OBJECTIVE：To investigate expression and mechanism of STMN3 in malignancy of esophageal squamous cell carcinoma (ESCC). METHODS：Expression of STMN3 in different tumors was analyzed in the TCGA database and its correlation with clinicopathological parameters was evaluated. CCLE database and Western blot were used to detect expression of STMN3 mRNA and protein levels in ESCC cell lines in vitro. Knockdown efficiency of STMN3 siRNA transfection in the cell lines was detected by RT-qPCR and Western blot，respectively. Effects of STMN3 on malignant phenotype of ESCC cells were detected by cell proliferation，colony formation，and cell apoptosis assays. The changes in the expression of proliferation-related proteins regulated by STMN3 were detected by Western blot. RESULTS：Compared with control group，STMN3 mRNA and protein were highly expressed in ESCC tissues and cell lines. STMN3 expression was associated with prognosis of patients，and those with high STMN3 expression had significantly worse prognosis (P<0.05). Knocking down STMN3 significantly inhibited the proliferation and colony formation of ESCC cells (P<0.01). In addition，the protein levels of EGFR and the phosphorylation levels of EGFR，S6，and SRC were significantly reduced. CONCLUSION：STMN3 was highly expressed in ESCC tissues，and STMN3 promoted the proliferation of ESCC cells by activating the EGFR-related signaling pathway.

OBJECTIVE：To investigate biological function and prognostic value of lysyl hydroxylase 2 (PLOD2) in hepatocellular carcinoma (HCC)，as well as its impact on tumor immune infiltration. METHODS：PLOD2 expression in HCC was analyzed using data from The Cancer Genome Atlas (TCGA) database. Correlations between PLOD2 expression and clinicopathological characteristics and its prognostic value were evaluated by statistical analysis. Signaling pathways associated with PLOD2 expression were identified using gene set enrichment analysis (GSEA). A stable knockdown of PLOD2 in a human hepatocellular carcinoma cell line，Huh7，was achieved using lentivirus-mediated shRNA technology，and the knockdown effect was verified by Western blot. Effects of PLOD2 on biological behavior of Huh7 cells were assessed using CCK-8，Transwell，and scratch assays. Correlations between PLOD2 expression and HCC immune infiltration was investigated using single-sample gene set enrichment analysis (ssGSEA). RESULTS：Bioinformatics analysis revealed that PLOD2 was significantly overexpressed in HCC compared to normal liver tissues (P<0.05 or P<0.01) and was positively correlated with patient age，T stage，pathological stage，tumor status，histological grade，and AFP level (P<0.05). Survival analysis showed that the median survival time of the PLOD2 high-expression group was significantly shorter than that of the low-expression group (P<0.05). The area under the receiver operating characteristic (ROC) curve for PLOD2 in diagnosing HCC was 0.656，indicating that PLOD2 had reference value in the diagnosis and prognosis prediction of HCC. GSEA revealed that several signaling pathways were associated with high PLOD2 expression：cancer pathways，cell cycle，and adhesion junctions. Cell line validation experiments demonstrated that knockdown of PLOD2 significantly inhibited proliferation，invasion，and migration of Huh7 cells compared to control cells (P<0.05). Migration ability of Huh7 cells with PLOD2 knockdown was significantly reversed upon re-expression of PLOD2 (P<0.05). Expression of PLOD2 was positively correlated with Th2 cell infiltration (r=0.372，P<0.01) and negatively correlated with Th17 and pDC infiltration (r=-0.310，P<0.01). CONCLUSION：This study elucidated the critical role of PLOD2 in HCC progression，with high PLOD2 expression associated with poor prognosis in HCC patients. PLOD2 may play a key role in the immunosuppressive microenvironment by regulating immune cell infiltration and function.

OBJECTIVE：To investigate the role of E2F transcription factor 5 (E2F5) in X-ray-induced ferroptosis of human keratinocyte，HaCaT. METHODS：HaCaT cells were irradiated with 0，2.5，5，10 and 20 Gy X-rays. Cell survival rates were detected by the CCK-8 method at 24，48 and 72 h after irradiation. Expression levels of E2F5，ferroptosis marker cyclooxygenase-2 (COX-2) and related protein glutathione peroxidase 4 (GPX4) were detected by Western blot at 24 h. For cells irradiated with 10 Gy X-ray，expression levels of E2F5，COX-2 and related protein GPX4 were detected by Western blot at 24，48 and 72 h，levels of ferrous ion and lipid peroxidation were detected by flow cytometry at 24 and 48 h，GSH (glutathione) levels were measured by GSH kit at 48 and 72 h，and colony formation was measured by colony formation assay at 12 d. Small interfering RNA (siRNA) transfection technique was used to silence E2F5 expression in the 10 Gy irradiated cells. Four groups of cells were set up：unirradiated blank control，irradiated control，negative control，and E2F5 silencing. Survival rate of these cells at 72 h after irradiation was detected by CCK-8，expression levels of COX-2 protein at 24 h was detected by Western blot，levels of ferrous ion 20 h and lipid peroxidation levels at 24 h were detected using flow cytometry，and colony formation ability was detected by colony formation assay at 12 d. RESULTS：The survival rates of the HaCaT cells decreased at 24 h after 2.5-20 Gy X-ray irradiation and 48 and 72 h after 5-20 Gy X-ray irradiation (P<0.05 or P<0.01). The protein expression of E2F5 increased at 24 h after 2.5-10 Gy X-ray irradiation. At 24 h after 2.5-20 Gy X-ray irradiation，COX-2 protein expression increased，GPX4 protein expression decreased (P<0.05 or P<0.01). At 48 and 72 h after 10 Gy X-ray irradiation，the expression of E2F5 and COX-2 protein increased，the expression of GPX4 protein decreased，and the intracellular GSH content decreased. At 24 and 48 h after 10 Gy X-ray irradiation，the levels of ferrous ions and lipid peroxidation increased，and the colony-forming ability decreased at 12 d (P<0.05 or P<0.01). After 10 Gy X-ray irradiation and compared with the negative control group，the survival rates of the E2F5 silenced group increased at 72 h after irradiation，the expression levels of COX-2 protein and lipid peroxidation decreased 24 h after irradiation，the level of ferrous ion decreased 20 h after irradiation，and the colony formation ability increased at 12 days after irradiation (P<0.05 or P<0.01). CONCLUSION：X-ray irradiation promoted ferroptosis by up-regulating E2F5 protein expression in HaCaT cells，thereby reducing cell viability.

OBJECTIVE：To investigate the mechanism of acute lung injury (ALI) induced by phosgene in rats and the protective effect of caffeic acid phenethyl ester (CAPE). METHODS：Thirty SD rats were randomly divided into a negative control group，a positive control group，a phosgene exposure group and a CAPE intervention group. The phosgene concentration was 10 mg/L and the dynamic exposure lasted for 3 min. After 6 h of exposure，pathological changes of lung tissue were determined，lung functions were detected，and lung coefficient was calculated. Total antioxidant capacity (T-AOC)，MDA concentration，MPO concentration，GSH concentration，CAT activity and SOD activity of lung tissues were detected using kits. Protein concentrations of bronchoalveolar lavage fluid (BALF) were detected by BCA method. Relative expression levels of TNF-α and IL-1β mRNA were detected by fluorescence quantitative PCR (qPCR). Concentrations of TNF-α and IL-1β in rat serum were detected by ELISA. Relative expression levels of DJ-1，Nrf2，Keap1 and SOD in rat lung tissue were detected by Western blot. RESULTS：Compared with the control group，the lung structure of rats exposed to 10 mg/L phosgene was significantly altered. The concentrations of MDA and MPO，activities of SOD，and protein concentrations of BALF were higher than those in the control group (P<0.01). The concentrations of GSH，activities of CAT and T-AOC were lower than those in the control group (P<0.01). The concentrations of TNF-α and IL-1β in the serum of the phosgene-exposed group were higher than those in the control group (P<0.01). The mRNA levels of TNF-α and IL-1β in lung tissues were higher than those in the control group (P<0.01). The expression levels of DJ-1 protein in the phosgene-exposed group were lower than that in the control group (P<0.05). Under the the CAPE intervention and compared with the phosgene exposure group lung injury was reduced，the concentrations of MDA and MPO，and the activities of SOD in lung tissue were lower (P<0.01)；the concentrations of GSH，the activities of CAT and T-AOC were higher (P<0.01)；the concentrations of TNF-α and IL-1β in the serum were lower (P<0.01)；the relative expression levels of TNF-α and IL-1β mRNA in lung tissues were lower (P<0.01)；the expression levels of DJ-1 protein in lung tissues were upregulated (P<0.05). CONCLUSION：The asphyxiating agent phosgene exposure caused lung injury in rats，and expression of DJ-1 molecule was decreased. CAPE intervention reduced lung injury，inhibited lung inflammation and oxidative stress，probably via antioxidant and anti-inflammatory mechanisms.

OBJECTIVE：To investigate the effect and molecular mechanism of leflunomide on the anti-inflammatory differentiation of macrophages. METHODS：Mouse mononuclear macrophages (RAW264.7) were treated with 200 mmol/L leflunomide for 0，1，3，6，12 and 24 h. From these cells，mRNA expression levels of anti-inflammatory factors (IL-10，Arg-1，CD206)，pro-inflammatory factors (TNF-α，IL-6) and mitochondrial fusion genes (MFN1 and MFN2) in cell pellet were detected by real-time quantitative PCR (qPCR). ELISA was used to detect the content of anti-inflammatory factors in cell supernatants after flumid treatment for different years. DCFH-DA and Mito-LX staining were used to detect levels of total reactive oxygen species (ROS) and mitochondrial ROS. Mitotracker-Green staining laser confocal was used to determine cell mitochondrial morphology. Kits were used to determine content of malondialdehyde (MDA)，an intracellular oxidation product. The mRNA and protein expressions of peroxisome proliferator-activated receptor-γ (PPAR-γ) genes were determined by qPCR and Western blot，respectively. PPAR-γ inhibitor T0070907 (40 mmol/L) were pretreated for 4 h，and the changes of anti-inflammatory factors were detected by qPCR and Western blot，respectively. RESULTS：Results from the qPCR and ELISA analyses showed that the relative mRNA expression levels of anti-inflammatory factors IL-10，Arg-1 and CD206 treated with leflunomide were significantly higher than those in the control group (P<0.05). Expression of mitochondrial fusion genes MFN1 and MFN2 increased，mitochondrial length increased，and contents of ROS and MDA in cells and mitochondria decreased (P<0.05). Compared with the blank control group，mRNA and protein expression of PPAR-γ in the leflunomide group increased，and its inhibitors T0070907 reversed the increase of anti-inflammatory factors and the decrease of pro-inflammatory factors induced by leflunomide (P<0.05). CONCLUSION：The mitochondrial fusion agonist leflunomide may be useful for anti-inflammatory therapy by reducing ROS production and activating PPAR-γ to regulate the anti-inflammatory differentiation of macrophages.

OBJECTIVE：To explore the impacts of the long non-coding RNA (lncRNA) HOX transcript antisense RNA，myeloid-specific 1 (HOTAIRM1)，on the vitality and apoptosis of MN9D cells which were induced by 1-methyl-4-phenylpyridinium ion ([MPP⁺]). METHODS：MN9D cells were treated with different concentrations (0，0.25，0.5，1，1.25，2.5 mmol/L) of MPP⁺ for various time periods (0，24，48，and 72 h). Subsequently，cell viability of each group was evaluated using the CCK-8 assay. Based on the results，the optimal concentration of MPP⁺ and the most appropriate treatment duration were determined for subsequent experiments. siRNA transfections were conducted. The MN9D cells were then categorized into the control group，MPP⁺ group，si-NC+MPP⁺ group，and si-HOTAIRM1+MPP⁺ group. Cell viability of each group was measured by CCK-8 assay. Expression of lncRNA HOTAIRM1 was detected through real-time fluorescent quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Apoptosis status was analyzed by flow cytometry. RESULTS：In comparison with the control group，lncRNA HOTAIRM1 was highly expressed in MN9D cells induced by MPP⁺ (P<0.05). When treated with 1 mmol/L MPP⁺ for 24 h，the cell viability was significantly decreased，while the expression level of lncRNA HOTAIRM1 was remarkably elevated. Hence，1 mmol/L MPP⁺ for 24 h treatment was selected for subsequent experiments. After the induction and siRNA interference，expression of lncRNA HOTAIRM1 was significantly reduced (P<0.001)，and viability of the cells were significantly enhanced (P<0.01). Knockdown of lncRNA HOTAIRM1 inhibited the apoptosis of the induced cells (P<0.01). CONCLUSION：Interfering with the expression of lncRNA HOTAIRM1 augmented viability of the MN9D cells induced by MPP⁺ and suppressed cell apoptosis. Knockdown of lncRNA HOTAIRM1 exerted a protective effect on the induced cells.

OBJECTIVE：To analyze the incidence and mortality of malignant tumors and their epidemic characteristics in the Hulunbuir in 2019. METHODS：Data from 6 registries were qualified after assessment and accepted as pooled data for final analysis. From the collected data，the incidence rate，mortality rate，age-standardized rate by Chinese standard population of incidence/mortality，age-standardized rate by world standard population of incidence/mortality，the cumulative rate of 0-74 years old and the sequence of incidence/mortality of malignant tumors were calculated. RESULTS：In 2019，the crude incidence rate was 2.888 8×10^-3，ASR China of incidence was 1.687 1×10^-3，ASR world of incidence was 1.675 1×10^-3，and the cumulative incidence rate (0-74 years old) was 18.83%. The top five cancers with high incidence were lung，colorectal，liver，breast and stomach cancers. The mortality rate was 1.915 8×10^-3，the ASR China of mortality was 1.085 7×10^-3，the ASR world of mortality was 1.096 1×10^-3，and the cumulative mortality rate (0-74 years old) was 12.41%. The top five cancers with high mortality were lung，liver，colorectal，stomach and esophageal cancers. CONCLUSION：Lung cancer，colorectal cancer，liver cancer，breast cancer，and stomach cancer pose a great threat to the health of residents in Hulunbuir and are the focus of the city's cancer prevention and treament work.

OBJECTIVE：To identify nucleic acid aptamers that bind specifically to human CD8α molecules. METHODS：A recombinant human CD8α protein with a 6×histidine tag was used as the screening target，and a randomized library of 81 nt was designed for nucleic acid aptamer screening by SELEX technology. The enriched libraries were subjected to high-throughput sequencing and cluster analysis，and the highest abundance sequences were used to verify their specific binding to CD8α protein and CD8⁺ cells by spot hybridization and flow cytometry. The affinity constants (K_ds) of all aptamer candidates were determined using flow cytometry. Finally，normal human peripheral blood lymphocytes were used as a model，and flow cytometry was used to preliminarily evaluate the clinical application of these candidates by prior art to identify CD8⁺ lymphocytes in clinical samples，and to detect whether the binding site of Apt 1 and CD8⁺ lymphocytes was the same as that of CD8 antibody and CD8α protein. RESULTS：The enriched library was sequenced by high-throughput sequencing and 42 803 valid sequence reads were obtained，which were clustered and analyzed by Clustal Omega online software. The top 10 sequences were ranked in order of abundance as the candidate nucleic acid aptamers，which were named as Apt 1-Apt 10，respectively. A highly conserved concordant sequence was present in all candidate sequences (CGTGAGGAGCTTGAAATCC). Among them，Apt 1 had a typical stem-loop structure (ΔG=-12.61 kcal/mol) and bound with high specificity to CD8α protein and CD8⁺ cells. In addition，all 10 sequences bound with high affinity to CD8α protein and CD8⁺ cells，and the affinity constants (K_ds) were at the namor level. Compared with commercial CD8 antibodies and CD8 aptamers disclosed in prior art，the candidate aptamers effectively identified CD8⁺ lymphocytes in human peripheral blood samples，and the binding sites of Apt 1 and CD8⁺ lymphocytes in human peripheral blood samples were different from those of CD8 antibodies. CONCLUSION：Successful screening of nucleic acid aptamers that bound with high affinity and specificity to human CD8α molecules laid the foundation for the subsequent development of clinical diagnostic and therapeutic approaches against CD8 targets.

OBJECTIVE：To evaluate genotoxicity and teratogenicity of Moringa oleifera seeds in rats. METHODS：According to the toxicity test combination recommended by national food safety standards，bacterial reverse-mutation test，mammalian erythrocyte micronucleus test，mouse spermatocyte chromosome aberration test and rat teratogenicity test were carried out. The bacterial reverse-mutation test was set up into 5 dose groups，the highest dose was 5.0 mg/dish，and four doses were used. After inoculation and culture at 37 ℃ for 48 h，the number of reverted colonies in each dish was counted. For the verification test，the group distance was changed to 5 times，and the rest unchanged. Kunming mice were selected for micronucleus test of mammalian erythrocyte and chromosome aberration test of mouse spermatocyte. Three dose groups were set up，with the maximum intragastric volume as the highest dose，and two doses were set up below 0.5 times group distance. The incidence of micronuclei in polychromatic erythrocytes in bone marrow smear and the type and corresponding number of chromosome aberrations in testicular smear were observed. In the teratogenicity test of rats，unmated sexually mature SD rats were selected，and three doses were set up，the high dose of the 90-day test was used as the high dose of the experiment，and two doses were set at the lower interval of the double group. The growth of pregnant rats was observed. After the pregnant rats were killed on the 20th day of gestation，their uteri were weighed，and the number of luteal bodies，absorbent fetuses，stillbirths，live fetuses and implantation numbers were recorded. The weight，length and appearance of the live fetus were examined. The bones of half of the fetal mice in each litter were examined. Half the fetal mice were examined for internal organs. RESULTS：Moringa oleifera seeds showed negative reaction in bacterial retro mutation test，micronucleus test of mammalian erythrocyte and chromosome aberration test of mouse spermatocyte. In rat teratogenicity test，there was no obvious toxic effect on pregnant mice and fetal mice，and no teratogenicity was shown. CONCLUSION：There was no genotoxicity and the NOAEL of Moringa oleifera seeds was 8 g/kg under the experimental conditions.

OBJECTIVE：To establish a mouse model of intestinal microbiota dysbiosis and to observe whether Tianyuan manna and/or yoghurt would regulate colonization of 5 kinds of intestinal microflora.METHODS：The SPF Kunming mice were divided into five groups：blank control group，model (yoghurt) control group，Tianyuan manna (containing yoghurt) 4.2，8.3 and 16.6 g/kg groups，with 10 mice in each group. The model was established by oral administration of 200 mg/kg norfloxacin for 7 consecutive days. After the model was established，Tianyuan manna and/or yoghurt were given by gavage for 16 consecutive days. The number of intestinal bifidobacteria and Lactobacillus (2 beneficial bacteria)，Clostridium perfringens，Enterococcus and Enterobacter (3 harmful bacteria) were counted by pouring dish method，and compared with the blank control group and the model control group. RESULTS：After the norfloxacin treatment，the number of viable intestinal microflora was significantly reduced compared with the blank control group (P<0.01)，indicating that the model was successful. After administration of Tianyuan manna and/or yoghurt，two beneficial bacteria exceeded the number of intestinal viable bacteria in normal mice (P<0.01)，and three harmful bacteria were lower (P<0.05 or P<0.01). Compared with the model control group，the number of harmful bacteria in each dose group of Tianyuan manna was significantly reduced (P<0.05 or P<0.01)，and the number of beneficial bacteria (Bifidobacterium) in the 8.3 g/kg group was slightly reduced (P<0.05). CONCLUSION：In the process balance and imbalance of intestinal flora imbalance，Tianyuan manna demonstrated regulatory effect on the colonization of five kinds of microbial flora.

OBJECTIVE：To evaluate the immunological risk of a porcine-derived absorbable anti-adhesion material. METHODS：SPF-grade BALB/c mice were randomly divided into a blank control，a positive control，and a test material groups，with 20 mice in each group，equally divided by gender. The blank control group underwent surgical operations without the implantation of materials. The positive control group was subcutaneously injected with a mixture of bovine serum albumin and complete Freund's adjuvant in equal volumes，immunized once a week for a total of 4 times. The test material group was implanted subcutaneously with 7 cm² of a porcine-derived absorbable anti-adhesion material on the back. After 4 and 8 weeks of implantation，10 mice from each group were sacrificed (equally divided by gender)，and specimens such as mouse blood，spleen，thymus，and the implanted material and surrounding tissues were collected. Enzyme-linked immunosorbent assay (ELISA) was used to detect the total antibody levels in the serum. Cytokine Bead Assay detection technology was used to detect the levels of serum cytokines IFN-γ，IL-6，and IL-12p70. The CCK8 method was used to detect the proliferation level of lymphocytes. And an electronic balance was used to weigh the spleen and thymus，and HE staining was used to analyze the histopathology of the spleen，thymus，and local implanted material tissues. RESULTS：Compared with the blank control group，the test material group showed no statistically significant differences in total antibody levels，cytokine expression levels，lymphocyte proliferation，immune organ weight and coefficient (P>0.05). After 4 weeks of implantation，the tissue sections of the test material group mice showed some irregularly red-staining around the implants，with a histological score of 27.7 points. After 8 weeks of implantation，no implants were seen in the tissue sections of the test material group mice，with a histological score of 3.4 points. CONCLUSION：In this study，the subcutaneous implantation of the porcine-derived absorbable anti-adhesion material did not cause significant adverse effects on the immune system of mice，and the risk of immunotoxicity is low.

OBJECTIVE：This study aimed to develop a method for assessing uncertainty in microbial testing of medical devices using the black-box approach，and to bolster integrity reliability of test outcomes. METHODS：The black-box method and the reproducibility standard deviation formula were used to determine combined uncertainty in microbial testing for medical devices. Parallel experiments were performed on domestically produced transcutaneous electrical nerve stimulators and recombinant collagen medical products to evaluate the effects of varying operators，sampling techniques，dilution factors，and reading methods on the total bacterial count for different samples. RESULTS：The black-box method was both convenient and practical in facilitating the calculation of combined uncertainty. It enabled the dynamic updating of laboratory data in response to new uncertainties and data. For the transcutaneous nerve stimulator，the synthetic standard uncertainty was 3.69，with an expanded uncertainty of 7.38，reporting a final detection result of (54.8±7.38) CFU/mL. For the recombinant collagen medical product，the synthetic standard uncertainty was 2.42，with an expanded uncertainty of 4.84，reporting a final detection result of (20.8±4.84) CFU/mL. The synthetic uncertainty for in-laboratory bacterial detection was 8.81. CONCLUSION：The black-box method effectively evaluated and reported the uncertainty in microbial testing for medical devices，which is crucial for improving test quality，compliance assessment，and the mutual recognition of results across laboratories.