Identification of whole-genome mutations and structural variations of bile cell-free DNA in cholangiocarcinoma.

Bile cell-free DNA (cfDNA) has been reported as a promising liquid biopsy tool for cholangiocarcinoma (CCA), however, the whole-genome mutation landscape and structural variants (SVs) of bile cfDNA remains unknown. Here we performed whole-genome sequencing on bile cfDNA and analyzed the correlation between mutation characteristics of bile cfDNA and clinical prognosis. TP53 and KRAS were the most frequently mutated genes, and the RTK/RAS, homologous recombination (HR), and HIPPO were top three pathways containing most gene mutations. Ten overlapping putative driver genes were found in bile cfDNA and tumor tissue. SVs such as chromothripsis and kataegis were identified. Moreover, the hazard ratio of HR pathway mutations were 15.77 (95% CI: 1.571-158.4), patients with HR pathway mutations in bile cfDNA exhibited poorer overall survival (P = 0.0049). Our study suggests that bile cfDNA contains genome mutations and SVs, and HR pathway mutations in bile cfDNA can predict poor outcomes of CCA patients.

Amino Acids Biostimulants and Protein Hydrolysates in Agricultural Sciences.

The effects of different types of biostimulants on crops include improving the visual quality of the final products, stimulating the immune systems of plants, inducing the biosynthesis of plant defensive biomolecules, removing heavy metals from contaminated soil, improving crop performance, reducing leaching, improving root development and seed germination, inducing tolerance to abiotic and biotic stressors, promoting crop establishment and increasing nutrient-use efficiency. Protein hydrolysates are mixtures of polypeptides and free amino acids resulting from enzymatic and chemical hydrolysis of agro-industrial protein by-products obtained from animal or plant origins, and they are able to alleviate environmental stress effects, improve growth, and promote crop productivity. Amino acids involve various advantages such as increased yield and yield components, increased nutrient assimilation and stress tolerance, and improved yield components and quality characteristics. They are generally achieved through chemical or enzymatic protein hydrolysis, with significant capabilities to influence the synthesis and activity of some enzymes, gene expression, and redox-homeostasis. Increased yield, yield components, and crop quality; improved and regulated oxidation-reduction process, photosynthesis, and physiological activities; decreased negative effects of toxic components; and improved anti-fungal activities of plants are just some of the more important benefits of the application of phenols and phenolic biostimulants. The aim of this manuscript is to survey the impacts of amino acids, different types of protein hydrolysates, phenols, and phenolic biostimulants on different plants by presenting case studies and successful paradigms in several horticultural and agricultural crops.

Heterologous Expression and Function of Cholesterol Oxidase: A Review.

Cholesterol was first found in gallstones as an animal sterol; hence it is called cholesterol. Cholesterol oxidase is the chief enzyme in the process of cholesterol degradation. Its role is obtained by the coenzyme FAD, which catalyzes the isomerization and oxidation of cholesterol to produce cholesteric 4-ene-3-ketone and hydrogen peroxide at the same time. Recently, a great advance has been made in the discovery of the structure and function of cholesterol oxidase, and it has proven added value in clinical discovery, medical care, food and biopesticides development and other conditions. By recombinant DNA technology, we can insert the gene in the heterologous host. Heterologous expression (HE) is a successful methodology to produce enzymes for function studies and manufacturing applications, where Escherichia coli has been extensively used as a heterologous host because of its economical cultivation, rapid growth, and efficiency in offering exogenous genes. Heterologous expression of cholesterol oxidase has been considered for several microbial sources, such as Rhodococcus equi, Brevibacterium sp., Rhodococcus sp., Streptomyces coelicolor, Burkholderia cepacia ST-200, Chromobacterium, and Streptomyces spp. All related publications of numerous researchers and scholars were searched in ScienceDirect, Scopus, PubMed, and Google Scholar. In this article, the present situation and promotion of heterologous expression of cholesterol oxidase, the role of protease, and the perspective of its possible applications were reviewed.

Treatment of Chronic Aplastic Anemia with Chinese Patent Medicine Pai-Neng-Da Capsule () for Replacing Androgen Partially: A Clinical Multi-Center Study.

OBJECTIVE: To evaluate the efficacy and safety of Pai-Neng-Da Capsule (, panaxadiol saponins component, PNDC) in combination with the cyclosporine and androgen for patients with chronic aplastic anemia (CAA). METHODS: A total of 79 CAA patients was randomly divided into 2 groups by a random number table, including PCA group [43 cases, orally PNDC 320 mg/d plus cyclosporine 5 mg/(kg·d) plus andriol 80 mg/d] and CA group [36 cases, orally cyclosporine 5 mg/(kg·d) plus andriol 160 mg/d]. All patients were treated and followed-up for 6 treatment courses over 24 weeks. The complete blood counts, score of Chinese medical (CM) symptoms were assessed and urine routine, electrocardiogram, hepatic and renal function were observed for safety evaluation. Female masculinization rating scale was established according to the actual clinical manifestations to evaluate the accurate degree of masculinization in female CAA patients treated by andriol. RESULTS: The effective rates were 88.1% (37/42) in the PCA group and 77.8% (28/36) in the CA group based on the standard for the therapeutic efficacy evaluation of hematopathy. There was no significant difference in the white blood cell (WBC) counts, platelet counts and hemoglobin concentration of peripheral blood between two groups after 6 months treatment. The masculinization score of female patient in the PCA group was significantly lower than the CA group (P<0.05). The mild abdominal distention was observed in 1 cases in the PCA group. In CA group, the abnormalities in the hepatic function developed in 2 cases and the renal disfunction was found in 1 case. CONCLUSION: The PNDC possesses certain curative effects in the treatment of CAA without obvious side-effects and can partially replace andriol thereby to reduce the degree of masculinization [Registried at Chinese Clinical Trial Registry (ChicTR1900028153)].

Synthesis of graphene oxide/polyethyleneimine sponge and its performance in the sustainable removal of Cu(II) from water.

Modification of graphene oxide (GO) with polyethyleneimine (PEI) has been studied to develop a GO/PEI sponge material that not only performs well in the adsorption of Cu(II) but also is easily separated from water. The results showed that GO had excellent affinity for PEI, and GO/PEI prepared at pH 9.0 using PEI with a MW of 70,000 was shown to be a good adsorbent for Cu(II). This GO/PEI was characterized with SEM, XRD, XPS, FTIR and TG analyses and was investigated for Cu(II) adsorption further. The adsorption isotherm data of Cu(II) were fitted well with the Langmuir model, from which the maximum adsorption of GO/PEI was calculated to be 150.9 mg/g at pH 5.5. This was much higher than that of GO and two commercial resins. GO/PEI showed high selectivity towards Cu(II). In GO/PEI, the contributions of amino groups on PEI and negative charges on GO were 79.2% and 20.8%, respectively. Cu(II) adsorption on GO/PEI decreased with decreasing pH, and 1 M HCl caused nearly complete desorption of the adsorbed Cu(II). Experimental results of five cycles of adsorption-desorption indicated that this material could be reused. Column studies showed that GO/PEI performed well in terms of both Cu(II) adsorption and stability in water.

Early Death and Survival of Patients With Acute Promyelocytic Leukemia in ATRA Plus Arsenic Era: A Population-Based Study.

Most randomized trials for acute promyelocytic leukemia (APL) have investigated highly selected patients under idealized conditions, and the findings need to be validated in the real world. We conducted a population-based study of all APL patients in Zhejiang Province, China, with a total population of 82 million people, to assess the generalization of all-trans retinoic acid (ATRA) and arsenic as front-line treatment. The outcomes of APL patients were also analyzed. Between January 2015 and December 2019, 1,233 eligible patients were included in the final analysis. The rate of ATRA and arsenic as front-line treatment increased steadily from 66.2% in 2015 to 83.3% in 2019, with no difference among the size of the center (≥5 or <5 patients per year, p = 0.12) or age (≥60 or <60 years, p = 0.35). The early death (ED) rate, defined as death within 30 days after diagnosis, was 8.2%, and the 3-year overall survival (OS) was 87.9% in the whole patient population. Age (≥60 years) and white blood cell count (>10 × 109/L) were independent risk factors for ED and OS in the multivariate analysis. This population-based study showed that ATRA and arsenic as front-line treatment are widely used under real-world conditions and yield a low ED rate and a high survival rate, which mimic the results from clinical trials, thereby supporting the wider application of APL guidelines in the future.

Coupled influence of pH and dissolved organic carbon on the immobilization of phosphorus by lanthanum-modified zeolite.

Wind-driven waves and currents in shallow lakes frequently trigger the resuspension of sediments in the photic layer, which is characterized with a high pH and high dissolved organic carbon (DOC) concentration. The mechanism of phosphorus-inactivating agents (PIAs) immobilizing phosphorus under the coupled influence of pH and DOC is not clarified, and the applicability of PIAs in eutrophic shallow lakes is thus still doubtful. We found that, under the coupled influence of pH and DOC, the uptake of phosphate by LMZ was affected mainly by pH at low DOC concentrations and by DOC at high DOC concentrations. A high pH (9.3) and high DOC concentration (24.7 mg/L) greatly increased the release of phosphorus from sediment to water. However, the addition of LMZ substantially reduced the P concentrations in water, mainly via capture of dissolved inorganic phosphorus. The results of the reversibility of the adsorption of phosphates and DOC showed that phosphate had much higher affinity than DOC towards LMZ. The phosphate once adsorbed on LMZ was resistant to release when exposed to conditions of either a high pH (9.5), high DOC concentration (250 mg/L) or both; i.e., only <5% of the adsorbed phosphate is releasable. Therefore, we proposed that, to avoid the coupled influence of pH and DOC in the photic layer of eutrophic shallow lakes, LMZ could be applied in multiple low doses in the season when the growth of algae is minimal (a low pH and low DOC concentration).

Interactions of phosphate and dissolved organic carbon with lanthanum modified bentonite: Implications for the inactivation of phosphorus in lakes.

Lanthanum-modified bentonite (LMB) is a widely used phosphorus-inactivating agent in lakes. However, dissolved organic carbon (DOC) exists ubiquitously in lakes, and its influence on phosphate binding is still not adequately understood. Our results showed that both phosphate and DOC can be adsorbed by LMB. The Langmuir adsorption maxima of phosphate and DOC were 9.06 mg P/g and 5.31 mg C/g, respectively, generating a C/P molar ratio ∼1.5. When phosphate and DOC coexisted at this ratio, the adsorption of phosphate was not influenced by DOC and vice versa. However, the phosphate capture by LMB was significantly reduced by raising the ratio above ∼9, and the reduction was increased with increasing the ratio. Once adsorbed by LMB, phosphate was essentially not desorbed by DOC, while adsorbed DOC can be mostly liberated by phosphate. It is deemed that phosphate can interact preferentially with La on LMB. However, DOC can still be adsorbed by LMB, even after LMB was saturated with phosphate, which was attributed to (i) the high coordination capacity of La; (ii) the interaction of DOC with the hydroxyl group(s) of the adsorbed phosphate via hydrogen bonding; and (iii) the interaction of DOC with the La sites unoccupied by phosphate. We proposed that LMB can be applied in the season (time) when the DOC/P ratio in lakes is low enough to facilitate the adsorption of phosphate, which will no longer be released into water, even after the C/P ratio is raised later.

Competitive adsorption of phosphate and dissolved organic carbon on lanthanum modified zeolite.

Lanthanum (hydr)oxide-based materials are attractive as highly efficient adsorbents for phosphate removal from both sewage and lake environment. However, dissolved organic carbon (DOC) coexists in the waters and exact information is still lacking on how DOC influence the phosphate adsorption process. In this study, competitive adsorption of phosphate and DOC on lanthanum modified zeolite (LMZ) was investigated using humic acid as the representative. In LMZ, lanthanum hydroxide was shown to be the active ingredient accounting for >98% of the binding sites of both phosphate and DOC. Without competition, the maximum adsorption capacity of phosphate and DOC estimated from the Langmuir isotherm model was 52.25 and 41.32 mg/g, respectively. When coexisted, DOC did not affect the adsorption of phosphate while phosphate reduced the adsorption of DOC by ~40%. In addition, preloading LMZ with DOC had little effect on phosphate adsorption while coating with phosphate substantially lowered DOC adsorption. Furthermore, phosphate can release most of the adsorbed DOC (>60%), while DOC can not replace adsorbed phosphate (<2%). The adsorption kinetics of both phosphate and DOC was best described by the psudo-second-order model (r2 > 0.999). The adsorption of both phosphate and DOC increased with decreasing pH or increasing ionic strength. We proposed that phosphate was competitive than DOC for the ligand exchange sites of singly-coordinated hydroxyls, but DOC can be solely adsorbed onto the uncharged hydroxyls via hydrogen bonding.

The recovery of phosphorus from source-separated urine by repeatedly usable magnetic Fe3O4@ZrO2 nanoparticles under acidic conditions.

The separation of urine at source for phosphorus (P) recovery is attractive taking into account the high P concentration and small volume. However, the treatment of urine is still challenging due to its unpleasant odor and hygiene problems. Because the above problems could be solved by acidification to keep the pH of urine below 4, we propose a novel strategy to recover P from acidified urine using tailored hydrous zirconia-coated magnetite nanoparticles (Fe3O4@ZrO2). This strategy involves the selective adsorption of phosphate by easily separable and reusable Fe3O4@ZrO2, the desorption of adsorbed phosphate, and the precipitation of desorbed phosphate as calcium phosphate fertilizer. The results indicated that at pH 4, the P in synthetic urine was selectively adsorbed and could be exhausted using Fe3O4@ZrO2. Nearly all (>97.5%) of the sequestered P on the Fe3O4@ZrO2 nanoparticles was stripped using ≥1 M NaOH solution and ~100% of the stripped P was then successfully transformed into calcium phosphate, upon adding CaCl2 at pH >12 and a Ca/P molar ratio of 3. The liquid/solid (Fe3O4@ZrO2 particles) mixture could be conveniently separated for reuse using an external magnetic field. The reusability of the Fe3O4@ZrO2 nanoparticles in the extraction of P from synthetic urine was confirmed using five cycles of the adsorption-desorption process and their performance validated using real urine samples. The mechanism of phosphate adsorption was investigated using XPS, FTIR and zeta potential measurements, showing that phosphate was chemically adsorbed on the surface through direct coordination to zirconium atom via ligand exchange.

Surfactant-loaded graphene oxide sponge for the simultaneous removal of Cu2+ and bisphenol A from water.

The objective of this study was to develop a novel graphene oxide (GO)-based adsorbent by loading the cationic surfactant hexadecyltrimethylammonium bromide (HDTMA) to simultaneously scavenge copper ion, a charged species, and bisphenol A, an uncharged organic compound, from water. The HDTMA modification process was studied and the GO/HDTMA composites characterized using SEM (scanning electron microscopy), XRD (X-ray diffraction), XPS (X-ray photoelectron spectroscopy) and FTIR (Fourier Transform Infrared) spectroscopy. Within the concentration range of 6.4-11.5%, HDTMA caused the 2D GO sheets to form into solid 3D networks by reducing the repulsive forces and increasing the hydrophobic interactions between the adjacent GO sheets. The unique feature of this material is the simultaneous uptake of charged heavy metal ions and uncharged organic contaminants. The negative charges on GO results in the retention of heavy metal ions, while the hydrophobic phase created by the alkyl chain in HDTMA enables the adsorption of organic contaminants. The adsorption capacity of Cu2+ and bisphenol A reached 59.7 mg/g and 141.0 mg/g, respectively. The adsorption processes for both Cu2+ and bisphenol A were rapid, attaining ∼100% removal in 1 h and 2 h, respectively. Increasing the pH favored the adsorption of the two solutes. The presence of NaCl reduced the retention of Cu2+, but was beneficial for the adsorption of bisphenol A. The results demonstrate that the 3D structure and the adsorption of the target species can be achieved by tailoring the surface coverage of HDTMA on GO.

Multiple risks analysis for aplastic anemia in Zhejiang, China: A case-control study.

To understand the risks associated with aplastic anemia (AA) in 4 cities of Zhejiang Province, China, with special focus on the joint contributions of multiple risks.Based on an Electronic Data Capture (EDC), a case control study was carried out. Data regarding socio-demographic, diseases history, living habits, and exposures to toxic substances, etc., were collected through survey questionnaires. t Test, chi-square test, or non-parametric rank sum test, and univariate and multivariate Logistic regression analysis were conducted to analyze data.The univariate logistic regression analysis results indicated that among all study participants (n = 1802), AA was associated with over 30 risks, in terms of their individual behaviors, daily and environmental exposures, diseases history, and family history. Multivariate logistic regression analysis further confirmed that the independent risks related to AA included presence of chemical factory within 3 km of living residence (odds ratio [OR] = 8.73, 95% CI: 1.42-53.74, P = .019), living in a newly decorated house/apartment (OR = 25.37, 95% CI: 4.44-144.81, P < .001), vegetarian diet (OR = 131.60, 95% CI: 3.45-5020.16, P = .009), preference of sugar (OR = 89.38, 95% CI: 7.22-1106.44, P < .001), preference of oily food (OR = 55.68, 95% CI: 5.12-605.26, P = .001), drinking lake water or pond water (OR = 58.05, 95% CI: 3.21-1049.81, P < .001), habit of staying up late (OR = 11.87, 95% CI: 3.43-41.02, P < .001), infection history (OR = 10.08, 95% CI: 2.75-36.93, P < .001). Result of receiver operating characteristic curve (ROC) analysis on the joint contribution of multiple risks indicated that AA was 13.835 times likely to occur when exposed to ≥1 risks than those exposed to 0 risks (95% CI: 9.995-19.149).Our study results demonstrated a comprehensive epidemiological pattern, in which the joint contributions of individual inherited health status, environment exposure, and individual behaviors lead to the occurrence of AA.

Clinical study of pai-neng-da capsule in the treatment of chronic aplastic anemia.

OBJECTIVE: To evaluate the efficacy and safety of Pai-Neng-Da Capsule (panaxadiol saponins component, PND), a new Chinese patent medicine, on patients with chronic aplastic anemia (CAA) and to explore the optimal therapeutic regimen for CAA. METHOD: A total of 36 patients with CAA were enrolled and divided into three groups: the AP group (20 cases, andriol 120 mg/day + PND 240 mg/day), the ACP group (13 cases, andriol 120 mg/day + cyclosporine 3-6 mg kd(-1) day(-1) + PND 240 mg/day), and the PND group (3 cases, PND 240 mg/day). All patients were treated and followed up for 6 months. Peripheral blood counts, renal and hepatic function and Chinese medical (CM) symptoms of patients were assessed and all indices were gathered at the beginning and end of the study. RESULT: In the AP group, no significant hematologic difference was observed at the end of 6-month treatment comparing with the beginning. In the ACP group, the blood counts were maintained at the same level after the 6-month treatment. In the PND group, trilineage hematologic improvement was displayed at the end of 6-month treatment comparing with the beginning. No significant difference was showed in renal and hepatic function in all patients. All patients' clinical symptom improved according to CM symptom score. The effective rates were 95%, 73% and 100%, respectively. CONCLUSION: PND improved the efficacy and decreased side effects by cutting down the dosage of andriol, and it could also improve patients' clinical symptom and quality of life. PND were effective and safe in the treatment of CAA, it could be used alone or in combination with pharmacological agents such as andriol and cyclosporine.

Molecular cloning and mRNA profile of insulin-like growth factor type 1 receptor in orange-spotted grouper, Epinephelus coioides.

The insulin-like growth factor type 1 receptor (IGF-IR) belongs to the tyrosine kinase (TK) receptor family. Besides being mitogenic, IGF-IR plays a crucial role in cell survival, transformation and maintenance of the malignant phenotype. In this study, we cloned the cDNA from the hypothalamus of the orange-spotted grouper (Epinephelus coioides) using reverse transcription PCR (RT-PCR) and the rapid amplification of cDNA ends (RACE) method. The deduced amino acid sequence showed that the receptor comprises 1413 amino acid residues. It contains cysteine-rich domains in its alpha-subunit, and a conserved transmembrane domain and TK domains in its beta-subunit. Comparison of the amino acid sequence with those of other species showed that the grouper IGF-IR shares 90.2%, 89.6%, 71.9% and 72% similarity with the IGF-IR of the Japanese flounder, turbot, zebrafish-a and zebrafish-b, respectively. When compared with its mammalian homologue, grouper IGF-IR contains a large insertion at its C-terminus. Phylogenetic analysis has revealed that the grouper IGF-IR belongs to the b-type IGF-IRs and has a higher similarity with flounder and turbot IGF-IR, and a lower similarity (<70%) with human, mouse and avian IGF-IR. Grouper IGF-IR transcripts were detected in the brain, peripheral tissues, embryos and early development larvae by semi-quantitative RT-PCR assay. It was observed that IGF-IR mRNA expression was greater in the brain than in peripheral tissues. The level of IGF-IR mRNA expression was much higher in retina, gonad, skeletal muscle and gill tissues than in liver, heart and thymus tissues. The expression of IGF-IR can be visualized as a ubiquitous signal in unfertilized eggs, embryos and early development larvae. The distribution pattern of IGF-IR mRNA in grouper development suggests that IGF-IR plays an important role in the embryo and early larval development stages.