ObjectiveTo explore the potential mechanism of Weicanqing Formula （微残清方， WF） combined with chemotherapy in treating acute myeloid leukemia （AML） based on malic enzyme 2 （ME2）-mediated bone marrow immune-metabolic homeostasis and . MethodsSixty-five male C57BL/6N mice were randomly divided into control group， model group， WF group， chemotherapy group， and the combination group （WF plus chemotherapy）， with 13 mice per group. Except for the control group， mice were injected with C1498 cells （appro-ximately 1×10⁶ cells per mouse） via the tail vein on day 1 to establish an AML model. Starting from day 8， mice in the chemotherapy group and the combination group were administered with cytarabine at 30 mg/（kg·d） via subcutaneous injection， once daily for 3 consecutive days； WF group and the combination group received WF at 7.54 g/（kg·d） by gavage， once daily for 21 consecutive days. On days 1， 7， 14， 21 and 28， hemoglobin （Hb） and white blood cell （WBC） levels were determined in each group. On day 28， Giemsa staining was used to observe morphological changes in bone marrow cells. Flow cytometry was employed to detect the expression levels of bone marrow T lympthocyte subsets， including CD8⁺， CD4⁺， and regulatory T lymphocytes （Treg）. The contents of glutamine （Gln）， nicotinamide adenine dinucleotide phosphate （NADP⁺）， and reduced nicotinamide adenine dinucleotide phosphate （NADPH） in bone marrow were determined using visible spectrophotometry. Adenosine triphosphate （ATP） concentration in bone marrow was measured by chemiluminescence assay. The mRNA expression of malic enzyme 2 （ME2） in bone marrow was detected by RT-qPCR， and the protein expression level of ME2 was determined by Western blotting. ResultsCompared to the control group， the model group showed decreased Hb levels on day 14， 21， and 28， and increased WBC levels on day 7， 14， 21， and 28 （P＜0.05）. On day 28， bone marrow CD8⁺ and CD8⁺/CD4⁺ levels decreased， while Treg cells， ATP， Gln， NADP⁺， NADPH， ME2 mRNA， and their corresponding protein levels increased （P＜0.05）. Compared to the model group， the chemotherapy group and the combination group both exhibited reduced Hb level on day 14， 21 and 28， as well as the increased WBC level； the combination group showed increased CD8⁺ level， decreased Treg， ATP， Gln， and NADPH content， as well as reduced ME2 mRNA expression and protein levels （P＜0.05）. Compared to the chemotherapy group， the combination group showed significantly increased Hb level on days 21 and 28， and increased WBC level on day 28 （P＜0.05）. Bone marrow smear pathology revealed that， in the model group， myeloid blast cells exhibited cytoplasmic vacuoles indicative of abnormal metabolic activity. In contrast， both the chemotherapy group and the combination group showed large numbers of metamyelocytes and band neutrophils， with only a few immature granulocytic blast cells observed. ConclusionWF may improve the prognosis of AML when used as an adjunct to chemotherapy by modulating ME2 expression， inhibiting metabolic energy competition within the bone marrow microenvironment， and reshaping the distribution of T lymphocyte subsets in the bone marrow.

Objective To compare the colonoscopy results of patients with positive fecal SDC2,ADHFE1,and PPP2R5C gene methylation tests to those with positive fecal occult blood tests,and analyze the effectiveness of colorectal cancer(CRC)screening.This study aims to provide a scientific basis for risk assessment in CRC screening.Methods From December 2023 to May 2024,9 284 combined test kits for SDC2,ADHFE1,and PPP2R5C gene methylation were distributed to high-risk individuals aged 40～80 years.Among them,841 patients(9.1％)tested positive.These patients were encouraged via telephone to undergo colonoscopy,with colonoscopy combined with pathological diagnosis as the gold standard,a total of 495 positive patients completed electronic colonoscopy.Among them,the 251 patients who tested positive for fecal SDC2,ADHFE1,and PPP2R5C gene methylation and completed electronic colonoscopy were the observation group;concurrently,244 patients who tested positive for fecal occult blood tests and underwent electronic colonoscopy were selected as the control group.Compare two groups of patients with polyp,number,shape,pathological changes and pathological types.Results There was no statistically significant difference in number and lesion location of polyps between the two groups of patients(P>0.05).The proportion of Yamada type Ⅰ in the observation group was lower than that in the control group,while the proportion of Yamada type Ⅱ was higher than that in the control group.The difference was statistically significant(P<0.05).In the observation group,1 case(0.4％)of CRC,62 cases(24.7％)of advanced adenomas,78 cases(31.1％)of non-advanced adenomas,20 cases(8.0％)of hyperplastic polyps,and 90 cases(35.9％)with no dysplastic lesions were identified.In the control group,6 cases(2.5％)of CRC,38 cases(15.6％)of advanced adenomas,53 cases(21.7％)of non-advanced adenomas,19 cases(7.8％)of hyperplastic polyps,and 128 cases(52.5％)with no dysplastic lesions were identified.The proportions of non-advanced adenomas and advanced adenomas were lower in the control group than those in the observation group,while the no dysplastic lesions rate was higher in the control group,the differences were statistically significant(P<0.05).Conclusion The detection rate of colorectal non-advanced adenomas and advanced adenomas is higher with fecal SDC2,ADHFE1,and PPP2R5C gene methylation testing compared to the fecal occult blood test.

Objective To explore the correlation between serum chemokine C-C ligand 21(CCL21),NOD like receptor protein 3(NLRP3)and hemorrhagic transformation(HT)after mechanical thrombectomy in patients with acute cerebral infarction(ACI).Methods A total of 210 ACI patients who underwent mechanical thrombectomy were selected and divided into the HT group(19 cases)and the non HT group(191 cases)based on the re-examination of head CT 24-hours after treatment.The differences in clinical data and serum levels of CCL21 and NLRP3 were compared between two groups.Multivariate Logistic regression analysis was used to analyze the influencing factors of HT occurrence.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of serum CCL21 and NLRP3 for HT.Results The proportion of atrial fibrillation,anterior circulation infarction and preoperative National Institutes of Health Stroke Scale(NIHSS)scores were higher in the HT group than those in the non HT group(P＜0.05).The preoperative serum levels of CCL21 and NLRP3 were higher in the HT group than those in the non HT group(P＜0.05).The Logistic regression model analysis results showed that atrial fibrillation,anterior circulation infarction,preoperative NIHSS score,CCL21 and NLRP3 were risk factors for postoperative HT in ACI patients(P＜0.05).The application of receiver operating characteristic(ROC)curve analysis results showed that the area under the curve(AUC)for predicting HT occurrence by CCL21 was 0.850(95％CI:0.763-0.936).The AUC predicted by NLRP3 was 0.787(95％CI:0.703-0.871),and the combined detection of the two further improved the predictive value(AUC=0.921,95％CI:0.867-0.976).Conclusion CCL21 and NLRP3 are closely related to the occurrence of HT in ACI patients after mechanical thrombectomy,and which are influencing factors for the occurrence of HT after mechanical thrombectomy.

Objective:To study the consistency between post-processing bone mineral density(BMD)values of multi-slice spiral computed tomography(MSCT)scan and the BMD value of quantitative computed tomography(QCT)for lumbar vertebra,so as to explore the feasibility of utilizing MSCT scan-based post-processing BMD values for lumbar vertebra in clinical practice.Methods:The MSCT equipment and QCT equipment were respectively adopted to conduct imaging scan for the L2-L4 of lumber vertebra of QRM-ESP145 European Spine Phantom(ESP),and L2-L4 of lumbar vertebra of adult sheep,and L2-L4 of lumbar vertebra of adult volunteer.The L2-L4 of ESP lumber vertebra and L2-L4 of lumbar vertebra of adult sheep were scanned respectively MSCT and QCT for three times,so as to measure BMD values.The L2-L4 of lumbar vertebrae of volunteers were scanned respectively by the two methods for one time according to the standard of clinical examination,which were reconstructed by three times so as to obtain mean of them.The BMD values of QCT scan were set as control group,and the BMD values of MSCT scan were set as experiment group.The experiment group was further divided into experiment 1 group[two dimension(2D)regional volumetric BMD values of the lumbar vertebra]and experiment 2 group[three dimension(3D)global volumetric BMD post-processing of the lumbar vertebra]according to the reliability of experiment.Then,the consistency between the MSCT 3D post-processing BMD values of three groups and QCT-measured BMD values was compared and analyzed.Results:The MSCT 3D post-processing BMD values of L2-L4 of ESP lumbar vertebra of three groups were respectively(120.83±0.97),(199.57±0.54)and(119.19±1.04)mg/cm3,and that of L2-L4 of lumbar vertebra of adult sheep of three groups were respectively(414.89±1.72),(410.50±0.77)and(420.25±2.71)mg/cm3,and that of L2-L4 of lumbar vertebra of volunteer were respectively(141.22±0.09),(137.38±0.37)and(152.03±1.03)mg/cm3.There were not statistically significant differences in BMD values between MSCT examination and QCT examination(P＞0.05).Conclusion:MSCT 3D post-processing BMD values on lumbar vertebra has high consistency with that of QCT measurements,which post-processing technique can replace QCT to conduct BMD examination,and reduce unnecessary radiation exposure and examination costs for patients.

BACKGROUND: Asthma is a common chronic inflammatory airway disease and intermittent hypoxia is increasingly recognized as a factor that may impact disease progression. The present study investigated whether intermittent hypoxia (IH) could aggravate asthma by promoting hypoxia-inducible factor-1α (HIF-1α)/nucleotide-binding oligomerization domain (NOD)-like receptor pyrin domain-containing protein 3 (NLRP3)/interleukin (IL)-1β-dependent pyroptosis and the inflammatory response and further elucidated the underlying molecular mechanisms involved. METHODS: A total of 49 patients diagnosed with severe bronchial asthma and diagnosed by polysomnography were enrolled at Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, between January 2022 and December 2022, and their general data and induced sputum were collected. BEAS-2B cells were treated with IL-13 and subjected to IH. An ovalbumin (OVA)-treated mouse model was also used to assess the effects of chronic intermittent hypoxia (CIH) on asthma. Pyroptosis, the inflammatory response, and related signalling pathways were assessed in vivo and in vitro . RESULTS: In this study, as the apnoea and hypopnea index (AHI) increased, the proportion of patients with uncontrolled asthma increased. The proportions of neutrophils and the levels of IL-6, IL-8, HIF-1α and NLRP3 in induced sputum were related to the AHI. NLRP3-mediated pyroptosis, which could be mediated by the HIF-1α signalling pathway, was activated in IL-13 plus IH-treated BEAS-2B cells and in the lungs of OVA/CIH mice. HIF-1α downregulation significantly reduced lung pyroptosis and ameliorated neutrophil inflammation by modulating the NLRP3/IL-1β pathway both in vitro and in vivo . Similarly, pretreatment with LW6, an inhibitor of HIF-1α, effectively blocked the generation of inflammatory cytokines in neutrophils. In addition, administration of the NLRP3 activator nigericin obviously increased lung neutrophil inflammation. CONCLUSIONS Obstructive sleep apnoea-hypopnea syndrome (OSAHS) is a risk factor for asthma exacerbation. IH aggravates neutrophil inflammation in asthma via NLRP3/IL-1β-dependent pyroptosis mediated by the HIF-1α signalling pathway, which should be considered a potential therapeutic target for the treatment of asthma with OSAHS.
NLR Family, Pyrin Domain-Containing 3 Protein/metabolism* ; Humans ; Asthma/metabolism* ; Animals ; Pyroptosis/physiology* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Mice ; Signal Transduction/physiology* ; Male ; Hypoxia/metabolism* ; Female ; Interleukin-1beta/metabolism* ; Adult ; Inflammation/metabolism* ; Middle Aged ; Mice, Inbred C57BL

Objective To evaluate the efficacy,safety and patient compliance of a quadruple therapy containing minocycline compared with the traditional quadruple therapy in the treatment of Helicobacter(H.)pylori.Methods This study included 200 H.pylori positive patients,with 100 assigned to the minocycline-containing bismuth quadruple therapy group(LBMC group)and the other 100 to the amoxicillin-containing bismuth quadruple therapy group(LBAC group).After matching the two groups of patients using the propensity score matching(PSM)method,there were 86 cases in each group.Telephone follow-up was conducted on the 14th day after the start of treatment to record patient medication compliance and adverse drug reactions.A 13C urea breath test was performed for re-examination at least one month after completing the treatment plan and discontinuing medication.The intention-to-treat(ITT)and per-protocol(PP)analyses were used to compare the H.pylori eradication rates between the two groups,and Chi-square test and t-test were used for intergroup comparison.Results In the ITT analysis,the eradication rates of the LBMC group and the LBAC group were 89.5%(77/86,95%CI:82.9%～96.1%)and 82.6%(71/86,95%CI:74.4%～90.7%),respectively.In the PP analysis,the eradication rates were 92.6%(75/81,95%CI:86.8%～98.4%)and 88.8%(71/80,95%CI:81.7%～95.8%),respectively.The adverse reaction rate of the LBMC group was 27.9%(24/86),and that of the LBAC group 31.4%(27/86),showing no statistically significant difference(P>0.05).In terms of compliance,the LBMC group was 94.2%(81/86),and the LBAC group 93.0%(80/86),revealing no statistically significant difference(P>0.05).Conclusion As a first-line treatment for eradicating H.pylori,regimens containing minocycline demonstrate equivalent eradication rates to those containing amoxicillin,with similar safety and compliance.They can be used as an alternative treatment for patients allergic to penicillin.

Objective To compare functional differences of CD8+T cells in lung tissues between young and aged C57BL/6J mice during the contraction phase and memory immune response phase after infection with influenza A(H1N1)virus.Methods Lung tissues from young(3-month-old)and aged(24-month-old)C57BL/6J female mice without influenza virus infection were collected to prepare single-cell suspensions,which were stimulated with phorbol 12-myristate 13-acetate(PMA)/ionomycin or cluster of differentiation(CD)3/CD28 antibodies(T-cell antigen receptor/co-stimulatory signals)respectively(non-specific antigens stimulation).Flow cytometry intracellular cytokine staining(ICS)was performed on lung CD8+T cells to detect their secretion capacity of tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ).Young and aged C57BL/6J mice were infected intranasally with 490 PFU PR8 influenza virus,and reinfected with homologous influenza virus 28 days later.Lung tissues were isolated on day 28(the contraction phase)and day 32(the memory immune response phase)after primary infection.Influenza virus-specific MHC-Ⅰ tetramer staining was used to detect the proportion of virus-specific CD8+T cells in lung tissue CD8+T cells,and ICS was used to analyze TNF-α,IFN-γ,and granzyme B expression in CD8+CD44high T cell subset.Results After non-specific antigen stimulation,TNF-α and IFN-γ secretion capacity in lung tissue CD8+T cells of aged group mice was significantly higher than that of young group(P＜0.05).After virus-specific antigen stimulation,there were no statistically significant differences in the proportion of virus-specific CD8+T cells and the expression levels of TNF-α,IFN-γ,and granzyme B between the two groups of mice during the contraction phase(P＞0.05),while during the memory immune response phase,the proportion of virus-specific CD8+T cells and the expression levels of TNF-α,IFN-γ,and granzyme B in the aged group mice were significantly lower than those in the young group(P＜0.05).Conclusion CD8+T cells in aged mice maintain normal immune-related factor expression function under non-specific antigen stimulation,but show impaired immune-related factor expression function during antigen-specific memory immune response phase,suggesting that aging leads to defects in the formation or maintenance of CD8+T cell immune memory.

Objective To compare the colonoscopy results of patients with positive fecal SDC2,ADHFE1,and PPP2R5C gene methylation tests to those with positive fecal occult blood tests,and analyze the effectiveness of colorectal cancer(CRC)screening.This study aims to provide a scientific basis for risk assessment in CRC screening.Methods From December 2023 to May 2024,9 284 combined test kits for SDC2,ADHFE1,and PPP2R5C gene methylation were distributed to high-risk individuals aged 40～80 years.Among them,841 patients(9.1％)tested positive.These patients were encouraged via telephone to undergo colonoscopy,with colonoscopy combined with pathological diagnosis as the gold standard,a total of 495 positive patients completed electronic colonoscopy.Among them,the 251 patients who tested positive for fecal SDC2,ADHFE1,and PPP2R5C gene methylation and completed electronic colonoscopy were the observation group;concurrently,244 patients who tested positive for fecal occult blood tests and underwent electronic colonoscopy were selected as the control group.Compare two groups of patients with polyp,number,shape,pathological changes and pathological types.Results There was no statistically significant difference in number and lesion location of polyps between the two groups of patients(P>0.05).The proportion of Yamada type Ⅰ in the observation group was lower than that in the control group,while the proportion of Yamada type Ⅱ was higher than that in the control group.The difference was statistically significant(P<0.05).In the observation group,1 case(0.4％)of CRC,62 cases(24.7％)of advanced adenomas,78 cases(31.1％)of non-advanced adenomas,20 cases(8.0％)of hyperplastic polyps,and 90 cases(35.9％)with no dysplastic lesions were identified.In the control group,6 cases(2.5％)of CRC,38 cases(15.6％)of advanced adenomas,53 cases(21.7％)of non-advanced adenomas,19 cases(7.8％)of hyperplastic polyps,and 128 cases(52.5％)with no dysplastic lesions were identified.The proportions of non-advanced adenomas and advanced adenomas were lower in the control group than those in the observation group,while the no dysplastic lesions rate was higher in the control group,the differences were statistically significant(P<0.05).Conclusion The detection rate of colorectal non-advanced adenomas and advanced adenomas is higher with fecal SDC2,ADHFE1,and PPP2R5C gene methylation testing compared to the fecal occult blood test.

Objective To explore the correlation between serum chemokine C-C ligand 21(CCL21),NOD like receptor protein 3(NLRP3)and hemorrhagic transformation(HT)after mechanical thrombectomy in patients with acute cerebral infarction(ACI).Methods A total of 210 ACI patients who underwent mechanical thrombectomy were selected and divided into the HT group(19 cases)and the non HT group(191 cases)based on the re-examination of head CT 24-hours after treatment.The differences in clinical data and serum levels of CCL21 and NLRP3 were compared between two groups.Multivariate Logistic regression analysis was used to analyze the influencing factors of HT occurrence.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of serum CCL21 and NLRP3 for HT.Results The proportion of atrial fibrillation,anterior circulation infarction and preoperative National Institutes of Health Stroke Scale(NIHSS)scores were higher in the HT group than those in the non HT group(P＜0.05).The preoperative serum levels of CCL21 and NLRP3 were higher in the HT group than those in the non HT group(P＜0.05).The Logistic regression model analysis results showed that atrial fibrillation,anterior circulation infarction,preoperative NIHSS score,CCL21 and NLRP3 were risk factors for postoperative HT in ACI patients(P＜0.05).The application of receiver operating characteristic(ROC)curve analysis results showed that the area under the curve(AUC)for predicting HT occurrence by CCL21 was 0.850(95％CI:0.763-0.936).The AUC predicted by NLRP3 was 0.787(95％CI:0.703-0.871),and the combined detection of the two further improved the predictive value(AUC=0.921,95％CI:0.867-0.976).Conclusion CCL21 and NLRP3 are closely related to the occurrence of HT in ACI patients after mechanical thrombectomy,and which are influencing factors for the occurrence of HT after mechanical thrombectomy.

Objective To compare functional differences of CD8+T cells in lung tissues between young and aged C57BL/6J mice during the contraction phase and memory immune response phase after infection with influenza A(H1N1)virus.Methods Lung tissues from young(3-month-old)and aged(24-month-old)C57BL/6J female mice without influenza virus infection were collected to prepare single-cell suspensions,which were stimulated with phorbol 12-myristate 13-acetate(PMA)/ionomycin or cluster of differentiation(CD)3/CD28 antibodies(T-cell antigen receptor/co-stimulatory signals)respectively(non-specific antigens stimulation).Flow cytometry intracellular cytokine staining(ICS)was performed on lung CD8+T cells to detect their secretion capacity of tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ).Young and aged C57BL/6J mice were infected intranasally with 490 PFU PR8 influenza virus,and reinfected with homologous influenza virus 28 days later.Lung tissues were isolated on day 28(the contraction phase)and day 32(the memory immune response phase)after primary infection.Influenza virus-specific MHC-Ⅰ tetramer staining was used to detect the proportion of virus-specific CD8+T cells in lung tissue CD8+T cells,and ICS was used to analyze TNF-α,IFN-γ,and granzyme B expression in CD8+CD44high T cell subset.Results After non-specific antigen stimulation,TNF-α and IFN-γ secretion capacity in lung tissue CD8+T cells of aged group mice was significantly higher than that of young group(P＜0.05).After virus-specific antigen stimulation,there were no statistically significant differences in the proportion of virus-specific CD8+T cells and the expression levels of TNF-α,IFN-γ,and granzyme B between the two groups of mice during the contraction phase(P＞0.05),while during the memory immune response phase,the proportion of virus-specific CD8+T cells and the expression levels of TNF-α,IFN-γ,and granzyme B in the aged group mice were significantly lower than those in the young group(P＜0.05).Conclusion CD8+T cells in aged mice maintain normal immune-related factor expression function under non-specific antigen stimulation,but show impaired immune-related factor expression function during antigen-specific memory immune response phase,suggesting that aging leads to defects in the formation or maintenance of CD8+T cell immune memory.