strength of 36.58 MPa, tearing energy of 81.2 kJ m-2, and toughness of 125.82 MJ m-3) and fast room-temperature self-healing ability with the aid of ethanol (≥90% within 8 h) originated from hierarchical hydrogen bonds. These properties are superior to those of most of the reported room-temperature self-healing polymer materials. Benefiting from plentiful hydrogen bonds, the WPU matrix achieves excellent adhesive properties without heating or adding curing agents. Interestingly, WPU-CHZ2-NAGA3 film emits inherent blue fluorescence due to the aggregation-induced emission effect of tertiary amine groups, and its potential applications in information encryption and anticounterfeiting are further demonstrated. Specially, a eutectic gel strain sensor is also fabricated with WPU-CHZ2-NAGA3 and deep eutectic solvent by a simple physical blending method, which can be used to monitor the movement of human fingers and wrists as well as the change in body temperature. In summary, this work provides new insight into the design and synthesis of multifunctional WPU with fast room-temperature self-healing and high mechanical properties.

[Effect of needle-knife on chondrocyte apoptosis of knee joint in rabbits with knee osteoarthritis based on CircSERPINE2-miR-1271-5P-ERG axis].

OBJECTIVE: To observe the effect of needle-knife on the chondrocyte apoptosis of knee joint in rabbits with knee osteoarthritis (KOA) based on the CircSERPINE2-miR-1271-5P-E26 specific transformation-related gene (ERG) axis, and to explore the mechanism of needle-knife for KOA. METHODS: Thirty-six New Zealand white rabbits were randomly divided into a normal group, a model group, a needle-knife group and a sham needle-knife group, 9 rabbits in each group. The rabbits in the model group, the needle-knife group and the sham needle-knife group were treated with modified Videman method to prepare KOA model. After successful modeling, the rabbits in the needle-knife group were treated with needle-knife at cord adhesion and nodules near quadriceps femoris tendon and internal and external collateral ligament on the affected knee joint; the rabbits in the sham needle-knife group were treated with sham needle-knife baside the needle insertion point of the needle-knife group (needle-knife was only inserted, without any operation). The treatment was given once a week, 3 times in total. The Lequesne MG behavioral score was used to evaluate the knee joint damage in each group before and after intervention. After intervention, HE staining and transmission electron microscopy were used to observe the cartilage tissue morphology and ultrastructure of chondrocytes in the knee joint in each group; TUNEL method was used to detect the level of chondrocyte apoptosis in the knee joint; real-time fluorescence quantitative PCR was used to detect the expression of CircSERPINE2, miR-1271-5P and ERG mRNA in knee cartilage tissue in each group. RESULTS: After intervention, compared with the normal group, the Lequesne MG behavioral score in the model group was increased (P<0.01). Compared with the model group and the sham needle-knife group, the Lequesne MG behavioral score in the needle-knife group was decreased (P<0.01). In the model group and the sham needle-knife group, the number of chondrocytes and organelles was decreased, the cell nucleus was shrunk, mitochondria was swelling or disappeared; in the needle-knife group, the number of chondrocytes and organelles was increased, the cell nucleus was not obviously shrunk and the mitochondria was not obviously swelling. Compared with the normal group, the level of chondrocyte apoptosis in the model group was increased (P<0.01); compared with the model group and the sham needle-knife group, the level of chondrocyte apoptosis in the needle-knife group was decreased (P<0.01, P<0.05). Compared with the normal group, the expression of CircSERPINE2 and ERG mRNA in the model group was decreased (P<0.01), and the expression of miR-1271-5P mRNA was increased (P<0.01); compared with the model group and the sham needle-knife group, the expression of CircSERPINE2 and ERG mRNA in the needle-knife group was increased (P<0.01), and the expression of miR-1271-5P mRNA was decreased (P<0.01). CONCLUSION: Needle-knife could reduce the knee joint damage and chondrocyte apoptosis in KOA rabbits, which may be related to up-regulating the expression of CircSERPINE2 and ERG mRNA, and inhibiting the expression of miR-1271-5P mRNA.

Flame-Retardant and Self-Healing Waterborne Polyurethane Based on Organic Selenium.

Waterborne polyurethane has drawn extensive attention due to its environmental friendliness and is widely used in many areas. However, it is still a great challenge to synthesize waterborne polyurethanes with flame retardancy and fast room-temperature self-healing ability, along with excellent mechanical performance and emulsion stability due to the mutually contradictory nature of these properties. Herein, waterborne polyurethanes containing organic selenium (SWPU-x) from 0.67 to 3.28 wt % were synthesized, which could simultaneously realize flame retardancy and self-healing ability based on the ability to scavenge active free radicals at high temperature and the dynamic switch of diselenide. All these SWPU-x films self-extinguished within 1 s after the ignition in the vertical combustion tests. The limiting oxygen index of SWPU-4 was improved to 28.5% with excellent UL-94 level (V-0) and self-healing efficiency (91.25%, after being healed in the photoreactor for 30 min at room temperature), together with high mechanical properties (tensile strength was 18.5 MPa and elongation at break was 869.63%), and the total heat release (THR) for SWPU-4 (49.28 MJ/m2) could decrease to 23.80% of the THR for the original waterborne polyurethane WPU (64.67 MJ/m2). This work discovered a new flame-retardant element (organic selenium) and studied its flame-retardant behaviors and self-healing function simultaneously, which would extremely extend the application of waterborne polyurethanes.

Mitigation of amplified spontaneous emission noise for an all-fiber coaxial aerosol lidar with different single-photon detectors.

For a coaxial single-photon lidar system, amplified spontaneous emission (ASE) noise from the fiber amplifier is inevitable. The ASE backscattering from specular reflection annihilates the far-field weak signal, resulting in low signal-to-noise ratio, short measurement distance, and even misidentification. We propose a method for calibrating and mitigating ASE noise in all-fiber coaxial aerosol lidar and demonstrate the method for a lidar system with different single-photon detectors (SPDs). The accuracy of the coaxial aerosol lidar is comparable to that of the biaxial one. We conducted an experiment using three different detectors, namely, InGaAs/InP SPD, up-conversion SPD, and superconducting nanowire SPD in the same coaxial lidar system. Compared with the biaxial system, the three different detectors we used have achieved more than 90% ASE noise suppression, the measured visibility percent errors of InGaAs/InP SPD data, up-conversion SPD data, and superconducting nanowire SPD data all within 20%, and the percent error within 10% are 99.47%, 100%, and 95.12%, respectively. Moreover, time-sharing optical switching allowed to obtain background noise with high accuracy.

Liquid-Free Ion-Conducting Elastomer with Environmental Stability for Soft Sensing and Thermoelectric Generating.

Ionic conductors are promising candidates for fabricating soft electronics, but currently applied ionic hydrogels and organogels suffer from liquid leakage and evaporation issues. Herein, we fabricated a free-liquid ionic conducting elastomer (LFICE) with dry lithium bis(trifluoromethane sulfonimide) and elastomeric waterborne polyurethane. The resultant versatile LFICE exhibits superior tensile strength (∼4.5 MPa), satisfactory stretchability (>900%), excellent ionic conductivity (8.32 × 10-4 S m-1 at 25 °C), and sensitive strain (3.21) and temperature (2.22% °C-1) response. The LFICE also presents durable environmental stability due to the all-solid-state feature. In the exploration of application prospects, the as-assembled LFICE sensor can precisely and repeatedly detect human motion and temperature changes, demonstrating its potentials in digital medical diagnosis and monitoring; the as-assembled LFICE thermoelectric generator (TEG) shows a high ionic thermovoltage of 4.41 mV K-1, paving a bright path for the advent of self-powered soft electronics. It is believed that this research boosts the facile fabrication of environmental stable stretchable ionic conductors holding great promise in next-generation soft electronics integrated with dual thermo- and strain-response and energy harvesting.

Redox-responsive carrier based on fluorinated gemini amphiphilic polymer for combinational cancer therapy.

Polymeric micelle has emerged as an efficient implement to overcome the shortcomings of conventional cancer chemotherapy due to its superior solubility of hydrophobic drugs and less side effects of drugs. However, insufficient dilution resistance and ordinary therapeutic effect severely restrict the further translation of current drug-loaded polymeric micelles. Here, we showed that well-defined G-Fn (n = 5, 9, 13) polymeric micelles possessed excellent capabilities as a drug carrier in light of high drug loading content, high stability and precise drug release combined with wonderful endocytosis efficiency to tumors. The representative G-F13 exhibited an excellent dilution resistance, outstanding high drug loading content (22 wt%) and drug loading efficiency (82%), which might be attributed to the extremely low critical micelle concentration conferred by its special Gemini structure and the superhydrophobicity of the fluorocarbon chain. Furthermore, the "cross-linked" internal fluoride membrane consisted of the two chains of the Gemini structure made G-F13 stable even after 24 h of incubation in 10% fetal bovine serum (FBS). The camptothecin (CPT) release was selectively triggered by glutathione (GSH) and H2O2, reaching 75% and 85% after 24 h respectively, in which only 15% of drugs leak under physiological conditions. The CCK-8 assays of Hela cells showed that CPT-loaded G-F13 micelles had high cell compatibility (200 µg/mL, 93% cell viability, 48 h) and high cancer cytotoxicity (IC50 0.1 µg/mL). Notably, a tenfold lower dosage of loaded CPT had an higher tumor growth inhibition than the free CPT. This result was attributed to the combined treatment of fluorinated drug carriers were more likely to penetrate the cell membrane to enter tumor cells, the cytotoxicity of selenic acid generated after the oxidation of G-F13 and the large amounts of CPT after redox release. Excellent physical and chemical properties as well as good therapeutic effects reveal that G-F13 can act as a promising drug carrier to widely use in cancer chemotherapy.

Fetal Familial Cerebral Cavernous Malformation With a Novel Heterozygous KRIT1 Variation.

BACKGROUND AND OBJECTIVES: To identify fetal familial cerebral cavernous malformation (CCM) and a novel variation. METHODS: A 37-year-old pregnant woman (G4P0) presented with right-handed numbness since 2 weeks at 31 weeks of gestation. Evaluation with brain MRI revealed multiple CCMs. As a result, fetal MRI, fetal whole exome sequencing, and maternal Sanger sequencing were performed. RESULTS: The mother's brain MRI demonstrated numerous CCMs involving the brain stem, cerebral hemispheres, and cerebellum. Fetal MRI showed a CCM located in the left frontal lobe in susceptibility-weighted imaging (SWI). The neuroimaging characteristics of the mother and the fetus suggested that their CCMs may be familial. Genetic analysis revealed a novel variation in KRIT1 (c.1A>G, p.0?), also called CCM1, in the mother and the baby. The mother delivered a daughter at 32 weeks of gestation with an Apgar score of 10 by cesarean section. DISCUSSION: This variation of the initial codon in the KRIT1 gene leads to a phenotype with an early-onset. To our knowledge, this is the first-ever reported case of fetal familial CCM and this novel variation. Brain MRI has excellent sensitivity and specificity, providing the best option for detecting CCMs, even in utero, primarily when SWI is used.

Tightening the Crosslinking Distance Restraints for Better Resolution of Protein Structure and Dynamics.

Chemical crosslinking coupled with mass spectrometry (CXMS) has been increasingly used in structural biology. CXMS distance restraints are usually applied to Cα or Cß atoms of the crosslinked residues, with upper bounds typically over 20 Å. The incorporation of loose CXMS restraints only marginally improves the resolution of the calculated structures. Here, we present a revised format of CXMS distance restraints, which works by first modifying the crosslinked residue with a rigid extension derived from the crosslinker. With the flexible side chain explicitly represented, the reformatted restraint can be applied to the modification group instead, with an upper bound of 6 Å or less. The short distance restraint can be represented and back-calculated simply with a straight line. The use of tighter restraints not only afford better-resolved structures but also uncover protein dynamics. Together, our approach enables more information extracted from the CXMS data.

The incidence of placenta related disease after the hysteroscopic adhesiolysis in patients with intrauterine adhesions.

OBJECTIVE: To analyze the correlation between placenta related disease of pregnant women with antecedent hysteroscopic adhesiolysis due to intrauterine adhesions (IUA). MATERIALS AND METHODS: This is a single center, non-randomized, open-label, retrospective cohort Study. 74 patients who had adhesiolysis and hormone therapy for IUA and progressed into the third trimester were group A and 296 without IUA were group B. The main outcome measure is the incidence of placenta related disease including placenta accreta spectrum, placenta previa, placental abruption, intrauterine growth restriction (IUGR), and pregnancy-induced hypertension (PIH). The second outcome is the perinatal, and intrapartum complications. RESULTS: Patients in group A had a higher frequency of prior pregnancy times (2.51 ± 1.56 vs.1.84 ± 1.06, p = 0.001) and lower frequency of prior delivery times (0.20 ± 0.41 vs. 1.30 ± 0.51, p < 0.05) than group B at baseline. At delivery, there is no difference between the incidence of PIH and IUGR between two groups. However, a significantly higher frequency of placenta accreta (17.6% vs. 1.4%, OR = 15.56, 95% CI 4.91-49.34), placenta increta (5.4% vs. 0.7%, OR = 8.4, 95% CI 1.51-46.78), placenta previa (8.1% vs. 2.0%, OR = 4.265, 95%CI1.33-13.63) and postpartum hemorrhage (21.6% vs. 3.4%, OR = 7.890, 95% CI 3.41-18.26) were observed in group A than in group B. CONCLUSIONS: Compared to general population, the rates of placenta accreta, placenta increta, placenta previa, postpartum hemorrhage are higher among the IUA patients after hysteroscopic adhesiolysis, and special attention is needed at the termination of the pregnancy.

The Conformational Preference of Chemical Cross-linkers Determines the Cross-linking Probability of Reactive Protein Residues.

Chemical cross-linking mass spectrometry (XLMS) is an emerging technique in structural biology. Providing the cross-linked peptides are identified by mass spectrometry with high confidence, a distance restraint can be applied between the two reactive protein residues, with the upper bound corresponding to the maximal span of the cross-linker. However, as the upper bound is typically over 20 Å, cross-link distance restraints are unrestrictive and provide a marginal improvement in protein structural refinement. Here we analyze the experimental cross-links for lysine or acidic residues and show that the distribution of Cß-Cß' distances can be described with two overlapping Gaussian species. In addition to the pairwise occurrence probability of the reactive protein residues, we show that the distribution profile of the cross-link distances is determined by the intrinsic conformational propensity of the cross-linker. The cross-linker prefers either a compact or extended conformation and, once attached to a reactive protein residue, predominantly an extended conformation. Consequently, the long-distance Gaussian species occurs at a much higher probability than the short-distance species in the observed cross-links. Together, the probabilistic distribution of the cross-link distance allows the construction of a more restrictive restraint for structural modeling and better use of the XLMS data.

A novel and nonderivatization method for the determination of valproic acid in human serum by two-dimensional liquid chromatography.

A novel and robust two-dimensional liquid chromatography with ultraviolet detection method (2D-LC-UV) was developed and validated for high-throughput determination of the concentrations of valproic acid (VPA) in human plasma. This 2D-LC system was composed of a first-dimensional LC column, a second-dimensional LC column and an intermediate transfer column. The sample was directly injected into the 2D-LC system after an easy protein precipitation treatment. After online preconcentration and primary separation by the first-dimensional column, the target was captured by an intermediate column and then transferred to second-dimensional column for analysis. The system transferred the target through "central cutting" mode whereby the drug peak was not subject to interference from the matrix. The analysis cycle time was completed within 7.0 min. Compared with other methods that have been developed, the analysis time was reduced and the operation was much easier without any derivatization. The calibration curve was linear over the 5.90-188.94 µg/ml range for the VPA concentrations. The intra-day and inter-day precisions were <5.6%. The recoveries were in the range from 95.2 to 98.0%. This method appears to be sensitive, precise, rapid and low-cost for the quantification of VPA in serum sample.

Estrogen therapy before hysteroscopic adhesiolysis improves the fertility outcome in patients with intrauterine adhesions.

PURPOSES: To describe the fertility outcomes after hysteroscopic adhesiolysis combined with preoperative hormone treatment in women with intrauterine adhesion (IUA). METHODS: This study is a retrospective cohort study. A total of 230 patients with IUA underwent hysteroscopic adhesiolysis combined with hormone treatment from Jan 2012 to Jun 2018. 148 patients who received preoperative estrogen treatment were enrolled into group A and 82 patients without preoperative estrogen treatment were enrolled into group B. All the patients underwent hysteroscopic adhesiolysis and received postoperative estrogen therapy, intrauterine indwelling device. Second or third look was performed after 2-3 months. RESULTS: 90.87% (209/230) patients complete the study. The AFS scores at baseline were higher in the group A than group B. After the preoperative E2 treatment, group A achieved the comparable AFS score to group B before the surgery. The cumulative fertility rate in group A was comparable in group B, both in the ITT analysis (49.32% vs. 52.44%, p = 0.651) and the PP analysis (54.07% vs.58.10%, p = 0.575).The mean conception time was also similar in group A and group B (8.30 ± 6.47 vs. 8.54 ± 5.68 months, p = 0.837). Besides, the surgery times in group A were less than group B. There was no difference in the rate of adverse events between two groups. CONCLUSIONS: Hysteroscopic adhesiolysis combined with preoperative oestrogen could reduce the preoperative AFS scores and the times of surgery which yield a similar conception rate in women with less severe intrauterine adhesions.

Effect of Heat Input on Porosity Defects in a Fiber Laser Welded Socket-Joint Made of Powder Metallurgy Molybdenum Alloy.

Porosity defects are still a challenging issue in the fusion welding of molybdenum and its alloys due to the pre-existing interior defects associated with the powder metallurgy process. Fiber laser welding of end plug and cladding tube made of nanostructured high-strength molybdenum (NS-Mo) alloy was performed in this work with an emphasis on the role of welding heat input. The distribution and morphology of porosity defects in the welded joints were examined by computed tomography (CT) and scanning electron microscopy (SEM). Preliminary results showed that laser welding of NS-Mo under low heat input significantly reduced the porosity defects in the fusion zone. The results of computed tomography (CT) showed that when the welding heat input decreased from 3600 J/cm (i.e., 1200 W, 0.2 m/min) to 250 J/cm (i.e., 2500 W, 6 m/min), the porosity ratio of the NS-Mo joints declined from 10.7% to 2.1%. Notable porosity defects under high heat input were related to the instability of the keyhole, expansion and the merging of bubbles in the molten pool, among which the instability of the keyhole played the dominant role. The porous defects at low heat input were generated as bubbles released from the powder metallurgy base metal (BM) did not have enough time to overflow and escape.