pH and redox dual response nano-suppository for the treatment of ulcerative colitis.

To improve treatment compliance and reach sustained and controlled drug release in the colon, we developed a hollow mesoporous silica nano-suppository that responded to both pH and redox stimuli. Firstly, we prepared hollow mesoporous silica nanoparticles containing disulfide bonds (HMSN-SS) and loaded them with 5-ASA. Secondly, we modified the surface of HMSN-SS with polydopamine (PDA) and chitosan (CS) and molded the suppository, which we named 5-ASA@HMSN-SS-PDA-CS (5-ASA@HSPC). By administering 5-ASA@HSPC rectally, it acted directly on the affected area. CS helped the nanoparticles adhere to the colon's surface, while PDA dissociates from HMSN-SS due to protonation in the acidic environment of the ulcerative colon. The disulfide bonds were destroyed by the reducing environment of the colon, leading to a stable and slow release of encapsulated 5-ASA from the pores of HMSN. Finally, in vitro release experiments and in vivo pharmacokinetic and pharmacodynamic experiments had demonstrated that 5-ASA@HSPC exhibited a slow and steady action at the colonic site, with an excellent safety profile. This novel approach showed great potential in the treatment of ulcerative colitis.

Accelerating the speed of innovative anti-tumor drugs to first-in-human trials incorporating key de-risk strategies.

Pharmaceutical companies have recently focused on accelerating the timeline for initiating first-in-human (FIH) trials to allow quick assessment of biologic drugs. For example, a stable cell pool can be used to produce materials for the toxicology (Tox) study, reducing time to the clinic by 4-5 months. During the coronavirus disease 2019 (COVID-19) pandemic, the anti-COVID drugs timeline from DNA transfection to the clinical stage was decreased to 6 months using a stable pool to generate a clinical drug substrate (DS) with limited stability, virus clearance, and Tox study package. However, a lean chemistry, manufacturing, and controls (CMC) package raises safety and comparability risks and may leave extra work in the late-stage development and commercialization phase. In addition, whether these accelerated COVID-19 drug development strategies can be applied to non-COVID projects and established as a standard practice in biologics development is uncertain. Here, we present a case study of a novel anti-tumor drug in which application of "fast-to-FIH" approaches in combination with BeiGene's de-risk strategy achieved successful delivery of a complete CMC package within 10 months. A comprehensive comparability study demonstrated that the DS generated from a stable pool and a single-cell-derived master cell bank were highly comparable with regards to process performance, product quality, and potency. This accomplishment can be a blueprint for non-COVID drug programs that approach the pace of drug development during the pandemic, with no adverse impact on the safety, quality, and late-stage development of biologics.

Alternaria alternata F3, a Novel Taxol-Producing Endophytic Fungus Isolated from the Fruits of Taxus cuspidata: Isolation, Characterization, Taxol Yield Improvement, and Antitumor Activity.

In this study, a novel taxol-producing endophytic fungus, strain F3, was isolated from the fruits of Taxus cuspidata and identified as Alternaria alternata according to its macroscopic and microscopic traits and sequence analysis of internal transcribed spacer (ITS). The presence of taxol was detected by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) and confirmed by ultra-high-performance liquid chromatography-electrospray coupled to tandem mass spectrometry (UPLC-ESI-MS/MS) and nuclear magnetic resonance (NMR). The fermentation parameters of strain F3 were then optimized for high taxol production. The maximum taxol yield of 195.4 µg L-1 by A. alternata F3 was observed in 200-mL yeast peptone dextrose (YPD) broth, at an initial pH value of 6.0, supplemented with 0.1 g L-1 sodium acetate, 0.25 g L-1 salicylic acid, and 0.00125 g L-1 silver nitrate and inoculum size 2%, and incubated at 28 °C and 150 rpm for 8 days, which was 2.12-fold compared with the initial yield of taxol. Also, fungal taxol exhibited antitumor activity towards human lung carcinoma (A549) cell line and human cervical carcinoma (Hela) cell line with IC50 values of 3.98 µg mL-1 and 0.35 µg mL-1. Overall, this is the first report on taxol-producing endophytic fungus isolated from the fruits of Taxus. This study offers a novel source for the production of taxol for anticancer treatment.

Preparation of multifunctional mesoporous SiO2nanoparticles and anti-tumor action.

A targeted drug delivery system was developed to accumulate specific drugs around tumor cells based on the redox, temperature, and enzyme synergistic responses of mesoporous silica nanoparticles. Mesoporous silica nanoparticles (MSN-NH2) and Doxorubicin (DOX) for tumor therapy were prepared and loaded into the pores of MSN- NH2 to obtain DOX@MSN(DM NPs). Hyaluronic acid (HA) was used as the backbone and disulfide bond was used as the linker arm to graft carboxylated poly (N-isopropylacrylamide)(PNIPAAm-COOH) to synthesize the macromolecular copolymer (HA-SS-PNIPAAm), which was modified to DM NPs with capped ends to obtain the nano-delivery system DOX@MSN@HA-SS-PNIPAAm(DMHSP NPs), and a control formulation was prepared in a similar way. DMHSP NPs specifically entered tumor cells via CD44 receptor-mediated endocytosis; the high GSH concentration (10 mM) of cells severed the disulfide bonds, the hyaluronidase sheared the capped HA to open the pores, and increased tumor microenvironment temperature due to immune response can trigger the release of encapsulated drugs in thermosensitive materials.In vitroandin vivoantitumor and hemolysis assays showed that DMHSP NPs can accurately target hepatocellular carcinoma cells with a good safety profile and have synergistic effects, which meant DMHSP NPs had great potential for tumor therapy.

Effects of cadmium (Cd) on fungal richness, diversity, and community structure of Haplic Cambisols and inference of resistant fungal genera.

Cadmium (Cd) is one of the most toxic and widely distributed pollutants in mining sites of Northeast China, and how Cd contamination may affect the fungal characteristics of the zonal Haplic Cambisols is still unknown. The study aims to investigate the richness and diversity of fungal community in Haplic Cambisols in response to Cd treatments and to infer Cd-resistant fungal genera. Haplic Cambisol was treated with different concentrations of CdCl2·2.5H2O solution (0 mg kg-1, 1 mg kg-1, 5 mg kg-1, 25 mg kg-1, and 50 mg kg-1, expressed as CK, T1, T2, T3, and T4, respectively), and fungal community was analyzed by high-throughput sequencing technology at 30 days, 60 days, or 80 days after Cd treatment (expressed as d30, d60, and d80, respectively). The results showed that Cd treatment usually increased the richness and diversity indices, the variation of diversity index under different Cd concentrations was not obvious, and different Cd incubation times had an inhibitory effect on fungal richness, but the diversity first increased and then decreased. Besides, Ascomycota and Mortierellomycota having the highest abundance in Haplic Cambisols showed the most pronounced changes under Cd treatment. Accordingly, Cd-resistant fungi were also found, such as Aspergillus, Fusarium, Penicillium, and Trichoderma, especially Aspergillus, which had relatively high abundance. The results obtained in this study had potentially significant findings for soil biodiversity and Cd bioremediation.

A pH-responsive mesoporous silica nanoparticle-based drug delivery system for targeted breast cancer therapy.

In order to make the drug specifically aggregate at the tumor site, we had developed a targeted drug delivery system based on pH responsive mesoporous silica nanoparticles. Mesoporous silica nanoparticles (MSN-COOH) were prepared and doxorubicin (DOX) was loaded into the pores of MSN-COOH, and then polyethyleneimine (PEI) and anisamide (AA) were modified on the surface of mesoporous silica, named DOX@MSN-PEI-AA(DMPA). DMPA specifically entered tumor cells through AA-mediated receptor endocytosis; PEI dissociated from the surface of the MSN in the acidic environment of cellular lysosomes/endosomes due to protonation of PEI, resulting in steady release of the encapsulated DOX from the pores of MSN in the cytoplasm of the target cells. In vitro and in vivo anti-tumor experiments and hemolytic experiments indicated that DMPA can accurately target breast cancer cells and show excellent safety at the same time, showing great potential for tumor therapy.

Graphene oxide affects growth and physiological indexes in Larix olgensis seedlings and the soil properties of Haplic Cambisols in Northeast China.

Changbai larch (Larix olgensis A. Henry) seedlings growing in a Haplic Cambisol and receiving 0 (Ck), 25, 50, 100, 250, or 500 mg L-1 graphene oxide (GO) were incubated for 30, 40, or 50 days, and the effects of applying GO on the growth and physiological characteristics of the seedlings and soil chemical properties and enzyme activities were investigated. The superoxide anion (except for 25 mg L-1 at 40 days and 50 mg L-1 at 50 days) and hydrogen peroxide contents of the leaves increased at 25-100 mg L-1 GO; however, superoxide dismutase (SOD) and peroxidase (POD) (except for 100 mg L-1 at 50 days) activities, soluble protein (except for 100 mg L-1 at 30 and 40 days), proline (except for 100 mg L-1 at 50 days), as well as seedling biomass (except for stems at 25-100 mg L-1, and leaves and roots at 50-100 mg L-1 for 30 days) all decreased. However, when the seedlings were exposed to 250-500 mg L-1 GO, especially at 40 and 50 days, these trends for tree growth and physiological parameters were reversed, suggesting the beneficial effect of GO at high concentrations on the seedlings. GO decreased the organic matter, alkali-hydrolyzale nitrogen, available phosphorus, and potassium contents of the soil at 40 and 50 days (except for available phosphorus at 50 days), as well as the acid phosphatase, urease (except for 30 days), dehydrogenase, and catalase activities (except for 30 and 40 days); thus, GO may inhibit nitrogen and phosphorus cycling in Haplic Cambisols (except for nitrogen at 30 days).

Exogenous succinic acid mediates responses of Larix olgensis A. Henry to cadmium stress.

Trace metal contamination of soil is an increasing problem. Organic acid application can restore trace metal elements such as cadmium (Cd) in contaminated soil. Changbai larch (Larix olgensis A. Henry) is an economically important forestry species in northeast China; however, growth is inhibited by severe Cd contamination. We investigated the effects of different concentrations of exogenous succinic acid (SA) on Cd tolerance and physiological and morphological toxicity in L. olgensis seedlings. Seedlings were planted in pots containing Cd-contaminated or uncontaminated Haplic Cambisol. Seedlings in Cd-contaminated soil were treated daily with SA solution at 0, 0.04, 0.2, 1.0, and 2.0 mmol kg-1 of soil for 10, 20 or 30 days. Cd treatment induced seedling damage and significantly increased the relative conductivity and malondialdehyde content of the leaves, inhibiting soluble protein and proline contents, superoxide dismutase and peroxidase activity, chlorophyl fluorescence and pigment content. Decreases in the length, surface area, volume of roots and leaves, and specific root length were also observed. Effects increased in control plants with time. SA treatment also reduced the Cd content of the fine roots and leaves and Mg, K, and Ca contents. Moreover, plant growth was significantly promoted and damage was reversed, especially at 5.0 and 10.0 mmol L-1 SA for 30 days. SA therefore alleviated Cd-induced injury, improving tolerance to Cd stress. SA application combined with afforestation could therefore help restore Cd-contaminated soil in northeast China. Further studies aimed at determining the detoxification mechanism of L. olgensis seedlings are now required.

Exogenous spermidine elevating cadmium tolerance in Salix matsudana involves cadmium detoxification and antioxidant defense.

In this study, exogenous spermidine role on Salix matsudana tolerance to cadmium was evaluated. Spermidine and cadmium presented antagonistic effects on the biomass, copper and zinc concentrations in S. matsudana. cadmium mainly distributed in the cell wall of subcellular fraction; 46.97%-60.43% of cadmium existed in a sodium chloride-extracted form. Cadmium contents in roots, leaves, and twigs ranged from 2002.67 to 3961.00, 111.59 to 229.72, and 102.56 to 221.27 mg/kg, respectively. Spermidine application elevated cadmium concentrations in the roots, cuttings, and cell wall and the ratio of deionized water-extracted cadmium, but decreased cadmium levels in the twigs and leaves and the fractions of cadmium extracted by ethanol and sodium chloride, respectively. Putrescine and malondialdehyde were important indicators of cadmium-induced oxidative damage. Exogenous spermidine alleviated the accumulation of superoxide anion, hydrogen peroxide, malondialdehyde via promoting the levels of spermidine, soluble protein, superoxide dismutase, reductive ascorbate, glutathione reductase, and glutathione peroxidase in S. matsudana leaves under the corresponding cadmium stress. The results indicated that S. matsudana was a candidate for cadmium rhizoremediation and extraction in leaves; the spermidine application enhanced the cadmium tolerance of S. matsudana through promoting cadmium accumulation in roots, cell wall, and less bioactive chemical forms and the antioxidative ability.

Effects of exogenous organic acids on the resistance of Changbai larch (Larix olgensis) seedlings to mixed Pb and Cd stress.

Mixed Pb and Cd soil contamination is an issue in Northeast China. We examined the effects of exogenous organic acids on the resilience of Changbai larch (Larix olgensis) seedlings, a pioneering forestry species in afforestation and vegetation restoration in Northeast China, under such stress. Mixed Pb and Cd stress led to significantly higher Pb and Cd content in the leaves and fine roots, malondialdehyde content in the leaves, superoxide dismutase activity, and soluble protein content in the leaves. Lower biomass of the roots, stems, and leaves was observed, with the roots showing the sharpest reduction in biomass. However, the application of organic acids mitigated or reversed these effects. This was most pronounced following treatment with 0.2 mmol·L-1 or 1.0 mmol·L-1 organic acids for 20 days. Citric acid had the greatest positive effect compared with succinic acid and oxalic acid. We suggest that exogenous organic acids have the potential to alleviate Pb and Cd-induced oxidation injury symptoms in Changbai larch seedlings, and may enhance resilience to mixed Pb and Cd stress.

Single and competitive adsorption of Cd(II) and Pb(II) from aqueous solution by activated carbon prepared with Salix matsudana Kiodz.

In this study, Salix matsudana activated carbon (SAC) was prepared by phosphoric acid activation, and the adsorption characteristics of Cd(II) and Pb(II) on SAC in single- and double-component solutions were investigated. In both systems, the adsorption capacities of both ions on SAC increased with the increasing initial pH value and temperature in the solutions, and the adsorption equilibrium was approached at 10 min. The adsorption process was spontaneous, endothermic, and depicted well by the pseudo-second-order adsorption model, and the equilibrium adsorption fitted reasonably well with the Langmuir isotherm. The maximum adsorption capacity (Qm) of Cd(II) and Pb(II) was 58.48 and 59.01 mg/g, respectively, in the single-element systems. However, it reduced to 25.32 and 31.09 mg/g, respectively, in the double-element system. The physicochemical property analysis showed that the specific surface area, total pore volume, and average pore diameter of SAC was 435.65 m2/g, 35.68 mL/g, and 3.86 nm, respectively. The SAC contained groups of -OH, C = O, and P = O. Results suggest that SAC had a good performance for the adsorption of Cd(II) and Pb(II) from solution, and the adsorption selectivity sequence was Pb(II) > Cd(II).

[Determination of four organophosphorus pesticides and their metabolites in vegetables using solid-phase extraction coupled with liquid chromatography tandem mass spectrometry].

metabolites were 76.9%- 102.8%, the precision ranged from 5.92% to 10. 19%, the limits of quantification ranged from 0. 001 to 0. 01 mg/L, and the linear correlation coefficients were 0. 998 2-0. 999 9 in the range of 0. 01 -1. 00 mg/L. This method was applied in the analysis of Chinese cabbages, hot peppers, tomatoes and onions from markets. This method can be applied in rapid detection of the organophosphorus pesticides and their metabolites in vegetables. metabolites were 76.9%-102.8%, the precision ranged from 5.92% to 10.19%, the limits of quantification ranged from 0.001 to 0.01 mg/L, and the linear correlation coefficients were 0.9982-0.9999 in the range of 0.01-1.00 mg/L. This method was applied in the analysis of Chinese cabbages, hot peppers, tomatoes and onions from markets. This method can be applied in rapid detection of the organophosphorus pesticides and their metabolites in vegetables.

[Clinical application of low hemoglobin density in screening for iron deficiency and the differential diagnosis of anemia].

OBJECTIVE: To investigate the clinical usefulness of the low hemoglobin density (LHD) and other red blood cell parameters in screening for iron deficiency (ID) and distinguishing iron deficiency anemia (IDA) from other kinds of anemia. METHOD: Whole blood count (CBC) and parameters of iron metabolism of the patients with ID (n = 251), IDA(n = 155), anemia of chronic disease(n = 90), renal anemia(n = 184), acute hemorrhagic anemia(n = 27) and non-anemia subjects(n = 82) were analysed by Advia 2120 hematology analyzer, Hitachi 7180 automatic biochemical analyzer, Beckman-Coulter DxI 800 immunoassay analyzer. SPSS 13.0 software was applied to anylyze data. RESULTS: The sensitivity of LHD in screening for ID was 67.2%, which was significantly higher than MCHC (38.8%) . There was a sensitivity of 91.2% for ID screening by combination of LHD and RDW. LHD only increased greatly in IDA (14.9%) group, while had no increase in other anemia. CONCLUSION: LHD is a reliable new marker with certain clinical usefulness. Combination of LHD and RDW is more suitable for screening ID. LHD is also a useful marker to distinguish IDA from other kinds of anemia.

[Effect of soil water potential on hydraulic parameters of Fraxinus mandshurica seedlings].

In this paper, soil water potential was controlled in the ranges of 0-(-)20 kPa (W1), -20-(-)40 kPa (W2), -40-(-)60 kPa (W3), -60-(-)80 kPa (W4) and -80-(-)160 kPa (W5) by a suite of specially designed root-sphere osmotic irrigation technique, and the dynamics of transpiration, water suction, tissue water potential and hydraulic resistance of Fraxinus mandshurica seedlings grown under these soil water conditions were studied. The results indicated that under sub-saturated soil water condition (W1), fine root water potential fluctuated at high levels from -0.08 MPa to -0.19 MPa, which brought about the lowest soil-root hydraulic resistance (35.52 MPa x s(-1) x m(-2) x mol(-1) in diurnal average), the highest water suction rate (4.44 mmol x m(-2) x s(-1) in average from 10:00 to 14:00), and the strongest transpiration rate (5.11 mmol x m(-2) x s(-1) in average from 10:00 to 14:00). Under field capacity (W2), soil-root hydraulic resistance increased several times above that under W1, water suction rate and transpiration rate declined significantly, while the diurnal single-peak pattern of transpiration was not altered. Under orderly increased soil water stress (W3-W5), soil-root hydraulic resistance (the average from 10:00 to 14:00) increased approximately 10-24 times above that under W1, which caused the consequent low water suction rate and low fluctuation levels of leaf water potential (-1.25 MPa-(-)1.94 MPa for W5), and transpiration was seriously restricted (the transpiration rate being only 0.91-1.55 mmol x m(-2) x s(-1) at midday for W3-W5). It is concluded that soil water was not equally available to Fraxinus mandshurica seedlings in the test ranges of soil water potential, and significant water stress could occur when the soil water potential was lower than -40 kPa.

[Residual of exogenous iodine in forest soils and its effect on some wild-vegetable plants].

To cultivate iodine-enriched forest vegetables in the mountainous regions of northeast China, a series of iodine application experiments were conducted. Iodine fertilizer (potassium iodide, KI) was added to the forest soils supporting a variety of wild-vegetable plants, with a dosage gradient of 1.00, 3.33, 10.00, 16.67 and 50.00 mg.kg-1. The results showed that iodine application was an effective approach to obtain iodine-rich vegetable materials, and the iodine contents in treated plants could be 2-40 times higher than control. But, the residual of added iodine in soil was generally not high, only occupying 40%-5% of the application dosage. The accumulation of iodine in plants did not consistently increase with the dosage, which tended to slow down when the application amount increased to 10.00 mg.kg-1. In a comprehensive consideration of soil iodine residual, plant iodine accumulation and application cost, the proper iodine dosage should be approximately 10 mg.kg-1. Soil conditions had significant influence on the efficiency of iodine application, and the cultivated lands should be chosen at fine-textured, fertile, and upper-sloped sites, while sandy soils and down-sloped sites were unfavorable to the preservation of added iodine and to plant absorption.