Expression characteristics of peripheral lymphocyte programmed death 1 and FoxP3+ Tregs in gastric cancer during surgery and chemotherapy

BACKGROUND Programmed death 1(PD-1)and CD4^(+)CD25^(+)FoxP3^(+)expression in peripheral blood T-cells has been previously reported in various types of cancer.However,the specific variation tendency during surgery and chemotherapy,as well as their relationship in gastric cancer patients,still remain unclear.Understanding this aspect may provide some novel insights for future studies on tumor recurrence and tumor immune escape,and also serve as a reference for determining the optimal timing and dose of clinical anti-PD-1 antibodies.AIM To observe and analyze the expression characteristics of peripheral lymphocyte PD-1 and FoxP3^(+)regulatory T cells(FoxP3^(+)Tregs)before and after surgery or chemotherapy in gastric cancer patients.METHODS Twenty-nine stomach cancer patients undergoing chemotherapy after a D2 gastrectomy provided 10 mL peripheral blood samples at each phase of the perioperative period and during chemotherapy.This study also included 29 agematched healthy donors as a control group.PD-1 expression was detected on lymphocytes,including CD4^(+)CD8^(+)CD45RO^(+),CD4^(+)CD45RO^(+),and CD8^(+)CD45RO^(+)lymphocytes as well as regulatory T cells.RESULTS We observed a significant increase of PD-1 expression on immune subsets and a larger number of FoxP3^(+)Tregs in gastric cancer patients(P<0.05).Following D2 gastrectomy,peripheral lymphocytes PD-1 expression and the number of FoxP3^(+)Tregs notably decrease(P<0.05).However,during postoperative chemotherapy,we only observed a decrease in PD-1 expression on lymphocytes in the CD8^(+)CD45RO^(+)and CD8^(+)CD45RO^(+)populations.Additionally,linear correlation analysis indicated a positive correlation between PD-1 expression and the number of CD4^(+)CD45RO^(+)FoxP3high activated Tregs(aTregs)on the total peripheral lymphocytes(r=0.5622,P<0.0001).CONCLUSION The observed alterations in PD-1 expression and the activation of regulatory T cells during gastric cancer treatment may offer novel insights for future investigations into tumor immune evasion and the clinical application of anti-PD-1 antibodies in gastric cancer.

Single-cell transcriptome profiling of sepsis identifies HLA-DR^(low)S100A^(high)monocytes with immunosuppressive function

Background Sustained yet intractable immunosuppression is commonly observed in septic patients,resulting in aggravated clinical outcomes.However,due to the substantial heterogeneity within septic patients,precise indicators in deciphering clinical trajectories and immunological alterations for septic patients remain largely lacking.Methods We adopted cross-species,single-cell RNA sequencing(scRNA-seq)analysis based on two published datasets containing circulating immune cell profile of septic patients as well as immune cell atlas of murine model of sepsis.Flow cytometry,laser scanning confocal microscopy(LSCM)imaging and Western blotting were applied to identify the presence of S100A9^(+)monocytes at protein level.To interrogate the immunosuppressive function of this subset,splenic monocytes isolated from septic wild-type or S100a9^(–/–)mice were co-cultured with naive CD4^(+)T cells,followed by proliferative assay.Pharmacological inhibition of S100A9 was implemented using Paquinimod via oral gavage.Results scRNA-seq analysis of human sepsis revealed substantial heterogeneity in monocyte compartments following the onset of sepsis,for which distinct monocyte subsets were enriched in disparate subclusters of septic patients.We identified a unique monocyte subset characterized by high expression of S100A family genes and low expression of human leukocyte antigen DR(HLA-DR),which were prominently enriched in septic patients and might exert immunosuppressive function.By combining single-cell transcriptomics of murine model of sepsis with in vivo experiments,we uncovered a similar subtype of monocyte significantly associated with late sepsis and immunocompromised status of septic mice,corresponding to HLA-DR^(low)S100A^(high)monocytes in human sepsis.Moreover,we found that S100A9^(+)monocytes exhibited profound immunosuppressive function on CD4^(+)T cell immune response and blockade of S100A9 using Paquinimod could partially reverse sepsis-induced immunosuppression.Conclusions This study identifies HLA-DR^(low)S100A^(high)monocytes correlated with immunosuppressive state upon septic challenge,inhibition of which can markedly mitigate sepsis-induced immune depression,thereby providing a novel therapeutic strategy for the management of sepsis.

Repetitive transcranial magnetic stimulation promotes neurological functional recovery in rats with traumatic brain injury by upregulating synaptic plasticity-related proteins

Studies have shown that repetitive transcra nial magnetic stimulation(rTMS)can enhance synaptic plasticity and improve neurological dysfunction.Howeve r,the mechanism through which rTMS can improve moderate traumatic brain injury remains poorly understood.In this study,we established rat models of moderate traumatic brain injury using Feeney's weight-dropping method and treated them using rTMS.To help determine the mechanism of action,we measured levels of seve ral impo rtant brain activity-related proteins and their mRNA.On the injured side of the brain,we found that rTMS increased the protein levels and mRNA expression of brain-derived neurotrophic factor,tropomyosin receptor kinase B,N-methyl-D-aspartic acid receptor 1,and phosphorylated cAMP response element binding protein,which are closely associated with the occurrence of long-term potentiation.rTMS also partially reve rsed the loss of synaptophysin after injury and promoted the remodeling of synaptic ultrastructure.These findings suggest that upregulation of synaptic plasticity-related protein expression is the mechanism through which rTMS promotes neurological function recovery after moderate traumatic brain injury.

Multidisciplinary discussion and management of synchronous colorectal liver metastases: A single center study in China

BACKGROUND The multidisciplinary team(MDT)has been carried out in many large hospitals now.However,given the costs of time and money and with little strong evidence of MDT effectiveness being reported,critiques of MDTs persist.AIM To evaluate the effects of MDTs on patients with synchronous colorectal liver metastases and share our opinion on management of synchronous colorectal liver metastases.METHODS In this study we collected clinical data of patients with synchronous colorectal liver metastases from February 2014 to February 2017 in the Chinese People’s Liberation Army General Hospital and subsequently divided them into an MDT+group and an MDT-group.In total,93 patients in MDT+group and 169 patients in MDT-group were included totally.RESULTS Statistical increases in the rate of chest computed tomography examination(P=0.001),abdomen magnetic resonance imaging examination(P=0.000),and preoperative image staging(P=0.0000)were observed in patients in MDT+group.Additionally,the proportion of patients receiving chemotherapy(P=0.019)and curative resection(P=0.042)was also higher in MDT+group.Multivariable analysis showed that the population of patients assessed by MDT meetings had higher 1-year[hazard ratio(HR)=0.608,95%confidence interval(CI):0.398-0.931,P=0.022]and 5-year(HR=0.694,95%CI:0.515-0.937,P=0.017)overall survival.CONCLUSION These results proved that MDT management did bring patients with synchronous colorectal liver metastases more opportunities for comprehensive examination and treatment,resulting in better outcomes.

Potent bromodomain and extraterminal domain inhibitor JAB-8263 suppresses MYC expression and exerts anti-tumor activity in colorectal cancer models

BACKGROUND The overexpression of the MYC gene plays an important role in the occurrence,development and evolution of colorectal cancer(CRC).Bromodomain and extraterminal domain(BET)inhibitors can decrease the function BET by recognizing acetylated lysine residues,thereby downregulating the expression of MYC.AIM To investigate the inhibitory effect and mechanism of a BET inhibitor on CRC cells.METHODS The effect of the BET inhibitor JAB-8263 on the proliferation of various CRC cell lines was studied by CellTiter-Glo method and colony formation assay.The effect of JAB-8263 on the cell cycle and apoptosis of CRC cells was studied by propidium iodide staining and Annexin V/propidium iodide flow assay,respectively.The effect of JAB-8263 on the expression of c-MYC,p21 and p16 in CRC cells was detected by western blotting assay.The anti-tumor effect of JAB-8263 on CRC cells in vivo and evaluation of the safety of the compound was predicted by constructing a CRC cell animal tumor model.RESULTS JAB-8263 dose-dependently suppressed CRC cell proliferation and colony formation in vitro.The MYC signaling pathway was dose-dependently inhibited by JAB-8263 in human CRC cell lines.JAB-8263 dose-dependently induced cell cycle arrest and apoptosis in the MC38 cell line.SW837 xenograft model was treated with JAB-8263(0.3 mg/kg for 29 d),and the average tumor volume was significantly decreased compared to the vehicle control group(P<0.001).The MC38 syngeneic murine model was treated with JAB-8263(0.2 mg/kg for 29 d),and the average tumor volume was significantly decreased compared to the vehicle control group(P=0.003).CONCLUSION BET could be a potential effective drug target for suppressing CRC growth,and the BET inhibitor JAB-8263 can effectively suppress c-MYC expression and exert anti-tumor activity in CRC models.

Artificial intelligence in small intestinal diseases:Application and prospects

The small intestine is located in the middle of the gastrointestinal tract,so small intestinal diseases are more difficult to diagnose than other gastrointestinal diseases.However,with the extensive application of artificial intelligence in the field of small intestinal diseases,with its efficient learning capacities and computational power,artificial intelligence plays an important role in the auxiliary diagnosis and prognosis prediction based on the capsule endoscopy and other examination methods,which improves the accuracy of diagnosis and prediction and reduces the workload of doctors.In this review,a comprehensive retrieval was performed on articles published up to October 2020 from PubMed and other databases.Thereby the application status of artificial intelligence in small intestinal diseases was systematically introduced,and the challenges and prospects in this field were also analyzed.

Automatic counting of retinal ganglion cells in the entire mouse retina based on improved YOLOv5

Glaucoma is characterized by the progressive loss of retinal ganglion cells (RGCs),although the pathogenic mechanism remains largely unknown.To study the mechanism and assess RGC degradation,mouse models are often used to simulate human glaucoma and specific markers are used to label and quantify RGCs.However,manually counting RGCs is time-consuming and prone to distortion due to subjective bias.Furthermore,semi-automated counting methods can produce significant differences due to different parameters,thereby failing objective evaluation.Here,to improve counting accuracy and efficiency,we developed an automated algorithm based on the improved YOLOv5 model,which uses five channels instead of one,with a squeeze-and-excitation block added.The complete number of RGCs in an intact mouse retina was obtained by dividing the retina into small overlapping areas and counting,and then merging the divided areas using a non-maximum suppression algorithm.The automated quantification results showed very strong correlation (mean Pearson correlation coefficient of 0.993) with manual counting.Importantly,the model achieved an average precision of 0.981.Furthermore,the graphics processing unit (GPU) calculation time for each retina was less than 1 min.The developed software has been uploaded online as a free and convenient tool for studies using mouse models of glaucoma,which should help elucidate disease pathogenesis and potential therapeutics.

Relationship between Ki-67 and CD44 expression and microvascular formation in gastric stromal tumor tissues

BACKGROUND A gastric stromal tumor(GST)is a mesenchymal tumor that occurs in the gastrointestinal tract;its biological characteristics are highly complex.Clinically,the severity of a GST is often evaluated by factors such as risk classification,tumor size,and mitotic figures.However,these indicators are not very accurate.Even patients classified as low risk are also at risk of metastasis and recurrence.Therefore,more accurate and objective clinical biological behavior evaluations are urgently needed.AIM To determine the relationship between Ki-67 and CD44 expression in GSTs and microvessel formation and prognosis.METHODS Eighty-six GST tissue specimens from our hospital were selected for this study.The immunohistochemical staining technique was used to detect Ki-67,CD44,and microvessel density(MVD)in the collected samples to analyze the different risk grades and mitotic figures.In addition,this approach was used to determine the differences in the expression of Ki-67 and CD44 in GST tissues with varying lesion diameters.RESULTS In GSTs with positive expression of the Ki-67 protein,the proportions of patients with medium-to-high risk and more than five mitotic counts were 24.07%and 38.89%,respectively.In GSTs with positive expression of the CD44 protein,the proportions of patients with medium-to-high risk and more than five mitotic counts were 23.73%and 38.98%,respectively.In GSTs with negative expression of the Ki-67 protein,these values were relatively high(3.70%and 11.11%,respectively).The MVD in GSTs with positive and negative expression of the CD44 protein was 15.92±2.94 and 13.86±2.98/Hp,respectively;the difference between the two groups was significant(P<0.05).CONCLUSION Ki-67 and CD44 expression in GSTs is correlated with the grade of tumor risk and mitotic figures.CD44 expression is correlated with microvessel formation in tumor tissues.

Inhibition of the Hedgehog Signaling Pathway Depresses the Cigarette Smoke-Induced Malignant Transformation of 16HBE Cells on a Microfluidic Chip

Background：The hedgehog signaling system （HHS） plays an important role in the regulation of cell proliferation and differentiation during the embryonic phases.However,little is known about the involvement of HHS in the malignant transformation of cells.This study aimed to detect the role of HHS in the malignant transformation of human bronchial epithelial （16HBE） cells.Methods：In this study,two microfluidic chips were designed to investigate cigarette smoke extract （CSE）-induced malignant transformation of cells.Chip A contained a concentration gradient generator,while chip B had four cell chambers with a central channel.The 16HBE cells cultured in chip A were used to determine the optimal concentration of CSE for inducing malignant transformation.The 16HBE cells in chip B were cultured with 12.25% CSE （Group A）,12.25% CSE ± 5 μmol/L cyclopamine （Group B）,or normal complete medium as control for 8 months （Group C）,to establish the in vitro lung inflammatory-cancer transformation model.The transformed cells were inoculated into 20 nude mice as cells alone （Group 1） or cells with cyclopamine （Group 2） for tumorigenesis testing.Expression of HHS proteins was detected by Western blot.Data were expressed as mean ± standard deviation.The t-test was used for paired samples,and the difference among groups was analyzed using a one-way analysis of variance.Results：The optimal concentration of CSE was 12.25%.Expression of HHS proteins increased during the process of malignant transformation （Group B vs.Group A,F =7.65,P 〈 0.05）.After CSE exposure for 8 months,there were significant changes in cellular morphology,which allowed the transformed cells to grow into tumors in 40 days after being inoculated into nude mice.Cyclopamine could effectively depress the expression of HHS proteins （Group C vs.Group B,F =6.47,P 〈 0.05） and prevent tumor growth in nude mice （Group 2 vs.Group 1,t=31.59,P〈 0.01）.Conclusions：The activity of HHS is upregulated during the CSE-induced malignant transformation of 16HBE cells.Cyclopamine can effectively depress expression of HHS proteins in vitro and prevent tumor growth of the transformed cells in vivo.