Bedside ultrasound-guided water injection assists endoscopically treatment in esophageal perforation caused by foreign bodies: A case report.

BACKGROUND: Fishbone migration from the esophagus to the neck is relatively uncommon in clinical practice. Several complications secondary to esophageal perforation after ingestion of a fishbone have been described in the literature. Typically, a fishbone is detected and diagnosed by imaging examination and is usually removed by a neck incision. CASE SUMMARY: Herein, we report a case of a 76-year-old patient with a fishbone in the neck that had migrated from the esophagus and that was in close proximity to the common carotid artery, and the patient experienced dysphagia. An endoscopically-guided neck incision was made over the insertion point in the esophagus, but the surgery failed due to having a blurred image at the insertion site during the operation. After injection of normal saline laterally to the fishbone in the neck under ultrasound guidance, the purulent fluid outflowed to the piriform recess along the sinus tract. With endoscopic guidance, the position of the fish bone was precisely located along the direction of liquid outflow, the sinus tract was separated, and the fish bone was removed. To the best of our knowledge, this is the first case report describing bedside ultrasound-guided water injection positioning combined with endoscopy in the treatment of a cervical esophageal perforation with an abscess. CONCLUSION: In conclusion, the fishbone could be located by the water injection method under the guidance of ultrasound and could be accurately located along the outflow direction of the purulent fluid of the sinus by the endoscope and was removed by incising the sinus. This method can be a nonoperative treatment option for foreign body-induced esophageal perforation.

The new progress in cancer immunotherapy.

The cross talk between immune and non-immune cells in the tumor microenvironment leads to immunosuppression, which promotes tumor growth and survival. Immunotherapy is an advanced treatment that boosts humoral and cellular immunity rather than using chemotherapy or radiation-based strategy associated with non-specific targets and toxic effects on normal cells. Immune checkpoint inhibitors and T cell-based immunotherapy have already exhibited significant effects against solid tumors and leukemia. Tumor cells that escape immune surveillance create a major obstacle to acquiring an effective immune response in cancer patients. Tremendous progress had been made in recent years on a wide range of innate and adaptive immune checkpoints which play a significant role to prevent tumorigenesis, and might therefore be potential targets to suppress tumor cells growth. This review aimed to summarize the underlying molecular mechanisms of existing immunotherapy approaches including T cell and NK-derived immune checkpoint therapy, as well as other intrinsic and phagocytosis checkpoints. Together, these insights will pave the way for new innate and adaptive immunomodulatory targets for the development of highly effective new therapy in the future.

In vivo evidence for GDP-fucose transport in the absence of transporter SLC35C1 and putative transporter SLC35C2.

Slc35c1 encodes an antiporter that transports GDP-fucose into the Golgi and returns GMP to the cytoplasm. The closely related gene Slc35c2 encodes a putative GDP-fucose transporter and promotes Notch fucosylation and Notch signaling in cultured cells. Here, we show that HEK293T cells lacking SLC35C1 transferred reduced amounts of O-fucose to secreted epidermal growth factor-like repeats from NOTCH1 or secreted thrombospondin type I repeats from thrombospondin 1. However, cells lacking SLC35C2 did not exhibit reduced fucosylation of these epidermal growth factor-like repeats or thrombospondin type I repeats. To investigate SLC35C2 functions in vivo, WW6 embryonic stem cells were targeted for Slc35c2. Slc35c2[-/-] mice were viable and fertile and exhibited no evidence of defective Notch signaling during skeletal or T cell development. By contrast, mice with inactivated Slc35c1 exhibited perinatal lethality and marked skeletal defects in late embryogenesis, typical of defective Notch signaling. Compound Slc35c1[-/-]Slc35c2[-/-] mutants were indistinguishable in skeletal phenotype from Slc35c1[-/-] embryos and neonates. Double mutants did not exhibit the exacerbated skeletal defects predicted if SLC35C2 was functionally important for Notch signaling in vivo. In addition, NOTCH1 immunoprecipitated from Slc35c1[-/-]Slc35c2[-/-] neonatal lung carried fucose detected by binding of Aleuria aurantia lectin. Given that the absence of both SLC35C1, a known GDP-fucose transporter, and SLC35C2, a putative GDP-fucose transporter, did not lead to afucosylated NOTCH1 nor to the severe Notch signaling defects and embryonic lethality expected if all GDP-fucose transport were abrogated, at least one more mechanism of GDP-fucose transport into the secretory pathway must exist in mammals.

The pathophysiologic significance of lymphocyte subset determination in children with infectious mononucleosis, mycoplasma pneumonia and Henoch-Schönlein purpura.

OBJECTIVE: This study aimed to explore lymphocyte subset determinations as an aid to understanding the pathophysiology of infectious mononucleosis (IM), pneumonia due to mycoplasma infection (P-MI) and Henoch-Schönlein purpura in children. METHODS: The peripheral blood lymphocyte subsets of 45 children with IM, 20 children with P-MI, and 31 children with Henoch-Schönlein purpura (HSP), who were treated in the pediatrics department of our hospital from April 2019 to February 2020, were determined by flow cytometry, and the number and percentage of lymphocyte subsets with CD3+, CD3 + CD4+, CD3 + CD8+, CD3 + CD4+/CD3 + CD8+, CD3-CD16 + CD56+, and CD3-CD19 + cells were observed, and the results were compared and analyzed. RESULTS: (1) The percentages of CD3+, CD3 + CD8 + lymphocyte subsets in children in IM group were significantly higher than those in children with P-MI and HSP, and the percentages of CD3-CD19 + lymphocyte subsets in children in IM group were significantly lower than those in children with P-MI and HSP. The percentages of CD3 + CD4 + lymphocyte subsets in children in the three groups were the lowest in children with IM, and the highest in children with P-MI.The differences in the percentages of CD3+, CD3 + CD4+, CD + CD8+, and CD3-CD19 + lymphocyte subsets among the IM, P-MI, and HSP groups were statistically significant (P < 0.01). (2) The results of CD3 + CD4+/CD3 + CD8 + in the three groups were the lowest in children with IM and the highest in children with P-MI. There was a significant difference among the three groups (P < 0.01); The ages of the children with IM and P-MI were lower than that of the children with HSP (p < 0.01), while there was no difference in the ages of the children with IM and P-MI (p > 0.05). (3) The difference in the percentage of CD3-CD16 + CD56 + lymphocyte subsets among the three groups was not statistically significant (P > 0.05). CONCLUSION: The determination of peripheral blood lymphocyte subsets is of significance for understanding the pathophysiology of IM, mycoplasma pneumonia, and HSP in children.

Evaluation and Comparison of Serological Methods for COVID-19 Diagnosis.

The worldwide pandemic of COVID-19 has become a global public health crisis. Various clinical diagnosis methods have been developed to distinguish COVID-19-infected patients from healthy people. The nucleic acid test is the golden standard for virus detection as it is suitable for early diagnosis. However, due to the low amount of viral nucleic acid in the respiratory tract, the sensitivity of nucleic acid detection is unsatisfactory. As a result, serological screening began to be widely used with the merits of simple procedures, lower cost, and shorter detection time. Serological tests currently include the enzyme-linked immunosorbent assay (ELISA), lateral flow immunoassay (LFIA), and chemiluminescence immunoassay (CLIA). This review describes various serological methods, discusses the performance and diagnostic effects of different methods, and points out the problems and the direction of optimization, to improve the efficiency of clinical diagnosis. These increasingly sophisticated and diverse serological diagnostic technologies will help human beings to control the spread of COVID-19.

Pulling-Force Spinning Top for Serum Separation Combined with Paper-Based Microfluidic Devices in COVID-19 ELISA Diagnosis.

The spread of Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2), resulting in a global pandemic with around four million deaths. Although there are a variety of nucleic acid-based tests for detecting SARS-CoV-2, these methods have a relatively high cost and require expensive supporting equipment. To overcome these limitations and improve the efficiency of SARS-CoV-2 diagnosis, we developed a microfluidic platform that collected serum by a pulling-force spinning top and paper-based microfluidic enzyme-linked immunosorbent assay (ELISA) for quantitative IgA/IgM/IgG measurements in an instrument-free way. We further validated the paper-based microfluidic ELISA analysis of SARS-CoV-2 receptor-binding domain (RBD)-specific IgA/IgM/IgG antibodies from human blood samples as a good measurement with higher sensitivity compared with traditional IgM/IgG detection (99.7% vs 95.6%) for early illness onset patients. In conclusion, we provide an alternative solution for the diagnosis of SARS-CoV-2 in a portable manner by this smart integration of pulling-force spinning top and paper-based microfluidic immunoassay.

Retroperitoneal parasitic fetus: A case report.

BACKGROUND: Fetus-in-fetu (FIF) is an extremely rare congenital abnormal mass, in which a normal fetus's vertebral axis frequently connected with malformed fetus around this axis. Here, we report the case of a male infant aged 26 d presenting with retroperitoneal parasitic fetus. CASE SUMMARY: In a prenatal examination, we first detected an abdominal mass measuring 7.8 cm × 5.1 cm × 6.8 cm in a mother's abdomen at 25 gestational weeks and teratoma was suspected. After the fetal was born, we did a magnetic resonance imaging (MRI) and ultrasonography on him and saw a distinctive limb with five-toes. According to the result of MRI, ultrasonography and postoperative pathology, he finally was diagnosed with FIF. CONCLUSION: A laparotomy was performed at 26 d of age with excision of the retroperitoneal cystic tumor, which measured about 10 cm in diameter. According to the result of imaging and histological test, FIF was confirmed.

IL-10 and IL-22 in Mucosal Immunity: Driving Protection and Pathology.

The barrier surfaces of the gastrointestinal tract are in constant contact with various microorganisms. Cytokines orchestrate the mucosal adaptive and innate immune cells in the defense against pathogens. IL-10 and IL-22 are the best studied members of the IL-10 family and play essential roles in maintaining mucosal homeostasis. IL-10 serves as an important regulator in preventing pro-inflammatory responses while IL-22 plays a protective role in tissue damage and contributes to pathology in certain settings. In this review, we focus on these two cytokines in the development of gastrointestinal diseases, including inflammatory bowel diseases (IBD) and colitis-associated cancer (CAC). We summarize the recent studies and try to gain a better understanding on how they regulate immune responses to maintain equilibrium under inflammatory conditions.

A modifier in the 129S2/SvPasCrl genome is responsible for the viability of Notch1[12f/12f] mice.

BACKGROUND: Mouse NOTCH1 carries a highly conserved O-fucose glycan at Thr466 in epidermal growth factor-like repeat 12 (EGF12) of the extracellular domain. O-Fucose at this site has been shown by X-ray crystallography to be recognized by both DLL4 and JAG1 Notch ligands. We previously showed that a Notch1 Thr466Ala mutant exhibits very little ligand-induced NOTCH1 signaling in a reporter assay, whereas a Thr466Ser mutation enables the transfer of O-fucose and reverts the NOTCH1 signaling defect. We subsequently generated a mutant mouse with the Thr466Ala mutation termed Notch1[12f](Notch1tm2Pst). Surprisingly, homozygous Notch1[12f/12f] mutants on a mixed background were viable and fertile. RESULTS: We now report that after backcrossing to C57BL/6 J mice for 11-15 generations, few homozygous Notch1[12f/12f] embryos were born. Timed mating showed that embryonic lethality occurred by embryonic day (E) ~E11.5, somewhat delayed compared to mice lacking Notch1 or Pofut1 (the O-fucosyltransferase that adds O-fucose to Notch receptors), which die at ~E9.5. The phenotype of C57BL/6 J Notch1[12f/12f] embryos was milder than mutants affected by loss of a canonical Notch pathway member, but disorganized vasculogenesis in the yolk sac, delayed somitogenesis and development were characteristic. In situ hybridization of Notch target genes Uncx4.1 and Dll3 or western blot analysis of NOTCH1 cleavage did not reveal significant differences at E9.5. However, qRT-PCR of head cDNA showed increased expression of Dll3, Uncx4.1 and Notch1 in E9.5 Notch1[12f/12f] embryos. Sequencing of cDNA from Notch1[12f/12f] embryo heads and Southern analysis showed that the Notch1[12f] locus was intact following backcrossing. We therefore looked for evidence of modifying gene(s) by crossing C57BL/6 J Notch1 [12f/+] mice to 129S2/SvPasCrl mice. Intercrosses of the F1 progeny gave viable F2 Notch1[12f/12f] mice. CONCLUSION: We conclude that the 129S2/SvPasCrl genome contains a dominant modifying gene that rescues the functions of NOTCH1[12f] in signaling. Identification of the modifying gene has the potential to illuminate novel factor(s) that promote Notch signaling when an O-fucose glycan is absent from EGF12 of NOTCH1.

Lunatic, Manic, and Radical Fringe Each Promote T and B Cell Development.

Lunatic, Manic, and Radical Fringe (LFNG, MFNG, and RFNG) are N-acetylglucosaminyltransferases that modify Notch receptors and regulate Notch signaling. Loss of LFNG affects thymic T cell development, and LFNG and MFNG are required for marginal zone (MZ) B cell development. However, roles for MFNG and RFNG in T cell development, RFNG in B cell development, or Fringes in T and B cell activation are not identified. In this study, we show that Lfng/Mfng/Rfng triple knockout (Fng tKO) mice exhibited reduced binding of DLL4 Notch ligand to CD4/CD8 double-negative (DN) T cell progenitors, and reduced expression of NOTCH1 targets Deltex1 and CD25. Fng tKO mice had reduced frequencies of DN1/cKit(+) and DN2 T cell progenitors and CD4(+)CD8(+) double-positive (DP) T cell precursors, but increased frequencies of CD4(+) and CD8(+) single-positive T cells in the thymus. In spleen, Fng tKO mice had reduced frequencies of CD4(+), CD8(+), central memory T cells and MZ B cells, and an increased frequency of effector memory T cells, neutrophils, follicular, and MZ P B cells. The Fng tKO phenotype was cell-autonomous and largely rescued in mice expressing one allele of a single Fng gene. Stimulation of Fng tKO splenocytes with anti-CD3/CD28 beads or LPS gave reduced proliferation compared with controls, and the generation of activated T cells by Concanavalin A or L-PHA was also reduced in Fng tKO mice. Therefore, each Fringe contributes to T and B cell development, and Fringe is required for optimal in vitro stimulation of T and B cells.

Opposite effects of high and low doses of interleukin-2 on T cell-mediated hepatitis in mice (interleukin-2 on hepatitis).

PURPOSE: Concanavalin A (Con A)-induced hepatitis is an extensively used animal model of T cell-mediated acute hepatitis. A variety of cytokines, including interleukin 4 (IL-4), interferon gamma (IFN-γ), and tumor necrosis factor alpha (TNF-α), have been shown to play important roles in Con A-induced liver injury. However, the role of IL-2, a critical cytokine in the development and function of T cells and a clinical therapeutics for virus infection and tumor, has not been carefully examined in this model. METHODS: In this study, we investigated the function of IL-2 in Con A-induced hepatitis by using various strategies of rhIL-2 pretreatment. We treated mice with two rhIL-2 administration strategies: a single injection of high dose of rhIL-2 (IL-2(hi), 50 × 10(3) U/mouse) and four injections of low dose of rhIL-2 (IL-2(4lo), 5 × 10(3) U/mouse). RESULTS: IL-2(hi) pretreatment ameliorated Con A-induced liver injury, while IL-2(4lo) aggravated Con A-induced liver injury. IL-2(hi) pretreatment reduced Con A-induced elevation of serum TNF-α while IL-2(4lo) pretreatment did not. Serum IL-4 and TNF-α were high 6 h after Con A injection in IL-2(4lo) mice, while it was undetectable in IL-2(hi) and non-pretreated mice. IL-2(hi) pretreatment reduced Con A-induced accumulation of T cells in liver while IL-2(4lo) pretreatment increased accumulation of NK cells. CONCLUSION: Various strategies of rhIL-2 administration play different roles in Con A-induced hepatitis, suggesting the importance of IL-2 administrative regime in clinical liver diseases.

CD4+ CD25+ Foxp3+ regulatory T cells protect against T cell-mediated fulminant hepatitis in a TGF-beta-dependent manner in mice.

Regulatory T cells (Tregs), which are characterized by expression of CD4, CD25, and Foxp3, play a crucial role in the control of immune responses to both self and non-self Ags. To date, there are only limited data on their role in physiological and pathological hepatic immune responses. In this study, we examined the role of hepatic Tregs in immune-mediated liver injury by using the murine Con A-induced hepatitis model. Con A treatment was associated with an increased number of Foxp3(+) Tregs in liver but not in spleen. Moreover, the expression levels of Foxp3, CTLA-4, glucocorticoid-induced TNF receptor, as well as the frequency of CD103 of Tregs were increased after Con A injection, being significantly higher in liver than in spleen. Depleting CD25(+) cells aggravated liver injury, whereas adoptively transferring CD25(+) cells or Tregs reduced liver injury in Con A-treated recipients. Con A treatment induced elevated serum levels and hepatic mononuclear mRNA expressions of TGF-beta, which were reduced by Tregs depletion. In addition, anti-TGF-beta mAbs blocked the suppressive function of Tregs from Con A-treated mice in vitro. Finally, TGF-beta receptor II dominant-negative mice, whose T cells express a dominant negative form of TGFbetaRII and therefore cannot respond to TGF-beta, had a higher mortality rate and severer liver injury than normal mice injected with the same dose of Con A. These results indicate that CD4(+)CD25(+) Tregs play an important role in limiting the liver injury in Con A-induced hepatitis via a TGF-beta-dependent mechanism.