OBJECTIVE To investigate the effects of calcitriol on sepsis-induced immunosuppression and its potential mechanism. METHODS A sepsis-induced immunosuppression mice model was established using cecal ligation and puncture （CLP）. The 7-day survival rate， serum levels of tumor necrosis factor- α （TNF- α）， interleukin-6 （IL-6）， and IL-1β were determined in sham operation group， CLP group and calcitriol group （1 μg/kg）； the morphological changes of lung tissue in mice were observed. Lipopolysaccharide （LPS） tolerance macrophage models （representing sepsis-induced immunosuppression） were established using mice macrophage cell line RAW264.7 cells. The levels of TNF-α and IL-6 in cell supernatants as well as mRNA expressions of IL-1β， nucleotide-binding domain leucine-rich repeat and pyrin domain-containing receptor 3 （NLRP3）， IL-18 and caspase-1 were assessed in culture medium group， LPS group， LPS tolerance group， and calcitriol （5 μmol/L） group. The following parameters were measured： propidium iodide （PI）-positive cell ratio， caspase-1 activity， lactate dehydrogenase （LDH） release， and Ca2+ levels. RESULTS Compared with CLP group， 7-day survival rate and serum levels of TNF-α， IL-6 and IL-1β were increased significantly in calcitriol group （P＜0.05）. Additionally， pulmonary tissue damage was markedly attenuated in calcitriol group. Compared with LPS tolerance group， the levels of TNF-α and IL-6 in cell supernatants， mRNA expressions of IL- 1β， NLRP3， IL-18 and caspase-1， PI-positive cell ratio， caspase-1 activity， LDH release， and Ca2+ levels were all increased significantly in calcitriol group （P＜0.05）. CONCLUSIONS Calcitriol can reverse sepsis-induced immunosuppression， and the mechanism of action may be E-mail：yarfwy@163.com achieved by the binding of calcitriol to vitamin D receptor，which promotes the release of Ca2+ from the endoplasmic reticulum， thereby driving the NLRP3/caspase-1-mediated pyroptosis pathway.

Objective:To observe the longitudinal changes of symptoms in the irradiation area of patients with postoperative radiotherapy for breast cancer,and investigate the correlative symptom indexes for predicting acute radiation dermatitis during radiotherapy.Methods:This study was designed as a prospective longitudinal study.A total of 103 patients with breast cancer who received three-dimensional con-formal radiotherapy in The Fourth Hospital of Hebei Medical University from May 2022 to December 2022 were enrolled using convenience and purposive sampling methods.The patients received a total of 50 Gy/25 radiotherapy sessions.We conducted weekly observations of five symptoms-itching,pain,swelling,burning,and tightness-in the irradiated area,for a total of six weeks.The incidence of symptoms associ-ated with radiation dermatitis across different severity levels was compared.We calculated the optimal number of symptoms for the occur-rence of grade 2 or higher radiation dermatitis,evaluated the predictive effect using the subject's work characteristic curve(ROC),and com-pared the risk of radiation dermatitis with the number of symptoms by applying a binary Logistic regression.Results:A total of 103 patients were included in the study.The total severity scores of symptoms were 0(0,0),0(0,0),0(0,1),1(0,2),2(1,3),3(2,4),respectively,show-ing a gradually increasing trend.From the 20 Gy/10-fraction radiotherapy point onward to the completion of radiotherapy,symptom scores exhibited statistically significant deviations from baseline values(P<0.001).From the beginning of 30 Gy/15 sessions to the end of radiother-apy,symptom scores were higher in patients with grade 2 and above dermatitis compared to those with dermatitis of less than grade 2(Z=2.12,2.81,4.08,P=0.034,0.005,0.001);at the end of 50Gy/25 sessions,the incidence rates of pruritus,pain,swelling,tightness,and burning were 68.9%(71 cases),68.9%(71 cases),46.6%(48 cases),36.9%(38 cases),and 15.5%(16 cases),respectively.At the 30 Gy/15 and 40 Gy/20 radiotherapy sessions,the optimal number of predicted symptoms were two and three,respectively.The corresponding areas under the ROC curve were 0.632 and 0.666.Sensitivity values were 48.6%and 43.2%,while specificity values reached 77.3%and 81.8%,re-spectively(95%CI:0.517-0.746,0.558-0.775,P=0.027,0.005).The subjects were assigned into low-risk and high-risk groups using the op-timal cut-off values,and binary Logistic regression showed that the risks of grade 2 and higher radiation dermatitis in the high-risk group were 3.39 and 3.58 times higher than that in the low-risk group(OR=3.388,95%CI:1.400-8.197,P=0.007;OR=3.584,95%CI:1.430-8.985,P=0.006).Conclusions:The symptoms of radiation dermatitis have appeared early and worsened with the severity of dermatitis.We should closely observe these symptoms throughout treatment.The risk of grade 2 and above radiation dermatitis increased when the number of symptoms was≥2 at 30 Gy/15 times and≥3 at 40 Gy/20 times.

Objective:To investigate the effects of anisodamine hydrobromide (654-1), 654-1+ norepinephrine and norepinephrine on early hemodynamic indexes of piglets with septic shock.Methods:A total of 38 healthy Bama pigs were selected as the study subjects, 32 of which were treated with lipopolysaccharide to create septic shock piglet model, and the other 6 were sham operation group. The animals were randomly divided into control group ( n=8), drug treatment group [654-1 group ( n=8), 654-1+ norepinephrine group ( n=8), norepinephrine group ( n=8)]. Hemodynamic parameters were recorded at T 0 (basic state), T 1 (successful shock modeling), T 2 (1 h after successful modeling), T 3 (2 h after successful modeling), T 4 (4 h after successful modeling), T 5 (6 h after successful modeling) and T 6 (8 h after successful modeling) respectively, including: Mean arterial pressure (MAP), cardiac index (CI), whole-heart end-diastolic volume index (GEDI), lactic acid (LAC). Results:Except for the sham operation group, MAP of all treatment groups at T 1 was significantly lower than that at T 0 (all P<0.05). MAP of all treatment groups at T 2-T 6 was significantly higher than that at T 1 (all P<0.05). T 1 MAP of all treatment groups was significantly lower than that of the sham operation group (all P<0.05). MAP at T 2-T 6 in the norepinephrine group and the 654-1+ norepinephrine group was higher than that in the control group (all P<0.05), and MAP at T 2-T 4 in the 654-1 group was significantly lower than that in the 654-1+ norepinephrine group (all P<0.05). LAC of all treatment groups at T 1-T 3 was significantly higher than that at T 0 (all P<0.05) except the sham operation group. LAC in the group 654-1 at T 4 to T 6 was significantly lower than that at T 1 (all P<0.05). LAC in the group 654-1 at T 4-T 6 was significantly lower than that in the norepinephrine group and the control group (all P<0.05). The CI of norepinephrine group at T 2, T 5 and T 6 was lower than that at T 0 (all P<0.05). There was no significant difference in CI between T 2 and T 6 compared with T 1 (all P>0.05). CI of the 654-1+ norepinephrine group at T 4 was significantly lower than that of T 0 ( P<0.05); The CI of the 654-1 group at T 2 was significantly higher than that of T 1 ( P<0.05). CI at T 1 in the 654-1+ norepinephrine group was significantly lower than that in the sham operation group (all P<0.05). The GEDI at T 1 to T 5 in the 654-1 group was significantly lower than that at T 0 in the 6541+ norepinephrine group (all P<0.05), and the GEDI at T 1 to T 2 was significantly lower than that at T 0 in the 6541+ norepinephrine group (all P<0.05), while the GEDI at T 2 and T 4 was higher than that at T 1 (all P<0.05). Conclusions:MAP decreased significantly in septic shock, LAC increased significantly in the early stage of shock. 654-1 can improve MAP in early stage of septic shock, and significantly reduce LAC level in early stage of septic shock.

Objective To investigate the anti-hepatocellular carcinoma mechanism of Chloranthus fortunei(A.Gray)Solms-Laub.via network pharmacology,molecular docking techniques and in vitro experiments.Methods Chemical composition of Chloranthus fortunei(A.Gray)Solms-Laub.was searched by literature.Swiss Target Prediction was used to find corresponding targets.STRING was used to construct protein-protein interactions network(PPI).DAVID was used to enrich GO analysis and KEGG pathway.AutoDock Vina 1.1.2 and Pymol visualisation was used for docking and validation.Results Chloranthus fortunei(A.Gray)Solms-Laub.had 61 active components,685 targets,and 279 intersections with disease targets.The PPI showed that the main active components were Luteolin,Chloranthalactone C,Shizukanolide H,Esculetin,7-Hydroxycoumarin.The key targets were GAPDH,VEGFA,STAT3,JUN,HSP90AA1,AKT1,CTNNB1,CASP3,and ALB.Biological process(BP)involved protein phosphorylation,signal transduction,regulation of RNA polymerase II promoter transcription,cell proliferation,apoptosis.Cellular component(CC)involved cytoplasm,nucleus,cell membrane,cellular exosome.Molecular function(MF)involves protein binding,ATPase,threonine kinase,protein kinase activity.KEGG involved cancer pathway,metabolic pathway,PI3K-Akt signalling pathway,cancer proteoglycans,lipids and atherosclerosis,cytomegalovirus infection,microRNAs in cancer,human T-cell leukaemia virus type 1,Ras signalling pathway,MAPK signalling pathway.Molecular docking showed that silverweed lactone H had a strong affinity for each of the other target proteins,indicating that this component plays a key role.The results of RT-qPCR assay and WB assay showed that there were significant differences in gene and protein expression levels before and after drug administration.Conclusion The Chinese medicine in Chloranthus fortunei(A.Gray)Solms-Laub.can treat hepatocellular carcinoma through the MAPK pathway,and the main active ingredients have good docking effects with the core target proteins of the disease.

Objective To analyze changes of preoperative T lymphocyte immunity and humoral immunity,and their relationship with pathological characteristics in patients with primary hepatocellular carcinoma(HCC).Methods A total of 150 patients with primary HCC underwent elective surgical treatment were enrolled as the HCC group,while 150 healthy controls during the same period were enrolled as the control group.Before surgery,the levels of peripheral blood CD3+,CD4+and CD8+were detected by flow cytometry,and CD4+/CD8+was calculated.The levels of IgA,IgG and IgM in peripheral blood were detected by immunoturbidimetry.The pathological characteristics of HCC patients were collected,and the differences of preoperative immune indexes in patients with different pathological characteristics were compared.The receiver operating curve(ROC)was used to evaluate the predictive value of immune indicators for pathological characteristics.Results The levels of CD3+,CD4+and CD4+/CD8+were lower in the HCC group than those in the control group,while levels of CD8+,IgA,IgG and IgM were higher in the HCC group than those in the control group(P＜0.05).The levels of CD3+,CD4+and CD4+/CD8+were lower in patients with lymph node metastasis than those of patients with distant lymph non-metastasis(P＜0.05).The levels of CD8+,IgA,IgG and IgM were higher in patients with lymph node metastasis than those of patients without metastasis(P＜0.05).The results of receiver operating characteristic(ROC)curves showed that area under the curve(AUC)values of CD3+,CD4+,CD8+,CD4+/CD8+,IgA,IgG,and IgM for predicting lymph node metastasis in HCC patients were 0.810,0.741,0.770,0.707,0.756,0.738 and 0.727.Conclusion There are abnormal changes of preoperative T lymphocyte immunity and humoral immunity in patients with primary HCC,which are closely related to lymph node metastasis.

Aimed to find a safe and effective drug to replace nitroimidazole drugs in aquaculture pro-duction for the prevention and treatment of Trichomoniasis in pigeons,which can improve the eco-nomic benefits of meat pigeon breeding and ensure food safety.Firstly,Trichomonas was isolated and cultured from the crop of diseased pigeons and identified.After stable passage,a quantitative method for in vitro detection of Trichomonas was established by combining an automated cell counter and quantitative real-time PCR technology.To prepared the drug,powders of Sophora fla-vescent and Philodendron were made into herbal water extracts(SFPA)and mixed with copper sulfate(CS)solution.Then added the drug to the culture medium of Trichomonas to determine the effective concentration of it.A total of 135 pairs each of Silver King and Mimas breeding pigeons in the same laying period were selected and randomly divided into three groups,with 6 replicates in each group and 15 pairs of breeding pigeons in each replicate.Four days before brooding,the three groups were fed with 200 mL of pure water,0.5 g/L metronidazole(MDZ)solution,and a mixed solution of 30 g/L SFPA and 0.5 g/L CS,respectively.The feeding experiment lasted for 26 d.Results showed that the mixed solution of SFPA and CS had a significant killing effect on Trichomonas in vitro(P＜0.05).Feeding the drug to breeding pigeons significantly reduced the in-fection rate of breeding pigeons by Trichomonas(P＜0.05).The drug had no significant effect on the serum biochemical indexes,antioxidant properties,immunoglobulin levels of breeding pigeons,the average cage weight,immune organ indexes,meat quality and slaughter performance of squabs(P＞0.05).The results suggested that adding SFPA and CS to pigeons can effectively prevent and treat Trichomoniasis and improve production performance.It can replace nitroimidazole drugs without affecting the immune level of breeding pigeons and the weight,immune level,slaughter performance and meat quality of squabs,thereby reduce drug residues in poultry products and en-hance the food safety.

Objective:To investigate the impact of postoperative complications on adverse outcomes following curative-intent resection for gallbladder cancer (GBC).Methods:The multi-center real-world study was conducted. The clinicopathological data of 629 patients with GBC, who were admitted to 14 medical centers including The First Affiliated Hospital of Army Medical University from the national multicenter database of Biliary Surgery Group of Elite Group of Chinese Journal of Digestive Surgery, from April 2020 to April 2024 were collected. There were 225 males and 404 females, aged (64±10)years. Patients underwent open curative-intent resection for GBC. Observation indicators: (1)surgery, postoperative complica-tions and adverse outcomes; (2) analysis of risk factors affecting postoperative adverse outcomes in patients and population attributable fraction (PAF). Missing data in predictor variables were addressed using multiple imputation with chained equations, while cases with missing outcome variables were addressed using the "multiple imputation then deletion (MID)" strategy. The severity of multicollinearity among independent variables was assessed using the variance inflation factor (VIF) test. Multivariable possion regression models with log link and robust error variance were construc-ted incorporating restricted cubic splines (3 knots) to address nonlinear relationships in continuous variables, calculating adjusted relative risk ( RR) with corresponding 95% confidence interval ( CI). Adjusted PAF was calculated for each imputed dataset using the AF package of R software, with subsequent pooling performed according to Rubin's rules. Results:(1) Surgery, postoperative complications and adverse outcomes. All 629 patients underwent curative-intent resection for GBC, of which 143 cases had postoperative complications, including 68 cases of intra-abdominal ascites, 39 cases of pulmonary infection, 21 cases of bile leakage, 12 cases of intra-abdominal hemorrhage, 11 cases of liver failure, 10 cases of pan-creatic fistula, 10 cases of wound infection, 10 cases of gastroparesis, 7 cases of cholangitis, 7 cases of sepsis. The same patient could have more than one kind of complication. Of 629 patients, there were 19 cases of postoperative 90-day death and 11 cases of missing data, 42 cases with post-operative 90-day reoperation and 7 cases with missing data, 44 cases with postoperative 90-day readmission and 3 cases with missing data, 155 cases with prolonged postoperative hospital stay and 3 cases with missing data. (2) Analysis of risk factors affecting the postoperative adverse outcomes in patients and PAF. Results of multivariate analysis showed that pulmonary infection and liver failure were independent risk factors for postoperative 90-day mortality ( RR=3.74, 12.15, 95% CI as 1.18-11.83, 1.98-74.48, P<0.05). Pulmonary infection demons-trated the highest PAF as 4.61% (95% CI as 3.94%-5.28%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, and intra-abdominal hemorrhage were independent risk factors for post-operative 90-day reoperation ( RR=4.80, 3.62, 3.46, 4.99, 95% CI as 2.49-9.26, 1.42-9.21, 1.34-8.92, 1.55-16.06, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 8.65% (95% CI as 8.22%-9.08%, P<0.05). Intra-abdominal ascites, bile leakage, and liver failure were independent risk factors for postoperative 90-day readmission ( RR=6.20, 3.33, 14.33, 95% CI as 3.21-11.95, 1.33-8.35, 3.72-55.28, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 9.11% (95% CI as 8.85%-9.37%, P<0.05). Intra-abdominal ascites, pulmonary infection, bile leakage, liver failure, and wound infection were independent risk factors for prolonged postoperative hospital stay ( RR=2.29, 2.21, 2.26, 2.14, 3.35, 95% CI as 1.63-3.23, 1.41-3.46, 1.32-3.86, 1.11-4.13, 1.70-6.60, P<0.05). Intra-abdominal ascites demonstrated the highest PAF as 6.03% (95% CI as 5.71%-6.35%, P<0.05). Conclusion:Pulmonary infection is the most significant risk factor for postoperative 90-day mortality after curative-intent resection for GBC, while intra-abdominal ascites is the most significant risk factor for postoperative 90-day reoperation, postoperative 90-day readmission, and prolonged postoperative hospital stay.

Multiple sclerosis(MS)is a complex autoimmune disorder characterized by inflammatory demyelination in the central nervous system.Prominent symptoms include damage to myelin sheaths in the brain,optic nerve,and spinal cord,as well as axonal dysfunction;however,the exact causes and mechanisms of MS remain unclear.Genetic and environmental factors are thought to interact via autoimmune mechanisms,potentially triggering the disease.Recent studies suggest that abnormal activation of the NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome may play a critical role in the pathogenesis of MS.In this context,this review summarizes the molecular mechanisms underlying NLRP3 activation and its connection to MS,considering relevant literature from the past decade.The findings aim to provide insights into the progression of MS and to identify potential therapeutic strategies by elucidating the underlying mechanisms.

CRM 197(cross-reacting material 197),a naturally occurring mutant of diphtheria toxin,is a safe and effective vaccine vector and extensively used on developing conjugate or combined vaccines.The mutant loses its enzymatic activity,but fully retains its receptor-binding ability and immunogenicity.In current work,the diphtheria toxin mutant CRM 197 and its fusion proteins with the receptor-binding do-main of botulinum neurotoxin serotype E(EHc)were developed using genetic engineering technology.These recombinant proteins were confirmed by Western blotting and SDS-PAGE.BALB/c mice were im-munized with the CRM197-EHc and EHc-CRM197 fusion proteins,and their immunogenicity was evalua-ted.These two fusion protein molecules,CRM197-EHc and EHc-CRM197,as subunit vaccines,elicited a robust humoral immune response targeting both CRM197 and EHc antigens in the immunized mice.Compared to the mixture of CRM197 and EHc,the mice vaccinated with the fusion proteins(CRM197-EHc and EHc-CRM197)induced higher levels of anti-CRM197 antibodies,and the mice vaccinated with EHc-CRM197 also generated strongest anti-EHc antibodies.Consequently,as a carrier molecule in the fusion protein vaccine,EHc enhances the immunogenicity of CRM197 molecules.Likewise,CRM197 boosts the immunogenicity of EHc in the EHc-CRM197 fusion protein.

Objective Piperazine derivatives are a group of emerging psychoactive substances with excitatory and hallucinogenic effects on the central nervous system.This study established a high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)screening method for the detection of 40 piperazine compounds in urine.Methods A 200 μL urine sample(spiked with an internal standard at 1 ng/mL)was subjected to liquid-liquid extraction with ethyl acetate.After nitrogen evaporation,the residue was redissolved in 200 μL methanol and injected for analysis.Separation was performed on a Waters Acquity UPLC? HSS T3 column(100 mm × 2.1 mm,1.8 μm).The mobile phase consisted of(A)20 mmol/L ammonium acetate buffer containing 0.1％formic acid and 5％acetonitrile,and(B)acetonitrile.Gradient elution was applied,and detection was carried out in multiple reaction monitoring(MRM)mode.Quantification was achieved using an internal standard calibration curve.Results The 40 piperazine substances demonstrated good linearity within the range of 1-50 ng/mL,with correlation coefficients of 0.995-0.998.The extraction recovery ranged from 51.51％to 104.1％.Intra-day precision was below 5％,while inter-day precision ranged from 1.61％to 10.17％.Accuracy was between-7.84％and 8.77％.The limits of detection were 0.2-1 ng/mL,and the limit of quantification was 1 ng/mL.Conclusion The proposed method requires only a small sample volume,exhibits high sensitivity,selectivity,and stability,and offers short run times.It is suitable for the qualitative and quantitative determination of piperazine derivatives in urine in forensic toxicology practice.