Objective To investigate the effects of the physical parameters of radiotherapy and intestinal flora on the risk of secondary radiation enteritis(RE)in patients with abdominal malignant tumors.Methods Ninety-eight patients with malignant abdominal tumors who were treated with radiotherapy from April 2020 to August 2023 at Anyang District Hospital in Puyang City were selected and assigned to the RE group(n=26)or the non-RE group(n=72).The clinical data,physical parameters of radiotherapy,and changes in the intes-tinal flora were compared between the two groups.Pearson's correlation analysis was used to determine the correlation between the phy-sical parameters of radiotherapy and changes in the intestinal flora,and logistic analysis was used to analyze the relationship between the physical parameters of radiotherapy,changes in the intestinal flora,and risk of RE.The receiver operating characteristic(ROC)curve and the area under the curve(AUC)were used to analyze the physical parameters of radiotherapy and changes in intestinal flora in predicting risk of RE.Results The doses of V20,V40,and D2cc in the small intestine,D2cc in the rectum,and D2cc in the colon were higher in the RE group than in the non-RE group(P＜0.05).The α diversity Chao1 index,Shannon index,and Simpson index of the intestinal flora in the RE group were lower than in the non-RE group,at the third week of radiotherapy,and the decrease in the α diversity Chao1 index,Shannon index,and Simpson index of the intestinal flora in the RE group was greater than in the non-RE group,before and after radiotherapy(P＜0.05).The doses of V20,V40,and D2cc in the small intestine,D2cc in the rectum,and D2cc in the colon were posi-tively correlated with decreases in the α diversity Chao 1 index,Shannon index,and Simpson index of the intestinal flora,before and after radiotherapy(P＜0.05).The logistic analysis showed that the decrease in V20,V40,and D2cc in the small intestine,D2cc in the rectum,D2cc in the colon,α diversity Chao1 index,Shannon index,and Simpson index of the intestinal flora were independent risk factors for occurrence of RE(P＜0.05).The AUC of the physical parameters of radiotherapy and changes in intestinal flora combined to predict the risk of RE as 0.920,which was greater than that of each indicator alone.Conclusion The physical parameters of radiotherapy are closely related to the changes of intestinal flora in patients with malignant abdominal tumors,which,in combination,can increase the risk of RE,and have a high predictive value for the risk of RE.

ObjectiveTo investigate the therapeutic effect of Baihe Wuyaotang （BWT） on non-alcoholic fatty liver disease （NAFLD） and elucidate its underlying mechanism. MethodC57BL/6J mice were randomly assigned to six groups： normal control， model， positive drug （pioglitazone hydrochloride 1.95×10^-3 g·kg^-1）， and low-， medium-， and high-dose BWT （1.3，2.5 and 5.1 g·kg^-1）. Following a 12-week high-fat diet （HFD） inducement， the mice underwent six weeks of therapeutic intervention with twice-daily drug administration. Body weight was monitored weekly throughout the treatment period. At the fifth week， glucose tolerance （GTT） and insulin tolerance （ITT） tests were conducted. Subsequently， the mice were euthanized for the collection of liver tissue and serum， and the subcutaneous adipose tissue （iWAT） and epididymal adipose tissue （eWAT） were weighed. Serum levels of total triglycerides （TG） and liver function indicators，such as alanine aminotransferase （ALT） and aspartate aminotransferase （AST）， were determined. Histological examinations， including oil red O staining， hematoxylin-eosin （HE） staining， Masson staining， and transmission electron microscopy， were performed to evaluate hepatic lipid deposition， pathological morphology， and ultrastructural changes， respectively. Meanwhile， Western blot and real-time quantitative polymerase chain reaction （Real-time PCR） were employed to analyze alterations， at both gene and protein levels， the insulin signaling pathway molecules， including insulin receptor substrate 1/2/protein kinase B/forkhead box gene O1 （IRS1/2/Akt/FoxO1）， glycogen synthesis enzymes phosphoenolpyruvate carboxy kinase （Pepck） and glucose-6-phosphatase （G6Pase）， lipid metabolism-related genes stearoyl-coA desaturase-1 （SCD-1） and carnitine palmitoyltransferase-1 （CPT-1）， fibrosis-associated molecules α-smooth muscle actin （α-SMA）， type Ⅰ collagen （CollagenⅠ）， and the fibrosis canonical signaling pathway transforming growth factor-β₁/drosophila mothers against decapentaplegic protein2/3（TGF-β₁/p-Smad/Smad2/3）， inflammatory factors such as interleukin（IL）-6， IL-8， IL-11， and IL-1β， autophagy markers LC3B Ⅱ/Ⅰ and p62/SQSTM1， and the expression of mammalian target of rapamycin （mTOR）. ResultCompared with the model group， BWT reduced the body weight and liver weight of NAFLD mice（P<0.05， P<0.01）， inhibited liver lipid accumulation， and reduced the weight of white fat： it reduced the weight of eWAT and iWAT（P<0.05， P<0.01） as well as the serum TG content（P<0.05， P<0.01）. BWT improved the liver function as reflected by the reduced ALT and AST content（P<0.05， P<0.01）. It improved liver insulin resistance by upregulating IRS2， p-Akt/Akt， p-FoxO1/FoxO1 expressions（P<0.05）. Besides， it improved glucose and lipid metabolism disorders： it reduced fasting blood glucose and postprandial blood glucose（P<0.05， P<0.01）， improved GTT and ITT（P<0.05， P<0.01）， reduced the expression of Pepck， G6Pase， and SCD-1（P<0.01）， and increased the expression of CPT-1（P<0.01）. The expressions of α-SMA， Collagen1， and TGF-β₁ proteins were down-regulated（P<0.05， P<0.01）， while the expression of p-Smad/Smad2/3 was downregulated（P<0.05）， suggesting BWT reduced liver fibrosis. BWT inhibited inflammation-related factors as it reduced the gene expression of IL-6， IL-8， IL-11 and IL-1β（P<0.01） and it enhanced autophagy by upregulating LC3B Ⅱ/Ⅰ expression（P<0.05）while downregulating the expression of p62/SQSTM1 and mTOR（P<0.05）. ConclusionBWT ameliorates NAFLD by multifaceted improvements， including improving IR and glucose and lipid metabolism， anti-inflammation， anti-fibrosis， and enhancing autophagy. In particular， BWT may enhance liver autophagy by inhibiting the mTOR-mediated signaling pathway.

Objective To determine the correlation between single-nucleotide polymorphisms of IL-17 with type 2 diabetes in Han population in southeastern Shanxi Provincein.Methods The basic information and related clinical data of the subjects were collected of normal healthy controls and T2DM.Whole blood DNA was extracted,and IL-17 polymorphisms(rs2275913,rs3819024,rs4711998 and rs8193036)were analyzed by using a polymerase chain reaction-high temperature ligase reaction.Results There was no significant difference in the genotype and allele frequency distribution of the four SNP loci of IL-17 gene between the two groups(P>0.05).IL-17 polymorphism was not linked with T2DM in multiple genetic models after controlling for age,BMI,WC,and dyslipidemia(P>0.05).Further analysis showed that the levels of AA genotype was substantially lower of FPG and HOMA-IR levels when compared with the AG and GG genotypes,and the differences among the three groups were statistically significant(P<0.05).However,BMI,FINS,HbA1c and HOMA-βwas no statistical significance among the three groups(P>0.05).Besides,WC and HOMA-IR of AA genotype(rs3819024)were notable higher than those of GG genotype in T2DM group(P<0.05).Conclusions The IL-17 gene polymorphisms rs2275913,rs3819024,rs4711998,and rs8193036 in the Han population of southeast Shanxi Province may not be associated with T2DM.In this region Han T2DM patients,AA genotype at rs2275913 of the IL-17 gene is associ-ated to FPG and HOMA-IR,while GG genotype at rs3819024 is related to WC and HOMA-IR,which could be the potential genotype of IL-17 impacting glucose metabolism.

Objective:To establish an antibody expression system to reduce the antibody-dependent enhancement (ADE) effect of target antibody.Methods:Site-directed mutagenesis was used to mutate the 234 and 235 sites of the Fc region of the mammalian cell antibody expression vector-L234A and L235A to establish the antibody expression vector pFRT-IgG1κ-FcM. An antibody Wt-WNV with significant ADE effect obtained in previous work was selected and expressed by the pFRT-IgG1κ-FcM system to obtain mutant antibody FcM-WNV. The binding ability of FcM-WNV to target antigen West Nile virus envelope protein-DⅢ (WNV E-DⅢ) was detected by ELISA, and the its binding ability to human high-affinity IgG Fc receptor hFcγRⅠ (hCD64 ) was analyzed by flow cytometry. The neutralizing activity of FcM-WNV in vitro was detected by pseudovirus infection of host cells (BHK21 and K562). Results:The expression levels of FcM-WNV and Wt-WNV were comparable, and FcM-WNV could recognize and bind to WNVE-DIII in a concentration-dependent manner. Compared with Wt-WNV, the binding ability of FcM-WNV to hCD64 was significantly weakened, showing a significant decrease in fluorescence intensity. Consistent with the previous experimental results, Wt-WNV at a concentration of 5 μg/ml significantly enhanced the infection of K562 by WNV pseudovirus, while FcM-WNV at a concentration of 5 μg/ml could effectively block pseudovirus infection in both K562 and BHK21 cells.Conclusions:The established antibody expression system can effectively reduce the ADE effect of the target antibody.

Objective To retrieve and evaluate relevant literatures on pain management for patients after hemorrhoidectomy and summarize the best evidence. Methods Based on the 6S evidence model, computer-based searches were conducted in relevant domestic and foreign databases for literatures related to pain after hemorrhoidectomy, including guidelines, expert consensus, systematic reviews, meta-analysis, and randomized controlled trial (RCT), from the establishment of the databases to March 12, 2023. The quality of evidence meeting the evaluation criteria was evaluated according to the corresponding literature evaluation criteria, and was summarized in combination with expert opinions. Results A total of 16 literatures were included (1 guideline, 6 expert consensus, 8 systematic reviews, and 1 RCT); the evidence included pain identification and assessment, drug management measures, traditional Chinese medicine therapy, and perioperative health education, eventually 21 pieces of evidence were extracted. Conclusion After hemorrhoidectomy, the most effective pain management plan should be developed based on the patient's condition and clinical status to relieve postoperative pain.

Hepatocellular carcinoma (HCC) is the most common primary hepatic malign-ancy in clinic. The prognosis of patients remains extremely poor because of the high malignancy and easy recurrence and metastasis of HCC. In recent years, the roles of platelets in promoting the malignant progression of HCC have increasingly attracted much attention. It is known that platelets could promote HCC cells proliferation and invasion through tumor microenvironment. On the other hand, platelets are capable to promote HCC cells distant metastasis by facilitating tumor cells evasion of immune surveillance. Besides, the platelet-derived growth factors and proangiogenic factors are also involved in the proliferation, invasiveness, and neovascularization of HCC. In addition, patients with HCC normally have a background of cirrhosis, and it is still controversial that whether the thrombocytopenia by portal hypertension and hypersplenism can promote the malignant progress of HCC. In view of this, the diagnostic and prognostic value of platelet levels, as well as platelet-associated scores in HCC have increasingly become research focus. The authors elaborate the detailed mechanisms of platelets in malignant progression of HCC, and discuss the recent research progress of platelets as effective diagnostic or prognostic tools for the assessment of HCC, which is of great importance to optimize the current treatment regimen and explore novel therapeutic strategies against HCC.

Objective:To establish an in vivo infection model of H5N1 pseudovirus and to detect the neutralizing activity of FHA3 antibody using this model. Methods:Based on the sequence information of hemagglutinin (HA) and neuraminidase (NA) of A/Anhui/1/2005/H5N1 strain, two recombinant plasmids of pcDNA3.1-HA5 and pcDNA3.1-NA1 were constructed. The two plasmids and plasmid pNL4-3.Luc.R-E- were co-transfected into 293T cells to prepare H5N1 pseudovirus supernatant. The morphology of pseudovirus particles in the supernatant was observed by electron microscopy. MDCK cells were infected with the pseudovirus supernatant and the virus titer was detected. BALB/c mice were injected with the pseudovirus supernatant by intraperitoneal injection and subjected to bioluminescence imaging at 2, 5, 8, and 12 d after infection to detect the pseudovirus infection in vivo. The functional activity of FHA3 antibody in vivo was evaluated using the established mouse infection model. Results:The recombinant plasmids pcDNA3.1-HA5 and pcDNA3.1-NA1 were correctly constructed and could be used to prepare pseudovirus supernatants of high titer by co-transfecting 293T cells with the plasmid pNL4-3.Luc.R-E-. The virus particles were round under electron microscope. H5N1 pseudovirus-infected mice exhibits strong fluorescence signals, which were attenuated by FHA3 treatment before challenge.Conclusions:The in vivo infection model of H5N1 pseudovirus was successfully constructed and FHA3 antibody was proved to be protective against the pseudovirus infection.

Objective:To explore the effects of accelerated rehabilitation surgery (ERAS) on visual analogue scale (VAS) and serum immunoglobulin in children undergoing low-temperature plasma radiofrequency tonsillectomy.Methods:A prospective study was conducted on 200 children who underwent bilateral tonsillectomy with low-temperature plasma radiofrequency from January 2022 to April 2023 at the Sixth Affiliated Hospital of South China University of Technology. They were randomly divided into an observation group ( n=100) and a control group ( n=100) using a random number table method. The control group received traditional perioperative treatment, while the observation group received perioperative treatment under ERAS mode. We compared the postoperative conditions, pain VAS at different time points, and incidence of complications between two groups, as well as serum immunoglobulins and inflammatory markers before and 2 days after surgery. Results:The observation group had shorter postoperative pseudo film detachment time time, white membrane formation time, dietary recovery time, and hospital stay than the control group (all P<0.05); The VAS of the observation group was lower than that of the control group at different time points after surgery (all P<0.05); Two days after surgery, the levels of immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) in both groups decreased, with the observation group showing a smaller decrease than the control group (all P<0.05); Two days after surgery, the levels of interleukin-8 (IL-8), interleukin-6 (IL-6), and C-reactive protein (CRP) significantly increased in both groups, with the observation group showing a lower increase than the control group (all P<0.05); The total incidence of complications in the observation group was 5.00%(5/100), significantly lower than the control group′s 13.00%(13/100, P<0.05). Conclusions:After undergoing ERAS intervention during the perioperative period, children undergoing low-temperature plasma tonsillectomy can accelerate their recovery process, shorten hospital stay, reduce pain VAS and inflammatory indicators, and reduce the impact on their serum immunoglobulin levels.

Aging-induced changes in the immune system are associated with a higher incidence of infection and vaccination failure. Lymph nodes, which filter the lymph to identify and fight infections, play a central role in this process. However, careful characterization of the impact of aging on lymph nodes and associated autoimmune diseases is lacking. We combined single-cell RNA sequencing (scRNA-seq) with flow cytometry to delineate the immune cell atlas of cervical draining lymph nodes (CDLNs) of both young and old mice with or without experimental autoimmune uveitis (EAU). We found extensive and complicated changes in the cellular constituents of CDLNs during aging. When confronted with autoimmune challenges, old mice developed milder EAU compared to young mice. Within this EAU process, we highlighted that the pathogenicity of T helper 17 cells (Th17) was dampened, as shown by reduced GM-CSF secretion in old mice. The mitigated secretion of GM-CSF contributed to alleviation of IL-23 secretion by antigen-presenting cells (APCs) and may, in turn, weaken APCs' effects on facilitating the pathogenicity of Th17 cells. Meanwhile, our study further unveiled that aging downregulated GM-CSF secretion through reducing both the transcript and protein levels of IL-23R in Th17 cells from CDLNs. Overall, aging altered immune cell responses, especially through toning down Th17 cells, counteracting EAU challenge in old mice.
Aging ; Animals ; Autoimmune Diseases ; Disease Models, Animal ; Granulocyte-Macrophage Colony-Stimulating Factor/metabolism* ; Mice ; Mice, Inbred C57BL ; Th17 Cells/metabolism* ; Uveitis/pathology* ; Virulence

Objective:To prepare and identify a broad-spectrum antibody FHA3 targeting influenza A virus hemagglutinin (HA).Methods:According to the single-chain antibody fragment (scFv) sequence, the heavy chain (VH) and light chain (VL) variable regions of FHA3 were amplified by PCR and a recombinant plasmid pFRT-IgG1κ-FHA3 was constructed by linking the expression vector pFRT-IgG1κ. FHA3 was expressed in the ExpiCHO system and purified by affinity purification. The binding activity of FHA3 to influenza A virus HA was detected by ELISA. The neutralizing activity of FHA3 was detected in vitro by infecting host cells with pseudovirus. Results:SDS-PAGE showed that high-purity FHA3 was obtained. FHA3 could bind to H1N1 HA, H2N2 HA, H3N2 HA, H5N1 HA, H7N9 HA and H9N2 HA in a concentration-dependent manner. FHA3 had good neutralizing activity in vitro that was it could effectively block the invasion of H5N1 and H7N9 pseudoviruses into target cells at a low concentration of 5 μg/ml and H1N1 pseudovirus at 0.012 5 μg/ml. Conclusions:A broad spectrum antibody targeting HA protein of influenza A virus with neutralizing activity in vitro was obtained.