Intestinal microbiota contributes to host defense against pathogens while avoiding the induction of inflammation in homeostatic conditions, but the mechanism is not fully understood. To investigate the potential role of the bacterial metabolite desaminotyrosine (DAT) in regulating host defense and inflammation, we pretreated mouse bone marrow-derived macrophages (BMDMs) with DAT for 12 hours and then challenged with bacterial lipopolysaccharide (LPS). We found that DAT priming-enhanced type I interferon response while selectively inhibiting proinflammatory interleukin (IL)-6 production after exposure to LPS. This is related to the fact that DAT is a natural antioxidant determined by radical scavenging assay in a cell-free system. DAT-primed cells had increased levels of the reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) upon LPS stimulation. Countering the increased NADPH by supplementing extra oxidized NADP+ to cells reversed DAT's effect on LPS-induced Il-6 and interferon-stimulated gene expressions. DAT-primed cells also were more resistant to oxidative stress-induced generation of reactive oxygen species and cell death. DAT promoted the production of antimicrobial effector nitric oxide in a cellular redox-dependent manner, leading to enhanced macrophage antimicrobial activity during Salmonella enterica infection. Our data suggest that DAT acts as a host-microbiota crosstalk signal in shaping host immune defense and inflammatory response.

IgA antibodies play an important role in mucosal immunity. However, there is still no effective way to consistently boost mucosal IgA responses, and the factors influencing these responses are not fully understood. We observed that colonization with the murine intestinal symbiotic protozoan Tritrichomonas musculis (T.mu) boosted antigen-specific mucosal IgA responses in wild-type C57BL/6 mice. This enhancement was attributed to the accumulation of free arachidonic acid (ARA) in the intestinal lumen, which served as a signal to stimulate the production of antigen-specific mucosal IgA. When ARA was prevented from undergoing its downstream metabolic transformation using the 5-lipoxygenase inhibitor zileuton or by blocking its downstream biological signaling through genetic deletion of the Leukotriene B4 receptor 1 (Blt1), the T.mu-mediated enhancement of antigen-specific mucosal IgA production was suppressed. Moreover, both T.mu transfer and dietary supplementation of ARA augmented the efficacy of an oral vaccine against Salmonella infection, with this effect being dependent on Blt1. Our findings elucidate a tripartite circuit linking nutrients from the diet or intestinal microbiota, host lipid metabolism, and the mucosal humoral immune response. Immunoglobulin A (IgA) is an important part of the mucosal immune response. Here, the authors show that an intestinal commensal protist can enhance mucosal IgA production upon vaccination and that this involves the accumulation of free arachidonic acid in the intestinal lumen.

Esophageal squamous cell carcinoma (ESCC) pathogenesis is influenced by both NFE2L2 (nuclear factor erythroid 2-related factor 2) and SQSTM1 (sequestosome 1), also known as p62. However, while there is evidence that these two proteins can interact with one another in a range of pathological contexts, whether these interactions govern the development or progression of ESCC remains unknown. In the present study, analyses of the GEPIA database revealed the simultaneous upregulation of both NFE2L2 and p62 in ESCC, as was further confirmed through biochemical analyses conducted with a human tumor microarray. Knocking down the expression of one or both of these factors demonstrated that both p62 and NFE2L2 mediate the progression of ESCC, as such downregulation altered the morphological characteristics of these cells and suppressed the epithelial-mesenchymal transition (EMT). Strikingly, these experiments revealed synergistic interactions between NFE2L2 and p62 in the promotion of ESCC invasivity and EMT induction. The treatment of cells with the autophagy inhibitors 3-MA, however, was sufficient to partially reverse the anti-metastatic effects of knocking down p62 and/or NFE2L2. Together, these data illustrate the ability of p62 and NFE2L2 to function in a synergistic manner, promoting ESCC cell metastatic progression and EMT induction through mechanisms linked to autophagic activity. As such, efforts to simultaneously target both of these proteins may represent a viable means of providing new treatment options to ESCC patients.

Objective:To explore the clinical application value of metagenomics next-generation sequencing (mNGS) in assisting the diagnosis of invasive fungal diseases, and to provide pathogenic evidence for the clinical diagnosis and treatment of mucormycosis.Methods:The alveolar lavage fluid of a patient with unexplained lung infection was detected by mNGS to determine the pathogen, and the treatment plan was adjusted according to the mNGS test report.Results:The patient had intermittent chest pain and cough for longer than 5 months. The chest CT showed multiple shadows and cavities in both lungs. There was no abnormality in routine laboratory and pathogenic examinations, and no significant improvement was found after various anti-infection treatments. The mNGS of alveolar lavage fluid submitted for examination revealed that the pathogen was Lichtheimia ramose. After switching to amphotericin B for anti-infection treatment, the patient’s infected lesions shrank and the symptoms relieved significantly.Conclusions:mNGS is an important auxiliary diagnostic method to identify rare pathogens, which is of great significance to assist the diagnosis of lung invasive mycosis.

Pseudothrombocytopenia (PTCP) is a condition in which the decreased platelet count does not agree with the clinical status of the patient, can lead to misdiagnose, unnecessary tests, unnecessary treatment. The present case describes a 73-year-old man suffered with pulmonary tuberculosis, treated with anti-tuberculosis therapy (isoniazid, rifampicin, pyrazinamide, ethambutol, 2HRZE/4HR). One month later, the patient had a significant decrease in platelets (101 to 56 x10 9 /L). Peripheral blood smear showed that 28% platelets were phagocytosis by neutrophils, 26% platelets were lack of granules and 6% platelets’ volume increased significantly. When the anticoagulant was changed from EDTA to sodium citrate, there was no change in the above phenomenon. By manual count, the value of platelets was 113 x 10 9 /L. After the completion of anti-tuberculosis therapy, platelet morphology gradually returned to normal. HRZE treatment may cause platelet morphology abnormal, resulting in PTCP. In such cases, we should regularly review the peripheral blood smear to ensure the accuracy of the results and avoid unnecessary examination and treatment. The emergence of PTCP may does not mean the presence of specific disorders.

Background: Sequestosome 1 (SQSTM1), also referred to as p62 and nuclear factor erythroid 2-related factor 2 (Nrf2) have implicated into the pathogenesis of esophageal squamous cell carcinoma (ESCC). While p62 and Nrf2 interact and influence each other in diverse pathological settings, in whether such interaction contributes to ESCC remains unclear. In this study, we aimed to explore the functional significance of p62 and Nrf2 in ESCC, and uncover a interaction between p62 and Nrf2 in mediating ESCC. Results: Herein, we show a critican interaction betwwen p62 and Nrf2 in ESCC. Huamn GEPIA database analysis revealed the expression of p62 and Nrf2 is upregulated in ESCC, which is further verified by biochemical assessements using a human tumor tissue bank. Genetic knockdown of p62 and/or Nrf2 demonstrated a mediator role of p62 and Nrf2 in the proliferation, migration and movement ability of ECSS cells. In addition, the downregulation of p62 or Nrf2 could change the morphology of ESCC cells, transforming them from shuttle to round shape, and inhibit their EMT. The knockdown approach also reveal a synergy between p62 and Nrf2 in promoting EMT and the invasive growth of ESCC cells. The xenograft assay in vivo revealed similar effects of p62 and/or Nrf2 knockdown on tumorigenesis and metastaisis of ESCC. Conclusions: These results indicate that p62 and Nrf2 promote EMT and metastasis of ESCC in a synergistic manner. Simultaneously targeting of p62 and Nrf2 may have a therapeutic potential in treating ESCC.

Rickettsia is the pathogen of Q fever, Brucella ovis is the pathogen of brucellosis, and both of them are Gram-negative bacteria which are parasitic in cells. The mixed infection of rickettsia and Brucella ovis is rarely reported in clinic. Early diagnosis and treatment are of great significance to the treatment and prognosis of brucellosis and Q fever. Here, we report a case of co-infection Rickettsia burneti and Brucella melitensis. The patient is a 49-year-old sheepherder, who was hospitalized with left forearm trauma. Three days after admission, the patient developed fever of 39.0°C, accompanied by sweating, fatigue, poor appetite and headache. Indirect immunofluorescence (IFA) was used to detect Rickettsia burneti IgM. After 72 hours of blood culture incubation, bacterial growth was detected in aerobic bottles, Gram-negative bacilli were found in culture medium smear, the colony was identified as Brucella melitensis by mass spectrometry. Patients were treated with doxycycline (100 mg bid, po) and rifampicin (600 mg qd, po) for 4 weeks. After treatment, the symptoms disappeared quickly, and there was no sign of recurrence or chronic infection. Q fever and Brucella may exist in high-risk practitioners, so we should routinely detect these two pathogens to prevent missed diagnosis.

Background Brain abscess due to the Nocardia genus is rarely reported that usually found in immunocompromised patients. Treatment of Nocardia brain abscess is troublesome and requires consideration of the severity of the underlying systemic disease, the difficulties in identifying the bacterium and the frequent delay in initiating adequate therapy. Case Presentation Here, we report a rare case of brain abscess caused by Nocardia farcinica. The patient’s medical history was complicated, bacterial was found in culture of brain abscess puncture fluid, the colony was identified as Nocardia farcinica by mass spectrometry. Targeted antibiotic treatment was implemented, brain abscess tended to alleviate, but the patient eventually developed fungal pneumonia and died of acute respiratory distress syndrome (ARDS). Conclusion Early diagnosis, reasonable surgical intervention, and targeted antibiotic treatment are critical for Nocardia brain abscess treatment. Any delay in diagnosis and appropriate therapy can have adverseconsequences.

BACKGROUND:Brain abscess due to the Nocardia genus is rarely reported and it is usually found in immunocompromised patients. Treatment of Nocardia brain abscess is troublesome and requires consideration of the severity of the underlying systemic disease. The difficulties in identifying the bacterium and the frequent delay in initiating adequate therapy often influence the prognosis of patients.CASE PRESENTATION:Here, we report a rare case of brain abscess caused by Nocardia farcinica. The patient's medical history was complicated: he was hospitalized several times, but no pathogens were found. At last, bacteria were found in the culture of brain abscess puncture fluid; the colony was identified as Nocardia farcinica by mass spectrometry. Targeted antibiotic treatment was implemented, brain abscess tended to alleviate, but the patient eventually developed fungal pneumonia and died of acute respiratory distress syndrome (ARDS).CONCLUSION:Brain abscess caused by Nocardia farcinica can appear in non-immunocompromised individuals. Early diagnosis, reasonable surgical intervention, and targeted antibiotic treatment are critical for Nocardia brain abscess treatment. In the treatment of Nocardia brain abscess, attention should paid be to the changes in patients' immunity and infection with other pathogens, especially fungi, avoided.

The global pandemic of novel coronavirus has brought great threat to the life and health of people all over the world. Currently, the prevention and treatment measures for novel coronavirus remain very limited. Nucleic acid testing is crucial for the rapid diagnosis of infected cases, the exclusion of suspected cases, and the promotion of epidemic prevention and control and clinical treatment. In this paper, the application and challenges of RT-PCR technology in novel coronavirus nucleic acid detection are reviewed, and the practical experience accumulated in our laboratory for a long time is summarized, such as increasing instruments and equipment, optimizing experimental process, strengthening information construction, and improving working conditions, to improve the ability and efficiency of nucleic acid testing, and provide a reference for nucleic acid detection of novel coronavirus in the laboratory.

Background: Brain abscess due to the Nocardia genus is rarely reported and it is usually found in immunocompromised patients. Treatment of Nocardia brain abscess is troublesome and requires consideration of the severity of the underlying systemic disease. The difficulties in identifying the bacterium and the frequent delay in initiating adequate therapy often influence the prognosis of patients. Case presentation: Here, we report a rare case of brain abscess caused by Nocardia farcinica. The patient’s medical history was complicated, he was hospitalized for several times, but no pathogens were found. At last, bacterial was found in culture of brain abscess puncture fluid, the colony was identified as Nocardia farcinica by mass spectrometry. Targeted antibiotic treatment was implemented, brain abscess tended to alleviate, but the patient eventually developed fungal pneumonia and died of acute respiratory distress syndrome (ARDS). Conclusion: brain abscess caused by Nocardia farcinica can be appear in non-immunocompromised individuals. Early diagnosis, reasonable surgical intervention, and targeted antibiotic treatment are critical for Nocardia brain abscess treatment. In the treatment of Nocardia brain abscess, we should pay attention to the changes of patients' immunity and avoid infection with other pathogens, especially fungi.

Objective:To investigate the diagnostic characteristics of facial abscess caused by Streptococcus constellatus.Methods:The clinical data of a case of facial abscess caused by Streptococcus constellatus admitted to the Oncology Department of Tianmen First People′s Hospital on June 27, 2020 were retrospectively analyzed, and the relevant literatures were reviewed.Results:The patient presented with a mass in the right maxillofacial region, with partial skin ulceration and purulent discharge. The patient had an anamnesis of nasopharyngeal carcinoma. Imaging indicated right middle ear mastoiditis and sinusitis. Laboratory examination found that IgA antibody of human herpesvirus was weakly positive, C-reactive protein level was 11.07 mg/L, procalcitonin level was 1.26 μg/L, blood culture was negative, and Streptococcus constellatus was detected in wound secretion. The pathogen was resistant to erythromycin and clindamycin.Conclusions:As a kind of opportunistic pathogen, Streptococcus constellatus can cause suppurative infection in various tissues and organs of human body, which should arouse clinical attention. We should improve the detection rate of pathogenic bacteria, so as to achieve early diagnosis and treatment.

Patients with severe coronavirus disease in 2019 (COVID-19 pneumonia) may have many sequelae, which seriously affect their quality of life and work. Here, we report a case of infection in China, reviewed the course, treatment, and rehabilitation of a patient suffering from severe COVID-19 pneumonia, and collected his examination reports, including chest CT, laboratory examination results, lung function examination, sleep monitoring report, sex hormones, sperm morphology and activity. The patient’s antiviral immunoglobulin G (IgG) continued to be positive for more than 11 months, and his small airway function was abnormal, and he suffered from respiratory problems (cough, chest pain, chest tightness, and shortness of breath), unstructured sleep apnea hypopnea syndrome, and nocturnal sleep hypoxemia. His abnormal sperm rate increased obviously, and sperm activity decreased obviously. Patients with severe COVID-19 pneumonia may have respiratory sequela, the abnormal sperm rate is obviously increased, and IgG positive can last for a long time.

目的:新冠肺炎患者应用核酸联合抗体检测的临床应用.方法:从我院2019年2月至2020年3月期间收治的新冠肺炎患者中,随机抽取50例作为实验的研究对象进行回顾性分析,选取2020年2月仅使用核酸检测,并将其这段时间患者的检测结果设置成为参照组,将其3月使用核酸联合抗体检测,并将其这段时间患者的检测结果设置成为对照组.研究新冠肺炎患者应用核酸联合抗体检测的临床应用.结果:新冠肺炎患者应用核酸联合抗体检测结果比单纯应用核酸检测结果更为准确,组间比较差异具有统计学意义(P<0.05).结论:新冠肺炎患者应用核酸联合抗体检测可以更好地筛选出病毒,提高检测者的诊断准确性,有着更好的推广价值.

Background: Qiliqiangxin (QLQX) is a preparation refined from a traditional Chinese medicine compound. It plays an important role in protecting cardiac function after myocardial infarction (MI). However, the underline mechanism of QLQX action is not clear. The purpose of this study was to detect the effects of QLQX on mitophagy after MI. Methods: Male FVB/NJ mice aged 8-10 weeks were underwent left coronary artery ligation and were orally administered either QLQX (0.25 g/kg/d) or saline. Twenty-eight days after surgical operation, the cardiac function of mice was detected by echocardiography. Electron Microscopy was used to observe the microstructure of cardiomyocytes. Myocardial apoptosis was examined by TdT-mediated dUTP Nick-End Labeling (TUNEL) and western blot. H9c2 cells were cultured in a hypoxic incubator chamber (5% CO2, 1% O-2, 94% N-2) for 12 h and pretreated with or without QLQX (0.5 mg/mL). The cell apoptosis, reactive oxygen species (ROS), mitochondrial membrane potential and mitophagy were detected. Results: When compared to sham group, the cardiac function of MI mice decreased significantly, and their cardiomyocyte apoptosis and mitochondrial damage were more serious. These MI-induced cardiac changes could be reversed by QLQX treatment. In vitro experiments also confirmed that QLQX could protect cardiomyocytes from hypoxia-induced apoptosis and mitochondrial damage. Further study indicated that QLQX could increase the expression of Pink1 and Parkin in cardiomyocytes. Conclusion: Qiliqiangxin could reduce cardiomyocytes apotosis and improved heart function in infarcted heart through Pink1-mediated mitochondrial autophagy.

目的 观察过氧乙酸在新型冠状病毒核酸检测实验室消毒应用效果,为新型冠状病毒检测实验室感染防控提供参考.方法 采用基因检测技术,对复方过氧乙酸消毒剂消毒新冠病毒检测实验室的效果进行评价.结果 该复方过氧乙酸含过氧乙酸约2.5 g/L与过氧化氢60 g/L,以其原液用气溶胶电动喷雾器按22 ml/m3 用量,对新型冠状病毒核酸检测实验室的标本制备间进行喷雾消毒并密闭作用60 min.该实验室已连续30 d,在每次工作结束时进行喷雾消毒,所有采集的实验室环境标本均未检测到新冠病毒核酸阳性,从未发生过工作人员感染新冠病毒的事件.结论 结果 证明用该复方过氧乙酸对新冠病毒检测实验室进行喷雾消毒,可有效果杀灭实验室内环境中新冠病毒,能有效保护工作人员的安全.

目的 观察p62基因敲除后对食管鳞状细胞癌EC109细胞生物学行为的影响.方法 使用CRISPR/Cas9技术构建p62敲除的EC109细胞株作为敲除组,选择野生型EC109细胞作为对照组.采用T7E1核酸内切酶检测打靶效果,Western blotting法检测细胞p62蛋白,倒置显微镜下观察细胞形态变化,实时无标记动态细胞分析技术观察细胞增殖能力,细胞划痕实验观察细胞的迁移能力,流式细胞仪检测细胞周期.结果 T7E1酶切后,敲除组在800 bp左右有敲除后的特异性条带,对照组只在870 bp左右有一条带.敲除组p62蛋白相对表达量低于对照组,细胞合胞体数量多于对照组(P均<0.01);敲除组细胞形态与对照组相比,细胞较圆且边界不清.敲除组10～80 h细胞增殖的CI值均低于同时点的对照组,细胞克隆数量少于对照组(P均<0.05);敲除组96 h划痕愈合率低于对照组,细胞周期中阻滞于G0/G1期的细胞比例高于对照组(P均<0.05).结论 p62基因敲除后食管鳞状细胞癌EC109细胞形成较多合胞体,且细胞的恶性行为受到抑制;p62有望成为治疗食管癌的有效基因靶点.

Cellular senescence, a process whereby cells enter a state of permanent growth arrest, appears to regulate cardiac pathological remodeling and dysfunction in response to various stresses including myocardial infarction (MI). However, the precise role as well as the underlying regulatory mechanism of cardiac cellular senescence in the ischemic heart disease remain to be further determined. Herein we report an inhibitory role of Nrf2, a key transcription factor of cellular defense, in regulating cardiac senescence in infarcted hearts as well as a therapeutic potential of targeting Nrf2-mediated suppression of cardiac senescence in the treatment of MI-induced cardiac dysfunction. MI was induced by left coronary artery ligation for 28 days in mice. Heart tissues from the infarct border zone were used for the analyses. The MI-induced cardiac dysfunction was associated with increased myocardial cell senescence, oxidative stress and apoptosis in adult wild type (WT) mice. In addition, a downregulated Nrf2 activity was associated with upregulated Keap1 levels and increased phosphorylation of JAK and FYN in the infarcted border zone heart tissues. Nrf2 Knockout (Nrf2(-/-)) enhanced the MI-induced myocardial, cardiac dysfunction and senescence. Qiliqiangxin (QLQX), a herbal medicine which could reverse the MI-induced suppression of Nrf2 activity, significantly inhibited the MI-induced cardiac senescence, apoptosis, and cardiac dysfunction in WT mice but not in Nrf2(-/-) mice. These results indicate that MI downregulates Nrf2 activity thus promoting oxidative stress to accelerate cellular senescence in the infarcted heart towards cardiac dysfunction and Nrf2 may be a drug target for suppressing the cellular senescence-associated pathologies in infarcted hearts.

目的 通过对HeLa细胞过表达TET1基因,探究TET1基因对宫颈癌细胞增殖迁移能力的影响.方法 采用TALE-VP64系统构建TET1过表达质粒,转染宫颈癌HeLa细胞,四唑盐(MTT)比色法监测细胞增殖状况,划痕试验检测细胞迁移能力的变化,Transwell试验检测HeLa细胞侵袭能力的变化.结果 MTT法检测HeLa细胞增殖状况,TET1过表达的Clone 1和Clone 2细胞增殖能力明显弱于野生型HeLa细胞(P＜0.05).Transwell试验检测TET1过表达Clone 1及Clone 2 HeLa细胞穿过magtrigel胶及小室的数量分别是(21±5)和(22±6)个/视野,而野生型HeLa细胞组为(38±7)个/视野,与野生型HeLa细胞组比较,差异有统计学意义(P＜0.05).划痕试验检测TET1过表达组(Clone 1、Clone 2)细胞和野生型HeLa组细胞24 h及48 h的迁移率,后者明显高于前者(P＜0.01).TET1过表达能够抑制HeLa细胞的增殖、侵袭、迁移等能力.结论 TET1基因过表达对宫颈癌HeLa细胞有抑癌基因的作用.

Objective To study the expression and role of TET1 in cervical cancer.Methods The proliferation gene of HeLa cells was tested by CRISPR/Cas9,and the proliferation status of cells was dynamically measured by real-time cell analyzer (RTCA).The effect of TET1 gene knockout on the migration ability of HeLa cells was detected by scratching test.Results The TET1 gene was successfully targeted by CRISPR/Cas9.TET1 knockout can increase the proliferation,invasion and migration of HeLa cells.Concltsion TET1 knockdown could increase the malignant behavior of cervical cancer HeLa cells.