Influence of non-pharmaceutical interventions on epidemiological characteristics of Mycoplasma pneumoniae infection in children during and after the COVID-19 epidemic in Ningbo, China.

Background: Since the outbreak of COVID-19, China has implemented a series of non-pharmaceutical interventions (NPIs), effectively containing the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as well as various respiratory pathogens. With the continuous relaxation of restrictions, China has entered a new phase of the post-pandemic era. However, the epidemiological differences of Mycoplasma pneumoniae (MP) between the two phases in Ningbo and even in China remain unclear. Methods: Data of children aged 0-14 years who visited the Ningbo Medical Center LiHuiLi Hospital due to acute respiratory tract infections from January 2020 to December 2023 were collected. PCR was used to detect 13 respiratory pathogens and the macrolide-resistance of Mycoplasma pneumoniae. Results: Among 10,206 children, 2,360 were infected with MP (23.12%). Among the total, the MP positive rate during the NPI phase (6.35%) was significantly lower than that during the non-NPI phase (34.28%), while the macrolide resistance rate increased from 62.5% (NPI phase) to 81.1% (non-NPI phase). The rate of MP co-infection increased from 11.2% (NPI phase) to 30.3% (non-NPI phase). MP infection exhibited obvious seasonality, with the highest prevalence in autumn (30.0%) followed by summer (23.6%). There were differences in MP positivity rates among different age groups, with the highest among school-age children at 39.5%. During the NPI phase, all age groups were less susceptible to MP, while during the non-NPI phase, the susceptible age for MP was 4-12 years, with 8 years being the most susceptible. The susceptible age for MP co-infection was 0-6 years. MP exhibited antagonistic effects against numerous pathogens. Compared to MP single infection, the proportion of pneumonia was higher in MP co-infection cases. Conclusion: The removal of NPIs significantly impacted the spread of MP, altering population characteristics including age, seasonality, macrolide resistance, and MP co-infection rates.

Association and prediction of subjective sleep quality and postoperative delirium during major non-cardiac surgery: a prospective observational study.

BACKGROUND: Postoperative delirium (POD) is an acute form of brain dysfunction that can result in serious adverse consequences. There has been a link between cognitive dysfunction and poor sleep. The present study aimed to determine the association and prediction of subjective sleep quality and postoperative delirium during major non-cardiac surgery. METHODS: One hundred and thirty-four patients, aged 60 years or older, were scheduled for elective laparotomy or orthopaedic procedures. The Pittsburgh Sleep Quality Index (PSQI) and sleep log were used to assess perioperative subjective sleep quality in participants. Nursing Delirium Screening Checklist (NU-DESC) was used for screening, and the Confusion Assessment Method (CAM) was used to diagnose POD during the first seven days following surgery. The association between subjective sleep quality and POD was assessed using a multivariate logistic regression model. Thereafter, the prediction performance of subjective sleep quality was evaluated using a receiver operating characteristic (ROC) curve. RESULTS: All assessments were completed on 119 patients who had an average PSQI score of 7.0 ± 2.4 before surgery. 23 patients (19.3%) suffered from POD. The multivariate logistic regression analysis showed that the occurrence of POD was closely related to age, BMI, PSQI and operation time. After adjusting for related factors, there was a statistically significant association between PSQI and POD occurrence (OR = 1.422, 95%CI 1.079-1.873, per 1-point increase in PSQI). The ROC curve analysis showed that the optimal PSQI cutoff value was 8.0 for predicting POD, and the area under the ROC (AUROC) value of PSQI was 0.741 (95%CI 0.635 to 0.817). The AUROC of the model developed by the multivariate logistic regression analysis was 0.870 (95%CI 0.797 to 0.925). CONCLUSIONS: The study found that preoperative subjective sleep quality was strongly associated with POD during major non-cardiac surgery. Additionally, PSQI combined with age, BMI, and operation time improved POD prediction.

Integrated fecal microbiota and metabolomics analysis of the orlistat intervention effect on polycystic ovary syndrome rats induced by letrozole combined with a high-fat diet.

BACKGROUND: This study aimed to compare the characteristics of the gut microbiota and their metabolite profiles between polycystic ovary syndrome (PCOS) and orlistat-treated PCOS rats (ORL-PCOS), which could help to better understand the underlying mechanism of the effect of orlistat on PCOS. METHODS: PCOS rat models were established using letrozole combined with a high-fat diet. Ten rats were randomly selected as a PCOS control group (PCOS). The other three groups (n = 10/group) were additionally supplemented with different doses of orlistat (low, medium, high). Then, fecal samples of the PCOS and ORL-PCOS groups were analysed by 16S rRNA gene sequencing and untargeted metabolomics. Blood samples were collected to detect serum sex hormones and lipids. RESULTS: The results showed that orlistat attenuated the body weight gain, decreased the levels of T, LH, the LH/FSH ratio, TC, TG and LDL-C; increased the level of E2; and improved estrous cycle disorder in PCOS rats. The bacterial richness and diversity of the gut microbiota in the ORL-PCOS group were higher than those in the PCOS group. The ratio of Firmicutes to Bacteroidetes was decreased with orlistat treatment. Moreover, orlistat treatment led to a significant decrease in the relative abundance of Ruminococcaceae and Lactobacillaceae, and increases in the abundances of Muribaculaceae and Bacteroidaceae. Metabolic analysis identified 216 differential fecal metabolites in total and 6 enriched KEGG pathways between the two groups, including steroid hormone biosynthesis, neuroactive ligand-receptor interaction and vitamin digestion and absorption. Steroid hormone biosynthesis was the pathway with the most significant enrichment. The correlations between the gut microbiota and differential metabolites were calculated, which may provide a basis for understanding the composition and function of microbial communities. CONCLUSIONS: Our data suggested that orlistat exerts a PCOS treatment effect, which may be mediated by modifying the structure and composition of the gut microbiota, as well as the metabolite profiles of PCOS rats.

Satellites reveal widespread decline in global lake water storage.

Climate change and human activities increasingly threaten lakes that store 87% of Earth's liquid surface fresh water. Yet, recent trends and drivers of lake volume change remain largely unknown globally. Here, we analyze the 1972 largest global lakes using three decades of satellite observations, climate data, and hydrologic models, finding statistically significant storage declines for 53% of these water bodies over the period 1992-2020. The net volume loss in natural lakes is largely attributable to climate warming, increasing evaporative demand, and human water consumption, whereas sedimentation dominates storage losses in reservoirs. We estimate that roughly one-quarter of the world's population resides in a basin of a drying lake, underscoring the necessity of incorporating climate change and sedimentation impacts into sustainable water resources management.

The protective effect of tanshinone IIa on endothelial cells: a generalist among clinical therapeutics.

INTRODUCTION: Tanshinone IIa (TSA) has been approved to treat cardiovascular diseases by the China State Food and Drug Administration. TSA has exhibited a variety of pharmacological effects, including vasodilator, antioxidant, anti-inflammatory, and anti-tumor properties. Endothelial cells play an important physiological role in vascular homeostasis and control inflammation, coagulation, and thrombosis. Accumulating studies have shown that TSA can improve endothelial function through various pathways. AREAS COVERED: The PubMed database was reviewed for relevant papers published up to 2020. This review summarizes the current clinical and pharmaceutical studies to provide a systemic overview of the pharmacological and therapeutic effects of TSA on endothelial cells. EXPERT OPINION: TSA is a representative monomeric compound extracted from Danshen and it exhibits significant pharmacological and therapeutic properties to improve endothelial cell function, including alleviating oxidative stress, attenuating inflammatory injury, modulating ion channels and so on. TSA represents a spectrum of agents that are extracted from plants and can restore the endothelial function to establish the beneficial and harmless molecular therapeutics. This also suggests the possible detection of endothelial cells for very early diagnosis of diseases. In future, precise therapeutic methods will be developed to repair endothelial cells injury and recover endothelial dysfunction.

Acquired multiple mutations ALK I1171N, L1196M and G1202R mediate lorlatinib resistance in EML4-ALK-rearranged malignant pleural mesothelioma: a case report.

EML4-ALK rearranged malignant pleural mesothelioma (MPM) is rare and its responses to anaplastic lymphoma kinase (ALK) inhibitors, including alectinib and lorlatinib, remain unexplored. In this case report, we describe a patient with EML4-ALK-rearranged stage IIIB MPM who was administered with alectinib and lorlatinib as first-line and fourth-line therapy, respectively. He had remarkable response evaluated as partial response on both regimens lasting approximately 3.5 months on each regimen. His plasma samples were collected during the treatment course and submitted for targeted sequencing to understand the molecular mechanisms of his therapeutic resistance. Sequencing analysis revealed the emergence of ALK I1171N and L1196M at alectinib progression. Meanwhile, ALK I1171N, L1196M, and G1202R mutations were identified at lorlatinib progression, wherein L1196M is confirmed to be in cis to G1202R. We speculate that these multiple mutations synergistically mediated his resistance to both alectinib and lorlatinib. Our report describes the detection of EML4-ALK rearrangement in a patient with MPM who had remarkable therapeutic response with ALK inhibitors. Moreover, our case also revealed acquired mechanisms of lorlatinib resistance mediated by multiple mutations ALK I1171N, L1196M, and G1202R, contributing an incremental step to our understanding of the complexity of acquired resistance mechanisms in sequential ALK inhibitor therapy. The reviews of this paper are available via the supplemental material section.

Toxicity Study of 28-Day Subcutaneous Injection of Arctigenin in Beagle Dogs.

Our previous studies have investigated the systematic pharmacokinetic characteristics, biological activities, and toxicity of arctigenin. In this research, the potential toxicities of arctigenin in beagle dogs were investigated via repeated 28-day subcutaneous injections. Beagle dogs were randomly divided into control, vehicle [polyethylene glycol (PEG)], and arctigenin 6, 20, 60 mg/kg treated groups. The whole experimental period lasted 77 days, including adaptive period (35 days), drug exposure period (animals were treated with saline, PEG, or arctigenin for 28 consecutive days), and recovery period (14 days). Arctigenin injection (60 mg/kg) affected the lymphatic hematopoietic, digestive, urinary, and cardiovascular systems, and all the impact on these tissues resulted in death in five dogs (three female and two male dogs); 20 mg/kg arctigenin injection resulted in toxic reactions of the lymphatic hematopoietic and digestive systems; and 6 mg/kg arctigenin and PEG injection did not lead to significant toxic reactions. Meanwhile, there were no sexual differences of drug exposure and accumulation when dogs underwent different dosages. As stated previously, the toxic target organs of arctigenin administration include lymphatic hematopoietic, digestive (liver and gallbladder), urinary (kidney), and cardiovascular (heart) systems, and the no observed adverse effect level (NOAEL) of arctigenin is less than 6 mg/kg.

Effect of Lactobacillus paracasei N1115 and fructooligosaccharides in nonalcoholic fatty liver disease.

INTRODUCTION: Nonalcoholic fatty liver disease (NAFLD) is a growing health concern worldwide. Administration of probiotics and prebiotics has been proposed as a convenient and effective treatment. Our study aims to evaluate the therapeutic benefits of Lactobacillus paracasei N1115 (N1115) and fructooligosaccharides (FOS) by examining the histopathogenesis and underlying molecular events of NAFLD. MATERIAL AND METHODS: An NAFLD mouse model was established by feeding C57BL/6 mice with a high-fat diet (HFD). N1115, FOS and synbiotics were administered for 16 weeks. RESULTS: N1115, FOS and synbiotics alleviated HFD-induced hepatic steato-sis and release of tumor necrosis factor-α, and slowed the progression of cirrhosis. Compared to the HFD group, these dietary supplements reduced serum total triglyceride and cholesterol, and appeared to decrease the fasting blood glucose and insulin. Intraperitoneal glucose tolerance tests, homeostatic model assessment of insulin resistance and real-time PCR showed that the regimens could overcome insulin resistance. These findings were associated with the transcriptional repression of inflammatory factors such as lipopolysaccharides, Toll-like receptor 4 and nuclear factor-κB. Lastly, N1115, FOS, and synbiotics improved the intestinal barrier functions and histologic integrity. This was accompanied by the restoration of the p38 MAPK pathway and in-creased expression of the tight junction components occludin-1 and claudin-1. CONCLUSIONS: N1115, FOS and synbiotics are effective in the prevention and treatment of NAFLD. Our data support the translation of these agents into clinical evaluation in human subjects with NAFLD and/or associated risk factors.

Dinitrophenyl hapten with laser immunotherapy for advanced malignant melanoma: A clinical study.

The present study aimed to evaluate the efficacy and safety of in situ immunotherapy with dinitrophenyl (DNP) hapten in combination with laser therapy for patients with malignant melanoma (MM). Between February 2008 and March 2012, 72 patients with stage III or IV MM were enrolled. Patients received in situ DNP alone (n=32) or in combination with laser therapy (n=32), and each group received dacarbazine chemotherapy. The levels of peripheral cluster of differentiation (CD)4+CD25+ regulatory T cells (Tregs), interleukin (IL)-10 and tumor growth factor (TGF)-ß were detected by ELISA. The association between delayed-type hypersensitivity (DTH) and survival time was evaluated. Although peripheral Treg levels significantly decreased over time in the two groups (P<0.001), there was no significant difference between the treatment groups (P=0.098). Patients receiving the combination treatment exhibited significantly higher interferon-γ production by CD8+ and CD4+ T cells (both P<0.001), as well as significantly reduced levels of IL-10, TGF-ß1 and TGF-ß2. In addition, patients in the combination treatment group experienced significantly longer overall survival (OS; P=0.024) and disease-free survival (DFS; P=0.007) times; a DTH response of ≥15 mm was also associated with increased OS time and DFS time (P≤0.001). Finally, no severe adverse events were observed in either treatment group. Overall, in situ immunization with DNP in combination with laser immunotherapy may activate focal T cells, producing a regional antitumor immune response that increases cell-mediated immunity and improves survival in MM patients. Thus, this may represent a novel therapeutic strategy for patients with unresectable, advanced MM.

Gut-lung axis: The microbial contributions and clinical implications.

Gut microbiota interacts with host immune system in ways that influence the development of disease. Advances in respiratory immune system also broaden our knowledge of the interaction between host and microbiome in the lung. Increasing evidence indicated the intimate relationship between the gastrointestinal tract and respiratory tract. Exacerbations of chronic gut and lung disease have been shown to share key conceptual features with the disorder and dysregulation of the microbial ecosystem. In this review, we discuss the impact of gut and lung microbiota on disease exacerbation and progression, and the recent understanding of the immunological link between the gut and the lung, the gut-lung axis.

Photosynthetic and growth responses of Schima superba seedlings to sulfuric and nitric acid depositions.

A continuing rise in acid deposition can cause forest degradation. In China, acid deposition has converted gradually from sulfuric acid deposition (SAD) to nitric acid deposition (NAD). However, the differing responses of photosynthesis and growth to depositions of sulfuric vs. nitric acid have not been well studied. In this study, 1-year-old seedlings of Schima superba, a dominant species in subtropical forests, were treated with two types of acid deposition SO4 (2-)/NO3 (-) ratios (8:1 and 0.7:1) with two applications (foliar spraying and soil drenching) at two pH levels (pH 3.5 and pH 2.5) over a period of 18 months. The results showed that the intensity, acid deposition type, and spraying method had significant effects on the physiological characteristics and growth performance of seedlings. Acid deposition at pH 2.5 via foliar application reduced photosynthesis and growth of S. superba, especially in the first year. Unlike SAD, NAD with high acidity potentially alleviated the negative effects of acidity on physiological properties and growth, probably due to a fertilization effect that improved foliar nitrogen and chlorophyll contents. Our results suggest that trees were damaged mainly by direct acid stress in the short term, whereas in the long term, soil acidification was also likely to be a major risk to forest ecosystems. Our data suggest that the shift in acid deposition type may complicate the ongoing challenge of anthropogenic acid deposition to ecosystem stability.

Neoadjuvant Therapy of DOF Regimen Plus Bevacizumab Can Increase Surgical Resection Ratein Locally Advanced Gastric Cancer: A Randomized, Controlled Study.

Locally advanced gastric cancer (LAGC) is best treated with surgical resection. Bevacizumab in combination with chemotherapy has shown promising results in treating advanced gastric cancer. This study aimed to investigate the efficacy of neoadjuvant chemotherapy using the docetaxel/oxaliplatin/5-FU (DOF) regimen and bevacizumab in LAGC patients.Eighty LAGC patients were randomized to receive DOF alone (n = 40) or DOF plus bevacizumab (n = 40) as neoadjuvant therapy before surgery. The lesions were evaluated at baseline and during treatment. Circulating tumor cells (CTCs) were counted using the FISH test. Patients were followed up for 3 years to analyze the disease-free survival (DFS) and overall survival (OS).The total response rate was significantly higher in the DOF plus bevacizumab group than the DOF group (65% vs 42.5%, P = 0.0436). The addition of bevacizumab significantly increased the surgical resection rate and the R0 resection rate (P < 0.05). The DOF plus bevacizumab group showed significantly greater reduction in CTC counts after neoadjuvant therapy in comparison with the DOF group (P = 0.0335). Although the DOF plus bevacizumab group had significantly improved DFS than the DOF group (15.2 months vs 12.3 months, P = 0.013), the 2 groups did not differ significantly in OS (17.6 ± 1.8 months vs 16.4 ± 1.9 months, P = 0.776. Cox proportional model analysis showed that number of metastatic lymph nodes, CTC reduction, R0 resection, and neoadjuvant therapy are independent prognostic factors for patients with LAGC.Neoadjuvant of DOF regimen plus bevacizumab can improve the R0 resection rate and DFS in LAGC. These beneficial effects might be associated with the reduction in CTC counts.

Expression of HSPA2 in human hepatocellular carcinoma and its clinical significance.

Hepatocellular carcinoma (HCC) remains one of the most common cancers worldwide. HSPA2 has been highlighted as an important marker in many types of cancers. However, little is known about the role of HSPA2 in HCC. The objective of the current study was to investigate the expression pattern and clinicopathological significance of HSPA2 in patients with HCC. Quantitative reverse-transcriptase ploymerase chain reaction (qRT-PCR) was applied to examine HSPA2 messenger RNA (mRNA) expression in 52 pairs of HCC tissues and adjacent noncancerous tissues. Immunohistochemistry (IHC) was performed to examine HSPA2 protein expression in paraffin-embedded tissues from 119 HCC patients. Statistical analyses were applied to evaluate the diagnostic value and associations of HSPA2 expression with clinicopathological characteristics. We identified abnormally elevated mRNA expression of HSPA2 in HCC tissues compared to paired adjacent noncancerous tissues (P < 0.001). Clinicopathological analysis showed that HSPA2 expression was significantly correlated with tumor size (P = 0.013), histological differentiation (P = 0.04), and tumor stage (P = 0.001). Patients with higher HSPA2 expression had shorter overall survival time, whereas those with lower HSPA2 expression had longer survival time. Furthermore, Cox regression analyses showed that HSPA2 expression was an independent predictor of overall survival. In conclusion, our findings provide evidences that positive expression of HSPA2 in HCC may be important in the acquisition of an aggressive phenotype and it is an independent biomarker for poor prognosis of patients with HCC.

Development of a DNA microarray method for detection and identification of all 15 distinct O-antigen forms of Legionella pneumophila.

Legionella is ubiquitous in many environments. At least 50 species and 70 serogroups of the Gram-negative bacterium have been identified. Of the 50 species, 20 are pathogenic, and Legionella pneumophila is responsible for the great majority (approximately 90%) of the Legionnaires' disease cases that occur. Furthermore, of the 15 L. pneumophila serogroups identified, O1 alone causes more than 84% of the Legionnaires' disease cases that occur worldwide. Rapid and reliable assays for the detection and identification of L. pneumophila in water, environmental, and clinical samples are in great demand. L. pneumophila bacteria are traditionally identified by their O antigens by immunological methods. We have recently developed an O serogroup-specific DNA microarray for the detection of all 15 distinct O-antigen forms of L. pneumophila, including serogroups O1 to O15. A total of 35 strains were used to verify the specificity of the microarray, including 15 L. pneumophila O-antigen standard reference strains and seven L. pneumophila clinical isolates as target strains, seven reference strains of other non-pneumophila Legionella species as closely related strains, and six non-Legionella bacterial species as nonrelated strains. The detection sensitivity was 1 ng of genomic DNA or 0.4 CFU/ml in water samples with filter enrichment and plate culturing. This study demonstrated that the microarray allows specific, sensitive, and reproducible detection of L. pneumophila serogroups. To the best of our knowledge, this is the first report of a microarray serotyping method for all 15 distinct O-antigen forms of L. pneumophila.

Effects of elevated O3 concentration on winter wheat and rice yields in the Yangtze River Delta, China.

The effects of a continuing rise of ambient ozone on crop yield will seriously threaten food security in China. In the Yangtze River Delta, a rapidly developing and seriously air polluted region in China, innovative open-top chambers have been established to fumigate winter wheat and rice in situ with elevated O(3). Five years of study have shown that the yields of wheat and rice decreased with increasing O(3) concentration. There were significant relationships between the relative yield and AOT40 (accumulated hourly O(3) concentration over 40 ppb) for both winter wheat and rice. Winter wheat was more sensitive to O(3) than rice. O(3)-induced yield declines were attributed primarily to 1000-grain weight and harvest index for winter wheat, and attributed primarily to grain number per panicle and harvest index for rice. Control of ambient O(3) pollution and breeding of O(3) tolerant crops are urgent to guarantee food security in China.

RUNX3 mediates suppression of tumor growth and metastasis of human CCRCC by regulating cyclin related proteins and TIMP-1.

Here we presented that the expression of RUNX3 was significantly decreased in 75 cases of clear cell renal cell carcinoma (CCRCC) tissues (p<0.05). Enforced RUNX3 expression mediated 786-O cells to exhibit inhibition of growth, G1 cell-cycle arrest and metastasis in vitro, and to lost tumorigenicity in nude mouse model in vivo. RUNX3-induced growth suppression was found partially to regulate various proteins, including inhibition of cyclinD1, cyclinE, cdk2, cdk4 and p-Rb, but increase of p27(Kip1), Rb and TIMP-1. Therefore, RUNX3 had the function of inhibiting the proliferative and metastatic abilities of CCRCC cells by regulating cyclins and TIMP1.

Use of a DNA microarray for detection and identification of bacterial pathogens associated with fishery products.

We established a microarray for the simultaneous detection and identification of diverse putative pathogens often associated with fishery products by targeting specific genes of Listeria monocytogenes, Salmonella, Shigella, Staphylococcus aureus, Streptococcus pyogenes, Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, and Yersinia enterocolitica and the 16S-23S rRNA gene internal transcribed spacer (ITS) region of Proteus mirabilis and Proteus vulgaris. The microarray contained 26 specific probes and was tested against a total of 123 target bacterial strains that included 55 representative strains, 68 clinical isolates, and 45 strains of other bacterial species that belonged to 8 genera and 34 species, and it was shown to be specific and reproducible. A detection sensitivity of 10 ng DNA or 10 CFU/ml for pure cultures of each target organism demonstrated that the assay was highly sensitive and reproducible. Mock and real fishery product samples were tested by the microarray, and the accuracy was 100%. The DNA microarray method described in this communication is specific, sensitive, and reliable and has several advantages over traditional methods of bacterial culture and antiserum agglutination assays.

[Preventive and therapeutic effects of quercetin on hyperuricemia and renal injury in rats].

OBJECTIVE: To study the preventive and therapeutic effects of Quercetin (Que) on hyperuricemia and renal failure in rats. METHODS: Seventy rats were divided into ten groups, normal control group, model control group, allopurinol group and seven Que groups (50, 40, 30, 20, 10, 5 and 2.5 mg/kg). Hyperuricemia and renal failure in rats were induced by adenine and ethambutol hydrochloride intragastrically, and then Que was given for 3 weeks. The items related to high serum uric acid and renal injury were observed. RESULTS: Levels of serum uric acid (SUA), serum urea nitrogen ( SUN), serum creatinine (SCr), malonaldehyde (MDA) and the activity of xanthine oxidase (XOD) in liver were lowered effectively by Que, and the activity of superoxide dismutase (SOD) in serum was increased. The better results of lowering serum uric acid and protecting against renal failure were at the dosage of Que between 10 and 20 mg/kg. CONCLUSION: Hyperuricemia and renal failure in rats could be intervened by Que. The possible mechanism might be inhibiting the activity of liver XOD and improving the ability of clearing oxygen free radicals and reducing lipid peroxidation, which might play an important role in reducing serum uric acid level and protecting kidney.

[Responses of Ilex integra Thunb. seedlings to elevated air ozone concentration].

One-year-old Ilex integra seedlings were exposed to charcoal-filtered (CF) and elevated ozone (E-O3, approximately 150 microL x L(-1)) for 84 days in six open-top chambers. Visible injury, growth parameters, pigments content, gas exchange, chlorophyll a fluorescence and antioxidant system were investigated during the growing season. At the end of experiment, foliage showed remarkable visible symptoms with dark-brown necrotic spots and patches which were concaved on the upper surface of the current-year leaves. Although relative height and diameter increment, total biomass and specific leaf weight (SLW) remained unaffected, E-O3 significantly decreased the percentage of stem biomass in total biomass. E-O3 induced significant decrease in net photosynthetic rate, chlorophyll a/b ratio and total phenolic compound content by 19%, 9% and 36%, respectively. However, stomatal conductance, intercellular CO2 concentration, chlorophyll a fluorescence parameters, pigment contents, MDA contents, total antioxidant capacity and total ascorbate content remained unaffected by E-O3. The results suggested that E-O3-induced change in components of chlorophyll contributed to the reduction of photosynthesis in Ilex integra seedlings. In addition, although visible symptom was found during the experiment, antioxidant system, most of the physiological parameters and growth were not significantly affected by E-O3.