M2-like tumor-associated macrophages-secreted Wnt1 and Wnt3a promotes dedifferentiation and metastasis via activating ß-catenin pathway in thyroid cancer.

BACKGROUND: Thyroid carcinoma (TC) has been a global issue for its rapid increasing incidence worldwide. Although most TC was not so aggressive with a good prognosis, treatment against anaplastic TC was relatively limited and the mechanisms are not well elucidated yet. METHODS: TC cell lines (IHH4 and TPC-1) were used. Flow cytometry was used to identify the surface marker of M2-like tumor-associated macrophages (TAMs) from cell culture. Quantitative real-time polymerase chain reaction, western blot analysis, immunostaining, and immunohistochemistry were used to detect the expression of Wnt1, Wnt3a, components of Wnt/ß-catenin pathway, and proliferation/epithelial-mesenchymal transition (EMT)-related proteins. Alkaline phosphatase activity assay, colony formation assay, and transwell assay were used to examine the roles of Wnt1, Wnt3a, and ß-catenin pathway in cell dedifferentiation, proliferation, migration, and invasion of TC cells, respectively. Subcutaneous tumor growth was monitored in nude mice. RESULTS: Coculture with M2-like TAMs facilitated dedifferentiation, proliferation, migration, and invasion in TC cells. EMT and proliferation-related proteins were also promoted in cocultured TC cells. The level of Wnt1 and Wnt3a was increased in the coculture system. Block of Wnt1 or Wnt3a suppressed malignant behaviors in cocultured tumor cells. Furthermore, Wnt1 or Wnt3a knockdown inhibited Wnt/ß-catenin signaling pathway, and suppressed EMT and proliferation-related signals in cocultured tumor cells. Knockdown of Wnt1 or Wnt3a inhibited tumor growth in xenograft model. CONCLUSION: M2-like TAMs promoted dedifferentiation, proliferation, and metastasis of TC by Wnt1 and Wnt3a secretion and ensuing ß-catenin activation.

Pharmacokinetics and safety of pegylated recombinant human granulocyte colony-stimulating factor in children with acute leukaemia.

AIMS: This open-label, phase I study evaluated the pharmacokinetics and safety of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) for the treatment of chemotherapy-induced neutropenia in children with acute leukaemia. METHODS: PEG-rhG-CSF was administered as a single 100 mcg/kg (3 mg maximum dose) subcutaneous injection at the end of each chemotherapy period when neutropenia occurred. Blood samples were obtained from patients treated with PEG-rhG-CSF. PEG-rhG-CSF serum concentrations were determined by an enzyme-linked immunosorbent assay. Population pharmacokinetic (PPK) analysis was implemented using the nonlinear mixed-effects model. Short-term safety was evaluated through adverse events collection (registered at clinicaltrials.gov identifier: 03844360). RESULTS: A total of 16 acute leukaemia patients (1.8-13.6 years) were included, of whom two (12.5%) had grade 3 neutropenia, six (37.5%) had grade 4 neutropenia, and eight (50.0%) had severe neutropenia. For PPK modelling, 64 PEG-rhG-CSF serum concentrations were obtainable. A one-compartment model with first-order elimination was used for pharmacokinetic data modelling. The current weight was a significant covariate. The median (range) of clearance (CL) and area under the serum concentration-time curve (AUC) were 5.65 (1.49-14.45) mL/h/kg and 16514.75 (6632.45-54423.30) ng·h/mL, respectively. Bone pain, pyrexia, anaphylaxis and nephrotoxicity were not observed. One patient died 13 days after administration, and the objective assessment of causality was that an association with PEG-rhG-CSF was "possible". CONCLUSIONS: The AUC of PEG-rhG-CSF (100 mcg/kg, 3 mg maximum dose) in paediatric patients with acute leukaemia were similar to those of PEG-rhG-CSF (100 mcg/kg) in children with sarcoma. PEG-rhG-CSF is safe, representing an important therapeutic option for chemotherapy-induced neutropenia in paediatric patients with acute leukaemia.

Implication of hsa_circ_0028007 in reinforcing migration, invasion, and chemo-tolerance of nasopharyngeal carcinoma cells.

BACKGROUND: Given the reliability of circRNAs in symbolizing cancer progression, this investigation was designed to expound the involvement of hsa_circ_0028007 in regulating chemosensitivity of nasopharyngeal carcinoma (NPC) cells. METHODS: Altogether, 241 pairs of NPC tissues and para-cancerous normal tissues were collected to identify NPC-symbolic circRNAs, which have been screened by circRNA microarray in advance. Expressions of the circRNAs were determined by means of real-time polymerase chain reaction (PCR). Besides, human NPC cell lines (ie, CNE2 and HONE1) were transfected by si-hsa_circ_0028007 and si-NC. Scratch assay, transwell assay, and MTT assay were performed to assess migration, invasion, and paclitaxel/cisplatin-resistance of NPC cell lines. RESULTS: Hsa_circ_0028007 expression was abnormally heightened within NPC tissues in comparison with matched non-tumor tissues (P < .05). Over-expressed hsa_circ_0028007 was strongly associated with advanced (III-IV) tumor stage, aggressive infiltration, and metastatic lymph nodes of NPC patients (P < .05). Regarding in vitro experiments, hsa_circ_0028007 expression was elevated in CNE2 and HONE1 cell lines as compared with HENE cell line (P < .05). Silencing of hsa_circ_0028007 not merely sensitized CNE2 and HONE1 cells against paclitaxel and cisplatin (P < .05), but also significantly repressed migration and invasion of the cell lines (P < .05). CONCLUSION: Hsa_circ_0028007 was involved in facilitating progression and chemo-resistance of NPC, which might offer an alternative for NPC treatment.

Ancient, highly heterogeneous mantle beneath Gakkel ridge, Arctic Ocean.

The Earth's mantle beneath ocean ridges is widely thought to be depleted by previous melt extraction, but well homogenized by convective stirring. This inference of homogeneity has been complicated by the occurrence of portions enriched in incompatible elements. Here we show that some refractory abyssal peridotites from the ultraslow-spreading Gakkel ridge (Arctic Ocean) have very depleted 187Os/188Os ratios with model ages up to 2 billion years, implying the long-term preservation of refractory domains in the asthenospheric mantle rather than their erasure by mantle convection. The refractory domains would not be sampled by mid-ocean-ridge basalts because they contribute little to the genesis of magmas. We thus suggest that the upwelling mantle beneath mid-ocean ridges is highly heterogeneous, which makes it difficult to constrain its composition by mid-ocean-ridge basalts alone. Furthermore, the existence of ancient domains in oceanic mantle suggests that using osmium model ages to constrain the evolution of continental lithosphere should be approached with caution.

Application of enhanced recovery after surgery in partial nephrectomy for renal tumors: A systematic review and meta-analysis.

Objectives: In recent years, enhanced recovery after surgery (ERAS) has been widely used in the field of urology, especially in radical cystectomy and radical prostatectomy, and has demonstrated its advantages. Although studies on the application of ERAS in partial nephrectomy for renal tumors are increasing, the conclusions are mixed, especially in terms of postoperative complications, etc, and its safety and efficacy are questionable. We conducted a systematic review and meta-analysis to assess the safety and efficacy of ERAS in the application of partial nephrectomy for renal tumors. Methods: Pubmed, Embase, Cohrance library, Web of science and Chinese databases (CNKI, VIP, Wangfang and CBM) were systematically searched for all published literature related to the application of enhanced recovery after surgery in partial nephrectomy for renal tumors from the date of establishment to July 15, 2022, and the literature was screened by inclusion/exclusion criteria. The quality of the literature was evaluated for each of the included literature. This Meta-analysis was registered on PROSPERO (CRD42022351038) and data were processed using Review Manager 5.4 and Stata 16.0SE. The results were presented and analyzed by weighted mean difference (WMD), Standard Mean Difference (SMD) and risk ratio (RR) at their 95% confidence interval (CI). Finally, the limitations of this study are analyzed in order to provide a more objective view of the results of this study. Results: This meta-analysis included 35 literature, including 19 retrospective cohort studies and 16 randomized controlled studies with a total of 3171 patients. The ERAS group was found to exhibit advantages in the following outcome indicators: postoperative hospital stay (WMD=-2.88, 95% CI: -3.71 to -2.05, p<0.001), total hospital stay (WMD=-3.35, 95% CI: -3.73 to -2.97, p<0.001), time to first postoperative bed activity (SMD=-3.80, 95% CI: -4.61 to -2.98, p < 0.001), time to first postoperative anal exhaust (SMD=-1.55, 95% CI: -1.92 to -1.18, p < 0.001), time to first postoperative bowel movement (SMD=-1.52, 95% CI: -2.08 to -0.96, p < 0.001), time to first postoperative food intake (SMD=-3.65, 95% CI: -4.59 to -2.71, p<0.001), time to catheter removal (SMD=-3.69, 95% CI: -4.61 to -2.77, p<0.001), time to drainage tube removal (SMD=-2.77, 95% CI: -3.41 to -2.13, p<0.001), total postoperative complication incidence (RR=0.41, 95% CI: 0.35 to 0.49, p<0.001), postoperative hemorrhage incidence (RR=0.41, 95% CI: 0.26 to 0.66, p<0.001), postoperative urinary leakage incidence (RR=0.27, 95% CI: 0.11 to 0.65, p=0.004), deep vein thrombosis incidence (RR=0.14, 95% CI: 0.06 to 0.36, p<0.001), and hospitalization costs (WMD=-0.82, 95% CI: -1.20 to -0.43, p<0.001). Conclusion: ERAS is safe and effective in partial nephrectomy of renal tumors. In addition, ERAS can improve the turnover rate of hospital beds, reduce medical costs and improve the utilization rate of medical resources. Systematic review registration: https://www.crd.york.ac.uk/PROSPERO, identifier CRD42022351038.

Archean cratonic mantle recycled at a mid-ocean ridge.

Basalts and mantle peridotites of mid-ocean ridges are thought to sample Earth's upper mantle. Osmium isotopes of abyssal peridotites uniquely preserve melt extraction events throughout Earth history, but existing records only indicate ages up to ~2 billion years (Ga) ago. Thus, the memory of the suspected large volumes of mantle lithosphere that existed in Archean time (>2.5 Ga) has apparently been lost somehow. We report abyssal peridotites with melt-depletion ages up to 2.8 Ga, documented by extremely unradiogenic 187Os/188Os ratios (to as low as 0.1095) and refractory major elements that compositionally resemble the deep keels of Archean cratons. These oceanic rocks were thus derived from the once-extensive Archean continental keels that have been dislodged and recycled back into the mantle, the feasibility of which we confirm with numerical modeling. This unexpected connection between young oceanic and ancient continental lithosphere indicates an underappreciated degree of compositional recycling over time.

An origin of ultraslow spreading ridges for the Yarlung-Tsangpo ophiolites.

As relics of ancient ocean lithosphere, ophiolites are the most important petrological evidence for marking the sutures and also play a key role in reconstructing plate configuration. They also provide valuable windows for studying crustal accretion and mantle processes occurring at modern ocean ridges. Abundant ophiolites are distributed along the Yarlung-Tsangpo suture and represent the relics of ocean lithosphere of the Neo-Tethys. They are characterized by an incomplete litho-stratigraphy, of which the mantle section is much thicker than the crustal section. Ocean crustal rocks outcropped in the Yarlung-Tsangpo ophiolites are much thinner than normal ocean crusts (~ 7 km) or even absent. Tectonic settings from which the Yarlung-Tsangpo ophiolites originated remain highly controversial, although an origin of the supra-subduction zone is prevailing. Moreover, their incomplete litho-stratigraphy has been commonly attributed to tectonic dismemberment during the late-stage emplacement after their formation. Nevertheless, such an incompleteness resembles the ocean lithosphere generated at modern ultraslow spreading ridges, such as the Southwest Indian Ridge (SWIR). In this paper, we present several lines of evidence that support the formation of the Yarlung-Tsangpo ophiolites at ultraslow spreading ridges, during which detachment faults were developed. This suggests that the Yarlung-Tsangpo ophiolites might represent the ocean core complexes (OCC) in the Neo-Tethys Ocean. The OCC with high topography in the seafloor were clogged in the trench and preserved as ophiolites through Indo-Eurasia collision. The clogging resulted in the demise of an old subduction and a new subduction was re-initiated beneath the clogged OCC.

Fermented Astragalus in diet improved laying performance, egg quality, antioxidant and immunological status and intestinal microbiota in laying hens.

In the era of increased antibiotic resistance and ever-stricter control on antibiotic use, it is urgent to develop green, safe, and non-residue alternatives to antibiotics applied to the poultry industry. To this end, we supplied the potential Lactobacillus plantarum (L. plantarum) fermented Astragalus in the diet of laying hens, with a final addition of 3‰. Its effects have been assessed on laying performance, egg quality, antioxidant and immunological status, and intestinal microbiota, and are compared to the control group, to the Astragalus group containing 3‰ unfermented Astragalus, and to the L. plantarum group containing 2% L. plantarum [5 × 108 colony-forming unit (CFU) per milliliter (mL)]. During the second half of the experimental period (15 to 28 days), the egg production rate was considerably higher in the fermented Astragalus group than that in the other groups, with the fermented Astragalus group having the lowest feed conversion ratio. No significant difference (P > 0.05) was noted among treatments on egg quality. Fermented Astragalus-treated hens exhibited significantly increased catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) in serum, and reduced malondialdehyde (MDA) in serum. Furthermore, fermented Astragalus supplementation resulted in a significant increase in ileal microbiota abundance relative to control. In conclusion, feeding laying hens with L. plantarum fermented Astragalus has beneficial effects on production, antioxidant potential, immunity, and ileal microbiota. L. plantarum fermented Astragalus is expected to be a novel feed additive used in poultry production.

Development of a reverse transcription recombinase polymerase amplification combined with lateral-flow dipstick assay for avian influenza H9N2 HA gene detection.

H9N2 avian influenza viruses (AIVs) have been detected from wild birds and domestic poultry worldwide. Serious diseases combined with secondary infection have caused high mortality and great economic losses to poultry industry. Therefore, simple, rapid, sensitive and accurate methods suitable for field detection of H9N2 AIVs are crucial to efficiently control virus infection and spread in time. In this study, an isothermal reverse transcription recombinase polymerase amplification with lateral-flow dipstick (RT-RPA-LFD) assay for detection of hemagglutinin (HA) gene of H9 subtype influenza viruses was developed. The optimal forward and reverse primers targeting HA gene of H9 subtype influenza viruses were labeled with fluorescein isothiocyanate (FITC) and biotin at the 5'-end, respectively. The amplification reaction could be finished in 20 min at a wide temperature range of 30-42°C, and then the products could be visualized with naked eyes. The developed H9 RT-RPA-LFD was able to detect 0.15 pg of H9N2 AIV RNA, which was 10 times more sensitive than that of conventional RT-PCR. The H9 RT-RPA-LFD assay did not detect nucleic acids extracted from H9 negative samples or from other poultry respiratory pathogens. The clinical performance of H9 RT-RPA-LFD was determined by testing 120 cloacal samples collected from chickens with respiratory syndromes. The coincidence rate of the detection results between RT-RPA-LFD and conventional RT-PCR was 95.8%. Therefore, the developed RT-RPA-LFD assay provides a rapid, reliable and sensitive method for field diagnosis of H9 subtype AIVs.

N,N-dimethyl-beta-alanine as an inexpensive and efficient ligand for palladium-catalyzed Heck reaction.

N,N-Dimethyl-beta-alanine was found to be a more powerful phosphine-free ligand than the previously reported ligand, N,N-dimethylglycine, in the Pd-catalyzed Heck reaction for a variety of aryl bromides, aryl iodides, and activated aryl chlorides with a practical turnover number of 10(3). Both kinetic and theoretical studies suggested that N,N-dimethyl-beta-alanine led to faster oxidative addition of an aryl halide to Pd than N,N-dimethylglycine. [reaction: see text]

Computational prediction of microRNA genes in silkworm genome.

MicroRNAs (miRNAs) constitute a novel, extensive class of small RNAs (approximately 21 nucleotides), and play important gene-regulation roles during growth and development in various organisms. Here we conducted a homology search to identify homologs of previously validated miRNAs from silkworm genome. We identified 24 potential miRNA genes, and gave each of them a name according to the common criteria. Interestingly, we found that a great number of newly identified miRNAs were conserved in silkworm and Drosophila, and family alignment revealed that miRNA families might possess single nucleotide polymorphisms. miRNA gene clusters and possible functions of complement miRNA pairs are discussed.

A 'hidden' 18O-enriched reservoir in the sub-arc mantle.

Plate subduction continuously transports crustal materials with high-δ(18)O values down to the mantle wedge, where mantle peridotites are expected to achieve the high-δ(18)O features. Elevated δ(18)O values relative to the upper mantle value have been reported for magmas from some subduction zones. However, peridotites with δ(18)O values significantly higher than the well-defined upper mantle values have never been observed from modern subduction zones. Here we present in-situ oxygen isotope data of olivine crystals in Sailipu mantle xenoliths from South Tibet, which have been subjected to a long history of Tethyan subduction before the India-Asia collision. Our data identify for the first time a metasomatized mantle that, interpreted as the sub-arc lithospheric mantle, shows anomalously enriched oxygen isotopes (δ(18)O = +8.03 ± 0.28 ‰). Such a high-δ(18)O mantle commonly does not contribute significantly to typical island arc basalts. However, partial melting or contamination of such a high-δ(18)O mantle is feasible to account for the high-δ(18)O signatures in arc basalts.

Perfluorooctane sulfonate blocked autophagy flux and induced lysosome membrane permeabilization in HepG2 cells.

Perfluorooctane sulfonate (PFOS) is an emerging persistent organic pollutant widely distributed in the environment, wildlife and human. In this study, as observed under the transmission electron microscope, PFOS increased autophagosome numbers in HepG2 cells, and it was confirmed by elevated LC3-II levels in Western blot analysis. PFOS increased P62 level and chloroquine failed to further increase the expression of LC3-II after PFOS treatment, indicating that the accumulation of autophagosome was due to impaired degradation rather than increased formation. With acridine orange staining, we found PFOS caused lysosomal membrane permeabilization (LMP). In this study, autophasome formation inhibitor 3-methyladenine protected cells against PFOS toxicity, autophagy stimulator rapamycin further decreased cell viability and increased LDH release, cathepsin inhibitor did not influence cell viability of PFOS-treated HepG2 cells significantly. These further supported the notion that autophagic cell death contributed to PFOS-induced hepatotoxicity. In summary, the data of the present study revealed that PFOS induced LMP and consequent blockage of autophagy flux, leading to an excessive accumulation of the autophagosomes and turning autophagy into a destructive process eventually. This finding will provide clues for effective prevention and treatment of PFOS-induced hepatic disease.

[The development of a rapid loop-mediated indirect PCR method for detection and differentiation of highly and lowly pathogenic porcine reproductive and respiratory syndrome virus].

The aim of this study is to establish the method of loop-mediated indirect PCR assay for detection of Reproductive and Respiratory Syndrome Virus (PRRSV) infection and differentiation of highly pathogenic PRRSV (HP-PRRSV) and lowly pathogenic PRRSV (LP-PRRSV). Based on the alignments of ORF2 gene sequences and ORFla gene sequences of PRRSV Chinese isolates deposited in GenBank, two pairs of specific probes were designed and labeled to both ends of the soybean Lectin gene fragment by PCR, respectively. The probe-labeled soybean Lectin genes were used to be reporter genes for detection and differentiation of PRRSV. After one round strand displacement reaction, the reporter genes were amplified by reverse PCR. The specific PCR products were 193bp, 355bp for HP-PRRSV and 193bp, 442bp for LP-PRRSV, respectively. The method could detect 5. 6 TCID50/mL LP-PRRSV RNA and 18 TCIDs0/ mL HP-PRRSV RNA, and co-infection did not affect detection sensitivity. No amplification was observed with other porcine originated pathogens including CSFV, PPV, PRV, PCV2, ETEC and Haemophilus parasui. Twenty clinical samples were used for comparative testing with conventional PCR. Fourteen samples were found positive for PRRSV by the loop-mediated indirect PCR, of which 4 were LP-PRRSV, 9 HP-PRRSV and 1 LP/HP-PRRSV co-infection, consistent with the conventional PCR test results. In conclusion, the loop-mediated indirect PCR is a simple, rapid, sensitive and specific etiologic diagnosis tool, and suitable for the differential diagnosis of HP/LP-PRRSV, especially for identification of mixed infection of HP/LP-PRRSV.

Abnormal splenic artery diameter/hepatic artery diameter ratio in cirrhosis-induced portal hypertension.

AIM: To determine an optimal cutoff value for abnormal splenic artery diameter/proper hepatic artery diameter (S/P) ratio in cirrhosis-induced portal hypertension. METHODS: Patients with cirrhosis and portal hypertension (n = 770) and healthy volunteers (n = 31) underwent volumetric computed tomography three-dimensional vascular reconstruction to measure the internal diameters of the splenic artery and proper hepatic artery to calculate the S/P ratio. The cutoff value for abnormal S/P ratio was determined using receiver operating characteristic curve analysis, and the prevalence of abnormal S/P ratio and associations between abnormal S/P ratio and major complications of portal hypertension were studied using logistic regression. RESULTS: The receiver operating characteristic analysis showed that the cutoff points for abnormal splenic artery internal diameter and S/P ratio were > 5.19 mm and > 1.40, respectively. The sensitivity, specificity, positive predictive value, and negative predictive value were 74.2%, 45.2%, 97.1%, and 6.6%, respectively. The prevalence of an abnormal S/P ratio in the patients with cirrhosis and portal hypertension was 83.4%. Patients with a higher S/P ratio had a lower risk of developing ascites [odds ratio (OR) = 0.708, 95%CI: 0.508-0.986, P = 0.041] and a higher risk of developing esophageal and gastric varices (OR = 1.483, 95%CI: 1.010-2.175, P = 0.044) and forming collateral circulation (OR = 1.518, 95%CI: 1.033-2.230, P = 0.034). After splenectomy, the portal venous pressure and maximum and mean portal venous flow velocities were reduced, while the flow rate and maximum and minimum flow velocities of the hepatic artery were increased (P < 0.05). CONCLUSION: The prevalence of an abnormal S/P ratio is high in patients with cirrhosis and portal hypertension, and it can be used as an important marker of splanchnic hemodynamic disturbances.

[Eukaryotic expression of NS1 major antigen region of PPV and development of an indirect ELISA based on the expressed protein].

To construct secretory expression vector of PPV NS1 gene, the fragment of PPV NS1 gene coding for major antigen region of the NS1 protein was amplified by PCR and inserted into multiple clone site of eukaryotic expression vector pPICZalpha-A. The recombinant pPICZalpha-A-NS1 plasmid was transferred into P. pastoris strain GS115 mediated by electro transform. Recombinant P. pastoris strain GS115 was induced to express the fusion protein by methanol. The expressed and purified protein was analyzed by SDS-PAGE and Western Blot. The recombinant protein was highly-expressed and showed a good immunoreactivity. The indirect ELISA method was developed for detecting antibodies against PPV by checkerboard titration assay. The result showed that the optimal concentration of coated antigen was 3.2 microg/mL and the best dilution of serum was 1 : 80. The positive cut-off value of the ELISA assay was OD450 > 0.4 and OD450 positive serum/OD450 negative serum > 2.0. Compared with HI and commercial ELISA kits, the assay revealed 94.2% and 92.1% agreement respectively. The assay demonstrates good specificity and sensitivity, and can be applied in the detection of porcine parvovirus.

[Influence of interpolation method and sampling number on spatial prediction accuracy of soil Olsen-P].

Different from the large scale farm management in Europe and America, the scattered farmland management in China made the spatial variability of soil nutrients at county scale in this country more challenging. Taking soil Olsen-P in Wuhu County as an example, the influence of interpolation method and sampling number on the spatial prediction accuracy of soil nutrients was evaluated systematically. The results showed that local polynomial method, ordinary kriging, simple kriging, and disjunctive kriging had higher spatial prediction accuracy than the other interpolation methods. Considering of its simplicity, ordinary kriging was recommended to evaluate the spatial variability of soil Olsen-P within a county. The spatial prediction accuracy would increase with increasing soil sampling number. Taking the spatial prediction accuracy and soil sampling cost into consideration, the optimal sampling number should be from 500 to 1000 to evaluate the spatial variability of soil Olsen-P at county scale.