Objective This study investigated the regulatory effect of high mobility group protein B1 (HMGB1) in the peripheral blood of patients with primary immune thrombocytopenia (ITP) on myeloid dendritic cells (mDC) and Th17/regulatory T cells (Treg) balance. Methods The study enrolled 30 newly diagnosed ITP patients and 30 healthy controls.Flow cytometry was used to measure the proportion of mDC, Th17, and Treg cells in the peripheral blood of ITP patients and healthy controls. ELISA was conducted to quantify the serum levels of HMGB1, interleukin 6 (IL-6), IL-23, IL-17, and transforming growth factor β(TGF-β). The mRNA levels of retinoic acid-related orphan receptor γt(RORγt) and forehead box P3(FOXP3) were detected by real-time PCR. The correlation between the abovementioned cells, cytokines, and platelet count was assessed using Pearson linear correlation analysis. Results The proportion of Th17 cells and the expression levels of HMGB1, IL-6, IL-23, IL-17 and the level of RORγt mRNA in the peripheral blood of ITP patients were higher than those in healthy controls. However, the Treg cell proportion and TGF-β level were lower in ITP patients than those in healthy controls. In patients with ITP, the proportion of mDC and the level of FOXP3 mRNA did not show significant changes. The proportion of mDC cells was significantly correlated with the expression of IL-6 and IL-23. Moreover, the expression of HMGB1 showed a significant correlation with the expression of mDC, IL-6, IL-23, RORγt mRNA, and IL-17. Notably, both the proportion of mDC cells and the expression of HMGB1 were negatively correlated with platelet count. Conclusion The high expression of HMGB1 in peripheral blood of ITP patients may induce Th17/Treg imbalance by promoting the maturation of mDC and affecting the secretion of cytokines, thereby potentially playing a role in the immunological mechanism of ITP.
Humans ; Th17 Cells/cytology* ; HMGB1 Protein/genetics* ; T-Lymphocytes, Regulatory/cytology* ; Female ; Male ; Dendritic Cells/metabolism* ; Adult ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic/genetics* ; Nuclear Receptor Subfamily 1, Group F, Member 3/genetics* ; Young Adult ; Interleukin-23/blood* ; Interleukin-17/blood* ; Interleukin-6/blood* ; Forkhead Transcription Factors/genetics* ; Myeloid Cells/cytology* ; Aged

Objective:To discuss the expression and clinical significance of inositol polyphosphate-4-phosphatase type Ⅱ(INPP4B)gene in hepatocellular carcinoma(HCC)based on The Cancer Genome Atlas(TCGA)database and experimental verification with clinical samples.Methods:Based on data from 424 clinical samples in the TCGA database(including 374 HCC tissues and 50 paracarcinoma tissues),Kaplan-Meier method and Cox regression analysis were used to evaluate the relationship between INPP4B gene and the clinical characteristics and survival prognosis of the HCC patients.The correlations between INPP4B gene and the number of 24 types of immune cells,matrix,immune cell infiltration and tumor purity in tumor tissue,and the expression level of the high-frequency mutant gene tumor protein 53(TP53)in HCC were analyzed.The clinicopathological data and paraffin-embedded tissue sections of 60 HCC patients treated with surgical resection from December 2022 to December 2023 were collected.According to clinical diagnosis,they were divided into poorly differentiated group(HCC-L group),moderately differentiated group(HCC-M group)and well-differentiated group(HCC-H group),with 20 cases in each group;20 patients during the same period who underwent biopsy and were pathologically diagnosed as non-tumor were selected as normal group,and their clinicopathologic data and liver tissue paraffin sections were collected.HE staining was used to observe the pathomorphology of HCC tissue and normal liver tissue of the subjects in various groups;immunohistochemistry method was used to detect the expressions of Ki-67 and INPP4B proteins in the HCC tissue and normal liver tissue of the subjects in various groups.Results:The TCGA database analysis results showed that compared with normal tissue,the expression level of INPP4B mRNA in HCC tissue was significantly increased(P<0.01).Compared with INPP4B low expression group,the overall survival(OS)of the patients in INPP4B high expression group was significantly prolonged(P<0.05).The univariate Cox regression analysis results showed that tumor stage,pathological stage,tumor status and residual tumor had impacts on OS of the HCC patients(P<0.05).The univariate regression analysis results showed that the INPP4B prognostic risk model score ratio was HR=0.781,95％confidence interval(CI):0.552-1.105,P=0.168.The AUC value for the impact of INPP4B on OS of the HCC patients was 0.558,indicating that the INPP4B gene prognostic risk model had certain predictive value in survival prognosis.The INPP4B mRNA expression level was not correlated with TNM stage,stage,patient gender,age,race or body mass index(BMI)(P>0.05).In tumor tissue with high and low INPP4B expression,22 types of immune cells showed statistically significant differences(P<0.05);the INPP4B mRNA expression level was positively correlated with the number of 23 types of immune cells except T helper(Th)17 cells(r>0),among which all Th cells except natural killer(NK)CD56+cells were statistically significant(P<0.01);INPP4B was significantly correlated with matrix(r=0.475),immune cell infiltration(r=0.641)and tumor purity(r=0.599)in tumor tissue(P<0.01).INPP4B was correlated with TP53(r=0.287,P<0.01).The HE staining results showed that clear and complete lobular structure,neatly arranged cells and slight inflammatory cell infiltration were observed in liver tissue of the subjects in normal group;completely destroyed lobular structure,significant hepatocellular steatosis,massive inflammatory cell infiltration,and lesions such as ballooning degeneration and small cell hyperplasia in some cells were observed in HCC tissue of the patients in HCC-L,HCC-M and HCC-H groups,and the lower the HCC differentiation degree,the more severe the tissue destruction;The immunohistochemistry results showed that compared with normal group,the expression levels of Ki-67 protein in HCC tissue of the patients in HCC-L,HCC-M and HCC-H groups were significantly increased(P<0.01),and the lower the differentiation degree of the HCC patients,the higher the Ki-67 positive rate.Brownish-yellow granules evenly distributed in the cells and INPP4B protein was highly expressed in liver tissue of the subjects in normal group;compared with normal group,the expression levels of INPP4B protein in HCC tissue of the patients in HCC-L,HCC-M and HCC-H groups were significantly decreased(P<0.01),and the lower the differentiation degree of the HCC tissue,the lower the INPP4B positive rate.Conclusion:INPP4B is a protective factor for the prognosis of HCC patients;as a new tumor suppressor gene,INPP4B may become a potential target for new drug screening in HCC treatment.

Objective To investigate the expression levels and clinical significance of type Ⅰ innate lymphoid cells(ILC1s),T-box transcription factor(T-bet),interleukin(IL)-12,IL-18 and interferon-gamma(IFN-γ)in peripheral blood of patients with primary immune thrombocytopenia(ITP).Methods Thirty-five ITP patients with their first episode were selected as the initial treatment group.Thirteen of these patients were followed up after receiving treatment.Additionally,20 healthy individuals underwent routine physical examinations during the same period were recruited as the control group.Peripheral blood samples were collected for analysis.The proportion of ILC1s was determined by flow cytometry.T-bet mRNA expression was measured using quantitative real-time PCR(qRT-PCR).Serum levels of IL-18,IL-12 and IFN-γ were quantified by enzyme-linked immunosorbent assay(ELISA).The differences in ILC1s proportion,T-bet mRNA expression and cytokine levels were compared between groups.Correlations between ILC1s proportion,T-bet mRNA,cytokine levels and platelet(PLT)counts were also analyzed.Results Compared with the control group,the initial treatment group exhibited significantly elevated levels of peripheral ILC1s,T-bet mRNA and serum IL-18,IL-12 and IFN-γ(P＜0.05).Among the 13 patients who were followed up,all these indices decreased significantly after treatment(P＜0.05).Correlation analysis revealed that the proportion of ILC1s in the initial treatment group was positively correlated with IL-12,IL-18,IFN-γ and T-bet mRNA levels(rs=0.666,0.647,0.677,and 0.750,respectively,P＜0.01),and negatively correlated with PLT count(rs=-0.637,P＜0.01).Conclusion Innate immunity may play a role in the pathogenesis and progression of ITP by regulating the expression levels of ILC1s,T-bet,IL-12,IL-18 and IFN-γ.

Objective To investigate the expression levels and clinical significance of type Ⅰ innate lymphoid cells(ILC1s),T-box transcription factor(T-bet),interleukin(IL)-12,IL-18 and interferon-gamma(IFN-γ)in peripheral blood of patients with primary immune thrombocytopenia(ITP).Methods Thirty-five ITP patients with their first episode were selected as the initial treatment group.Thirteen of these patients were followed up after receiving treatment.Additionally,20 healthy individuals underwent routine physical examinations during the same period were recruited as the control group.Peripheral blood samples were collected for analysis.The proportion of ILC1s was determined by flow cytometry.T-bet mRNA expression was measured using quantitative real-time PCR(qRT-PCR).Serum levels of IL-18,IL-12 and IFN-γ were quantified by enzyme-linked immunosorbent assay(ELISA).The differences in ILC1s proportion,T-bet mRNA expression and cytokine levels were compared between groups.Correlations between ILC1s proportion,T-bet mRNA,cytokine levels and platelet(PLT)counts were also analyzed.Results Compared with the control group,the initial treatment group exhibited significantly elevated levels of peripheral ILC1s,T-bet mRNA and serum IL-18,IL-12 and IFN-γ(P＜0.05).Among the 13 patients who were followed up,all these indices decreased significantly after treatment(P＜0.05).Correlation analysis revealed that the proportion of ILC1s in the initial treatment group was positively correlated with IL-12,IL-18,IFN-γ and T-bet mRNA levels(rs=0.666,0.647,0.677,and 0.750,respectively,P＜0.01),and negatively correlated with PLT count(rs=-0.637,P＜0.01).Conclusion Innate immunity may play a role in the pathogenesis and progression of ITP by regulating the expression levels of ILC1s,T-bet,IL-12,IL-18 and IFN-γ.

Objective:To explore the genetic etiology for a patient with Alstr?m syndrome (ALMS) presenting as dilated cardiomyopathy.Methods:A 41-year-old male patient who had presented at the Sixth Medical Center of PLA General Hospital on October 20, 2021 was selected as the study subject. Clinical and laboratory examinations were carried out. Whole exome sequencing (WES) was employed for genetic testing, and candidate variants were validated by Sanger sequencing and pathogenicity analysis.Results:The patient had a 14-year medical history characterized by dilated cardiomyopathy, complete atrioventricular block, visual impairment, sensorineural hearing loss, truncal obesity, insulin resistance, type 2 diabetes, hypertension, renal dysfunction, and paranoid delusions. Genetic testing revealed that he has harbored compound heterozygous variants of the ALMS1 gene, namely c. 6823C>T (p.Arg2275Ter) and c. 9442_9445dup (p.Ser3149LysfsTer2). Sanger sequencing confirmed that they were inherited from his father and mother, respectively. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), both variants were predicted to be pathogenic (PVS1_VeryStrong+ PM2_Supporting+ PM3+ PP3, PVS1_VeryStrong+ PM2_Supporting+ PM3). Literature review indicated that the complete atrioventricular block in the patient was a phenotype unreported previously. Conclusion:The c. 6823C>T (p.Arg2275Ter) and c. 9442_9445dup (p.Ser3149LysfsTer2) compound heterozygous variants of the ALMS1 gene probably underlay the pathogenesis in this patient. Above findings have expanded the phenotypic spectrum of ALMS and provided insights for clinicians dealing with similar cases.

ObjectiveThis study aims to examine the effect of Rhei Radix et Rhizoma-Coptidis Rhizoma on reducing insulin resistance in db/db mice by regulating the adenylate activated protein kinase （AMPK）/UNC-51-like kinase 1 （ULK1）/key molecule of autophagy， benzyl chloride 1 （Beclin1） pathway and elucidate the underlying mechanism. MethodSixty 6-week-old male db/db mice were studied. They were randomly divided into the model group， metformin group （0.26 g·kg^-1）， and low-， middle-， and high-dose groups （2.25， 4.5， 9 g·kg^-1） of Rhei Radix et Rhizoma-Coptidis Rhizoma. A blank group of db/m mice of the same age was set， with 12 mice in each group. After eight weeks of continuous intragastric administration， the blank group and model group received distilled water intragastrically once a day. The survival status of the mice was observed， and fasting blood glucose （FBG） was measured using a Roche blood glucose device. Fasting serum insulin （FINS） was measured using an enzyme-linked immunosorbent assay， and the insulin resistance index （HOMA-IR） was calculated. Hematoxylin-eosin （HE） staining was performed to observe the pathological changes in the liver of the mice. The protein expression levels of AMPK， Beclin1， autophagy associated protein 5 （Atg5）， and p62 in liver tissue were determined by using Western blot. The protein expression levels of autophagy associated protein 1 light chain 3B （LC3B） and ULK1 in liver tissue were determined using immunofluorescence. Real-time fluorescence quantitative PCR （Real-time PCR） was used to measure mRNA expression levels of AMPK， Beclin1， Atg5， ULK1， and p62. ResultCompared with the blank group， the model group exhibited a significant increase in body mass （P<0.01）. Additionally， the levels of FBG， FINS， and HOMA-IR significantly changed （P<0.01）. The structure of liver cells was disordered. The protein expression levels of AMPK， Beclin1， and Atg5 in liver tissue were significantly decreased （P<0.01）， while the expression level of p62 protein was significantly increased （P<0.01）. The expression levels of mRNA and proteins were consistent. Compared with the model group， the body mass of the metformin group and high and medium-dose groups of Rhei Radix et Rhizoma-Coptidis Rhizoma was significantly decreased （P<0.05）. FBG， FINS， and HOMA-IR were significantly decreased （P<0.05，P<0.01）. After treatment， the liver structure damage in each group was alleviated to varying degrees. The protein expressions of AMPK， Beclin1， Atg5， LC3B， and ULK1 were increased （P<0.05，P<0.01）， while the protein expression of p62 was decreased （P<0.01）. The expression levels of mRNA and proteins were generally consistent. ConclusionThe combination of Rhei Radix et Rhizoma-Coptidis Rhizoma can effectively improve liver insulin resistance， regulate the AMPK autophagy signaling pathway， alleviate insulin resistance in db/db mice， and effectively prevent the occurrence and development of type 2 diabetes.

Objective:To investigate the efficacy and safety of intravenous thrombolysis and antiplatelet therapy in patients with stroke warning syndrome (SWS), as well as influencing factors of the outcome in patients with SWS.Method:Patients with SWS admitted to the 521 st Hospital of Ordnance Group from June 1, 2018 to December 31, 2023 were retrospectively included. Some patients were treated with ateplase intravenous thrombolysis, followed by oral antiplatelet therapy; some patients only received antiplatelet therapy. The main outcome measure was the modified Rankin Scale score at 90 days after onset, with a score of 0-2 defined as good outcome. Results:A total of 35 patients with SWS were included, including 26 males (74.3%) with an age of 58.29±11.06 years. Nineteen patients (54.3%) received intravenous thrombolysis, and 27 (77.1%) had good outcome at 90 days. There was no statistically significant difference in demographic, baseline data, and good outcome between the intravenous thrombolysis group and the antiplatelet therapy group. One patient had new stroke and one had transient ischemic attack in the intravenous thrombolysis group. There were statistically significant differences in ABCD2 score, systolic blood pressure, low-density lipoprotein cholesterol, fasting blood glucose, highest National Institutes of Health Stroke Scale (NIHSS) score at onset, and symptom duration between the good outcome group and the poor outcome group (all P<0.05). Conclusions:The efficacy of intravenous thrombolysis is similar to that of antiplatelet drugs alone in treating SWS. ABCD2 score, systolic blood pressure, low-density lipoprotein cholesterol, fasting blood glucose, highest NIHSS score at onset, and duration of symptoms may be influencing factors for the outcome of patients with SWS.

Significant research progress has been made in the development of antibody-drug conjugates (ADCs) for the treatment of ad-vanced breast cancer (ABC),ushering new hope for patients with this refractory disease. Through the conjugation of specific antibodies with highly potent cytotoxic drugs,tumor cells can be precisely targeted and killed while minimizing damage to normal tissues. ADCs such as trastuzumab-emtansine (T-DM1) and trastuzumab deruxtecan (T-DXd),have shown excellent efficacy in the treatment of HER2-positive ABC,significantly prolonging patient survival. Furthermore,ADCs targeting triple-negative breast cancer (TNBC),such as sacituzumab govitecan (SG),have also achieved positive results in clinical trials. With the continuous research,development,and optimization of ADCs,as well as the exploration of combination treatment strategies,ADCs are expected to play an increasingly important role in the treatment of ABC in the future. This article provides an overview of the research progress of ADCs in the treatment of ABC,exploring their efficacy and safety. We aim to offer more treatment options and renewed hope for patients with ABC.

A 44-year-old female patient was treated with norethisterone 5 mg once every 8 hours due to atypical hyperplasia of endometrium. One month later, the patient developed asthma with cyanosis, and sudden cardiac arrest later. According to the results of electrocardiogram and echocardiogram examination, acute pulmonary embolism with cardiac arrest was diagnosed. Cardiac resuscitation was immediately implemented, and thrombolysis and anticoagulation therapies were performed, accompanied by veno-arterial extracorporeal membrane oxygenation (VA-ECMO) for life support. After 5 minutes of thrombolytic therapy, the patient′s heart beat and blood pressure gradually recovered. The results of pulmonary artery CT angiography supported the diagnosis of acute pulmonary embolism and effectiveness of thrombolysis. After 31 hours of thrombolytic therapy, the patient has stabilized vital signs and VA-ECMO was removed. After 47 hours of thrombolytic therapy, the patient′s consciousness fully recovered, the tracheal tube was removed, and anticoagulant therapy was continued. After 11 days of thrombolytic therapy, the echocardiography was re-performed, showing normal pulmonary artery pressure, and the exercise tolerance was checked to be good. At 1 month of follow-up, the cardiac function and structure were normal on echocardiography, no thrombus was found on lower limb vascular ultrasound.

A 44-year-old female patient was treated with norethisterone 5 mg once every 8 hours due to atypical hyperplasia of endometrium. One month later, the patient developed asthma with cyanosis, and sudden cardiac arrest later. According to the results of electrocardiogram and echocardiogram examination, acute pulmonary embolism with cardiac arrest was diagnosed. Cardiac resuscitation was immediately implemented, and thrombolysis and anticoagulation therapies were performed, accompanied by veno-arterial extracorporeal membrane oxygenation (VA-ECMO) for life support. After 5 minutes of thrombolytic therapy, the patient′s heart beat and blood pressure gradually recovered. The results of pulmonary artery CT angiography supported the diagnosis of acute pulmonary embolism and effectiveness of thrombolysis. After 31 hours of thrombolytic therapy, the patient has stabilized vital signs and VA-ECMO was removed. After 47 hours of thrombolytic therapy, the patient′s consciousness fully recovered, the tracheal tube was removed, and anticoagulant therapy was continued. After 11 days of thrombolytic therapy, the echocardiography was re-performed, showing normal pulmonary artery pressure, and the exercise tolerance was checked to be good. At 1 month of follow-up, the cardiac function and structure were normal on echocardiography, no thrombus was found on lower limb vascular ultrasound.