Objective:To detect the expression of Toll-like receptor 7(TLR7)in blood neutrophils of patients with allergic rhi-nitis(AR),allergic asthma(AA)and allergic rhinitis combined with allergic asthma(ARA)before and after allergen challenge.Methods:A total of 44 AR,36 AA,19 ARA patients and 19 healthy control(HC)were recruited,the extracts of Artemisia siever-siana wild allergen(ASWE),house dust mite allergen(HDME)and Platanus pollen allergen(PPAE)were used to challenge periph-eral blood of HC as well as the patients,the expression of TLR7 in neutrophils was detected by flow cytometry.Results:In a resting state,compared with HC,upregulated expression of TLR7 in neutrophils in patients with airway allergy:the percentage of TLR7+neu-trophils in AR,AA,ARA patients increased 6 times,3.18 times and 6.15 times,respectively,and the mean fluorescence intensity(MFI)of TLR7 expression in neutrophil in ARA was enhanced by 24%.The results of allergen challenge tests showed that the expres-sion of neutrophils TLR7 did not change in HC subjects after the allergen challenge,whereas the expression of TLR7 in neutrophils of patients with airway allergy was upregulated after the allergens challenge.After HDME and PPAE challenge,the percentage of TLR7+neutrophils in peripheral of AR patients increased by 8.40%and 33.03%,respectively,and the MFI of TLR7 enhanced by 29.62%and 35.72%,respectively.After ASWE and PPAE challenge.The percentage of TLR7+neutrophils in peripheral of AA pa-tients increased by 59.18%and 1.06 times,respectively,and the MFI of TLR7 enhanced by 68.93%and 47.33%,respectively.How-ever,after HDME stimulation only enhanced the MFI of neutrophil TLR7 in AA patients by 66.14%;after HDME challenge,the per-centage of TLR7 and MFI in peripheral blood neutrophils of patients with ARA increased by 21.05%and 35.61%,respectively,while after PPAE challenge,the percentage of TLR7+cells in patients with ARA increased by 20.07%.Conclusion:The expression of TLR7 in blood neutrophils is increased in patients with allergic airway diseases after allergen stimulation,suggesting that allergens may be involved in the occurrence and development of allergic airway diseases by inducing TLR7 expression changes in neutrophils.

Background Cooking oil fumes are closely related to immune response, and adipose tissue also plays an important role in immune regulation. At present, the biological effect and mechanism of inflammation of adipose tissue induced by oil fume exposure are not clear yet. Objective To investigate the inflammatory effect of different exposure duration of cooking fumes on adipose tissue in mice and explore the role of Nod-like receptor pyrin domain 3 (NLRP3)/cysteinyl aspartate specific proteinase 1 (Caspase 1)/interleukin (IL)-1β signaling pathway. Methods Forty 8-week-old female C57BL/6J mice were randomly divided into 3-day control group (CON₃ group), 7-day control group (CON₇ group), 3-day oil fume exposure group (COF₃ group), and 7-day oil fume exposure group (COF₇ group), with 10 mice in each group. The mice were exposed to oil fumes in a cooking oil fume formation and exposure equipment (COFFEE) for 20 min, followed by a 10-min pause, 1 h a day for consecutive 3 d or 7 d. General condition of mice was observed and body weight was measured every day. After exposure, blood was sampled from the eyeball. Serum levels of IL-6, IL-27, and IL-1β were detected by enzyme-linked immunosorbent assay (ELISA). The adipose tissue of mice was collected and observed after hematoxylin-eosin (HE) staining. The percentages of CD4⁺ and CD8⁺T cells in adipose tissue were detected by flow cytometry. Real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of nuclear factor-κB (NF-κB), NLRP3, Caspase 1, and IL-1β in adipose tissue. Western blot was used to detect the expression levels of NLRP3, Caspase 1, and IL-1β in adipose. Results Compared with the corresponding control group, serum IL-6, IL-27, and IL-1β contents in the COF₃ group and the COF₇ group were significantly increased (P<0.05) except IL-6 in the COF₃ group, and the levels in the COF₇ group were significantly higher than those in the COF₃ group (P<0.05). Vacuolar lipid droplets in adipocytes decreased, cytoplasm shrank, and inflammatory cells infiltrated in the COF₇ group after HE staining. The flow cytometry results showed that the proportions of CD4⁺ and CD8⁺T cells in adipocytes of the COF₃ group and the COF₇ group were increased compared to the corresponding control group, with a significant increase in the COF₇ group (P<0.05), and the CD4⁺/CD8⁺T ratio also significantly increased progressively in the two groups (P<0.05). The results of RT-qPCR showed that compared with the corresponding control group, the mRNA expression levels of NF-κB, NLRP3, Caspase 1, and IL-1β in adipose tissue of mice in the COF₃ group and the COF₇ group were significantly increased (P<0.05, P<0.01). The mRNA expression levels of mice in each exposure group gradually increased over time. The Western blot results showed that compared with the corresponding control group, the protein expressions of NLRP3 and Caspase 1 in the COF₃ group were significantly increased (P<0.01), and the expression of IL-1β protein also increased but without statistical significance. The protein expressions of NLRP3, Caspase 1, and IL-1β in the COF₇ group were significantly higher than those in the CON₇ group (P<0.05, P<0.01). Conclusion Acute exposure to cooking oil fumes can induce significant inflammatory response in adipose tissue, and the effect gradually increases with the extension of exposure time. The mechanism of action may be related to the activation of NLRP3 inflammasome signaling pathway.

Background It is unclear if there is any combined effect of air pollutants and non-optimal temperature on metabolic syndrome, or any molecular mechanisms of related signaling pathways in the process, which requires urgent systematic research. Objective To observe the effects of combined exposure to PM_2.5 and non-optimal temperature on metabolic damage at gene and protein levels in mice, and elucidate the role of related signaling pathway in crucial organs. Methods A total of 60 six-week-old male C57BL/6J mice were randomly divided into six groups: a normal temperature-filter air group (T_N-FA), a normal temperature-concentrated PM_2.5 group (T_N-PM), a heat-filter air group (T_H-FA), a heat-concentrated PM_2.5 group (T_H-PM), a cold-filter air group (T_C-FA), and a cold-concentrated PM_2.5 group (T_C-PM). The Shanghai Meteorological and Environmental Animal Exposure System (Shanghai-METAS) was used to provide combined exposure settings of air types [concentrated PM_2.5 and filter air (FA)] and temperatures [normal (22°C), cold (4°C), and heat (30°C)] for 4 weeks. Skeletal muscle and white adipose tissue (WAT) of the mice were sampled at the end of exposure, and transcriptomics and Western blot (WB) assay were adopted to observe selected gene and protein expression levels in the samples respectively. Results The transcriptomics results indicated that the PM_2.5 exposure enhanced the number of differentially expressed genes. Specifically, 4820 genes were differentially expressed in the T_N-PM mice compared to the T_N-FA mice at normal temperature, and 1143 genes were differentially expressed in the Tc-PM mice compared to the Tc-FA mice in the cold environment. The phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling pathway and the endoplasmic reticulum protein processing pathway were identified as the most significant pathways in metabolic injury resulting from combined exposure to PM_2.5 and non-optimal temperature exposure. The WB results showed that exposure to PM_2.5 in the normal temperature and the cold environments led to a significant increase in the expression of p-AKT in WAT (P<0.01, P<0.05) and a significant decrease in the expression of GLUT4 (P<0.05, P<0.01). In skeletal muscle, exposure to PM_2.5 led to a significant decrease in GLUT4 (P<0.05) in all environments, with a consistent trend of change as observed in WAT. Conclusion Cold/heat exposure might promote PM_2.5-induced metabolic disorder through suppression of the AKT/GLUT4 pathway, aggravating metabolic damage.

[摘 要] 目的：探讨橙花叔醇通过Wnt-β-catenin通路抑制黑色素瘤A-375和WM-115细胞恶性生物学行为的分子机制。方法: 体外培养黑色素瘤细胞A-375和WM-115，用不同浓度的橙花叔醇处理，采用SRB法和克隆形成实验、FCM术、Transwell实验和细胞划痕实验、DCFH-DA染色法、qPCR和WB法分别检测橙花叔醇对A-375和WM-115细胞的增殖能力、细胞周期和凋亡、迁移能力、活性氧（ROS）水平和Wnt-β-catenin通路及其下游相关基因和相关蛋白表达的影响。利用ULCAN和GEPIA2数据库分析黑色素瘤中Wnt-β-catenin通路的激活与患者预后的关系。结果：与对照组比较，橙花叔醇处理组A-375和WM-115细胞的增殖能力受明显抑制（均P<0.01）、细胞周期阻滞于G2/M期（P<0.05或P<0.01）、细胞凋亡率增加（均P<0.01）、迁移能力降低（P<0.05或P<0.01）、ROS水平升高（均P<0.01）、Wnt-β-catenin通路被抑制而其下游基因和蛋白表达明显上调（均P<0.01）。数据库数据分析显示，WNT1基因高表达患者OS低于低表达患者（P<0.01）。结论：橙花叔醇通过上调A-375和WM-115细胞中ROS水平影响Wnt-β-catenin通路，从而抑制其恶性生物学行为；Wnt-β-catenin通路可能是黑色素瘤治疗的潜在靶点。

【Objective】 To explore the emergency treatment of penoscrotal avulsion injury based on the American Association for Surgery and Trauma (AAST) penoscrotal injury grade. 【Methods】 Data of 30 patients with penoscrotal avulsion injury treated in our hospital with in-situ suture, skin grafting or skin flap during Oct.2003 and Dec.2017 were reviewed. 【Results】 Among the 30 patients, 29 received emergency surgery, including in-situ suturein 15 grade Ⅰ-Ⅲ cases, skin graft in 8 grade Ⅳ-Ⅴ cases, and skin flap in 6 cases;1 case received delayed operation due to wound infection on admission. In the perioperative period,25 cases had more than 90% wound healing, 3 cases had 70% wound healing after wound dressing change, and 2 cases had complete necrosis. Second-stage skin graft yielded satisfactory penoscrotal appearance. 【Conclusion】 According to the specific conditions of patients, reasonable surgical methods can be selected. For grade Ⅰ-Ⅲ patients, in-situ suture can be used, while for grade Ⅳ-Ⅴ patients, wound condition, age, fertility and other factors should be taken into consideration so as to preserve the function of testis to the maximum extent.

Objective:To study the expressions of heat shock protein (HSP) 90α and HSP90β in colorectal cancer and paracancer tissues, and to investigate the relationships between HSP90α, HSP90β and clinicopathological features of colorectal cancer patients, and to analyze their correlation.Methods:The tumor tissues and paracancer tissues of 117 patients with colorectal cancer were selected from the Department of Gastrointestinal Surgery, Third Affiliated Hospital of Shandong First Medical University from January 2016 to December 2020. The expression levels of HSP90α and HSP90β were detected by immunohistochemistry, and the relationships between the two proteins and clinicopathological features and the correlation of their expressions were analyzed.Results:The positive expression rates of HSP90α in colorectal cancer tissues and paracancer tissues were 74.4% (87/117) and 12.0% (14/117) , and there was a statistically significant difference ( χ2=92.83, P<0.001) . The positive expression rate of HSP90β in colorectal cancer tissues and paracancer tissues was 61.5% (72/117) and 10.3% (12/117) , and there was a statistically significant difference ( χ2=66.86, P<0.001) . The expression of HSP90α was correlated with tumor location ( χ2=8.67, P=0.003) , vascular invasion ( χ2=8.68, P=0.003) , lymph node metastasis ( χ2=8.52, P=0.004) , T stage ( χ2=21.07, P<0.001) , N stage ( χ2=11.94, P=0.003) , M stage ( χ2=5.37, P=0.020) , pathological stage ( χ2=25.64, P<0.001) . The expression of HSP90β was correlated with lymph node metastasis ( χ2=4.03, P=0.045) , T stage ( χ2=11.09, P=0.007) , N stage ( χ2=6.56, P=0.038) , M stage ( χ2=12.43, P<0.001) , pathological stage ( χ2=17.34, P=0.001) . There was a positive correlation between the expressions of the two proteins in colorectal cancer tissues ( r=0.42, P<0.001) . Conclusion:The expressions of HSP90α and HSP90β in colorectal cancer tissues are significantly higher than those in paracancer tissues, and they are related to lymph node metastasis and pathological stage. There is a positive correlation between the two proteins, which may be involved in the occurrence and development of colorectal cancer and are expected to become new tumor markers.

Due to the complexity of bioactive ingredients in biological samples,the screening of target proteins is a complex process.Herein,a feasible strategy for directing protein immobilization on silica magnetic beads for ligand fishing based on SpyTag/SpyCatcher(ST/SC)-mediated anchoring is presented.Carboxyl functional groups on the surface of silica-coated magnetic beads(SMBs)were coupled with SC using the 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysulfosuccinimide method,named SC-SMBs.The green fluorescent protein(GFP),as the capturing protein model,was ST-labeled and anchored at a specific orientation onto the surface of SC-SMBs directly from relevant cell lysates via ST/SC self-ligation.The characteristics of the SC-SMBs were studied via electron microscopy,energy dispersive spectroscopy,and Fourier transform infrared spectroscopy.The spontaneity and site-specificity of this unique reaction were confirmed via electrophoresis and fluorescence analyses.Although the alkaline stability of ST-GFP-ligated SC-SMBs was not ideal,the formed isopeptide bond was unbreakable under acidic conditions(0.05 M glycine-HCl buffer,pH 1-6)for 2 h,under 20％ethanol solution within 7 days,and at most temperatures.We,therefore,present a simple and universal strategy for the preparation of diverse protein-functionalized SMBs for ligand fishing,prompting its usage on drug screening and target finding.

A biosensor was constructed to detect the endotoxin.The amine-terminated aptamer was immobilized on the gold electrode surface covered in advance with a self-assembled monolayer (SAM) of 3-mercaptopropionic acid (MPA).The modification of the gold electrode was confirmed by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS).It was found that MPA assembly time at 6 h in the gold electrode surface to form a stable self-assembled monolayer.The biosensor has a good linear relationship with concentration of endotoxin in the range of 0.001 ～0.1 EU/ml,it is possible to be applied for quality control in the actual biological sample.

Objective:To investigate the extracellular signal-regulated kinase 5 (ERK5) and matrix metallo proteinase-9 (MMP-9) expres-sion levels in osteosarcoma tissues and their clinical significance. Methods:The ERK5 and MMP-9 expression levels in 71 specimens of osteosarcoma tissue and 40 specimens of normal bone tissue were detected by immunohistochemistry. The relationship between ERK5 and MMP-9 expression levels, their clinical characteristics, and prognosis of patients with osteosarcoma were analyzed. Results:The positive expression of ERK5 and MMP-9 in osteosarcoma tissues was 85.9%(61/71) and 74.65%(53/71), respectively, which were significantly higher than those in normal bone tissues at 12.5%(5/40) and 10.0%(4/40) (all P<0.05). The positive expression of ERK5 and MMP-9 was associated with Enneking stage and metastasis (all P<0.05). Kaplan-Meier analysis showed that the survival duration of patients with positive ERK5 and MMP-9 expression levels was shorter than those of the patients in the negative expression groups (all P<0.05). Univariate analysis of COX proportional hazards regression model revealed that tumor size, Enneking stage, metastasis, and positive ERK5 and MMP-9 expression levels are relevant to the overall survival of patients with osteosarcoma (all P<0.05). Multi-variate analysis of COX proportional hazards regression model confirmed that Enneking stage, metastasis, and positive ERK5 and MMP-9 expression levels can act as independent prognostic factors for osteosarcoma patients (all P<0.05). Conclusion:The ERK5 and MMP-9 expression levels are high in osteosarcoma tissues and are related to the clinical characteristics and prognosis of patients with osteo-sarcoma. Thus, ERK5 and MMP-9 expression levels may play important roles in osteosarcoma development and progression.

Objective] To summarize national famous doctor of traditional Chinese medicine-Wu Liangcun ’s experience about the clinical treatment of ovarian cancer. [Method] By studying and following Professor Wu Liangcun to diagnose, this paper discusses Professor Wu ’s academic views and clinical experience of diagnosis and treatment of ovarian cancer respectively from the aspects such as etiology and pathogenesis, and treatment based on syndrome differentiation. It expounds the characteristics of his prescription and also gives proven cases. [Result]Professor Wu Liangcun proposes that the pathogenesis of ovarian cancer is mainly congenital deficiency and physical weakness resulted from cold coagulation, qi stagnation, blood stasis and phlegmatic hygrosis. All these remain a standstill, gather together and become a disease. He uses the syndrome differentiation method to divide the common ovarian cancer into four types:blood stasis and vital energy retardation, phlegm dampness condensation, deficiency of liver-yin and kidney-yin, asthenia of qi and blood. In the treatment, he organically combines the dialectics and the differentiation of disease; In the selection of drugs, he streamlines the prescription and the drug, which maximizes the efforts of the drugs and has good efforts in clinic.[Conclusion] Professor Wu Liangcun is good at making a diagnosis and giving treatment of ovarian cancer in combination with disease and syndrome, and he uses medicine flexibly. It has a very good effect and practical value for guiding clinic, and it should be promoted.