Steroidal saponins and homoisoflavonoids from the fibrous roots of ophiopogon japonicus and their anti-pulmonary fibrosis activities.

The chemical investigation of the fibrous roots of Ophiopogon japonicus afforded two new steroidal saponins, named ophiojaponin F (1) and ophiojaponin G (2), together with twelve known steroidal saponins (3-14) and ten known homoisoflavonoids (15-24). The structures of the isolated compounds were established unambiguously via spectroscopic analyses (NMR and HR-ESI-MS). Ophiojaponin F (1) is a 23-hydroxylated spirostanol saponin, and this type of steroidal saponin rarely been reported in liriopogons. All isolates were evaluated for their anti-pulmonary fibrosis activities on TGF-ß1-actived NIH3T3 cells for the first time. Among them, compounds 3, 4, 11-13, 15-19, 21 and 24 showed potential anti-pulmonary fibrosis effects with IC50 values ranging from 3.61 ± 0.86 µM to 21.33 ± 1.82 µM, and the main component ophiopogonin D (4) displayed the best activity with an IC50 value of 3.61 ± 0.86 µM. Thus, ophiopogonin D may be a potent candidate for the treatment of pulmonary fibrosis.

Sexual Orientation Microaggression Experiences and Coping Responses of Lesbian, Gay, and Bisexual Individuals in Taiwan: A Qualitative Study.

This qualitative study explored the sexual orientation microaggression (SOM) experiences and coping strategies of lesbian, gay, and bisexual (LGB) individuals in Taiwan. In total, 30 LGB individuals (17 women and 13 men; 17 homosexual and 13 bisexual individuals) who experienced SOMs participated in qualitative, semistructured interviews, during which their SOM experiences were assessed. Through the interviews, several types of SOMs were identified, corresponding to three main types of microaggression (microassaults, microinsults, and microinvalidations) previously identified in Western studies. The participants reported various coping responses to SOMs, ranging from active responses to choosing not to respond, to protecting themselves or to minimizing the negative consequences of confrontation. The results provide mental health professionals with insight regarding the contexts of and coping responses to the SOMs experienced by LGB individuals.

Exploring Trauma and Resilience among NYS COVID-19 Pandemic Survivors.

The New York State (NYS) Office of Mental Health created the NYS COVID-19 Emotional Support Helpline and enlisted graduate students to provide phone-based emotional support initially to the NYS community. This NYS-funded initiative transformed into providing psychosocial support for callers across the United States. Four NYS doctoral students acted as the helpline agents and received 251 individual calls from May-August 2020. The agents documented the calls with clinical notes which cannot be traced back to specific callers. The purpose of this retrospective qualitative study was to explore the themes that emerged from the calls to give voice to the trauma that callers were reporting during the early phases of the pandemic, and the resilience they demonstrated as they engaged with the Helpline. The agents' clinical transcripts were converted into codes using a critical-constructivist grounded theory approach (Levitt, 2021) with the NVIVO qualitative data analysis software. A second research team audited the initial codes for construct clarity. Emergent themes detailed the unique traumas that helpline callers divulged, how the agents provided support, and the callers' capacities for resilience. Recommendations are suggested to inform clinicians working with pandemic survivors, to offer guidance on providing distance or virtual interventions as well as to enhance policymakers' understanding of addressing mental health needs across populations served via the NYS COVID-19 Emotional Support Helpline.

Lack of PTRHD1 mutation in patients with young-onset and familial Parkinson's disease in a Taiwanese population.

Mutations in the peptidyl-tRNA hydrolase domain containing 1 (PTRHD1) gene have been recently identified in consanguineous Iranian and African families with juvenile parkinsonism and intellectual disability. However, the pathogenicity of PTRHD1 mutations in the disease and their role in young-onset Parkinson's disease (PD) remains unclear. We aimed to investigate PTRHD1 mutations in a Taiwanese cohort with young-onset and familial PD. We enrolled 464 participants, including 178 probands from PD pedigrees without known PD-causative gene mutations and 286 patients with young-onset PD (age of onset <50 years). All exons and exon-intron boundary junctions of PTRHD1 were analyzed by Sanger sequencing. We did not find any pathogenic coding variants or previously reported mutations, suggesting that PTRHD1 mutations are rare in young-onset and familial PD in our population.

Terpenoid and phenolic derivatives from the aerial parts of Elsholtzia rugulosa and their anti-inflammatory activity.

Seven undescribed terpenoids, including three pairs of enantiomers, named (±)-rugulolides A-C, and one cyclopentenone derivative, named rugulolide D, together with twenty-six known compounds, were isolated from the aerial parts of Elsholtzia rugulosa. The chiral separation of rugulolides A-C was achieved by high-performance liquid chromatography using the chiral column. Their structures were elucidated unambiguously based on comprehensive spectroscopic analysis in conjunction with electronic circular dichroism (ECD) and single-crystal X-ray diffraction experiments. Rugulolides A-D are rare naturally occurring terpenoid derivatives featuring a methylated α,ß-unsaturated-γ-lactone or a cyclopent-2-en-1-one nucleus. All the isolates were evaluated for their inhibitory effects on lipopolysaccharide-induced nitric oxide production in RAW264.7 cell, among them, four compounds showed moderate inhibition with IC50 values ranging from 12.46 to 23.10 µM.

An M29 Aminopeptidase from Listeria Monocytogenes Contributes to In Vitro Bacterial Growth but not to Intracellular Infection.

Aminopeptidases that catalyze the removal of N-terminal residues from polypeptides or proteins are crucial for physiological processes. Here, we explore the biological functions of an M29 family aminopeptidase II from Listeria monocytogenes (LmAmpII). We show that LmAmpII contains a conserved catalytic motif (EEHYHD) that is essential for its enzymatic activity and LmAmpII has a substrate preference for arginine and leucine. Studies on biological roles indicate that LmAmpII is required for in vitro growth in a chemically defined medium for optimal growth of L. monocytogenes but is not required for bacterial intracellular infection in epithelial cells and macrophages, as well as cell-to-cell spreading in fibroblasts. Moreover, LmAmpII is found as dispensable for bacterial pathogenicity in mice. Taken together, we conclude that LmAmpII, an M29 family aminopeptidase, can efficiently hydrolyze a wide range of substrates and is required for in vitro bacterial growth, which lays a foundation for in-depth investigations of aminopeptidases as potential targets to defend Listeria infection.

Molecular investigation of infection sources and transmission chains of brucellosis in Zhejiang, China.

In the present study, a total of 7793 samples from 5 different types of hosts were collected and tested, with a seroprevalence of 2.4% (184/7793). Although the seroprevalence of human and animal brucellosis is relatively low, numbers of human brucellosis cases reported have increased continuously from 2004 to 2018. A total of 118 Brucella strains containing 4 biotypes were obtained, including Brucella melitensis bv.1 (n = 8) and bv.3 (n = 106), Brucella abortus bv.3 (n = 3) and bv.7 (n = 1). Twenty-one shared MLVA-16 genotypes, each composed of 2 to 19 strains obtained from different hosts, suggest the occurrence of a brucellosis outbreak epidemic with multiple source points and laboratory infection events. Moreover, 30 shared MLVA-16 genotypes were observed among 59.6% (68/114) B. melitensis isolates from Zhejiang and strains from other 21 different provinces, especially northern provinces, China. The analysis highlighted the imported nature of the strains from all over the northern provinces with a dominant part from the developed areas of animal husbandry. These data revealed a potential transmission pattern of brucellosis in this region, due to introduced infected sheep leading to a brucellosis outbreak epidemic, and eventually causing multiple laboratory infection events. It is urgent to strengthen the inspection and quarantine of the introduced animals.

Deletion of glutaredoxin promotes oxidative tolerance and intracellular infection in Listeria monocytogenes.

Thiol-disulfide glutaredoxin systems of bacterial cytoplasm favor reducing conditions for the correct disulfide bonding of functional proteins, and therefore were employed by bacteria to defend against oxidative stress. Listeria monocytogenes has been shown to encode a putative glutaredoxin, Grx (encoded by lmo2344), while the underlying roles remain unknown. Here we suggest an unexpected role of L. monocytogenes Grx in oxidative tolerance and intracellular infection. The recombinant Grx was able to efficiently catalyze the thiol-disulfide oxidoreduction of insulin in the presence of DTT as an election donor. Unexpectedly, the deletion of grx resulted in a remarkably increased tolerance and survival ability of this bacteria when exposed to various oxidizing agents, including diamide, and copper and cadmium ions. Furthermore, loss of grx significantly promoted bacterial invasion and proliferation in human epithelial Caco-2 cells and murine macrophages, as well as a notably increasing invasion but not cell-to-cell spread in the murine fibroblasts L929 cells. More importantly, L. monocytogenes lacking the glutaredoxin exhibited more efficient proliferation and recovery in the spleens and livers of the infected mice, and hence became more virulent by upregulating the virulence factors, InlA and InlB. In summary, we here for the first time demonstrated that L. monocytogenes glutaredoxin plays a counterintuitive role in bacterial oxidative resistance and intracellular infection, which is the first report to provide valuable evidence for the role of glutaredoxins in bacterial infection, and more importantly suggests a favorable model to illustrate the functional diversity of bacterial Grx systems during environmental adaption and host infection.

Detection of fetal subchromosomal aberration with cell-free DNA screening led to diagnosis of parental translocation: Review of 11344 consecutive cases in a university hospital.

BACKGROUND: Fetal chromosome aberrations and sub-chromosomal copy number variations (CNVs) are not rare. There are several ways to detect duplications and deletions; cell-free DNA screening (cfDNA screening) is nowadays an accurate and safe detection method. The objective of this study is to report the feasibility of cfDNA screening as an indicator of parental balanced chromosome translocation. RESULTS: From February 2015 to March 2016, cfDNA screening was offered to 11344 pregnant women. 137 out of 11344 individuals tested positive for aneuploidies using cfDNA screening were confirmed by karyotyping. 6 additional cases also tested positive for other deletion/duplication were confirmed by chromosomal microarray analysis (CMA). 11201 patients tested negative and 10342 of them were confirmed through interviews after delivery. Among the 137 cases that were screened positive in cfDNA screening, 91 were common trisomies (63 cases of trisomy 21, 25 cases of trisomy 18 and 3 cases of trisomy 13) and 46 cases were positive for sex-chromosomal abnormalities. In addition, 6 cases were positive for other deletion/duplication in which 2 were identified as terminal duplication and deletion on different chromosomes. The cfDNA screening findings were confirmed by CMA or karyotyping, and the origins of CNVs were validated afterward by karyotyping or fluorescence in situ hybridization (FISH) using parental blood samples. CONCLUSION: CfDNA screening may help identify deletions and duplications in fetus, which in some cases may indicate risk of a parent being a balanced rearrangement carrier, and that the diagnostic follow-up testing is necessary.

[Characterization of a recombinant aminopeptidase Lmo1711 from Listeria monocytogenes].

We aimed to obtain the recombinant aminopeptidase encoded by Listeria monocytogenes (L. monocytogenes) gene lmo1711, and characterized the enzyme. First, the amino acid sequences of Lmo1711 from L. monocytogenes EGD-e and its homologues in other microbial species were aligned and the putative active sites were analyzed. The putative model of Lmo1711 was constructed through the SWISS-MODEL Workspace. Then, the plasmid pET30a-Lmo1711 was constructed and transformed into E. coli for expression of the recombinant Lmo1711. The his-tagged soluble protein was purified using the nickel-chelated affinity column chromatography. With the amino acid-p-nitroaniline as the substrate, Lmo1711 hydrolyzed the substrate to free p-nitroaniline monomers, whose absorbance measured at 405 nm reflected the aminopeptidase activity. The specificity of Lmo1711 to substrates was then examined by changing various substrates, and the effect of metal ions on the catalytic efficiency of this enzyme was further determined. Based on the bioinformatics data, Lmo1711 is a member of the M29 family aminopeptidases, containing a highly conserved catalytic motif (Glu-Glu-His-Tyr-His-Asp) with typical structure arrangements of the peptidase family. The recombinant Lmo1711 with a size of about 49.3 kDa exhibited aminopeptidase activity and had a selectivity to the substrates, with the highest degree of affinity for leucine-p-nitroaniline. Interestingly, the enzymatic activity of Lmo1711 can be activated by Cd²âº, Zn²âº, and is strongly stimulated by Co²âº. We here, for the first time demonstrate that L. monocytogenes lmo1711 encodes a cobalt-activated aminopeptidase of M29 family.

Flagellar Basal Body Structural Proteins FlhB, FliM, and FliY Are Required for Flagellar-Associated Protein Expression in Listeria monocytogenes.

Listeria monocytogenes is a food-associated bacterium that is responsible for food-related illnesses worldwide. In the L. monocytogenes EGD-e genome, FlhB, FliM, and FliY (encoded by lmo0679, lmo0699, and lmo0700, respectively) are annotated as putative flagella biosynthesis factors, but their functions remain unknown. To explore whether FlhB, FliM, and FliY are involved in Listeria flagella synthesis, we constructed flhB, fliM, fliY, and other flagellar-related gene deletion mutants using a homologous recombination strategy. Then, we analyzed the motility, flagella synthesis, and protein expression of these mutant strains. Motility and flagella synthesis were completely abolished in the absence of flhB, fliM, or fliY. These impaired phenotypes were fully restored in the complemented strains CΔflhB, CΔfliM, and CΔfliY. The transcriptional levels of flagellar-related genes, including flaA, fliM, fliY, lmo0695, lmo0698, fliI, and fliS, were downregulated markedly in the absence of flhB, fliM, or fliY. Deletion of flhB resulted in the complete abolishment of FlaA expression, while it decreased FliM and FliY expression. The expression of FlaA was abolished completely in the absence of fliM or fliY. No significant changes were found in the expression of FlhF and two flagella synthesis regulatory factors, MogR and GmaR. We demonstrate for the first time that FlhB, FliM, and FliY not only mediate Listeria motility, but also are involved in regulating flagella synthesis. This study provides novel insights that increase our understanding of the roles played by FlhB, FliM, and FliY in the flagellar type III secretion system in L. monocytogenes.

A feasible diagnostic approach for the translocation carrier from the indication of products of conception.

BACKGROUND: Chromosome translocations are rare but frequently associated with infertility. The objective of this study is to investigate the feasibility of using chromosomal microarray analysis (CMA) on products of conception (POC) samples as an indicator of parental balanced translocation. From January 2011 to December 2016, CMA using Affymetrix Cytoscan™750K array was performed on 1294 POC samples in our hospital. Karyotyping and fluorescence in situ hybridization (FISH) using parental blood samples were performed to validate the origin of subchromosomal copy number variations (CNVs). RESULTS: In the 1294 cases of POCs, we detected CNVs of terminal duplication and deletion that imply unbalanced translocation derivatives in 16 cases, and accurate diagnosis with the parental study was made in all the cases by karyotyping and/or FISH. In 10/16 (62.5%) of these cases, CNVs were inherited from one carrier parent of balanced translocation (Cases 1 to 10), while 6/16 (37.5%) cases occurred de novo (Cases 11 to 16). CONCLUSION: This study clearly illustrated the importance of the utilization of CMA on POC, followed by parental karyotyping and FISH to better characterize CNVs. This approach is especially useful for couples in whom one partner carries a cryptic/submicroscopic balanced translocation but has an apparently normal karyotype.

Carboxyl-Terminal Residues N478 and V479 Required for the Cytolytic Activity of Listeriolysin O Play a Critical Role in Listeria monocytogenes Pathogenicity.

Listeria monocytogenes is a facultative intracellular pathogen that secretes the cytolysin listeriolysin O (LLO), which enables the bacteria to cross the phagosomal membrane. L. monocytogenes regulates LLO activity in the phagosome and minimizes its activity in the host cytosol. Mutants that fail to compartmentalize LLO activity are cytotoxic and have attenuated virulence. Here, we showed that residues N478 and V479 of LLO are required for LLO hemolytic activity and bacterial virulence. A single N478A mutation (LLON478A) significantly increased the hemolytic activity of LLO at a neutral pH, while no difference was observed at the optimum acidic pH, compared with wild-type LLO. Conversely, the mutant LLOV479A exhibited lower hemolytic activity at the acidic pH, but not at the neutral pH. The double mutant LLON478AV479A showed a greater decrease in hemolytic activity at both the acidic and neutral pHs. Interestingly, strains producing LLON478A or LLOV479A lysed erythrocytes similarly to the wild-type strain. Surprisingly, bacteria-secreting LLON478AV479A had barely detectable hemolytic activity, but exhibited host cell cytotoxicity, escaped from the phagosome, grew intracellularly, and spread cell-to-cell with the same efficiency as the wild-type strain, but were highly attenuated in virulence in mice. These data demonstrate that these two residues are required for LLO hemolytic activity and pathogenicity in mice, but not for escape from the phagosome and cell-to-cell spreading. The finding that the nearly non-hemolytic LLON478AV479A mutant grew intracellularly indicates that mutagenesis of a virulence determinant is a novel approach for the development of live vaccine strains.

Rescula as an alternative therapy for beta-blockers with long-term drift effect in glaucoma patients.

The purpose of this study was to evaluate both the intraocular pressure (IOP)-decreasing and neuroprotective effects of Rescula (0.12% unoprostone isopropyl) as an alternative therapy to betablockers with a long-term drift effect in patients with glaucoma. Twenty-eight patients with unilateral or bilateral glaucoma were treated with Rescula instead of the original beta-blocker therapy. IOP was measured using a Goldmann applanation tonometer, and visual field defects were evaluated quantitatively by Humphrey automatic perimetry central 30-2 threshold test. The mean follow-up time was at least 1 year. Rescula achieved a significant (p = 0.00001) and long-lasting reduction in IOP (from 20.78 +/- 2.71 to 17.14 +/- 2.70 mmHg) in patients with open-angle glaucoma after 12 months of follow-up. It also demonstrated a significant (p = 0.02) IOP-reducing effect (from 20.67 +/- 3.60 to 16.36 +/- 3.67 mmHg) in patients with angle-closure glaucoma 12 months later. The mean deviation of visual field defects changed from -13.27 dB baseline to -10.64 dB at 12 months as evaluated by Humphrey field analyzer II central 30-2 threshold test after Rescula; however, there was no statistical difference (p = 0.098). Our results showed that Rescula has a significant IOP-reducing effect as an alternative therapy to beta-blockers with long-term drift effect in patients with open-angle and angle-closure glaucoma. However, a neuroprotective effect to prevent further progression of the visual field defect in patients with glaucoma was not demonstrated in this study.

[Study on the adsorption behavior of Ga, In and Tl on nanometer-size titanium dioxide by ICP-AES].

The adsorption behavior of nanometer TiO2 towards Gallium (Ga) Indium (In) and Thallium (Tl) was investigated with inductively coupled plasma atomic emission spectrometry (ICP-AES). The optimum conditions for adsorption were studied in detail, and the test included sorption kinetics, effect of pH on adsorption ratio, enrichment factor, and adsorption capacity. Under the optimum conditions, Ga (III), In(III) and Tl(I) ions could be adsorbed and recovered quantitatively. The static adsorption capacities of Ga(III), In(III) and Tl(I) on nanometer TiO2 were 48.6, 46.6 and 23.4 mg x g(-1) respectively. For the elution of Ga(III), In (III) and Tl(I), a mixture of 0.1 mol x L(-1) EDTA solution and 1.0 mol x L(-1) HNO3 was used, and the recovery ratio was above 92%. According to the definition of IUPAC, the detection limits (3sigma) of this method for Ga, In and Tl with an enrichment factor of 12.5 are 3.0, 6.0 and 13 ng x mL(-1), respectively; and relative standard deviations (RSD) are 1.85%, 1.96% and 3.4%, respectively (n = 6). The proposed method has been applied successfully to the analysis of geological samples with satisfactory results.

A clinical and roentgenographic study of Little League elbow.

BACKGROUND: Comparisons of medial elbow injury rate and the incidence of clinical and radiographic findings among Little League baseball players have not been documented. HYPOTHESIS: Injury rate and clinical and radiographic findings in Little Leaguers of different positions may be similar. STUDY DESIGN: Survey and retrospective review. METHODS: Altogether, 343 Little Leaguers (120 pitchers, 40 catchers, and 183 fielders) participated in the study. The survey (including age, playing position, playing experience, and symptoms), clinical examination, and roentgenogram of both elbows were obtained for analysis using Pearson's chi-square test. RESULTS: Clinically, 58% of the pitchers presented with soreness compared to 63% in catchers and 47% in fielders. Radiographic examination revealed hypertrophy of the medial humeral epicondyle in all pitchers and catchers and 90% of the fielders. Separation of the medial epicondyle was found in 63% of pitchers, 70% of catchers, and 50% of fielders, while fragmentation was found in 19% of pitchers, 40% of catchers, and 15% of fielders. In subjects with separation or fragmentation, 49% and 56% complained of elbow soreness, respectively. CONCLUSION: Similar rates of medial elbow changes in Little Leaguers may imply that the roentgenological changes of the medial epicondyle could be an adaptive yet physiological reaction to the excessive valgus stress of throwing. The higher incidence of medial epicondylar separation and fragmentation in catchers that has not been reported before may be secondary to the increased stress placed on the elbow when throwing from the squatting position.

[Effects of rhein on the function of human mesangial cells in high glucose environment].

AIM: To study the mechanisms of anti-diabetic nephropathy of rhein on cultured human mesangial cells (HMCs). METHODS: To mimic the hyperglycemic (HG) environment of diabetic nephropathy, 30 mmol x L(-1) glucose were added to 10% FBS RPMI 1640. The HMCs were treated with rhein for 8, 24, 48 or 72 h, at these time, the bioactivity, total activity of transforming growth factor-beta1 (TGFbeta1), activity of p38MAPK (p38 mitogen-activated protein kinases, by using immunoprecipitate and Western blot), MMP-2 (matrix metalloproteinase-2), and MMP-9 (matrix metalloproteinase-9, by using gelatinase zymography) and the proliferation of HMCs in high glucose media were measured. Meanwhile the levels of secretion of FN in cultured HMCs were measured. RESULTS: The results showed that rhein markedly inhibit the proliferation of HMCs, significantly reduce the bioactivity of TGFbeta1 and FN secretion in HMCs, and decrease the increased activity of p38MAPK, but showed no action on the activities of MMP-2 and MMP-9. CONCLUSION: Rhein reduced the secretion of FN and inhibited the proliferation of HMCs may through inhibiting the bioactivities of TGFbeta1 and p38MAPK.

NH4OAc promoted cyclocondensation of 3-(o-allylphenyl)pentane-1,5-diones: synthesis of tetracyclic benzofused 1-azahomoisotwistanes.

A facile two-step synthetic route for preparing the novel tetracyclic skeleton of benzofused 2,6-diaryl-1-azahomoisotwistanes 2 had been developed. The route was carried out by a one-pot tandem cross-coupling reaction of o-allylbenzaldehydes 1 with aryl methyl ketones 3, and NH(4)OAc mediated the cascade cyclocondensation reaction of the resulting 1,5-diketones 4 with the 3-o-allylphenyl group in good yield in two steps.

[Molecular expression of peripheral lymphocytes of the patients after organ transplantation].

AIM: To investigate the time course of the gene expression after the organ transplantation, and try to identify the relationship of the gene and protein changes with the pathological progression and the immune rejection. METHODS: We systematically observed the gene expression of CD4, CD8, P56, P59 by real-time PCR. The time courses of gene expression were analyzed by Fluorescent Differential Display after the organ transplantation. At the same time points, the clinical outcomes of the patients were followed up so that we could identify the relationship of the gene expression and the immune rejection. RESULTS: Immunosuppressive drugs could not prevent the transplantation rejection completely. P59 gene and tyrosine phosphatase genes started to express positively within 24 h after organ transplantation. Between 24-48 h, PKC-theta and CD4 gene expression levels increased. The expressions of PKC-theta, MHC-II, serine/threonine kinase, tyrosine phosphatase, serine/threonine kinase 15, clotting factor VIII, interferon gamma-inducible protein 16 and HLA-DR-H increased by three times compared to pre-transplantation in patients with kidney and heart transplantations. The expression of P59 gene remained stable within 24 h after transplantation, and then decreased to the lowest level at day 10. P56 and CD8 levels were down-regulated within 72 h, and then up-regulated to the first peak at day 5. The expression of CD4 was also inhibited within 24 h, and then increased to the first peak at 48 h. CONCLUSION: The monitoring on the expressions of the proteins and genes on the peripheral lymphocytes may help to early find the clinical immune rejection after organ transplantation and judge the efficiency of the anti-rejection drugs.