The extracellular vesicular pseudogene LGMNP1 induces M2-like macrophage polarization by upregulating LGMN and serves as a novel promising predictive biomarker for ovarian endometriosis recurrence.

STUDY QUESTION: How does ectopic endometrial stromal cell (Ecto-ESC)-derived extracellular vesicular Legumain pseudogene 1 (EV-LGMNP1), a newly identified pseudogene of Legumain (LGMN), contribute to M2-phenotype macrophage polarization, and does it predict recurrence in patients with ovarian endometriosis (EMs)? SUMMARY ANSWER: EV-LGMNP1, which is abundant in Ecto-ESCs and serum from ovarian EMs, can direct macrophages towards an M2 phenotype by upregulating LGMN expression and is a promising biomarker for predicting ovarian EMs recurrence. WHAT IS KNOWN ALREADY: Extracellular vesicles (EVs) can mediate cell-to-cell crosstalk to promote disease progression via cargo molecule transport. Recently, LGMNP1, a newly identified pseudogene of LGMN, has been reported to promote cancer progression by upregulating LGMN. LGMN is a well-studied protein that can induce M2-like polarization. STUDY DESIGN, SIZE, DURATION: An in vitro study was conducted with Ecto-ESCs isolated from ectopic endometrial samples, collected from two patients with ovarian EMs (diagnosed by laparoscopy and histological analysis). A clinical retrospective cohort study of 52 ovarian EMs patients and 21 controls with available preoperative serum samples was carried out (2013-2017). The follow-up period ended either at the time of recurrence or on 31 December 2018. PARTICIPANTS/MATERIALS, SETTING, METHODS: Ecto-ESC-derived EVs (EV/Ecto-ESCs) were characterized by nanoparticle tracking analysis, transmission electron microscopy and western blotting. EV internalization by THP-1 cells, which are the most widely used primary human macrophages model, was detected by fluorescence labelling. After EV treatment, THP-1 cell polarization was detected by quantitative real-time PCR (qRT-PCR) and western blot analyses of CD86 (M1-related marker) and CD206 (M2-related marker). LGMNP1 mRNA expression level in EVs from both primary ectopic endometrioc stromal cells and serum was examined using qRT-PCR. Additionally, the expression of LGMN, the downstream target gene of LGMNP1, in THP-1 cells was evaluated using qRT-PCR and western blotting. Kaplan-Meier and multivariate Cox regression analyses were applied to evaluate the independent predictive factors of EMs recurrence-free survival. A novel nomogram model based on serum EV-LGMNP1 was then formulated to predict EMs recurrence. MAIN RESULTS AND THE ROLE OF CHANCE: In vitro assays demonstrated that EV/Ecto-ESCs drove macrophages towards an M2-like phenotype. Moreover, LGMNP1 contributed to EV/Ecto-ESC-induced M2 macrophage polarization by upregulating LGMN mRNA expression levels. Clinically, serum EV-LGMNP1 was more highly expressed in recurrent EMs patients than in controls and EMs patients without recurrence. Survival analysis and our novel nomogram reconfirmed that serum EV-LGMNP1 was a novel promising and meaningful non-invasive biomarker for predicting EMs recurrence. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: In vitro experiments were only performed on samples from two patients with ovarian endometriosis, and a larger sample size is needed. ESCs isolated from the eutopic endometrium of EMs and non-EMs patients should be studied in the future. Additionally, in vitro experiments should be performed using endometrial epithelium cells and further in vivo experiments, such as using mice endometriotic models to investigate whether EV/Ecto could induce M2 macrophage polarization, should be conducted. Moreover, multicentre, large-sample data are needed to validate our predictive nomogram model. WIDER IMPLICATIONS OF THE FINDINGS: Our study provides novel insights into the mechanism of M2 polarization involved in ovarian EMs progression mediated by an 'EV-shuttled pseudogene LGMNP1' mode. In addition, serum EV-LGMNP1 may serve as a novel non-invasive biomarker for predicting recurrence, providing a new therapeutic target for ovarian EMs. STUDY FUNDING/COMPETING INTEREST(S): This project was supported by funding from the National Natural Science Foundation of China (81971361), the Natural Science Foundation of Shanghai Science and Technology (19ZR1406900), the Shanghai 'Rising Stars of Medical Talent' Youth Development Program (AB83030002019004), the Clinical Research Plan of SHDC (SHDC2020CR4087), the Shanghai Municipal Health Commission (202040498), the Research and Innovation Project of the Shanghai Municipal Education Commission (2019-01-07-00-07-E00050) and the Clinical Research Plan of SHDC (SHDC2020CR1045B). There are no competing interests to declare.

Air quality improvement assessment and exposure risk of Shandong Province in China during 2014 to 2020.

As one of the most polluted provinces in China, air pollution events occur frequently in Shandong. Based on the hourly (or daily) concentrations of six air pollutants (PM2.5, PM10, O3, NO2, SO2 and CO), the situations of air quality improvement in three kinds of cities (key cities, coastal cities and general cities) are assessed comprehensively during 2014-2020. Contrary to the daily maximum 8-h average ozone (MDA8 O3), the annual average concentrations of other pollutants show the downward trends during 2014-2020. Therein, the improvement rates of annual average concentrations of air pollutants in key cities are highest. By 2020, the day proportions of O3 as the primary pollutant are up to 38% in three kinds of cities. Besides, due to the impact of COVID-19, the monthly average concentrations of PM2.5, PM10, NO2, SO2 and CO in February 2020 decrease by 32.1-49.5% year-on-year. There are still about 50% of population exposed to high-risk regions (R i > 2), which are mainly concentrated in main urban areas and industrial areas. Thus, the adjustment of industrial structure and energy composition in the context of carbon peak and carbon neutrality should be implemented in the future. Supplementary Information: The online version contains supplementary material available at 10.1007/s13762-022-04651-5.

[Genomic investigation of human Streptococcus suis infection in Shandong Province].

To investigate Streptococcus suis (S.suis) isolated from patients in Shandong province using genomic epidemiology and pathogenologic analysis. To provide the foundation to establish reasonable and accurate prevention and control measures of human S. suis infection. Molecular typing, whole genome phylogenetic tree, virulence gene typing, antibiotic resistance profile and mobile genetic elements carrying antibiotic resistance genes of isolated S. suis strains were investigated. The pathogenicity of isolated strains was also evaluated by comparing their capacity to induce pro-inflammatory cytokine production in vitro. S. suis infections in Shandong province were predominantly due to serotype 2 and sequence type 1 strains. The major symptoms were meningitis. The studied strains could be divided into five lineages. All strains belong to highly pathogenic type in Shandong province,Strains from lineage 2 possessed higher capacity to stimulate pro-inflammatory cytokine production than other strains did, even though other strains belong to highly pathogenic strains. In addition, multiple antibiotic resistance genes and corresponding mobile genetic elements werewidespread in S. suis strains from Shandong province, except strains from lineage 3. High diversities in genome, evolutionary path and pathogenicity of S. suis strains from Shandong province were revealed. It was necessary to surveillant the S. suis strain in genomic level.

Short communication: Influence of preserving factors on detection of ß-lactamase in raw bovine milk.

The refrigeration (4, 8, 24, or 48 d), freezing (1, 7, or 30 h), thawing (25, 40, or 60°C), and usage of preservatives (sodium azide, potassium dichromate, sodium thiocyanate, bronopol, and methanol), of raw bovine milk were investigated for ß-lactamase activity. The ß-lactamase activity in all samples was assessed by using the test strips, and the samples with the preservatives were further assessed for ß-lactamase activity using the cylinder plate method. Results showed that the refrigeration, freezing, and thawing of raw bovine milk samples had no influence on the assay of the ß-lactamase activity. The addition of sodium azide or potassium dichromate as a sample preservative failed the test of ß-lactamase activity, whereas the addition of sodium thiocyanate, bronopol, or methanol had no influence on the test. In conclusion, sodium azide and potassium dichromate were not suitable preservatives for assessing ß-lactamase activity in raw milk when the cylinder plate method was used.

[Establishment and validation of clinical prediction model for steroid-resistant nephrotic syndrome in children].

Objective: To identify the clinically relevant factors of steroid-resistant nephrotic syndrome (SSNS) in children and establish a predictive model followed by verifying its feasibility. Methods: A retrospective analysis was performed in a total of 111 children with nephrotic syndrome admitted to Children's Hospital of ShanXi from January 2016 to December 2021. The clinical data of general conditions, manifestations, laboratory tests, treatment, and prognosis were collected. According to the steroid response, patients were divided into SSNS and steroid resistant nephrotic syndrome (SRNS) group. Single factor Logistic regression analysis was used for comparison between the 2 groups, and variables with statistically significant differences were included in multivariate Logistic regression analysis. The multivariate Logistic regression analysis was used to identify the related variables of children with SRNS. The area under the receiver operating characteristic curve (ROC), the calibration curve and the clinical decision curve were used to evaluate its effectiveness of the variables. Results: Totally 111 children with nephrotic syndrome was composed of 66 boys and 45 girls, aged 3.2 (2.0, 6.6) years. There were 65 patients in the SSNS group and 46 in the SRNS group.Univariate Logistic regression analysis showed that the 6 variables, including erythrocyte sedimentation rate, 25-hydroxyvitamin D, suppressor T cells, D-dimer, fibrin degradation products, ß2-microglobulin, had statistically significant differences between SSNS and SRNS groups (85 (52, 104) vs. 105 (85, 120) mm/1 h, 18 (12, 39) vs. 16 (12, 25) nmol/L, 0.23 (0.19, 0.27) vs. 0.25 (0.20, 0.31), 0.7 (0.6, 1.1) vs. 1.1 (0.9, 1.7) g/L, 3.1 (2.3, 4.1) vs. 3.3 (2.7, 5.8) g/L, 2.3 (1.9,2.8) vs. 3.0 (2.5, 3.7) g/L, χ2=3.73, -2.42, 2.24, 3.38, 2.24,3.93,all P<0.05), were included in the multivariate Logistic regression analysis. Finally, we found that 4 variables including erythrocyte sedimentation rate, suppressor T cells, D-dimer and ß2-microglobulin (OR=1.02, 1.12, 25.61, 3.38, 95%CI 1.00-1.04, 1.03-1.22, 1.92-341.04, 1.65-6.94, all P<0.05) had significant correlation with SRNS. The optimal prediction model was selected. The ROC curve cut-off=0.38, with the sensitivity of 0.83, the specificity of 0.77 and area under curve of 0.87. The calibration curve showed that the predicted probability of SRNS group occurrence was in good agreement with the actual occurrence probability, χ2=9.12, P=0.426. The clinical decision curve showed good clinical applicability. The net benefit is up to 0.2. Make the nomogram. Conclusions: The prediction model based on the 4 identified risk factors including erythrocyte sedimentation rate, suppressor T cells, D-dimer and ß2-microglobulin was suitable for the early diagnosis and prediction of SRNS in children. The prediction effect was promising in clinical application.

Effects of essential oil/palygorskite composite on performance, egg quality, plasma biochemistry, oxidation status, immune response and intestinal morphology of laying hens.

The current study aimed to assess the effects of different levels of essential oil/palygorskite composite (EO-PGS) supplementation on performance, egg quality, oxidative status, immunity and intestinal morphology of laying hens. A total of 480 laying hens aged 65 wk were randomly assigned into 4 groups (6 replicates of 20 hens each). Hens were fed the basal diet supplemented with 0 (control diet), 0.5, 0.75 or 1.0 g/kg EO-PGS for 56 d. Data were analyzed by One-way ANOVA. Results showed that birds fed with diet supplemented with EO-PGS had increased the egg production (P < 0.05) more than birds fed with control diet. The yolk index and shell thickness were increased in 0.75 and 1.0 g/kg EO-PGS groups at d56 (P < 0.05). There was no significant difference in plasma biochemical parameters among all groups. Compared with the control group, supplementation of EO-PGS increased the immunoglobulin-G and interleukin-2 levels in plasma (P < 0.05). The total antioxidant capacity in plasma and liver, the plasma catalase concentration, the activity of total superoxide dismutase in the liver and the activity of glutathione peroxidase in the spleen were increased in the EO-PGS groups (P < 0.05). The concentration of malondialdehyde in the liver was decreased with the increasing level of EO-PGS (P < 0.05). The crypt depth of ileum and duodenum of birds fed with EO-PGS supplemented diet had a tendency to decrease (0.05

[Clinical characteristics and outcomes of patients newly diagnosed with multiple myeloma with extramedullary disease].

Objective: To compare the clinical characteristics and outcomes of patients newly diagnosed with multiple myeloma(NDMM)with bone-related extramedullary(EM-B)disease and those with extraosseous extramedullary(EM-E)disease and to address their prognostic factors. Methods: The clinical features, outcomes, and prognostic factors were retrospectively analyzed in 80 patients with NDMM with extramedullary disease. Results: Among 80 patients with extramedullary disease, 51 had EM-B and 29 EM-E. The level of ß(2)-microglobulin(5.82 mg/L vs 3.99 mg/L, P=0.030), lactate dehydrogenase(256 U/L vs 184 U/L, P=0.003), 1q21 amplification rate(78.6% vs 53.1%, P=0.035), and Ki-67 proliferation index(50% vs 25%, P=0.002)in the EME group were significantly higher than those in the EM-B group. The posieive rate of CD56(14.3% vs 66.7%)and overall response rate(60% vs 82.3%)in EM-E group were significantly lower than those in EM-B group. The median overall survival (OS)of patients with EM-E and EM-B was 14.5 and 49.5 months, and the median progression-free survival(PFS)of the two groups was 9.0 and 18.0 months. Patients with EM-E had a significantly shorter OS(P=0.035)and PFS(P < 0.001)than those of patients with EM-B, whereas the PFS did not significantly differ(P=0.263)when patients accepted proteasome inhibitor(PI)-based regimens for induction therapy. Multivariate analysis with Cox model showed the best response that did not achieve partial response(PR)was an independent poor prognostic factor for both OS and PFS in NDMM patients with EM-E(P=0.031, P=0.005), ISS-III, and the best response that did not achieve PR were independent prognostic factors for the shorter OS in patients with NDMM with EM-B(P=0.009, P=0.044). Conclusions: The clinical characteristics and outcomes of patients with NDMM with EM-E are different from patients with EM-B. Outcomes of patients with EM-E is significantly poor. PI induction therapy improved the PFS of patients with EM-E.

The effects and combinational effects of Bacillus subtilis and montmorillonite on the intestinal health status in laying hens.

This study was conducted to evaluate the effects and combinational effects of Bacillus subtilis (BS) and montmorillonite (MMT) on laying performance, gut mucosal oxidation status, and intestinal immunological and physical barrier functions of laying hens. Three hundred sixty laying hens (29-week-old) were randomly assigned to a 2 × 2 factorial arrangement of treatments (n = 6) for 10 wk as follows: (1) basal diet; (2) the basal diet plus 5 × 108 cfu BS/kg; (3) the basal diet plus 0.5 g MMT/kg; and (4) the basal diet plus 5 × 108 cfu BS/kg and 0.5 g MMT/kg. Dietary supplementation with BS increased egg production and egg mass, the activities of catalase (CAT) and total superoxide dismutase in the intestinal mucosa, and villus height and villus height-to-crypt depth ratio of the jejunum (P < 0.05) but downregulated the mRNA expression levels of toll-like receptor 4 and myeloid differentiation factor 88 (MyD88) in the duodenum and jejunum, interleukin 1 beta in the duodenum, and nuclear factor kappa B P65 (NF-κB P65) and tumor necrosis factor alpha in the jejunum (P < 0.05). Dietary supplementation with MMT increased egg production and egg mass, the concentration of secretory immunoglobulin A in the duodenum, and the occludin mRNA expression level in the jejunum (P < 0.05) but reduced feed conversion ratio, malondialdehyde concentration in the duodenum and jejunum, and the mRNA expression level of MyD88 in the jejunum (P < 0.05). In addition, there was an interaction effect between BS and MMT supplementation on the CAT activity and the MyD88 mRNA expression level in the duodenum and the mRNA expression level of occludin in the jejunum (P < 0.05). In conclusion, dietary BS and MMT and their combination may improve the intestinal health status of laying hens, which may contribute to the increase in hens' laying performance.

[Study on antibacterial properties and osteoblast activity of antimicrobial peptide coatings on titanium implants].

Objective: To investigate the antibacterial property and biological activity of Ti dental implant with antimicrobial peptide Pac-525 coatings, and to study the effect of peptide Pac-525 coatings on Porphyromonas gingivalis's antibacterial performance and osteoblast proliferation and adhesion. Methods: After ultrasonic micro arc oxidation, alkali treatment and silane treatment, forty-five pure titanium specimens were exposed to antibacterial peptide Pac-525 in different concentration (0.25, 0.50, 0.75 g/L). The titanium specimens in the control group were only treated with ultrasonic micro arc oxidation, alkali treatment and silane treatment. The morphologies of coatings were observed by scanning electron microscope (SEM), and the element changes were detected by energy spectrum analyzer. Orange acridine-ethidium bromide double staining was used to detect the average percentage of live bacteria and biofilm thickness, after the specimens in each group and Porphyromonas gingivalis were co-cultured for 72 hours. Cell counting Kit-8 method and immunofluorescence staining were used to test the proliferation of osteoblasts, the number and growth morphologies of adherent cells, respectively. Results: SEM and energy spectrum analysis showed that the Pac-525 particles loaded on the surface of the coating, and the C and N elements in the Pac-525 coating group were significantly more than those in the control group. The average percentage of living bacteria in the control group, 0.25, 0.50 and 0.75 g/L antimicrobial peptides were 0.58%, 0.45%, 0.34% and 0.28%, respectively, and the difference between each group was statistically significant (P<0.05). The biofilm thickness of Porphyromonas gingivalis in 0.50 and 0.75 g/L antibacterial peptide group were (98.3±1.2) and (94.5±2.5) µm respectively, which were significantly less than those in control group and 0.25 g/L antibacterial peptide group [(117.6±1.5) and (118.0±1.3) µm] (P<0.05), respectively. The number of bone cell adhesion and proliferation of all antimicrobial peptides were significantly greater than those in the control group (P<0.05), and the cells stretched better. Conclusions: The antibacterial peptide coating of titanium implants could inhibit the formation of bacterial biofilm. It had good antibacterial properties and could promote the adhesion and proliferation of osteoblasts.

[Efficacy and safety of CTD and PCD regimens in treatment of patients with newly diagnosed multiple myeloma].

Objective: To observe the efficacy and safety of CTD (cyclophosphamide, thalidomide, dexamethasone) and PCD (bortezomib, cyclophosphamide, dexamethasone) regimens in treatment of patients with newly diagnosed multiple myeloma (NDMM) . Methods: A retrospective analysis was carried out on 88 cases of NDMM patients admitted to our hospital from July 2013 to January 2016, including 49 cases in CTD group and 39 cases in PCD group. The outcomes of two different regimens were analyzed, including response, prognosis, and adverse events. Results: The total overall remission rates (ORR, better than PR) of CTD and PCD were 65.3% (32/49) and 84.6% (33/39) , while very good partial response (VGPR) were 30.6% (15/49) and 53.8% (21/39) , and differences were statistically significant (P=0.041, P=0.028) . The median follow-up was 11.5 (3-33) months. The median progression-free survival (PFS) was (23.0±4.5) months in CTD groups, but it was not achieved in PCD group, with statistically significant differences (P=0.050) . Medial overall survival was not achieved in both two groups, without statistically significant difference (P=0.257) . There were statistical differences between patients with minor response (MR) and patients without MR in medium OS in CTD group (P=0.005) , and there were statistical difference between patients with VGPR and without VGPR in medium OS in CTD group (P=0.042) . Infection was a common adverse event in two groups. The incidences of peripheral neuropathy and herpes zoster were markedly higher in PCD group than CTD group, and the incidences of thrombus, palpation and rash, etc., were higher in CTD group. Conclusion: Both CTD and PCD regimens were effective first-line induction chemotherapy choice for NDMM. PCD regimen is better than CTD in treatment power and deep remission.

Simultaneous determination of 38 veterinary antibiotic residues in raw milk by UPLC-MS/MS.

A selective and rapid method has been developed to determine, simultaneously, 38 veterinary antibiotic residues in raw milk by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). One milliliter of raw milk was diluted with 0.5 mL water and 3 mL acetonitrile, then purified using an Oasis HLB cartridge. The eluates were evaporated by nitrogen drying and then reconstituted to 4 mL with water/acetonitrile (8:1) before being injected into the UPLC-MS/MS system. The results indicated recoveries of 68-118% for 14 ß-lactams, 79-118% for eight quinolones, 71-106% for eight sulfonamides, 76-116% for four tetracyclines, 78-106% for three macrolides, and 88-103% for one lincosamides, with coefficients of variation less than 15% for intraday and interday precisions. The limit of quantification for all antibiotics was 0.03-10 µg kg(-1). This methodology was then applied to field-collected real raw milk samples and trace levels of four antibiotics were detected.

Analysis of a 69-kb contiguous genomic sequence at a putative tumor suppressor gene locus on human chromosome 6q27.

Multiple neoplasias including B-cell non-Hodgkin's lymphoma, breast carcinoma, and ovarian carcinoma, have been associated with frequent deletions of the distal region on the long arm of human chromosome 6, suggesting the presence of one or more tumor suppressor gene(s) at this locus. Loss of heterozygosity analysis of breast and ovarian tumors has further restricted the minimal region of loss within 6q27. To further characterize this genomic region for gene content including putative tumor suppressor genes as well as other elements that may contribute to tumorigenesis, a 68940-bp contiguous sequence, encompassing markers D6S193 and D6S297, was generated by random shotgun sequencing of a cosmid, P1, and PAC contig. In addition, exon trapping was performed utilizing a subset of these clones. Sixteen trapped exons, ranging in size from 44 to 399 bp, span this approximately 69-kb region. Many other putative exons have been identified computationally. Further analysis has identified 13 potential promoters and 13 putative polyadenylation sites in the region. Northern analysis identified a transcript mapping within this interval that is expressed in ovarian, breast, and lymphoid-derived tumor cell lines. Consideration of these data, together with the demonstration of several regions of high CpG content, suggests the possibility of several genes at this locus.

[A rapid FIA-spectrophotometric method with online extraction for the content uniformity test of atropine sulfate tablets].

A rapid spectrophotometric method based on FIA for the content uniformity test of atropine sulfate tablets has been developed. By means of the self-designed FIA system with an on-line extraction device, many difficulties resulting from extraction, such as slow analytical rate, environmental pollution, poor reproducibility, etc, were overcome, and the routine procedure of the official content uniformity test was tremendously simplified. Results obtained with the proposed method agree well with those obtained with the official test. Furthermore, the proposed method attained a sampling rate of 60 samples per hour, a recovery of 100.0%, a relative standard deviation of 0.7%, and a detection limit of 0.24 micrograms/ml. Uniform design proves effective to optimize complex FIA conditions, and FIA promises a new approach to test the content uniformity of pharmaceutical tablets.