[Application of ultrasound-guided endoscopic retrograde appendicitis therapy in children with appendix-related chronic abdominal pain]. / è¶ å£°å¼•å¯¼å† é•œä¸‹é€†è¡Œé˜‘å°¾ç‚Žæ²»ç–—æœ¯åœ¨é˜‘å°¾ç›¸å ³æ ¢æ€§è ¹ç—›æ‚£å„¿ä¸­çš„åº”ç”¨ä»·å€¼.

OBJECTIVES: To study the clinical efficacy of ultrasound-guided endoscopic retrograde appendicitis therapy in children with appendix-related chronic abdominal pain. METHODS: A retrospective analysis was performed on the medical data of 30 children with the chief complaint of chronic abdominal pain who were admitted from August 2019 to May 2021. All the children were found to have inflammation of the appendix or intracavitary stool and fecalith by ultrasound and underwent ultrasound-guided endoscopic retrograde appendicitis therapy. The medical data for analysis included clinical manifestations, endoscopic findings, white blood cell count, neutrophil percentage, length of hospital stay, and cure rate. RESULTS: Among the 30 children with chronic abdominal pain, there were 13 boys (43%) and 17 girls (57%), with a mean age of (9±3) years (range 3-15 years) at diagnosis. The median duration of the disease was 12 months, and the median length of hospital stay was 3 days. The children had a median white blood cell count of 6.7×109/L and a neutrophil percentage of 50%±13%. Fecalith and a large amount of feces were flushed out of the appendix cavity for 21 children (70%) during surgery. The follow-up rate was 97% (29/30), and the median follow-up time was 11 months (range 5-26 months). Of the 29 children, abdominal pain completely disappeared in 27 children (93%). CONCLUSIONS: Ultrasound-guided endoscopic retrograde appendicitis therapy is effective in children with chronic abdominal pain caused by feces or fecalith in the appendix cavity.

Three New Truffle Species (Tuber, Tuberaceae, Pezizales, and Ascomycota) from Yunnan, China, and Multigen Phylogenetic Arrangement within the Melanosporum Group.

Based on a multi-locus phylogeny of a combined dataset of ITS, LSU, tef1-α, and rpb2 and comprehensive morphological analyses, we describe three new species from the Melanosporum group of genus Tuber and synonymize T. pseudobrumale and T. melanoexcavatum. Phylogenetically, the three newly described species, T. yunnanense, T. melanoumbilicatum and T. microexcavatum, differ significantly in genetic distance from any previously known species. Morphologically, T. yunnanense is distinctly different from its closest phylogenetically related species, T. longispinosum, due to its long shuttle-shape spores (average the ratio of spore length to spore width for all spores (Qm) = 1.74). Tuber melanoumbilicatum differs from the other species in having a cavity and long shuttle-shaped spores (Qm = 1.65). Although T. microexcavatum sampled ascomata have relatively low maturity, they can be distinguished from its closely related species T. pseudobrumale by the ascomata size, surface warts, and spore number per asci; additionally, phylogenetic analysis supports it as a new species. In addition, molecular analysis from 22 newly collected specimens and Genebank data indicate that T. pseudobrumale and T. melanoexcavatum are clustered into a single well-supported clade (Bootstrap (BS) = 100, posterior probabilities (PP) = 1.0); and morphological characteristics do not differ. Therefore, based on the above evidence and publication dates, we conclude that T. melanoexcavatum is a synonym of T. pseudobrumale. By taking into account current knowledge and combining the molecular, multigene phylogenetic clade arrangement and morphological data, we propose that the Melanosporum group should be divided into four subgroups. Diagnostic morphological features and an identification key of all known species in the Melanosporum group are also included. Finally, we also provide some additions to the knowledge of the characterization of T. pseudobrumale, T. variabilisporum, and T. pseudohimalayense included in subgroup 1 of the Melanosporum group.

[The extracellular segment of mouse soluble SIRPα enhances the phagocytosis of macrophages to L1210 leukemia cells].

Objective To study the role of mouse extracellular segment protein of signal-regulatory protein α gene (mSIRPαext) of mice in tumor immune regulation by cloning mSIRPαext, constructing its prokaryotic expression vector and achieving the soluble expression of SIRPα. Methods The mSIRPαext gene was amplified from mouse lymph node and the prokaryotic expression vector of pET32a-SIRPαext was further constructed. After the soluble expression of recombinant Trx-mSIRPαext fusion protein containing thioredoxin (Trx) tag was achieved, mouse bone marrow-derived monocytes were cultured and induced to differentiate into macrophages. Then the macrophages were co-cultured with L1210 leukemia cells labeled with 5(6)-carboxyfluorescein diacetate succinimide ester (CFSE). Trx-mSIRPαext protein and Trx control protein were added into the co-culture system. The role of recombinant protein in the macrophage phagocytosis of tumor cells was observed by immunofluorescence cytochemical staining and confocal microscopy. Results Purified soluble Trx-mSIRPαext protein was obtained and it was showed that it could enhance the phagocytosis of macrophages in mouse L1210 leukemia cells in vitro phagocytosis experiments. Conclusion Prokaryotic expression of Trx-mSIRPαext protein can effectively enhance the phagocytosis of macrophages in leukemia cells, thus playing the role of anti-tumor immunotherapy.

Effect of protein S-nitrosylation on autolysis and catalytic ability of µ-calpain.

The effect of S-nitrosylation on the autolysis and catalytic ability of µ-calpain in vitro in the presence of 50µM Ca(2 +) was investigated. µ-Calpain was incubated with different concentrations of nitric oxide donor S-nitrosoglutathione (GSNO) and subsequently reacted with purified myofibrils. Results showed that the amount of 80kDa µ-calpain subunit significantly decreased as GSNO increased from 0 to 300µM, but increases of GSNO to 300, 500 and 1000µM did not result in further inhibition. The catalytic ability of nitrosylated µ-calpain to degrade titin, nebulin, troponin-T and desmin was significantly reduced when the GSNO concentration was higher than 300µM. The cysteine residues of µ-calpain at positions 49, 351, 384, and 592 in the catalytic subunit and at 142 in small subunit were S-nitrosylated, which could be responsible for decreased µ-calpain activity. Thus, S-nitrosylation can negatively regulate the activation of µ-calpain resulting in decreased proteolytic ability on myofibrils.

[Expression and Activity Regulation of Indoleamine 2,3-Dioxygenase in Acute Myeloid Leukemia Cells].

OBJECTIVE: To investigate the expression and activity regulation of indoleamine 2,3-dioxygenase(IDO) in acute myeloid leukemia cells. METHODS: Expression of IDO and TLR9 in HL-60 and K562 cells cocultured with or without IFN-γ,Tα1,IFN-γ+Tα1 and chloroquine were determined by reverse transcription-polymerase chain reaction(RT-PCR). Then, the IDO activity in HL-60 and K562 cells cocultured with or without IFN-γ,Tα1,IFN-γ+ Tα1 was assayed by coomassie brilliant blue staining and modified colorimetric method. RESULTS: Both IDO and TLR9 mRNA were expressed in HL-60 and K562 cells; IFN-γ increased the expression and activity of IDO in a concentration-dependent manner; Tα1 decreased the expression and activity of IDO in a concentration-dependent manner; the up-regulation of IFN-γ on IDO induced expression and activity had been weakened by Tα1(P<0.01); Chloroquine had no effect on the expression of IDO. The expression of TLR9 in HL-60 cells and K562 cells cocultured with IFN-γ,Tα1,IFN-γ+Tα1 and chloroquine was not significantly changed. CONCLUSION: IDO can be expressed in acute myeloid leukemia cells and possesses the activity. IDO may play an important role in immune tolerance induced by leukemia cells, and become a new predictor of AML prognosis. Tα1 decreases the expression and activity of IDO, which can weaken the induction of IFN-γ on IDO expression and activity, thus Tα1 as an immune modulator may be a new agent for AML immunotherapy.

Activity and expression of nitric oxide synthase in pork skeletal muscles.

The objective of this study was to investigate the biochemical changes of nitric oxide synthase (NOS) in pork skeletal muscles during postmortem storage. Longissimus thoracis (LT), psoas major (PM) and semimembranosus (SM) muscles of pork were removed immediately after slaughter and stored under vacuum condition at 4°C for 0, 1 and 3d. Results showed that all three muscles exhibited NOS activity until 1d while SM muscle retained NOS activity after 3d of storage. The content of nNOS in SM muscle was stable across 3d of storage while decreased intensity of nNOS was detected at 1 and 3d of aging in PM and LT muscles due to the degradation of calpain. Immunostaining showed that nNOS was located at not only sarcolemma but also cytoplasm at 0 and 1d of storage. Our data suggest that postmortem muscles possess NOS activity and nNOS expression depends on muscle type.

Effect of nitric oxide on µ-calpain activation, protein proteolysis, and protein oxidation of pork during post-mortem aging.

The aim of the current research was to examine the influence of nitric oxide (NO) on calpain activation, protein proteolysis, and oxidation in post-mortem pork. Five longissimus muscles were removed from carcass after slaughter, and samples were incubated with water, nitric oxide synthase (NOS) inhibitor, or NO donor for 24 h at 4 °C. The samples were taken out and then stored under 4 °C for 1, 4, and 7 d. Results showed that autolysis of µ-calpain increased by incubation with NOS inhibitor after storage for 1 d (P<0.05). Degradation of titin and nebulin increased by treatment of NOS inhibitor among three treatments (P<0.05). Higher levels of protein oxidation were observed after samples incubated with NO donor than treatment of NOS inhibitor (P<0.05). These data indicated that NO could participate in regulating calpain activation and its proteolysis activity during post-mortem aging.

Suppressive effects of electrolyzed reduced water on alloxan-induced apoptosis and type 1 diabetes mellitus.

Electrolyzed reduced water, which is capable of scavenging reactive oxygen species, is attracting recent attention because it has shown improved efficacy against several types of diseases including diabetes mellitus. Alloxan produces reactive oxygen species and causes type 1 diabetes mellitus in experimental animals by irreversible oxidative damage to insulin-producing ß-cells. Here, we showed that electrolyzed reduced water prevented alloxan-induced DNA fragmentation and the production of cells in sub-G1 phase in HIT-T15 pancreatic ß-cells. Blood glucose levels in alloxan-induced type 1 diabetes model mice were also significantly suppressed by feeding the mice with electrolyzed reduced water. These results suggest that electrolyzed reduced water can prevent apoptosis of pancreatic ß-cells and the development of symptoms in type 1 diabetes model mice by alleviating the alloxan-derived generation of reactive oxygen species.