Candida albicans  infection was constructed to observe the effects of cinnamaldehyde (CA) on the C. albicans cell wall structure and cell wall -β-D-glucan contents. This study provides a theoretical basis for CA treatment to target invasive C. albicans infection.The incidence rate of invasive candidiasis is high, its treatment is difficult, and the prognosis is poor. In this study, an immunosuppressive mouse model of invasive C. albicans infection were given an oral dosage of CA (240 mg.kg− 1.d− 1) for 14 days. Then, mouse lung tissue samples were collected for detection of the levels of -β-D-glucan and transmission electron microscopy observations, using fluconazole as a positive control and 2% Tween 80 saline as a negative control.Immunosuppressed mice with invasive C. albicans infection was successfully established. The levels of -β-D-glucan in the CA treatment group, fluconazole positive control group, invasive C. albicans infection immunosuppressive mouse model group, and 2% Tween 80 normal saline control group were 86.55 ± 126.73 pg/ml, 1985.13 ± 203.56 pg/ml, 5930.57 ± 398.67 pg/ml and 83.36 ± 26.35 pg/ml, respectively. Statistically, the CA treatment group, fluconazole positive control group and invasive C. albicans infection immunosuppressive mouse model group were compared with each other (P < 0.01) and compared with the 2% Tween 80 saline group (P < 0.01), showing that the differences were very significant. Comparison of the CA treatment group with the fluconazole positive control group (P < 0.05) displayed a difference as well. Electron microscopy showed that CA destroyed the cell wall of C. albicans, where the outer layer of the cell wall fell off and became thinner and the nuclei and organelles dissolved, but the cell membrane remained clear and intact.The immunosuppressive mouse model of invasive C. albicans by interfering with the synthesis of -β-D-glucan to kill C. albicans. However, CA does not affect the cell membrane. This study provides a theoretical basis for CA treatment to target invasive C. albicans infection.CA destroys the cell wall structure of  Candida. Invasive candidiasis is a serious infection that affects the blood, heart, brain, eyes, bones and/or other parts of the body [Candida usually lives inside the body  and on the skin without causing any problems [Candida can enter the blood or internal organs and cause infection. Candida blood infection  is the most common form of invasive candidiasis. In the United States, candidaemia is one of the most common causes of blood infections in hospitalized patients [6H5CHCHCHO and a relative molecular mass of 132.6. A large number of pharmacological studies on CA in China and other countries have shown that it has anti-inflammatory, hypoglycaemic, antitumour, antibacterial and other pharmacological activities [C. albicans infection in mice [Candida in vivo. The results are reported below.Invasive candidiasis is an infection caused by a yeast, which is a type of fungus, called the body . Candidaproblems . Howeverpatients , 4, whicpatients . Clinicapatients . For exapatients . These dpatients –10. A nepatients , 12. Howtivities ; in parttivities , 16. How in mice , and thiC. albicans (ATCC 90028) and Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. provided 6- to 8-week-old male BALB/c mice weighing 22–25 g with an animal quality certificate , 2016–0001).The Institute of Microbiology, Chinese Academy of Sciences, provided CA (purity greater than 98%) was provided by Jiangxi Cedar Medical Oil Co., Ltd. (batch number: 20171002); cyclophosphamide was provided by Jiangsu Hengrui Pharmaceutical Co., Ltd. (batch number: 20170830); fluconazole was provided by Pfizer Pharmaceutical Co., Ltd. (batch number: 20170630); Gram-negative bacteria lipopolysaccharide (LPS) was provided by Zhanjiang Anders Biotechnology Co., Ltd. (batch number: 20170801); Dynamic test tube detector provided by Shenzhen Ryder Power Technology Co., Ltd. (LKM02–32). Microscope, model: Olympus CX43 ; Transmission electron microscope (JEOL JEM-1200EX type); Ultra-clean workbench (Beijing Semiconductor No. 1 Factory).Experiments were performed at the Animal Experiment Center of the Second Hospital of Hebei Medical University. The feeding conditions were as follows: IVR laminar flow cabinet, temperature 22 °C, humidity 50–70%, 5 mice/cage. Protocols were approved by the Animal Care and Use Committee of Hebei Medical University .C. albicans infection (model group) and 10 mice in the normal saline control group; and (3) 90 mice in the CA experimental groups, including 30 immunosuppressive model mice with invasive C. albicans infection treated with CA (CA group), 30 mice in the fluconazole positive control group (Fluconazole group), and 30 mice treated with the negative control 2% Tween 80 saline . The study used a double-blind approach and samples were collected for testing.The experimental study of 170 mice is divided as follows using a random number table: (1) immunosuppressive group 40 ; (2) 30 immunosuppressive model mice with invasive − 1.d− 1 cyclophosphamide for two consecutive days. Blood was collected from the tail 1.0 day before treatment and 2.0, 4.0, 6.0, and 8.0 days after treatment. White blood cells and neutrophils in each sample were counted using a Beckman Coulter Hmx five-stage automatic blood analyser [Thirty BALB/c mice were injected intraperitoneally with 200 mg.kganalyser . As a noC. albicans (1 × 107 cfu/ml). Then, the mice were observed for 14 days. On the day that each mouse died or on the 14th day after treatment, an overdose of pentobarbital (150 mg/kg) was injected intraperitoneally to each mouse to stop the heartbeat and initiate respiratory failure and death. Then, kidney, liver and lung tissue specimens were collected for fungal microscopic examination, culture, pathological examination, -β-D-glucan detection and electron microscopic observations [On the fourth day of immunosuppression, 30 mice were intraperitoneally injected with 0.25 ml of rvations . On the C. albicans (1 × 107 cfu/ml) and 12 h later, 240 mg.kg− 1.d− 1 was orally administered followed by continued administration for 14 consecutive days (human effective dose × 10 = mouse dose), for a total dose of 3360 mg/kg [− 1.d− 1) of the 30 mice in the positive control group over 14 consecutive days was 3360 mg/kg. In the negative control group , 30 mice orally received 0.25 ml of 2% Tween 80 in normal saline for 14 consecutive days. On the 15th day, all surviving mice were intraperitoneally injected with sodium pentobarbital solution (150 mg/kg) and euthanized. Then, a mouse lung tissue specimen (0.5 g) was collected from each mouse under aseptic conditions for -β-D-glucan detection and electron microscopy observation.On the fourth day of immunosuppression, 30 mice were intraperitoneally injected with 0.25 ml of 60 mg/kg . The totTwo grams of Tween 80 was added to 97.76 ml of 0.9% saline to prepare a 2% Tween 80 solution.Then, 240 mg of CA and 240 mg of fluconazole were gradually added to the 2% Tween 80 solution to obtain 0.24% CA and fluconazole solutions, respectively.C. albicans yeast suspension was prepared in 0.1% Tween 80 saline at a concentration of 1 × 107 cfu/ml for later use. Then, the yeast suspension was diluted 10-fold with 0.1% Tween 80 saline, and 100 μl of the diluted solution was inoculated into a glucose protein agar plate. The cells were cultured at 37 °C, and the colonies were counted to determine the survival rate of the Candida strain.A Candida infection model group, CA treatment group and fluconazole control group were prepared with 10% potassium hydroxide, examined under a microscope and cultured on glycoprotein one agar. The agar was observed for the growth of C. albicans.Lung tissue specimens of the mice in the invasive At the end of the experiment, all mice were euthanized by anaesthesia. Then, lung tissue specimens were collected and completely fixed in 4% paraformaldehyde solution. They were conventionally dehydrated, cleared and embedded to make 5 μm thick sections, and the sections were subjected to periodic acid-Schiff (PAS) staining. Histopathological changes were observed under a microscope.C. albicans infection model group, CA treatment group and fluconazole control group were collected, and the contents of -β-D-glucan in lung tissue were determined by the velocity turbidimetric method. Usually, 500 mg of tissue homogenate was centrifuged at 400×g for 10 min. Then, 100 μg of the supernatant was added to a sample dilution bottle and mixed gently with a vortex, and then the sample was placed in a test tube thermostat (75 °C) for 10 min. Then, a pipette was used to add 0.25 μg of reagent to the LPS reagent to prepare a solution. After 100 μg of the prepared sample test solution was added to the reaction tube, 50 μg of the reagent solution was added. Then, the solutions were mixed, and each test tube was inserted into the LKM dynamic tube tester. Before reading the results, the reaction was carried out at 37 °C for 75 min.Lung tissue specimens of the mice in the Lung tissue specimens were collected from mice in the model group, CA group and fluconazole group, fixed with 2% glutaraldehyde and 1% phosphotungstic acid, and dehydrated in an ethanol series  followed by acetone and embedded in epoxy resin. Ultra-thin sections were prepared, stained with uranyl acetate and lead citrate, and observed under a transmission electron microscope.After 14 days, the surviving mice in the model group, CA group and fluconazole group were given an overdose of 1% sodium pentobarbital (150 mg/kg) intraperitoneally. The mice quickly lost consciousness without struggling, their heartbeat and respirations gradually stopped, and the mice eventually died of respiratory failure.P < 0.05.SPSS 21.0 statistical software was used for statistical analysis. The measurement data results are expressed as the mean ± standard deviation and comparison of the count data between groups was performed with the chi-square test. Comparative analysis between groups was performed by analysis of variance, and pairwise comparison of data was performed by the LSD t test. At α = 0.05, all differences were regarded as statistically significant when − 1.d− 1 cyclophosphamide for 2 consecutive days. The total numbers of white blood cells and granulocytes in the immunosuppressed mice on day 4 were 1.48 × 109/L and 16 × 109/L, respectively, which were significantly lower than those in the normal saline control group (P < 0.01). The white blood cell count gradually returned to normal on the 8th day. After 28 days of observation, the mice did not die naturally. These results show that intraperitoneal injection of cyclophosphamide for 2 consecutive days can effectively induce immunosuppression in mice.Table 7 cfu/ml C. albicans suspension, 2 mice died on the 2nd day, 6 mice died on the 3rd day, 6 mice died on the 4th day, 2 mice died on the 5th day, 2 mice died on the 6th day, 2 mice died on the 7th day, and 1 mouse died on the 8th day. A total of 21 mice died within 14 days for a mortality rate of 70%. The mice in the saline control group did not die, and there was a significant difference between the two groups (P < 0.01)  was observed, as shown in , 1985.13 ± 203.56 (pg/ml), 5930.57 ± 398.67 (pg/ml) and 83.36 ± 26.35 (pg/ml), respectively. The values of the CA treatment group and the fluconazole positive control group were compared with those of the invasive C. albicans infection model group and the 2% Tween 80 normal saline negative control group and a significant difference was found (P < 0.01). Comparing the value in the CA treatment group with that of the fluconazole positive control group, P < 0.05, indicating a significant difference -β-D-glucan detected in the CA treatment group, fluconazole positive control group, invasive -β-D-glucC. albicans.Explanation: CA has a significant effect on -β-D-glucan content in the cell wall of C. albicans in the infection model group were plate-like multilayer thick-walled structures with transparent cell membranes and complete inclusions, as shown in  Electron microscopy showed that the cell membrane of C. albicans treated with CA was still intact. This indicates that CA destroys only the fungal cell wall in the body without affecting the cell membrane. Currently, the main clinical drugs used to treat C. albicans infections are azoles, which target fungal cell membranes, have strong toxicity and side effects and are prone to drug resistance. Therefore, CA is an antifungal drug that acts on fungal cell walls for use in the treatment of severe fungal infections, especially Candida infections, and is unlikely to have toxic side effects on the human body.CA has antimicrobial properties, and determination of its minimum inhibitory concentration indicated that it is a powerful natural antimicrobial agent . It has C. albicans and affects the synthesis of -β-D-glucan but it does not affect the cell membrane or exhibit toxic side effects. This study provides a theoretical basis for the use of CA treatment against invasive C. albicans infection.As invasive fungal infections have become more common and severe, they have also become an increasingly serious public health problem , 28. The


The complement system, an innate immune system, may either play an antitumor role, or promote tumorigenesis and cancer progression in different kinds of cancer. The function of complement in hepatocellular carcinoma (HCC) is unclear.The gene expressions of the complement system were based on data obtained from TCGA and GEO. We explored gene expressions, mutation, enrichment analysis, clinicopathology, patients' outcome, and immune infiltration via Gepia2, cBioPortal, Metascape, UALCAN, Kaplan–Meier Plotter, and TIMER 2.Five complement genes, including C1R, C6, C7, CFP, and CFHR3, were not only found to be significantly downregulated in HCC samples compared with normal liver samples, but also found to be significantly associated with overall survival, disease‐free survival, and progress‐free survival in HCC patients. In addition, lower mRNA expression of C1R, C6, C7, and CFHR3 were found correlated with advanced cancer stages and higher tumor grades in HCC patients. Also, the expression levels of CFP were correlated with many sets of immune markers of tumor immune cells, such as those of CD8+ T cells, CD4+ T cells, B cells, M2 macrophages, neutrophils, DCs, Th1 cells, Th2 cells, and T cell exhaustion in HCC. Based on that, we developed a prognostic model for HCC patients—Riskscore = (−0.0053)*C6+(−0.0498)*C7+(−0.1045)*CFHR3.C1R, C6, C7, CFP, and CFHR3 could be prognostic biomarkers for patients with HCC. C1R, C6, C7, CFP, and CFHR3 could be prognostic biomarkers for patients with HCC. Based on that, we developed a prognostic model for HCC patients—Riskscore = (−0.0053)*C6+(−0.0498)*C7+(−0.1045)*CFHR3. Study shows that its morbidity has elevated to 6th in cancer, while it also ranks the third death cause of cancer.The complement system is a fundamental branch of innate immunity of human body, containing more than 50 inherent components and membrane binding receptors and regulators. Three main pathways of complement activation depending on C3It is reported that the complement system has a complicated relationship with human cancer.To address this, we thoroughly explored the expressions of the complement‐related genes and their correlation with prognosis and immune infiltration in patients with HCC via The Cancer Genome Atlas (TCGA) public and GEO database for the first time.22.1TCGA is a comprehensive database containing information on more than 30 kinds of human cancers, including mRNA sequencing, DNA sequencing, pathological data, and clinical pathological information.http://www.ncbi.nlm.nih.gov/geo/). This data set contains 268 HCC samples and 243 normal controlled samples from HCC patients.As there are only 50 normal liver samples in TCGA database, we also collected HCC data set GSE25097 from Gene Expression Omnibus  patients in the TCGA and GSE25097 databases were diagnosed with HCC. (2) There were clear criteria for the diagnosis and staging of HCC. (3) The study data provided OR (odds ratio)/HR (hazard ratio) and its 95% confidence interval (CI), or could be converted to OR and its 95% CI. The exclusion criterion was as follows: (1) Repeat recorded cases. (2) Data was incomplete.2.3http://timer.cistrome.org/) is an online analysis tool that can not only analyze gene expression profiles in different kinds of cancers, but also provide an advanced algorithm to evaluate the abundance of tumor‐infiltrating immune cells by gene expression profiles based on TCGA database.First, differences in mRNA expressions of the whole complement system between cancer samples and normal controlled samples were analyzed by TIMER2.TIMER2 (www.gepia.cancer-pku.cn) is a web server analyzing RNA sequencing expression data of tumors and normal samples from both TCGA and Genotypic Tissue Expression (GTEx) projects using standard processing pipelines.Then, we used the pair‐controlled samples for HCC samples in GEPIA database and datasets GSE25097 to further verify the different mRNA expressions between HCC samples and their normal counterparts. Gene Expression Profiling Interactive Analysis , a web tool based on level 3 RNA‐seq and clinical data of 31 cancer types from TCGA database, can be used to explore the association of the transcriptional expression of a specific gene with cancer patients' clinicopathologic parameters.Next, UALCAN database was used to examine the relationship between mRNA expressions of the complement system and patients' clinicopathologic information, such as tumor stage and grade. UALCAN (2.4www.kmplot.com), which is a web tool for survival analysis by product‐limit method based on gene expression data and survival information of patients with different kinds of cancers based on TCGA database.p‐value.In this study, we evaluated the prognostic value of mRNA expressions of the complement system in patients with HCC by calculating overall survival (OS), disease‐free survival (DFS), and progress‐free survival (PFS) using the Kaplan–Meier plotter , mRNA expression 2.6www.string-db.org), an online analysis tool that collects, scores and integrates all publicly available sources to predict PPI between various proteins.We performed a protein–protein interaction (PPI) network analysis on the prognostic genes of the complement system to explore their interactions by STRINGS , a free and well‐maintained online bioinformatics database for GO and KEGG enrichment analysis.p < .01, minimum count > 3, and enrichment factor > 1.5.Next, we used the list of co‐expressed genes to conduct Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis to predict the function of these prognostic genes in HCC by Metascape , by spearsmans' correlation coefficients. Then, we examined the relationships between the prognostic complement genes and immune gene marker sets in HCC sample from TCGA using TIMER2. The gene markers of tumor‐infiltrating immune cells, including T cells, monocytes, tumor‐associated macrophages (TAMs), M1 and M2 macrophages, neutrophils, DCs, Tregs, and T cell exhaustion, were referenced from prior studies.2.8A prognostic model was constructed based on our results of the complement expressions and OS prognostic information in the cluster. The method of the least absolute shrinkage and selection operator (LASSO) regression algorithm is applied to find a model that can best explain the data but contains the fewest free parameters.First, timeROC(v 0.4) analysis was performed to compare the predictive accuracy of each gene and score. Then, LASSO regression algorithm for feature selection, using tenfold cross‐validation. The K–M survival analysis with log‐rank test was also used to test the constructed model. All analytical methods above were performed using R software version 4.0.3.2.9t‐test in UALCAN and GSE25097. A fold change >2 or <0.5, or log2(FC) > 1 or <−1 with p‐values < .05 in both Gepia database and GSE25097 were considered statistically significant in mRNA expression in this study. The K–M curve diagrams of patients' survival were displayed as HR and log‐rank p‐values. In Spearman's correlation coefficients, we defined the strength of correlation as follows: 0.00–0.1, negligible correlation; 0.10–0.39, weak correlation; 0.40–0.69, moderate correlation; 0.70–0.89, strong correlation; and 0.90–1.0, very strong correlation. In this study, p‐values < .05 were considered statistically significant.The differential expression of complements was explored using the Wilcoxon test in TIMER2, ANOVA in GEPIA, and Students' 33.1n = 371) and normal tissues (n = 50) in patients (Table p < .05). In contrast, nine complement genes, including C2, C5AR2, C4BPB, CD46, CD59, ITGAM, ITGAX, C1QBP, and C1QR, were significantly upregulated in HCC tissues compared with normal samples  than in normal samples (n = 160), while the mRNA expression of C1QA was significantly higher in HCC samples (n = 368) than in normal samples (n = 160). The expression of the other 27 complements showed no difference between HCC samples and normal tissues, with a log2(FC) > 1 or <−1 and p‐value < .05. We then used data set (GSE25097) to confirm the mRNA expression of all the complement genes. Our result revealed that the mRNA expressions of C1R, C6, C7, C9, CFP, CFHR3, and MASP1 were significantly downregulated in HCC tissues (n = 268) compared with normal liver tissues (n = 243), with a fold change >2 or <0.5 and p‐value < .05 , since they contained more paired normal liver samples. According to the result from Gepia database  but did not affect OS in HCC patients.As shown in Figure 3.5Then, we used String database to build a PPI network between the five complements genes found above, including C1R, C6, C7, CFP, and CFHR3, as they were not only downregulated in HCC, but also correlated with the prognosis of patients with HCC. We found that these genes generally interacted with other complement genes to moderate the immune system and hormone system Figure . There wSo, we calculated co‐expressed genes of these five complement genes in HCC samples from TCGA database. Table Next, we conducted GO and KEGG pathway enrichment analysis by these co‐expressed genes to further predict the function of the five prognostic complement genes in HCC patients. As shown in Figure 3.6Given the relationship of complements with the immune system, we further studied the correlations between complement genes and marker genes of tumor‐infiltrating immune cells, including T cells, TAMs, M1 and M2 macrophages, monocytes, T cell exhaustion, Tregs, and DCs, from HCC tissues.First, we examined C1R, C6, C7, CFP, and CFHR3 as they were considered to have prognosis values in HCC patients. Among them, we found only CFP had a statistical correlation with the immune cells in HCC samples. The mRNA expression of CFP had a weak positive correlation with CD8+ T cells and DCs , T cells (CD3E and CD2), B cells CD79A), M2 macrophages (CD163), neutrophils (CCR7), DCs , Th1 cells (IFNG and TBX21), Th2 cells (GATA3), and T cell exhaustion , CR1, CR2, CR3 (ITGAM), and CR4 (ITGAX) were positively associated with almost all kinds of tumor immune cells, including CD8+ T cells, CD4+ T cells, B cells, macrophages, monocytes and DCs *C6+−0.0498)*C7+(−0.1045)*CFHR3, with a lambda.min = 0.0296 . Also, the time‐dependent ROC analysis showed that the 1‐year AUC of this model was 0.705  were considered to have prognostic value. We used KEGG analysis to further speculate the biological function of C1R, C6, C7, CFP, and CFHR3 in HCC. We found that the lower mRNA expressions of C1R, C6, C7, CFP, and CFHR3 were correlated with pathways like Fanconi anemia pathway, cell cycle, MicroRNAs in cancers, and so on. These pathways were reported to promote proliferation, migration, and invasion in HCC.C1R, a complement protein, combined with C1q and C1S to activate the classical pathway of the complement system. Our study found its prognostic value in patients with HCC. A previous study had shown the serum concentration of C1R had prognostic value in non‐small cell lung cancer.C6 is a complement protein composing the membrane attack complex (MAC). Our study showed its prognostic value in HCC patients, which was consistent with other studies like Mu et al.C7 is also a complement protein composing MAC. Our study showed the prognostic value of mRNA expression of C7 in HCC. Lisha et al.CFP, also known as properdin—activating the alternative pathway of the complement system by combining C3bBb, was also reported to have prognostic value in HCC, lung adenocarcinoma, and pancreatic adenocarcinoma.CFHR3, a negative regulator of the complement system, was reported to have prognostic value in patients with HCC according to our study. Other studies, like Liu et al.p < .05, but not according to GSE25097 . Similarly, the mRNA expression of MASP1 was significantly downregulated in HCC samples than in normal liver tissues in GSE25097 with a fold change >2 or <0.5 and p < .05, but not according to Gepia database . So, we did not consider their mRNA expressions were significantly different between HCC samples and normal liver tissues in this study, but we believed further research are needed to confirm the mRNA expressions of MBL2, C8A, and MASP1 in HCC as they might have an important biological function in HCC. Transcription levels of other complement genes, like CD46, CD59, and C1QBP, were found up‐ or downregulated in HCC samples only in one data set—TIMER2, without a fold change >0.5 and <2. So, we did not believe the expression of these genes was significantly altered in HCC.The mRNA expressions of MBL2 and C8A were significantly downregulated in HCC samples than in normal liver tissues in Gepia database with a fold change >2 or <0.5 and Here, we reported that five complement genes showed prognostic values in patients with HCC, including C1R, C6, C7, CFP, and CFHR3. As shown in our result, the mRNA expressions of these five genes were significantly lower in HCC samples than in normal controlled samples. Moreover, lower expression of C1R, C6, C7, CFP, and CFHR3 was associated with poorer OS, DFS, and PFS in HCC patients. Also, we found that the mRNA expression of C1R, C6, C7, and CFHR3 was associated with tumor grade, while the mRNA expression of C1R, C6, and CFHR3 was associated with the cancer stage. So, we thought C1R, C6, C7, and CFHR3 could be prognostic biomarkers for patients with HCC. Although CFP was not associated with tumor grade or cancer stage, it was associated with the mRNA expressions of many immune marker genes in HCC, including CD8+ T cells , CD4+ T cells , B cells (CD79A), M2 macrophages (CD163), neutrophils (CCR7), DCs , Th1 cells , Th2 cells (GATA3) and T cell exhaustion , and so on. Thus, we thought CFP could also be a biomarker for HCC patients. As higher CFP expression was associated with better OS in HCC patients, we speculated it might be associated with the immune system regulating HCC and might be a new target or biomarker for immune therapy. Furthermore, we developed a prognostic model for HCC patients using these complement genes.Moreover, mRNA expressions of many complement genes were not altered or unclear according to our results. However, these genes were also found associated with OS, DFS, and PFS in HCC patients, like C2, C3, C5, and so on. Thus, we further examined these genes and found that the mRNA expressions of C3, C5, CFB, and CLU were correlated with cancer stage and prognosis in patients with HCC. In addition, the mRNA expressions of C1S, C8B, MBL2, CFI, and C4BPA were correlated with tumor grade and prognosis in HCC patients. Other studies, like Wang et al.Apart from complement genes mentioned above, the mRNA expressions of C2, CFH, CFHR1, CFHR2, CFHR4, SERPING1, MASP2, MASP1, C4BPB, VTN, CPN1, and CR2 were associated with OS, DFS, and PFS in patients with HCC. However, their mRNA expressions were neither altered in HCC, nor associated with cancer stage or tumor grade in HCC. As result, we did not consider these genes had prognostic value in HCC.Generally, it is believed that the activation of the complement system is associated with tumorigenesis, progression, and so on, by causing inflammation in the tumor microenvironment (TME).There were some limitations in our study. First, all the data was retrieved from online public databases (TCGA and GEO), further studies with larger sample sizes are required to validate and to explore the application of the five complement members in the prognosis and treatment of HCC. Second, we did not explore the possible mechanisms of these five complement genes in HCC via basic experiments. Finally, 27 complement genes other than C1R, C6, C7, CFP, and CFHR3 were also associated with better OS and might have an important biological function in HCC. We were intended to confirm their role in HCC and investigate the mechanism between these genes and HCC in further study.5In summary, we assessed the gene expressions of the complement system in HCC. We have found C1R, C6, C7, CFP, and CFHR3 have prognostic value as they were not only significantly downregulated in HCC but also correlated with better OS, DFS, and PFS in HCC patients. Also, C1R, C6, C7, and CFHR3 presented correlations with tumor grades and cancer stage in HCC patients, while CFP presented correlations with the immune markers of tumor immune cells in HCC. Furthermore, we constructed a prognostic model based on our results: Riskscore = (−0.0053)*C6+(−0.0498)*C7+(−0.1045)*CFHR3.The data of the patients in this study were obtained from the public database data sets.Xinye Qian and Zhoujing Yang conceived and designed the study; Lu Gao and Yipiao Liu downloaded and organized The Cancer Genome Atlas data; Xinye Qian and Zhoujing Yang performed data analysis and wrote the paper; Jun Yan critically revised the article for essential intellectual content and administrative support. All authors read and approved the final manuscript.Supporting information.Click here for additional data file.


The solution was modified with the addition of Zn at a dose of 1 µg/L and Zn: 1 µg/L with prolactin (PRL): 0.1 µg/L. After 2 h and 48 h of storage, the levels of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, sodium, potassium, creatinine and total protein were determined. Zinc added to the Biolasol® composition at a dose of 1 µg/L showed minor effectiveness in the protection of nephrons. In turn, Zn2+ added to Biolasol + PRL (PRL: 0.1 µg/L) acted as a prolactin inhibitor. We do not recommend the addition of Zn(II) (1 µg/L) and Zn(II) (1 µg/L) + PRL (0.1 µg/L) to the Biolasol solution.Zinc is an effective anti-inflammatory and antioxidant trace element. The aim of this study was to analyse the protective effect of zinc and zinc–prolactin systems as additives of preservation solutions in the prevention of nephron damage caused during ischemia. The study used a model for storing isolated porcine kidneys in Biolasol It is a component of enzymes, including DNA and RNA polymerases. It is a part of superoxide dismutase, carbonic anhydrase, lactate dehydrogenase, malate dehydrogenase and glutamate dehydrogenase. As a component of superoxide dismutase, it reduces the production of reactive oxygen species and protects against oxidative damage. In turn, being part of carbonic anhydrase, it affects the acid–base balance, regulating it in the lungs and renal tubules  was from POCh S.A Gliwice, Poland. Pig prolactin was from “FZNP” Biochefa, Sosnowiec, Poland. All substances used in the study were of analytical grade.We used Biolasol solution . The composition and functions of the individual components are shown in Forty kidneys from 20 Polish Large White pigs, weighing 90–110 kg, aged 175–180 days, were included in the study. The animals were slaughtered at the Meat Plant H.A.M in Radzionków/Poland in a special room using 220 V electricity.The research was conducted with the consent of the II Local Ethics Commission for Animal Experiments in Cracow (No. 1046/2013) and in accordance with the European Union regulations (Directive 86/609 CEE) on the protection of animals during slaughter or killing.The kidneys were collected according to the previously described procedure . The graPerfusion, preservation and * reperfusion of kidneys in the Biolasol solution/control; n = 10 kidneys;Perfusion, preservation and * reperfusion of kidneys in the Biolasol solution modified with PRL (0.1 µg/L); n = 10 kidneys;2+ ; n = 10 kidneys;Perfusion, preservation and * reperfusion of kidneys in the Biolasol solution modified with Zn2+  and PRL (0.1 µg/L); n = 10 kidneys.Perfusion, preservation and * reperfusion of kidneys in the Biolasol solution modified with ZnPerfusion, preservation and * reperfusion of kidneys in the Ringer’s solution/sham; n = 10 kidneys.* reperfusion—flushing the graft after 48 h of preservation with the same solution that was used for previous perfusion and preservation.2O) ensuring a continuous flow of solution stream. Perfusate samples were collected into Eppendorf tubes from the renal vein at two time points: 0 and 30 min of perfusion. After 30 min, the kidneys were stored under simple hypothermia for 48 h (acceptable cold ischemic time for the kidney). After this time, renal reperfusion was performed. The test samples were taken at 0 and 30 min of reperfusion. The collected samples (after perfusion and reperfusion) were centrifuged at 3000 rpm for 15 min at T = 4 °C and stored at T = −20 °C until the tests were completed. Then, the biochemical markers of renal function were determined. Diagnostic tests were applied, allowing for indirect assessment of renal function [The isolated porcine kidneys were placed in 500 mL of Biolasol (Group A), Ringer (Group C) or modified Biolasol  chilled to 4 °C and transported in thermostable containers (4–6 °C) to the Biochefa FZNP laboratory. The grafts were stored under simple hypothermia for 2 h. The kidneys were then cannulated  and perfused  was determined using reagent kits  at 340 nm (linearity: 0–500 U/L) and expressed as U/L. The analyses were performed in accordance with the manufacturer’s instructions.The activity of aspartate aminotransferase (AST) was determined using reagent kits  at 340 nm (linearity: 0–500 U/L) and expressed as U/L. The analyses were performed in accordance with the manufacturer’s instructions.The activity of lactate dehydrogenase (LDH) was determined using reagent kits  at 340 nm (linearity: 1000 U/L) and expressed as U/L. The analyses were performed in accordance with the manufacturer’s instructions.Sodium concentration was determined using reagent kits  at 405 nm (linearity: 0–300 mmol/L) and expressed as mmol/L. The analyses were performed in accordance with the manufacturer’s instructions.Potassium concentration was determined using reagent kits  at 600 nm (linearity: 0–50 mmol/L) and expressed as mmol/L. The analyses were performed in accordance with the manufacturer’s instructions.Creatinine concentration was determined using reagent kits  at 490 nm (linearity: 0–25 mg/dL) and expressed as mg/dL. The analyses were performed in accordance with the manufacturer’s instructions.Total protein concentration was determined using reagent kits  at 540 nm (linearity: 1 to 15 mg/dL) and expressed as mg/dL. The analyses were performed in accordance with the manufacturer’s instructions.A Marcel S330 spectrophotometer  was used for biochemical tests. The photometric accuracy of the spectrophotometer was ±0.005 Abs.Kidney samples were collected after completion of reperfusion (48 h 30 min). The samples were homogenized in chilled 0.1 M phosphate buffer (pH = 7). Biochemical determinations were performed in supernatants obtained by centrifuging the homogenates at 15,000 rpm for 3 min. The renal tissue supernatant was used to evaluate the activity of ALT, AST, LDH, as well as creatinine and protein concentrations.p < 0.05 were considered statistically significant.The test results are shown as mean ± SEM (standard error mean). The parameters between the groups were compared by one-way analysis of variance (ANOVA) followed by post hoc Bonferroni test for means comparison (n = 10 for each group) . STATISTWe do not recommend the addition of Zn(II) (1 µg/L) and Zn(II) (1 µg/L) + PRL (0.1 µg/L) to the Biolasol solution.


Cell experiments demonstrated that FCPN was taken up by SMMC-7721 cells and significantly inhibited the proliferation of SMMC-7721 cells, which illustrated that FCPN had good targeting ability compared with PN and CPN. According to the results of in vivo animal experiments, FCPN significantly inhibited tumor growth. Tissue distribution experiments proved that FCPN could accumulate significantly in tumor tissues, which further explained why FCPN had good targeting ability. These results clearly suggested that folate-functionalized homotypic CM bionic nanosystems might represent a very valuable method for liver cancer treatment in the future.In this study, we prepared a folic acid-functionalized SMMC-7721 liver cancer cell membrane (CM)-encapsulated paclitaxel nanocrystals system (FCPN) for hepatoma treatment. Transmission electron microscopy (TEM) characterization showed that FCPN was irregular spherical shapes with a particle size larger than 200 nm and a coated thickness of approximately 20 nm. In an  Folic acid, N-hydroxysuccinimide (NHS), dicyclohexyl carbodiimide (DCC), dimethyl sulfoxide (DMSO), sodium dodecyl sulfate (SDS), ethanol, methanol, paraformaldehyde, and Pluronic 127 (F127) were purchased from Aladdin Chemical Reagent Co. . Thiazolyl blue tetrazolium bromide (MTT), propidium iodide (PI), Annexin V-FITC apoptosis detection kit, trypsin, Triton X-100, and bovine serum albumin were supplied by Nanjing Jiancheng Bioengineering Institute . SMMC-7721 cell lines were obtained from the National Experimental Cell Resource Platform . RPMI 1640 medium and fetal bovine serum (FBS) were purchased from Beijing Dingguo Changsheng Biotech Co. Ltd. . Phenylmethanesulfonyl fluoride (PMSF), 4′,6-diamidino-2-phenylindole (DAPI), mitochondrial membrane potential assay kit (JC-1), calcein-AM/PI were provided by Beyotime Biotechnology Co. . Bcl-2, Bax, and cleaved caspase-3 were obtained from Abcam . N-Cadherin, galectin-3, EpCAM, CD147, and EGFR were purchased from Wanleibio .2.2.The liver cancer cell line SMMC-7721 was cultured with a nutrient solution composed of 90% RPMI-1640 medium, 1% penicillin, 1% streptomycin, and 10% FBS. Cell digestion was performed using 0.25% trypsin during cell passage. FBS (90%) and DMSO (10%) were used for cryopreservation. The cells were finally stored at −80 °C.2.3.2.3.1.3 and 0.7 mM EDTA was added to the centrifuge tube. In addition, 30 µL of PMSF was replenished, and then the cell suspension was transferred to a homogenizer for homogenization in an ice bath. Finally, CM was obtained by gradient centrifugation at 4 °C and stored at −20 °C.First, the liver cancer cell line SMMC-7721 was cultured in a 15 mL cell culture flask. When the cells were overgrown, after digestion and centrifugation, 15 mL of 0.2× PBS was added to the centrifuge tube to swell and burst the cells for 24 h. Then, after centrifugation at 1250 r/min for 10 min, the supernatant was discarded. A solution (3 mL) containing 1 mM NaHCO2.3.2.First, 500 mg of FA was dissolved in 10 mL of DMSO with 0.25 mL of triethylamine. At room temperature, 0.25 g of DCC and 0.26 g of NHS were added to the FA solution and stirred overnight. Then, the solution was centrifuged to obtain the supernatant. A mixed solution containing 30% acetone and 70% ether was added to the above solution to obtain a yellow precipitate. The precipitate was washed three times with ether and dried under a vacuum to obtain afaint yellow solid powder, which was an FA active ester  were dissolved in methanol and diluted, and the content of the drugs was determined by HPLC  solution, flow rate 1.0 mL/min, wavelength 227 nm). The drug loading was calculated by the following formula:2.5.2.5.1.A scanning electron microscope (SEM)  was used to characterize the morphology of the unsheared nanocrystals. Transmission electron microscopy (TEM)  was used to observe the morphological structures of PN, CPN, and FCPN.2.5.2.The sizes and zeta potentials of PN, CPN, and FCPN were obtained using a laser dispersion particle size analyzer .2.5.3.2 flow of 150 mL/min at a rate of 10 °C/min. XRD was carried out by an X-ray refractometer with Cu-K radiation .The solid state of F127, pure PTX, PN, and FCPN was detected by DSC and XRD. DSC was performed using a differential scanning calorimeter  from 70 °C to 300 °C under N2.5.4.The targeting effects of CPN and FCPN were achieved by CM surface proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to characterize whether the encapsulation was successful. Membrane proteins were extracted from CM, CPN, and FCPN with RIPA lysis buffer and further measured with BCA analysis. Then, the protein sample and marker were added, and electrophoresis was performed in an electrophoresis device. After staining with Coomassie Brilliant Blue, image development was performed under a microscope and analyzed by Quantity One 1-D Analysis Software . In addition, western blotting was used to analyze SMMC-7721 CM proteins, including CD147  containing 0.05% SDS was used as the release medium at 37 °C. PTX, PN, and FCPN  were added to 200 mL of dissolution medium. Within a specified period of time, 2 mL samples were removed and filtered with a 0.22 µm organic filter membrane. At the same time, 2 mL of fresh release medium was replenished. The released drug content was detected by HPLC.2.7.2.7.1.The preparation process of FITC-labeled (PN) was as follows. First, PTX was dissolved in 1 mL of ethanol to form an organic phase. At room temperature, the surfactants F127 and FITC were dissolved in 10 mL of water as the water phase. Then, 1 mL of the organic phase was injected into the water phase under rapid stirring (1200 rpm) and sonicated for 5 min to obtain FITC-labeled PN. FITC-labeled PN was dried for 2 h under vacuum to remove ethanol. FITC-labeled PN was mixed with CM and FA-CM, and then the systems were sonicated and incubated for 2 h at 37 °C to obtain FITC-labeled CPN and FITC-labeled FCPN.Cell uptake was detected by a confocal laser scanning microscope . SMMC-7721 cells were inoculated in Petri dish. FITC-labeled PN, CPN, and FCPN were added and incubated for 1 h. Cells were fixed in 4% paraformaldehyde solution for 10 min. Hoechst 33342 and rhodamine phalloidin were successively used to treat the cells for staining. Next, the cells were immersed in 1 mL of PBS and observed by CLSM.Cell uptake was also detected by flow cytometry. SMMC-7721 cells were seeded in six-well plates and cultured for 24 h. FITC-labeled PN, CPN, and FCPN were added to the cells for 1 h. The cells were quantitatively analyzed by flow cytometry .To verify the targeting specificity of FCPN, FITC-labeled FCPN was cocultured with the human liver cancer cell line SMMC-7721, the human breast cancer cell line SK-BR-3 and the mouse glioma cell line C6 for 1 h. The uptake of CMs by these three types of cells was observed by CLSM.2.7.2.The toxicity of PTX, PN, CPN, and FCPN was determined by the MTT test. SMMC-7721 cells were counted and seeded into a 96-well plate with 5000 cells per well for 48 h. Then, pharmaceutical preparations with different drug concentrations  were added into a 96-well plate and incubated for 48 h. Thiazolyl blue tetrazolium bromide (MTT) at 5 mg/mL was added and incubated for 4 h under dark conditions. DMSO (150 μL) was added to 96 wells. The plate was placed in a shaker and shaken for 10 min under dark conditions. The absorbance value (OD) of formazan was measured at 492 nm with a microplate reader . Cell viability was calculated using the following formula:t represents the absorbance of cells in the drug preparation treatment group and ODc represents the absorbance of cells in the control group.OD2.7.3.SMMC-7721 cells were cultured in 12-well plates. After treatment with PTX and PTX preparations  for 24 h, 500 μL of calcein-AM/PI detection working solution was added to each well and incubated at 37 °C in the dark for 30 min. Then, the cells were observed under a fluorescence microscope  .2.7.4.2 incubator. PTX, PN, CPN, and FCPN  were added to six-well plates. After culturing for 48 h, the cells were digested with trypsin. Then, the collected cells were placed in another centrifuge tube, and 500 μL of binding buffer was added for resuspension. Five microliters of Annexin V-FITC and PI were added and mixed gently in dark conditions. Finally, the cell samples were tested with a flow cytometer .SMMC-7721 cells were seeded in a six-well plate and incubated for 24 h at 37 °C in a 5% CO2.7.5.SMMC-7721 cells were cultured in 12-well plates. The cells were treated with PTX, PN, CPN, and FCPN  for 24 h. JC-1 staining working solution was added and incubated at 37 °C for 30 min. The cells were immediately observed under a fluorescence microscope .2.7.6.The cells were cultured in 24-well plates for 24 h and then treated with different pharmaceutical products  for 24 h. The cells were fixed with 4% paraformaldehyde solution, permeabilized with Triton X-100, and blocked with goat serum for 1–2 h in turn. Then, we added cleaved caspase-3 to the cells overnight at 4 °C. The cells were incubated for 2 h with the secondary antibody, and the nuclei were stained with DAPI. Fluorescence microscopy was used to observe cell immunofluorescence.2.7.7.SMMC-7721 cells were cultured in a 15 mL cell culture flask. The cells were treated with different preparations  for 48 h. The adherent cells were scraped off on ice. The collected cells were treated with 300 µL of lysis buffer. After centrifugation, the total protein concentration of the collected supernatant was determined by BCA analysis.Then, the protein samples were prepared and added to the sample well for polyacrylamide gel electrophoresis. Next, the polyacrylamide gel was moved onto the nitrocellulose membrane and blocked with BSA for blocking solution. After incubation with specific antibodies  and the secondary antibody at 4 °C, the film was processed with ECL developer. Quantity One 1-D Analysis Software  was used to analyze the expression.2.8.2.8.1.6) were injected into the vicinity of the right front leg of nude mice. A week later, there were noticeable tumor masses, which indicated that the tumor model was successfully established.Balb/c nude mice  were purchased from Liaoning Changsheng Biotechnology Co., Ltd.  and fed in an SPF-free environment. The experiment was carried out under the Animal Management Regulations of Jinzhou Medical University (2020). SMMC-7721 cells . Normal saline, PTX, PN, CPN, and FCPN  were injected by the tail vein. Nude mice were injected once every three days for a total of seven times. Before each dose, we measured the body weight and the longest and shortest diameters of the tumor. The volume of the tumors was obtained according to the following formula:When the tumor volume reached 300 mmWc is the weight of the tumor in the saline group and Wt is the average weight of the tumor in each drug treatment group.The tumor inhibition rate was obtained according to the following formula:in vivo. The sections were observed with a fluorescence microscope .The mice were sacrificed after the last dose, and we removed the tumor tissue and the main organs . After that, the tumors and the main organs were fixed with paraformaldehyde and embedded in paraffin. The samples were sliced and were stained using ki67 for immunohistochemistry study. Hematoxylin and eosin (H&E) also were used to stain the sections to assess necrosis of tumor cells and safety of FCPN 2.8.3.3, the tumors were randomly divided into four groups (n = 9). PTX, PN, CPN, and FCPN  were injected into mice by the tail vein. Then, the tissues  of three mice were collected at each time point . The heart, liver, spleen, lung, kidney, and tumor tissues were washed with saline, weighed, and then homogenized with DMSO. Two hundred microliters of tissue samples were transferred to 1.5 mL microcentrifuge tubes, and 20 µL of butyl paraben was added to each tube as an internal standard. Each mixture was vortexed for 5 min with 1 mL of methyl tert-butyl ether. After centrifugation at 12,000 rpm for 10 min, the organic layer was transferred to another test tube and evaporated in vacuum. Thirty microliters of methanol were added to each tube to redissolve the samples. PTX content was determined by HPLC.When the tumor volume reached 300 mm2.8.4.In vivo fluorescence signals of different tissues  were observed using an in vivo imaging system  at excitation wavelengths of 518 nm and 494 nm.To further verify the targeting ability of FCPN, FITC-labeled PN, CPN, and FCPN were injected into the tail veins of tumor-bearing nude mice. After 3 h, the mice were sacrificed by cervical dislocation. 2.8.5.Male SD rats were purchased (180–220 g) from the Department of Laboratory Animal Science of Jinzhou Medical University for experiments. Twelve rats were randomly divided into three groups and fasted for 12 h. PTX, PN, and FCPN  were injected into the rat body by tail vein. After administration, 1.5 mL of blood was taken from the fundus venous plexus at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 h. Whole blood was placed in a sodium heparin tube and centrifuged at 12,000 r/min for 5 min to separate the plasma. Butyl p-hydroxybenzoate was used as an internal standard, and methyl tert-butyl ether was used to extract PTX. The plasma drug concentration was determined by HPLC.2.9.p<.05 was considered statistically significant.All data results were analyzed using GraphPad Prism  and plotted using the mean ± SD. The data were analyzed to determine the difference in means between groups. 3.3.1.in vivo, allowing the PN coated in SMMC-7721 CMs to be more targeted to tumor tissue. It is well known that FRs are highly expressed in SMMC-7721 cells. Folic acid-functionalized CPN (FCPN) should further improve the active targeting of CPN and improve the antitumor effect of PTX. The prepared nanocrystals under a low shear state were characterized by SEM. The images showed a rod-like form with a diameter of approximately 200 nm and a length of approximately 1000 nm, as observed in Figure S1 proved that the freeze-drying process did not affect the particle size and of CPN and FCPN. The seven days stability of CPN and FCPN in Figure S2 also indicated the particle size and electric potentials did not change significantly, which illustrated the preparation had good stability. The SDS-PAGE experiment in t1/2) of FCPN, PN, and PTX were 6.07 ± 0.85 h, 2.48 ± 0.31 h, and 1.27 ± 0.33 h, respectively. It is obvious that FCPN prolonged the action time of the drug, which effectively avoided being captured by the reticuloendothelial system. The AUCs(0–∞) of FCPN, PN, and PTX were 166.23 ± 23.81, 34.64 ± 0.89, and 15.06 ± 2.31, respectively. The AUC(0–∞) of FCPN was 11.04 times and 4.80 times that of free PTX and PN, respectively, which proved that FCPN effectively enhanced the bioavailability of PTX.The synthetic schematic diagram of FCPN is drawn in 3.2.Confocal laser scanning microscopy and flow cytometry were used to verify the targeting of FCPN at the cellular level. In the cellular uptake study, the CLSM images are shown in 3.3.in vitro. To more fully verify the proapoptotic effect, the immunofluorescence and western blot analyses of the expression of apoptotic proteins were performed. The cleaved caspase-3 green fluorescence intensity of the FCPN group in The toxicity of PTX, PN, CPN, and FCPN to SMMC-7721 liver cancer cells was evaluated by the MTT test. The cytotoxicity of different treatment groups is shown in 3.4.in vivo safety of FCPN are shown in 3, which was significantly reduced compared with the 584.30 ± 29.04 mm3, 876.42 ± 83.72 mm3, and 1129.21 ± 59.27 mm3 of the CPN, PN, and PTX groups in in vivo. In in vivo.The antitumor effect and 4.In vivo animal experiments further proved that FCPN significantly improved bioavailability and increased drug accumulation in tumor tissues, which effectively inhibited tumor growth. This indicated that FA-CM was a promising active targeting material for the functionalization of particles.In this study, a bionic nanodrug system (FCPN) for targeted therapy of liver cancer was successfully prepared by CM coating technology. It had high drug loading and exhibited sustained release. Cell experiments showed that FCPN increased cell uptake, promoted cell apoptosis, and demonstrated that it had a good targeting effect on SMMC-7721 cells. Click here for additional data file.


Preservation of IPLs during left upper lobectomy may be beneficial for LLL expansion and induces less movement and positional change in the left main bronchus.The benefits of dissecting inferior pulmonary ligament (IPL) during upper lobectomy using video-assisted thoracoscopic surgery (VATS) for early-stage lung cancer remains controversial. This study evaluates the effect of IPL dissection by comparing the lung volume, bronchial angle, and bronchial tortuosity of the left lower lobe (LLL) during VATS upper lobectomy. Medical records of all patients who underwent VATS left upper lobectomy for early-stage lung cancer were evaluated. Patients were divided into group P (preservation) and group D (dissection). Pre- and post-surgery lung volumes, bronchial angles , and bronchial tortuosity (curvature index of the left main bronchus) were measured using computed tomography images for comparison. Forty patients were included in each group. Patient characteristics such as age, gender, body mass index, and smoking status, and preoperative lung volume, bronchial angles, and tortuosity were not significantly different between the two groups, and there was no statistically significant difference in the axial and vertical angulations; however, the change in pre- and postoperative bronchial tortuosity (0.03 ± 0.03 vs. 0.06 ± 0.03) and lung volume (−558.1 ± 410.0 mL vs. −736.3 ± 382.7 mL) showed a significant difference ( Several thoracic surgeons believe that performing inferior pulmonary ligament (IPL) dissection during upper lobectomy leads to positive postoperative outcomes with respect to lung expansion, reduction in dead space and its accompanying pleural effusion, reduced chest tube indwelling time, and prevention of atelectasis ,2,3,4,5.In addition, some surgeons believe that IPL preservation may be beneficial in terms of postoperative pleural fluid drainage by providing normal lymphatic drainage and channel ,7,8,9. THowever, it is important to consider that IPL preservation may lead to insufficient lung expansion, causing pleural effusion and atelectasis ,10,11. DTherefore, this study attempted to analyze the impact of IPL dissection and/or preservation on the expansion, bronchial angle change, and bronchial tortuosity change of the left lower lobe after left upper lobectomy.This study was approved by the ethics committees/institution review boards of both institutions . The requirement for written informed consent was waived due to the retrospective nature of this study. All methods were performed in accordance with the relevant guidelines and regulationsn = 24), emphysematous destruction or interstitial lung disease (n = 6), cancer-related pneumonia on preoperative chest computed tomography (CT) (n = 8), and inadequate/unsuitable CT images (n = 26)  . All incOne surgeon from each institution performed surgeries for this study. Depending on the preference of each surgeon, IPL preservation was performed at one institution and IPL dissection in another. However, at both institutions, both surgeons adhered to the same oncological principle, performing complete anatomical lobectomy and mediastinal lymph node dissection. In all patients, double-lumen endotracheal intubation was used for administering general anesthesia, and single-lung ventilation was used for performing the surgery. Three incisions were made, and fissure-based technique was used for performing VATS lobectomy. Moreover, endoscopic staplers were used for the division of hilar structures, including the pulmonary vessels and bronchus. All patients were extubated after the surgery in the operating room. The chest tube was removed if there was no air leakage and when the drain volume was in the range between 200 and 250 mL/day based on the patient’s weight.Chest CT scans were performed by various multi-detector CT machines including: Somatom Sensation 16, Somatom Sensation 64 and Somatom Definition AS+ . Participants were scanned from the lung apex to the adrenal glands in supine position during breath-holding at the end of inspiration. Volumetric CT scans were obtained from all patients with the following parameters: 120 kVp, 100–150 mAs, pitch of 1, 1.0 mm section thickness, and contiguous or overlapping section interval.™ Ver.4.4.6; TeraRecon, Foster City, CA, USA) according to the previously reported threshold-based, three-dimensional auto-segmentation technique : the curve length of the left main bronchus divided by the straight length of the left main bronchus. With commercially available reconstruction software, the full lengths of the left main bronchus were semi-automatically traced using the curved multiplanar technique; the CI was then calculated as the midline curve length of the left the main bronchus divided by the straight length using an electronic caliper after precise manual correction .t-test: height, weight, and body mass index; the preoperative right lung volume, postoperative right lung volume, and pre- and post-operative right lung volume difference; the preoperative left lung volume, post-operative left lung volume, and pre- and post-operative left lung volume difference; the preoperative horizontal angle of the left main bronchus, post-operative horizontal angle of the left main bronchus, and pre- and post-operative horizontal angle difference; the preoperative vertical angle of the left main bronchus, post-operative vertical angle of the left main bronchus, and pre- and post-operative vertical angle difference; the preoperative CI of the left main bronchus, post-operative CI of the left main bronchus, and pre- and post-operative CI difference. The interobserver reproducibility for measurements  was assessed via intraclass correlation coefficients. A p-value of < 0.05 was considered statistically significant. All statistical analyses were performed with commercially available software SPSS 23 .Continuous variables were expressed as mean  ±  standard deviation and categorical variables as frequency or percentage. Normality of the variables was confirmed using the Shapiro–Wilk test. The following variables were compared between the two groups using the independent In this retrospective study, 80 patients were included; they were divided into two groups: 40 patients in the IPL preservation group (group P) and the remaining 40 patients in the IPL dissection group (group D). The average age in group P and group D was 63.6 ± 11.1 years and 64.2 ± 9.7 years, respectively, with no statistically significant difference. Moreover, there was no statistically significant difference between the two groups with respect to the number of men (%), height, weight, smoking status (%), chronic obstructive pulmonary disease (%), preoperative forced expiratory volume in 1 s (FEV1), preoperative forced vitality capacity (FVC), preoperative carbon monoxide lung diffusion capacity (DLCO), tumor size, and pathologic cancer stage. Regarding lung fissure during the operation, seven patients (17.5%) in group P and eight patients (20.0%) in group D had complete fissure, with no statistically significant difference. Chest tube indwelling time was 4.5 ± 3.8 days in group P and 4.8 ± 3.4 days in group D, with no statistically significant difference .p = 0.622). The postoperative right lung volume was 2619.1 ± 654.0 mL in group P and 2622.7 ± 505.8 mL in group D (p = 0.978). The difference between the preoperative and postoperative right lung volumes was 209.3 ± 454.2 mL in group P and 147.6 ± 344.1 mL in group D; there was no statistically significant difference between the two groups (p = 0.496) (The preoperative right lung volume was 2409.8 ± 665.0 mL in group P and 2475.0 ± 500.0 mL in group D (= 0.496) .p = 0.458). The postoperative left lung volume was 1412.8 ± 420.6 mL in group P and 1325.6 ± 383.3 mL in group D (p = 0.335). The difference between preoperative and postoperative left lung volume was −558.1 ± 410.0 mL (decrease of 28.3%) in group P and −736.3 ± 382.7 mL (decrease of 35.7%) in group D, which was statistically significant (p = 0.043) (The preoperative left lung volume was 1970.9 ± 574.3 mL in group P and 2061.9 ± 515.3 mL in group D (= 0.043) .p = 0.497). The postoperative angle 1 was 10.6 ± 5.7° in group P and 8.5 ± 8.2° in group D (p = 0.198). The difference between the pre- and postoperative angle 1 was −2.9 ± 5.5° in group P and −5.7 ± 7.7° in group D. Although the difference was lower in group P, the difference was not statistically significant (p = 0.065). The preoperative angle 2  was 27.9 ± 5.5° in group P and 28.5 ± 6.9° in group D (p = 0.689). The postoperative angle 2 was 12.2 ± 9.0° in group P and 9.3 ± 9.8° in group D (p = 0.170). The difference between the pre- and postoperative angle 2 was −15.7 ± 8.4° in group P and −19.2 ± 11.0° in group D, with no statistically significant difference (p = 0.114)  was 13.5 ± 5.1° in group P and 14.3 ± 4.6° in group D (= 0.114) .p = 0.142). The postoperative left main bronchus CI was 1.08 ± 0.03 in group P and 1.11 ± 0.05 in group D (p = 0.003). The difference between pre- and postoperative left main bronchus CIs was 0.03 ± 0.03 in group P and 0.06 ± 0.03 in group D, with statistically significant difference (p < 0.001) (The preoperative left main bronchus curvature index (CI) was 1.05 ± 0.02 in group P and 1.04 ± 0.02 in group D (< 0.001) .According to our findings, the degree of left lung volume loss was less in group P than in group D, i.e., the expansion of the remaining lung was greater in the IPL preservation group. Although there was no statistically significant change in the postoperative bronchial angle, there was less variation in the postoperative bronchial angle in group P. Moreover, for the postoperative bronchial tortuosity, there was statistically significantly less bending in group P than in group D. The IPL maintains the structural integrity of the bronchus by minimizing the risk of torsion and kicking by fixating the residual (left lower) lung. We believe that this may play a role in residual lung expansion. According to several previous studies, the benefits of IPL dissection during upper lobectomy may be to enhance the range of motion of the inferior lobe and to improve the re-expansion of the inferior lobe, ultimately reducing postoperative complications, including atelectasis and pleural effusion, by improving the residual lung capacity of the superior lobe ,2,4,5. HIn our study, a quantitative comparison was attempted between IPL preservation and IPL dissection during upper lobectomy; this was achieved by developing a bronchial tortuosity index. This newly developed index was devised to overcome the existing ambiguous method of bronchial angle measurement and unclear absolute reference when determining the angulation change. The bronchial tortuosity index appears to provide greater objectivity in measuring the bronchial angle and angulation change. Using this method, the physiological anatomy was better preserved (less change to the shape of the bronchus) in the IPL preservation group than in the IPL dissection group. We believe that IPL preservation promotes a positive impact on postoperative outcome, including reduced cough and improved pulmonary function. We also believe that IPL preservation allows for the maintenance of bronchial structure integrity, as indicated by the bronchial tortuosity index, and thereby positively influences postoperative lung volume. In our study, patients who underwent video-assisted thoracoscopic surgery (VATS) lobectomy were only included and those who underwent open thoracotomy lobectomy were excluded because, when compared with open thoracotomy, VATS tends to result in less pain and minimal alteration to the chest wall shape, thus having a relatively less negative impact on pulmonary function than open thoracotomy ,14,15. MAdvocates of IPL dissection when performing upper lobectomy consider the oncological aspects of IPL preservation as a disadvantage, i.e., they assert that without IPL dissection, there cannot be a clear dissection of the inferior mediastinal nodes, specifically the paraesophageal lymph node or the pulmonary ligament lymph node of the mediastinal lymph node in the lower zone ,7,9. IPLThere are several limitations to this study. First, this was a retrospective study with a small sample size. Second, two surgeons from two institutions—one surgeon per institution—performed surgeries, which may have impacted the results owing to factors based on the surgeon. However, the surgical techniques utilized are very similar between the two institutions, because surgeons from both of these institutions were trained at the same institution and there are highly shared activities. Third, the postoperative pulmonary function test was not performed in all patients on a regular basis; even if it was performed, we believe that it would have been difficult to compare the results between the two institutions. Moreover, postoperative evaluation and comparison of clinical symptoms were not performed because evaluating clinical symptoms via medical records review would have been inaccurate. Therefore, the results of this study may not provide an exact clinical implication. Fourth, we only included patients undergoing left upper lobectomy. This is because the right side of the lung is more complex than the left side, which would make IPL factors, including lung volume, bronchial angulation, and tortuosity, insufficient for analysis. However, our study may be noteworthy in that it attempted to present a newly developed parameter—bronchial tortuosity index—for evaluating the benefits of IPL preservation.IPL preservation during left upper lobectomy may be beneficial for left lower lobe expansion and induces less movement and positional change to the left main bronchus . NonetheInferior pulmonary ligament preservation during left upper lobectomy may be beneficial for left lower lobe expansion with less movement and positional change of the left main bronchus.


Ulva ohnoi, some of which may be used for food or natural products but could disturbing coastal ecosystems due to large scale green-tide, to check values of U. ohnoi oil through experimental results. U. ohnoi oil was extracted from bulk of Ulva biomass to confirm its antioxidant and antibacterial activity, and the efficacy of U. ohnoi oil in the state of inflammation was confirmed through animal experiments. To confirm the anti-inflammatory effect, a mouse model induced with DSS was used. As a result of measuring NO using plasma after induction of inflammation, the amount of NO produced in the U. ohnoi oil group was decreased compared to the control group. Expression of inflammatory cytokines TNF-α, IL-6, and IL-1β was decreased compared to the control group. As a result of observing H&E staining, lower crypt loss and inflammatory cell infiltration were found in the U. ohnoi oil group compared to the control group. Consequently, U. ohnoi oil appears to have great anti-inflammatory properties.This study was conducted to examine the physiological activity of  ROS is also produced by external stimulation such as ozone and air pollution2. Oxidative stress caused by free radicals causes non-selective and irreversible modifications to the DNA, proteins, sugars, and lipids in living bodies, leading to aging and disease such as cancer, brain disease, heart disease, autoimmune disease, and arteriosclerosis4. Antioxidants are needed for protection against side effects because of the relative instability of free radicals and the possibility of damage to cells and tissues5. Therefore, many natural antioxidants, such as tocopherol and vitamin C, and synthetic antioxidants, such as dibutyl-1-hydroxytoluene (BHT) and butylhydroxyanisole (BHA), are used in response to such problems7. Synthetic antioxidants are effective and economical, but high doses are known to reduce phosphatide, blood enzymes, catalase, and peroxide10.All living organisms produce reactive oxygen species (ROS), such as superoxide radical anion (O11. Interleukin-6 (IL-6) is a multifunctional cytokine produced by B cells, T cells, and fibroblasts and is involved in immune and inflammatory responses12. Interleukin-1β (IL-1β) is a potent inflammatory cytokine that is crucial for host-defense responses to infection and injury and regulates various physiological responses such as cell recruitment, appetite, sleep, and body temperature13. In macrophages, various stimuli such as cytokines, TNF-α, and lipopolysaccharide (LPS) activate NF-κB, a transcription factor of inflammatory responses, resulting in the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) to produce nitric oxide (NO) and cause inflammation15. NO is synthesized from L-arginine by the activity of iNOS and performs various biological functions16, acting as a physiological regulator and signaling molecule. However, excessive production may contribute to tissue damage from stroke and ulcerative colitis17.Tumor necrosis factor-α (TNF-α) is an inflammatory cytokine that was first discovered as an endotoxin-induced serum factor that causes necrosis of tumors in 1975 and is known as a derivative of inflammatory responses to various infections or tumors18, extracting EPA and DHA from fish involves problems related to marine pollution19.Omega-3 fatty acids are polyunsaturated fatty acids, namely, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), and are known to be effective against cardiovascular disease, dementia prevention, insulin resistance, and inflammation. As omega-3 fatty acids cannot be produced from the body and must be obtained from dietary sourcesEuphausia superba) is a zooplankton found in the Antarctic water that feeds directly on minute phytoplankton. This species is receiving the spotlight as a food resource in the future due to its excellent nutritional value20, as it contains large amounts of unsaturated fatty acids and antioxidants such as EPA and DHA21. However, krill contains a large amount of fluoride, which is harmful to the human body22. There are also reports that global warming is accelerating due to the decline in krill populations. Therefore, it is urgent to find a replacement because overfishing of krill threatens the survival of other marine life such as whales, penguins and seals23.The Antarctic krill  widely distributed around the North America, South Korea, Japan, and Mediterranean sea, usually attached to hard bottom on the rocky shore. The rise of sea surface temperature and nutritive salts due to global warming causes mass reproduction of U. ohnoi30, causing large scare green-tide in some contaminated costal ecosystem, such as Japan31, and South Korea30. U. ohnoi has a fast growth rate, so no problem of material depletion is raised, and it is thought that it is valuable as a new marine natural product while solving environmental problems. About 700 tons of U. ohnoi were collected in blooming area in Japan, and some algal biomass were used as food ingredients31. In addition, morphologically similar green alga, U. lactuca has been used for food in Europe for a long time, and multiple studies have been performed on its various food and physiological activities. It contains a large amount of polysaccharides containing sulfuric acid, and this acid polysaccharide has been reported to have anti-viral, immunity-boosting, and anti-cancer effects32. Some studies in South Korea reported anti-cancer effects and immune activities by extracting glycoproteins from single-cell layered green algal, Monostroma nitidum32.U. ohnoi using an organic solvent extraction method to find out the utility of U. ohnoi oil for its physiological activity. There have been various efficacy studies using U. ohnoi, but studies using U. ohnoi oil are insufficient. Conventionally, oily ingredients such as krill oil and omega-3 are exerting the function of maintaining human health. We would like to consider the aspect of recycling discarded U. ohnoi through the use as a new material by proving the functionality of U. ohnoi, the subject of our research.The purpose of this study is to extract oil from In this experiment, DPPH  , folin&ciocalteu’s reagent , bacto agar , bacto tryptic soy broth , NO (nitric oxide) kit , ELISA (enzyme-linked immunosorbent assay) kit , TRIzol reagent , chloroform , isopropyl alcohol , ethanol , DEPC (diethyl pyrocarbonate) water , agarose , RT-PCR premix kit , Hematoxylin , Eosin Y , centrifuge , enzyme-linked immunosorbent assay (ELISA) reader , fluoro box , and Permount  were used.U. ohnoi used in this study was collected from the north-east part of Jeju Island  in 2019 . We have transported small amount of Ulva biomass collected from the blooming area to the laboratory, thus sample collection was not threatened with endangered species and have not destroyed natural seaweed habitats. In addition, sampling was carried out after permission of National Research Foundation (NRF) of South Korea, thus government is permitting the collection of Ulva because it is working on a research project to manage Ulva biomass. Sample were washed with tap water and distilled water several times, and dried in an oven at 60℃. Organic solvent extraction was performed to extract U. ohnoi oil. The weight of the dried sample in the beaker was 29.3 g, 33.5 g, and 30.7 g of A, B, and C, respectively, and 200 ml of hexane was added per 10 g. Thereafter, extraction was performed at room temperature for 48 h and the organic solvent was removed using speed vacuum. The yields are 0.19%, 0.23%, and 0.21% for A, B, and C, respectively. All the experiments involving plants adhered to relevant ethical guidelines on plant usage. Krill oil is manufactured by Neptune , and omega-3 is manufactured by Now Foods . The composition of krill oil, omega-3, and U. ohnoi oil used in the experiment is shown in Table The 33. DPPH was dissolved in ethanol at a final concentration of about 0.2 mM. U. ohnoi oil (100 µl) by concentration was added to 0.1 ml of ethanol solution of DPPH. After blocking the light and reacting for 20 min, the absorbance was measured at 517 nm. DPPH scavenging effect was calculated from [1—(Abssample – Absblank)/Abscontrol] × 100, where Abssample is the absorbance of the sample, and Absblank is the absorbance of the color control, and Abscontrol is the absorbance of the negative control. As a control group, ethanol was added instead of the sample. L-ascorbic acid, which is known to have high antioxidant capacity, was used as a control to compare antioxidant activity.DPPH radical scavenging activity was used to eliminate absorption characteristics when the compound was stabilized by receiving electronic or hydrogen radicals and measured in accordance with using 1,1-diphenyl-2-picryhydrazyl (DPPH)34. 200 µl of the U. ohnoi oil was added to 1 ml of 0.2 N folin-ciocalteu reagent and 800 µl of 7.5% Na2CO3 was added. After blocking the light and reacting at room temperature for 2 h, the absorbance was measured at 620 nm using a spectrophotometer. Gallic acid was used as a standard reagent, and polyphenol content was calculated using a standard curve. The TPA was expressed as gallic acid equivalents (GAE) in mg/1 g material.The total polyphenol content assay (TPA) was determined using the Folin-Denis reagentU. ohnoi oil was performed by URDA  method35. The sample was prepared at a concentration of 10 mg/ml using 0.01% acetic acid (HAc). The strains used were gram-positive bacteria Bacillus subtilis KCTC1021, gram-negative bacteria Escherichia coli D31, and fungus Candidia albicans KCTC76965. Each strain was inoculated with 1 platinum loop in tryptic soy broth (TSB) medium and pre-culture was performed at 37℃ for 24 h. The activated bacteria measured absorbance at 630 nm to make the O.D value of the bacteria 0.01 (≒ 0.5 McF). 0.03% TSB, sabouraud’s dextrose broth (SDB), 1% agarose, and 10 mM phosphate buffer (pH 6.5) were added to the underlay gel at 9.5 ml concentrations and then poured into the plate to solidify. The plate was pierced with a diameter of 2.5 mm and 5 µl of sample was injected into each well. All samples were tested with 0.01% acetic acid 5 µl, which was not affected by solvent. After the first culture at 37℃ for 3 h, 10 ml overlay gel containing 6% TSB, SDB, 1% agarose, and 20 mM phosphate buffer (pH 6.5) was poured into the cell, and then solidified at room temperature, and then the clear zone was confirmed after the second culture at 37℃ for 24 h.The antibacterial activity measurement using U. ohnoi oil, various enzymes were treated to determine whether there was any change in antibacterial activity. The enzymes used in the experiment are trypsin, chymotrypsin, lipase, α-amylase, pronase, and proteinase. 5 µl of U. ohnoi oil at a concentration 10 mg/ml was mixed with 1 µl of each enzyme and incubated for 3 h at 37 ℃, followed by secondary culture at 37 ℃ for 24 h. The cultured extracts were tested for URDA against B. subtilis, E. coli, and C. albicans.In order to confirm the properties of the antibacterial activity in the U. ohnoi oil (UO). Krill oil, ω-3, and U. ohnoi oil are prepared using distilled water at concentrations of 100 mg/kg, 100 mg/kg, and 25 mg/kg and administered orally at 100 µl each for 10 days  dissolved in distilled water for 7 days after 4 days of sample administration. The body weight of the experimental animals was measured at the same time every day and is shown in Figure Male ICR mouse aged 4 weeks were obtained from Orient bio . They were housed in a controlled environment  with a 12 h light/dark cycle. The animals allowed to free access to food and filtered water. They were quarantined and acclimatized for 1 week before use. The animals were kept in groups of 6 per cage. After 1 week habituation period, the experimental animals were classified into five groups as follows: one is normal, another is control and the others is DSS + krill oil (KO), DSS + omega-3 (ω-3), and DSS + ys Table . U. ohno36. Blood was collected to obtain plasma and centrifuged at 7000 rpm for 10 min at 4 ℃. NO concentration was then determined by the addition of 50 µl each Griess reagents 1 and 2. After incubation for 10 min at room temperature, the absorbance was read at 540 nm.The amount of NO production was measured using plasma of 6 mice per group, and Griess reagent was usedIn this study, colon tissue from 6 mice per group was removed and used, 1 ml of 1 × phosphate-buffered saline (PBS) was added, crushed, and centrifuged at 7000 rpm for 10 min, and then the supernatant was taken. The amount of inflammatory cytokine TNF-α, IL-6, and IL-1β was measured using an enzyme-linked immunosorbent assay (ELISA) kit. After adding 100 µl of antigen to the antibody-coated plate, incubated at 37℃ for 2 h, and washed 3 times using a washing buffer. After the addition of 100 µl of the secondary antibody, it was incubated at 37℃ for 1 h and washed 3 times. After adding 100 µl of the TMB solution, incubate at 37℃ for 30 min, and reacted at room temperature for 10 min after adding 100 µl of stop solution, absorbance was measured at 450 nm.RNA used in the experiment was extracted from the colon tissue of the mouse using TRIzol reagent, and it was dissolved in 50 µl diethyl pyrocarbonate (DEPC) water. Reverse transcription-polymerase chain reaction (RT-PCR) is performed using and RT-PCR premix kit, and 300 ng of total RNA, 20 pmol of each of forward and reverse primers are added to use a total of 20 µl of the reaction solution. The cDNA was synthesized by reacting at 50℃ for 30 min and 94℃ for 5 min, followed by 35 cycles of denaturation at 94℃ for 40 s, annealing at 58℃ for 40 s, and extension at 72℃ for 90 s. The PCR product was confirmed by electrophoresis on 1.2% agarose gel, and GAPDH was used as control. The primer sequence used in the experiment is as follows Table 37. H&E stains are still essential for recognizing various tissue types and the morphologic changes that form the basis of contemporary cancer diagnosis38. Each specimen was sectioned into a thickness of 7 µm and histochemical staining was performed using hematoxylin and eosin. For deparaffinization, xylene was added and repeated three times for 5 min, and xylene was removed in the order of 100%, 95%, and 70% ethanol. For nuclear staining, it was stained with hematoxylin solution for 5 min, and then washed with flowing water. Hematoxylin was removed using 1% HCl solution and 1% ammonia solution, followed by immersion in eosin solution for 2 min to stain the cytoplasm. Then, it was dehydrated in the order of 70%, 95%, and 100% ethanol. To remove alcohol, it was put in xylene and repeated three times for 3 min, and then the cover glass was and sealed.Hematoxylin & eosin (H&E) staining is primarily used in histology to visualize the structure of tissuesU. ohnoi oil was measured using DPPH and Folin-Denis reagent. The radical scavenging ability was measured by concentrations of 0.1, 0.3, 0.5, 0.7, and 1 mg/ml, and ascorbic acid was used as a positive control  Fig. B. The toU. ohnoi oil was measured against B. subtilis, E. coli, and C. albicans strains using the URDA method. In general, B. subtilis and E. coli showed antibacterial activity, but not against the fungus C. albicans. The size of the clear zone was 10.36 mm in B and 7.35 mm in A in the B. subtilis strain, indicating that the clear zone size was 1.41 time higher than that of B U. ohnoi oil. As a result of measuring the activity against E. coli strains, all three types of U. ohnoi oil showed high antibacterial activity of 10 mm or more  is known as a mediator of the inflammatory response secreted when cells stimulated by inflammatory substances such as LPS or TNF-α induce inflammationice Fig. A. The NOThe amount of TNF-α, IL-6, and IL-1β production were measured by ELISA method. All three cytokines were increased in the control group. In the case of TNF-α and IL-6, the amount of production of 43.9 ± 20.2 pg/ml and 17.41 ± 2.68 pg/ml, respectively, in the UO group and significantly decreased compared to the control group  staining, and observed using an optical microscope. The normal group represents typical colon tissue, but the control group lost crypt and destroyed the nucleus Fig. . On the U. ohnoi, a species of green algae known for its immune activity, based on the results of prior research using U. ohnoi extracts to investigate its effects on physiological activity. This study conducted experiments to evaluate the antioxidant, antibacterial activities of the U. ohnoi oil, and measured its anti-inflammatory activity by using a DSS-induced inflammatory bowel disease mouse model. As a result of measuring DPPH radical scavenging activity to evaluate the antioxidant activity of U. ohnoi oil, the radical scavenging activity of A and B U. ohnoi oil was 77.85 ± 5.39% and 86.71 ± 8.36%, respectively, at 0.7 mg/ml, showing the highest activity. In particular, B U. ohnoi oil showed similar activity to the control substance, ascorbic acid. When the total polyphenol was measured, it was confirmed that among the three types of U. ohnoi oil, U. ohnoi oil of B contained the highest polyphenol as 107.45 ± 0 mg GAE/g. This confirms that U. ohnoi oil exhibits a function that can replace krill oil, an animal oil. In addition, the efficacy of antioxidants was repeatedly confirmed through additional experiments. Since the existing oils with excellent antioxidant efficacy are being used in abundance in the food and cosmetics industry, it is possible to industrially use U. ohnoi oil, which has antioxidant properties.This study recognized the superiority of U. ohnoi oil using URDA method, B. subtilis and E. coli strains showed strong antibacterial activity, but no antibacterial activity was observed for C. albicans. In the B. subtilis strain, the U. ohnoi oil from B showed higher activity than the U. ohnoi oils from other regions, and there was no significant difference in the C. albicans strain. The change in antibacterial activity was measured by treating six enzymes to investigate the properties of the antibacterial active ingredients of U. ohnoi oil, but there was no significant change compared with before treating enzymes. Ulva ohnoi oil showed some antibacterial activity, and this is a result that can prove the wide utility of U. ohnoi oil. In addition, it can be suggested as one of the many causes of reduced inflammation in the research results related to colitis.Meanwhile, as a result of evaluating the antibacterial activity of U. ohnoi oil. Compared with the normal group, weight loss and diarrhea were observed in all groups administered with DSS. In terms of proof, the U. ohnoi oil administered group and the normal group produced a similar amount of NO (an inflammation mediator) compared to the control group, indicating the effect of U. ohnoi oil to inhibit NO production. The inhibition of production of TNF-α IL-6, and IL-1β of U. ohnoi oil was measured through ELISA. The production of cytokines increased significantly the control group compared to the normal group, and the production was inhibited in the U. ohnoi group. The exact mechanism for the effectiveness of Ulva ohnoi oil to inhibit inflammatory cytokines will need to be investigated through future studies.After inducing inflammatory bowel disease with DSS, this study examined the anti-inflammatory effect by administering krill oil, ω-3, and U. ohnoi oil group showed a significant decrease compared to the control group. In case of the mRNA expression of inflammatory factors  in the colon tissue, the U. ohnoi oil group showed similar expression levels to the krill oil administered group. These results support the results of measuring inflammatory cytokines in colon tissue.In case of TNF-α and IL-6, the U. ohnoi. These results show that U. ohnoi oil has a higher anti-inflammatory effect than krill oil in mice with DSS-induced inflammatory bowel disease, and that it was effective in inhibiting inflammatory expression.As a result of analyzing the inflammatory response of the colon tissue using H&E staining, the group that induced bowel disease with DSS showed crypt loss and inflammatory cell infiltration, but this was mitigated in the mice treated with DSS U. ohnoi oil used in this study, and the one-minute efficacy is more beneficial. The U. ohnoi is being multiplied due to the warming of the climate, and the part that can be used in the industry is extremely weak. As the result of this study, it is expected that if the usefulness of U. ohnoi oil is utilized in industry, there will be a synergistic effect of reducing environmental pollution and utilizing biomass.Krill oil, which has previously been helping health, needs an alternative material due to the decrease in krill shrimp. It is similar to the efficacy of krill oil, which is the *p < 0.05.Data were analyzed using the predictive analytics software (PASW 18 for Windows) and presented as mean ± S.D. A one-way analysis of variance (ANOVA) test was performed to assess the NO production in mouse plasma and the generation of inflammatory cytokines in the colon. The significance level was set at Supplementary Information.