Establishment of a Diamond-Blackfan anemia like model in zebrafish.

BACKGROUND: Anemia is defined as a lack of erythrocytes, low hemoglobin levels, or abnormal erythrocyte morphology. Diamond-Blackfan anemia (DBA) is a rare and severe congenital hypoplastic anemia that occurs due to the dominant inheritance of a ribosomal protein gene mutation. Even rarer is a case described as Diamond-Blackfan anemia like (DBAL), which occurs due to a loss-of-function EPO mutation recessive inheritance. The effective cures for DBAL are bone marrow transfusion and treatment with erythropoiesis-stimulating agents (ESAs). To effectively manage the condition, construction of DBAL models to identify new medical methods or screen drugs are necessary. RESULTS: Here, an epoa-deficient mutant zebrafish called epoaszy8 was generated to model DBAL. The epoa-deficiency in zebrafish caused developmental defects in erythroid cells, leading to severe congenital anemia. Using the DBAL model, we validated a loss-of-function EPO mutation using an in vivo functional analysis and explored the ability of ESAs to alleviate congenital anemia. CONCLUSIONS: Together, our study demonstrated that epoa deficiency in zebrafish leads to a phenotype resembling DBAL. The DBAL zebrafish model was found to be beneficial for the in vivo assessment of patient-derived EPO variants with unclear implications and for devising potential therapeutic approaches for DBAL.

Transcriptome analysis reveals diverse Curvularia tsudae strategies in response to cadmium stress.

Cadmium (Cd) is a highly toxic heavy metal that poses significant threats to living organisms. Curvularia tsudae has demonstrated remarkable survival capabilities in the presence of high Cd concentrations, exhibiting its exceptional Cd tolerance. Although some physiological studies have been conducted, the molecular mechanisms underlying Cd tolerance in C. tsudae is largely unknown. In this study, a comparative transcriptome analysis was performed to explore the molecular mechanisms of C. tsudae under Cd stress. Among the 10,498 identified unigenes, 2526 differentially expressed genes (DEGs) were identified between the Cd-free and Cd-treated samples. Functional annotation and enrichment analysis of these DEGs identified several key biological processes involved in coping with Cd stress. Genes related to cell wall modification and organic acid metabolism contributes to Cd binding or chelation. Fourier transform infrared spectroscopy (FTIR) analysis further highlighted the modifications in functional groups with the cell wall under Cd stress. Furthermore, the transporters tended to be modulated in response to Cd stress, and up-regulated genes involved in antioxidants likely contributes to high Cd tolerance. The processes from DNA to protein metabolism appeared to responsive to the presence of Cd stress as well. These results contribute to the advance of the current knowledge about the response of C. tsudae to Cd stress and lay the foundation for further advancements in using fungi for the remediation of Cd-polluted environments.

Identification and characterization of human hematopoietic mesoderm.

The embryonic mesoderm comprises heterogeneous cell subpopulations with distinct lineage biases. It is unclear whether a bias for the human hematopoietic lineage emerges at this early developmental stage. In this study, we integrated single-cell transcriptomic analyses of human mesoderm cells from embryonic stem cells and embryos, enabling us to identify and define the molecular features of human hematopoietic mesoderm (HM) cells biased towards hematopoietic lineages. We discovered that BMP4 plays an essential role in HM specification and can serve as a marker for HM cells. Mechanistically, BMP4 acts as a downstream target of HDAC1, which modulates the expression of BMP4 by deacetylating its enhancer. Inhibition of HDAC significantly enhances HM specification and promotes subsequent hematopoietic cell differentiation. In conclusion, our study identifies human HM cells and describes new mechanisms for human hematopoietic development.

Hyperhomocysteinemia potentiates megakaryocyte differentiation and thrombopoiesis via GH-PI3K-Akt axis.

Hyperhomocysteinemia (HHcy) is closely associated with thrombotic diseases such as myocardial infarction and stroke. Enhanced platelet activation was observed in animals and humans with HHcy. However, the influence of HHcy on thrombopoiesis remains largely unknown. Here, we reported increased platelet count (PLT) in mice and zebrafish with HHcy. In hypertensive patients (n = 11,189), higher serum level of total Hcy was observed in participants with PLT ≥ 291 × 109/L (full adjusted ß, 0.59; 95% CI 0.14, 1.04). We used single-cell RNA sequencing (scRNA-seq) to characterize the impact of Hcy on transcriptome, cellular heterogeneity, and developmental trajectories of megakaryopoiesis from human umbilical cord blood (hUCB) CD34+ cells. Together with in vitro and in vivo analysis, we demonstrated that Hcy promoted megakaryocytes (MKs) differentiation via growth hormone (GH)-PI3K-Akt axis. Moreover, the effect of Hcy on thrombopoiesis is independent of thrombopoietin (TPO) because administration of Hcy also led to a significant increase of PLT in homozygous TPO receptor (Mpl) mutant mice and zebrafish. Administration of melatonin effectively reversed Hcy-induced thrombopoiesis in mice. ScRNA-seq showed that melatonin abolished Hcy-facilitated MK differentiation and maturation, inhibited the activation of GH-PI3K-Akt signaling. Our work reveals a previously unrecognized role of HHcy in thrombopoiesis and provides new insight into the mechanisms by which HHcy confers an increased thrombotic risk.Trial Registration clinicaltrials.gov Identifier: NCT00794885.

Effects of arbuscular mycorrhizal fungus inoculation on the growth and nitrogen metabolism of Catalpa bungei C.A.Mey. under different nitrogen levels.

Evidence suggests that arbuscular mycorrhizal fungi (AMF) may promote the growth of woody plants. However, the effects of AMF on nitrogen (N) metabolism in plants, especially trees, and its regulatory mechanism are rarely reported. Here, the effects of AMF inoculation on the growth and N nutrition status of Catalpa bungei under different N levels were reported. Three N levels (low, medium, high) and two mycorrhizal inoculation treatments (inoculation with Rhizophagus intraradices or not) were used with factorial design. The results showed that medium N could significantly improve the physiological metabolism and growth of C. bungei seedlings. However, when N was excessive, growth was significantly inhibited whether inoculated AMF or not. Compared with non-inoculated treatments, AMF inoculation could promote the absorption of N and P, improve photosynthesis under low to medium N levels, thus promoting the growth of seedlings. AMF changed the biomass allocation in seedlings by reducing the stem mass ratio and root/shoot ratio, and increasing the leaf mass ratio. At medium N levels, compared with non-inoculated treatment, AMF inoculation could significantly promote root growth by changing root hormone levels and improving root architecture and root activity. Under N addition, AMF inoculation could improve the absorption and assimilation of N by regulating the expression of key enzyme genes of N metabolism and nitrate transporter genes (NRT2.4, NRT2.5, NRT2.7) in roots, and enhancing the activities of the key enzyme of N metabolism. This study may provide a reference for the application of AMF in the cultivation and afforestation technology of C. bungei in Northwest China.

Lactobacillus treatment as adjuvant for geographic tongue and fissured tongue with gastritis: case report.

Background: Geographic tongue and fissured tongue are both common oral mucous membrane diseases relatively. The treatment of this disease is typically causative therapy because of the unknown etiology, and other afflictions. Gastrointestinal disease is one of the factors that induce geographic tongue with a fissured tongue. As an adjuvant drug that can inhibit the growth of Helicobacter pylori, the most common pathogen of chronic gastritis, Lactobacillus is widely used in clinic. However, there are seldom studies about Lactobacillus used in the treatment of geographic tongue and fissured tongue. Therefore, this case we used typically causative therapy with Lactobacillus as adjuvant to treat patient with a geographic tongue and fissured tongue with gastrointestinal disease and explore the effectiveness for further application. Case Description: A 33-year-old female patient presented with a geographic tongue and fissured tongue with dysgeusia. At the first visit, there were smooth red lesions on the back of the tongue with the hyperplasia of the surrounding filiform papilla. The cracks in the middle of the tongue had a crack of about 2.5 cm long and 0.8 cm deep, and 5-6 light cracks on the tip of the tongue. The patient reported a history of chronic gastritis for 1 year, often with soft stool and gastroesophageal reflux disease. We administered drug treatment, including sodium bicarbonate tablets, Kangfuxin solution, compound chlorhexidine solution, and stomatitis spray. In addition, we suggested the patient use a yogurt machine to make and drink fresh yogurt with Lactobacillus as adjuvant everyday day and suggested a daily regimen of a light diet with no stimulating food, regular work, and sufficient rest. Ten days after the first visit, the symptoms were obviously alleviated. Twenty days after the initial diagnosis, the geographic tongue lesions had disappeared and the cracks had basically healed. The patient reported that the gastric symptoms had improved, and there was no soft stool or stomach discomfort. Conclusions: This study used Lactobacillus as the adjuvant with typically causative therapy in the treatment of geographic tongue and fissured tongue.

Insights from comparative transcriptome analysis in the responses of Pb-tolerant fungi Curvularia tsudae to Pb stress.

The fungus Curvularia tsudae can survive in environments that are extremely contaminated by heavy metals; however, the underlying molecular mechanisms of heavy metal tolerance are not clear. In this study, we determined the effects of lead (Pb) stress on the growth of C. tsudae and used RNA-Seq to identify significant genes and biological processes involved. The present study showed that C. tsudae had an outstanding resistant capacity to Pb stress and could survive at a concentration of 1600 mg L-1 Pb. Although an obvious inhibition on the growth was observed, the fungus exhibited tolerance as it continued to grow at a Pb concentration of 1600 mg L-1 for seven days. A total of 9997 (9020 up and 977 down) differentially expressed genes (DEGs) were detected in the mycelium of C. tsudae at Pb free (0 mg L-1) and Pb stressed samples. Pathway enrichment analysis identified several biological processes for managing Pb stress. Genes involved in carbohydrate metabolism tended to be modulated in response to Pb stress, while amino acids and the lipid metabolism would also be induced by Pb stress, and up-regulated genes involved in antioxidant substances and ABC transporters may be committed to high Pb tolerance. Our study contributes to the current literature on C. tsudae response to Pb stress and provides a useful reference for fungi as bioremediators in heavy metal-contaminated environments.

Clinical observation of allergic contact stomatitis treated with Kangfuxin solution as adjuvant: case report.

Background: Allergic contact stomatitis (ACS) is common among people with allergic constitution and who have allergy reaction to specific allergen such as drugs, food, and materials because of immune dysfunction. With the development of materials science, the increasing diversity of cosmetics and food additives has gradually raised the incidence rate of ACS. Now systemic and local therapy are adopted in the therapy of ACS. However, the systemic therapy would drop the drugs' concentration after it reaches the treatment area through the layers of human barriers, while the locally-used drugs such as collutory may not be suitable for patients with skin lesion. Kangfuxin contains a variety of biological extracts which is anti-inflammatory and curative and can produce connective tissues whether it's skin or mucous membrane. It can be used not only in non-oral diseases such as gastric ulcer or gynecological diseases, but also in the treatment of recurrent aphthous ulcer and many kinds of stomatitis and has shown good anti-inflammatory and curative effects. This study aimed to explore the effectiveness of Kangfuxin solution as a local-used adjuvant drug to treat ACS. Case Description: We present a 22-year-old male with ACS whose complaint at the first visit was severe pain, accompanied by salivation, tongue enlargement, bleeding, tonsil enlargement, and symptoms of difficulty in eating, fatigue, and dizziness. After the physical and laboratory examination, we found no abnormalities other than a history of eating the kiwi fruit, which is a common allergen. Thus, he was diagnosed as ACS. In this case, we provided a pharmacologic therapeutic intervention of chlorpheniramine (one tablet, three times a day) and Kangfuxin solution (gargle for 15 min, three times a day) with advice that no exposure to the allergen. On the third day, the patient felt no significant relief of symptoms, while one week after the first visit, the symptoms had obviously alleviated and most of the red lip erosion disappeared. The patient recovered completely with no discomfort in ten days after the initial visit. Conclusions: This study investigated the therapeutic effect of Kangfuxin solution combined with chlorpheniramine on ACS.

Generation of a thrombopoietin-deficient thrombocytopenia model in zebrafish.

BACKGROUND: The production of platelets is tightly regulated by thrombopoietin (THPO). Mutations in the THPO gene cause thrombocytopenia. Although mice lacking Thpo present with thrombocytopenia, predicting phenotypes and pathogenicity of novel THPO mutations in mice is limited. Zebrafish can be a powerful tool for fast validation and study of candidate genes of human hematological diseases and have already been used as a model of human thrombocytopenia. OBJECTIVES: We aim to investigate the role of Thpo in zebrafish thrombopoiesis and to establish a Thpo-deficient zebrafish model. The model could be applied for illustrating the clinically discovered human THPO variants of which the clinical significance is not known and to evaluate the effect of THPO receptor agonists (THPO-Ras), as well as a screening platform for new drugs. METHODS: We generated a thpo loss-of-function zebrafish model using CRISPR/Cas9. After disruption of zebrafish thpo, thposzy6 zebrafish presented with a significant reduction of thpo expression and developed thrombocytopenia. Furthermore, we performed in vivo studies with zebrafish with the thposzy6 mutation and found two human clinical point mutations (c.091C > T and c.112C > T) that were responsible for the thrombocytopenia phenotype. In addition, effects of THPO-RAs used as therapeutics against thrombocytopenia were evaluated in the Tg(mpl:eGFP);thposzy6 line. RESULTS AND CONCLUSIONS: Zebrafish with the mutation thposzy6 presented with a significant reduction of thpo expression and developed thrombocytopenia. Thpo loss-of-function zebrafish model can serve as a valuable preclinical model for thrombocytopenia caused by thpo-deficiency, as well as a tool to study human clinical THPO variants and evaluate the effect of THPO-RAs.

Establishment of a Bernard-Soulier syndrome model in zebrafish.

Platelets play an essential role in thrombosis and hemostasis. Abnormal hemostasis can cause spontaneous or severe post-traumatic bleeding. Bernard-Soulier syndrome (BSS) is a rare inherited bleeding disorder caused by a complete quantitative deficiency in the GPIb-IX-V complex. Multiple mutations in GP9 lead to the clinical manifestations of BSS. Understanding the roles and underlying mechanisms of GP9 in thrombopoiesis and establishing a proper animal model of BSS would be valuable to understand the disease pathogenesis and to improve its medical management. Here, by using CRISPR-Cas9 technology, we created a zebrafish gp9SMU15 mutant to model human BSS. Disruption of zebrafish gp9 led to thrombocytopenia and a pronounced bleeding tendency, as well as an abnormal expansion of progenitor cells. The gp9SMU15 zebrafish can be used as a BSS animal model as the roles of GP9 in thrombocytopoiesis are highly conserved from zebrafish to mammals. Utilizing the BSS model, we verified the clinical GP9 mutations by in vivo functional assay and tested clinical drugs for their ability to increase platelets. Thus, the inherited BSS zebrafish model could be of benefit for in vivo verification of patient-derived GP9 variants of uncertain significance and for the development of potential therapeutic strategies for BSS.

Identification of the atypical cadherin FAT1 as a novel glypican-3 interacting protein in liver cancer cells.

Glypican-3 (GPC3) is a cell surface heparan sulfate proteoglycan that is being evaluated as an emerging therapeutic target in hepatocellular carcinoma (HCC). GPC3 has been shown to interact with several extracellular signaling molecules, including Wnt, HGF, and Hedgehog. Here, we reported a cell surface transmembrane protein (FAT1) as a new GPC3 interacting protein. The GPC3 binding region on FAT1 was initially mapped to the C-terminal region (Q14517, residues 3662-4181), which covered a putative receptor tyrosine phosphatase (RTP)-like domain, a Laminin G-like domain, and five EGF-like domains. Fine mapping by ELISA and flow cytometry showed that the last four EGF-like domains (residues 4013-4181) contained a specific GPC3 binding site, whereas the RTP domain (residues 3662-3788) and the downstream Laminin G-2nd EGF-like region (residues 3829-4050) had non-specific GPC3 binding. In support of their interaction, GPC3 and FAT1 behaved concomitantly or at a similar pattern, e.g. having elevated expression in HCC cells, being up-regulated under hypoxia conditions, and being able to regulate the expression of EMT-related genes Snail, Vimentin, and E-Cadherin and promoting HCC cell migration. Taken together, our study provides the initial evidence for the novel mechanism of GPC3 and FAT1 in promoting HCC cell migration.

Famciclovir leads to failure of hematopoiesis, but may have the benefit of relieving myeloid expansion in MDS-like zebrafish.

Famciclovir (FCV) is an antiviral drug that is often utilized after bone marrow transplantation to prevent viral infection. Yet, its role in hematopoiesis is poorly understood. Here, by utilizing a zebrafish model, we found that FCV exposure led to hematopoietic failure by impairing the proliferation of hematopoietic stem and progenitor cell (HSPC) and inducing HSPC apoptosis. On the other hand, FCV treatment could effectively relieve myeloid malignancies in the c-mybhyper MDS-like fish model, and played a role not only in the embryonic stage but also in adult zebrafish. This study reveals that FCV functions as a double-edged sword, with hematotoxicity at a high level, but that appropriate FCV treatment may be beneficial for the treatment of MDS.

Angptl7 promotes insulin resistance and type 2 diabetes mellitus by multiple mechanisms including SOCS3-mediated IRS1 degradation.

Angptl7 is a secreted and circulating cytokine that belongs to Angiopoietin-like family. The current knowledge about the function of Angptl7 is still limited, and its biological role is only marginally known, such as in the promotion of angiogenesis and inflammation. Here, we demonstrated that Angptl7 promotes insulin resistance and type 2 diabetes mellitus (T2DM). We found that the circulating Angptl7 levels in T2DM patient and mouse models were significantly elevated. Artificial overexpression of Angptl7 in hepatic cells inhibited glucose uptake and impaired insulin signaling pathway. Furthermore, in vivo overexpression of Angptl7 in experimental healthy mice also caused insulin resistance-like characteristics. Mechanistic studies revealed that Angptl7 can upregulate SOCS3 expression, leading to the IRS1 degradation in proteasome. Furthermore, over-expressed Angptl7 inhibited the phosphorylation of Akt and promoted the phosphorylation of ERK1/2, which was known to be associated with insulin resistance. Taken together, our study provided strong evidence that Angptl7 promotes insulin resistance and T2DM by multiple mechanisms, which made Angptl7 a new potential therapeutic target for treatment of insulin resistance and T2DM.

Genome-wide identification and transcript profiles of walnut heat stress transcription factor involved in abiotic stress.

BACKGROUND: Walnut (Juglans regia) is an important tree cultivated worldwide and is exposed to a series of both abiotic and biotic stress during their life-cycles. The heat stress transcription factors (HSFs) play a crucial role in plant response to various stresses by regulating the expression of stress-responsive genes. HSF genes are classified into 3 classes: HSFA, HSFB, and HSFC. HSFA gene has transcriptional activation function and is the main regulator of high temperature-induced gene expression. HSFB gene negatively regulates plant resistance to drought and NaCl. And HSFC gene may be involved in plant response to various stresses. There are some reports about the HSF family in herbaceous plants, however, there are no reports about the HSFs in walnut. RESULT: In this study, based on the complete genome sequencing of walnut, the bioinformatics method was used and 29 HSF genes were identified. These HSFs covered 18 HSFA, 9 HSFB, and 2 HSFC genes. Phylogenetic analysis of these HSF proteins along with those from Arabidopsis thaliana showed that the HSFs in the two species are closely related to each other and have different evolutionary processes. The distribution of conserved motifs and the sequence analysis of HSF genes family indicated that the members of the walnut HSFs are highly conserved. Quantitative Real-Time PCR (qRT-PCR) analysis revealed that the most of walnut HSFs were expressed in the walnut varieties of 'Qingxiang' and 'Xiangling' under high temperature (HT), high salt and drought stress, and some JrHSFs expression pattern are different between the two varieties. CONCLUSION: The complex HSF genes family from walnut was confirmed by genome-wide identification, evolutionary exploration, sequence characterization and expression analysis. This research provides useful information for future studies on the function of the HSF genes and molecular mechanism in plant stress response.

Zebrafish for thrombocytopoiesis- and hemostasis-related researches and disorders.

Platelets play vital roles in hemostasis, inflammation, and vascular biology. Platelets are also active participants in the immune responses. As vertebrates, zebrafish have a highly conserved hematopoietic system in the developmental, cellular, functional, biochemical, and genetic levels with mammals. Thrombocytes in zebrafish are functional homologs of mammalian platelets. Here, we summarized thrombocyte development, function, and related research techniques in zebrafish, and reviewed available zebrafish models of platelet-associated disorders, including congenital amegakaryocytic thrombocytopenia, inherited thrombocytopenia, essential thrombocythemia, and blood coagulation disorders such as gray platelet syndrome. These elegant zebrafish models and methods are crucial for understanding the molecular and genetic mechanisms of thrombocyte development and function, and provide deep insights into related human disease pathophysiology and drug development.

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Mycorrhizae, formed through the colonization of soil mycorrhizal fungi into the roots of host plants, are common symbiosis in the terrestrial ecosystems. The establishment of mycorrhizae is mainly based on the bidirectional nutrient exchanges between the symbiotic partners. Mycorrhizal fungi can absorb mineral nutrients, such as nitrogen and phosphorus, from soil and transport them to the host plants for their growth. As an exchange, host plants supply mycorrhizal fungi with the carbohydrates in the form of lipids or sugars, which are essential for fungal growth. In recent years, the mechanism of nutrient exchange between the mycorrhizal fungi and host plants has been a hot research topic. Important progresses have been achieved in mechanisms of host plants nutrient uptake and transport mediated by the mycorrhizal fungi. In this review, recent advances in nutrient exchange between arbuscular mycorrhizal fungi, ectomycorrhizal fungi and host plants were summarized, especially in the absorption and bidirectional transfer mechanisms of important nutrients, such as carbon, nitrogen and phosphorus. The potential regulatory effects of nutrient exchange in the mycorrhizal development were also reviewed. In addition, key problems and prospects of related researches were analyzed. This paper would be meaningful for the establishment of mycorrhizal model and the optimization of mycorrhizal effects.

Bioaugmentation in a pilot-scale constructed wetland to treat domestic wastewater in summer and autumn.

In order to determine whether bioaugmentation is an effective technique in wetlands before the plants were harvested, the nitrogen (N) removal from a constructed wetland (CW) planted with Phragmites was evaluated after inoculating with Paenibacillus sp. XP1 in Northern China. The experiment was loaded with secondary effluent of rural domestic wastewater (RDW) using the batch-loaded method for over a 17-day period in summer and autumn. Chemical oxygen demand (CODcr), ammonia nitrogen (NH3-N), and total nitrogen (TN) decreased significantly in the CW with Phragmites inoculated with Paenibacillus sp. XP1. Four days after treatments were set up, the removal efficiencies were found to be 76.2 % for CODcr, 83 % for NH3-N, and 63.8 % for TN in summer and 69.5 % for CODcr, 76.9 % for NH3-N, and 55.6 % for TN in autumn, which were higher than the control group without inoculation during the entire 17-day experiment. The inoculated bacteria did not have a noticeable effect on total phosphorus (TP) removal in autumn. However, bioaugmentation still keep a low P concentration in the whole CW. First-order kinetic model represented well the CODcr, TN, and TP decay in CWs with bioaugmentation, resulting in very good coefficients of determination, which ranged from 0.97 to 0.99. It indicated that bioaugmentation would be an effective treatment for pollutant removal from RDW in the CWs.

Allelopathic effects of Ailanthus altissima extracts on Microcystis aeruginosa growth, physiological changes and microcystins release.

The use of allelochemicals has been proved an environmentally friendly and promising method to control harmful algal blooms. This study was conducted to explore the application potential of Ailanthus altissima (A. altissima) extracts in Microcystis aeruginosa (M. aeruginosa) control for the first time. Four treatments with A. altissima extractions (25mgL(-1), 50mgL(-1), 100mgL(-1), and 200mgL(-1) respectively) and a control group were built to investigate the effects of A. altissima on the growth, cellular microstructure and cell viability, physiological changes, and release of extracellular matters. Results showed that the cell density of M. aeruginosa was effectively inhibited by A. altissima extract, and the inhibition rates were dose-dependent within 5d. Especially for the treatment with 200mgL(-1) of extract, the inhibitory rates remains above 90% after 5d exposure. In addition, A. altissima effectively decreased the amount of extracellular cyanotoxin microcystins and destroyed the photosynthesis-related structure of algae cell during the experimental period. The results demonstrated the A. altissima extracts can be used as an effective and safe algicide to control algal blooms. However, it must be noted that specific compounds responsible for algicidal effect should be isolated and identified to explore inhibition mechanism of A. altissima in future study.

Optimization and interpretation of O3 and O3/H2O2 oxidation processes to pretreat hydrocortisone pharmaceutical wastewater.

Chemical synthetic pharmaceutical wastewater must be pretreated before traditional biological treatment to improve the biodegradability of pollutants due to their high concentration and complex composition. This study investigated O3 and O3/H2O2 systems to pretreat hydrocortisone wastewater. The optimum treatment efficiency of the O3 system was achieved under pH value of 5.76, dosage of O3 of 217.5 mg/L, and reaction time of 90 min. The removal efficiency of chemical oxygen demand (COD) and total organic carbon (TOC) was 94% and 92%, respectively. The ratio of biochemical oxygen demand (BOD) and COD (B/C) increased from 0.066 to 0.310. Moreover, the optimum H2O2/O3 molar ratio was 0.3 with the optimum pH of 3, and reaction time was reduced to 15 min to reach COD removal efficiency of 67.3% in the O3/H2O2 oxidation system. Interestingly, isobutanol was assumed to be a good hydroxyl radical trapping agent due to the great decrease of Fenton reagent reaction rate constant. By comparing two different kinetic models, oxidation mechanism could be interpreted by the pseudo first-order kinetic model with the high correlation coefficient R2 above 0.9787. It can be concluded that ozonation system was controlled by both direct oxidation and free radical oxidation, and particularly the later one played a dominant role.