Objective:To investigate the relationship between the BCL2 family protein BIM and ibrutinib resistance in chronic lymphocytic leukemia (CLL) and to analyze its regulatory mechanisms on apoptosis and autophagy.Methods:RNA sequencing (RNA-seq) was used to examine changes in the expression of BCL2 family proteins in samples from patients with CLL, MEC1 cell lines, and ibrutinib-resistant cell lines (MR). Western blot was used to analyze changes in BIM protein expression during apoptosis in MR. shRNA knockdown was used to assess the effects of BIM on cell proliferation and apoptosis. RNA-seq and the autophagy inhibitor chloroquine treatment were used to study autophagy-related changes in MR.Results:BIM expression was significantly downregulated before and after drug resistance in CLL primary cells and MEC1 cell lines ( P<0.0001). Knockdown of BIM in CLL cells inhibited ibrutinib-induced apoptosis and promoted cell proliferation ( P<0.05 for both). In addition, protective autophagy was increased in MR and apoptosis was increased after administration of chloroquine and small interfering RNA. The increased expression of LC3-Ⅱ protein in BIM-knockdown cell lines ( P<0.01) suggested that reduction of BIM may mediate autophagy activation. Conclusion:Downregulation of BIM may be a key factor in promoting ibrutinib resistance in CLL by activating protective autophagy. These findings provided a potential target for improving CLL treatment.

Objective:To investigate the relationship between the BCL2 family protein BIM and ibrutinib resistance in chronic lymphocytic leukemia (CLL) and to analyze its regulatory mechanisms on apoptosis and autophagy.Methods:RNA sequencing (RNA-seq) was used to examine changes in the expression of BCL2 family proteins in samples from patients with CLL, MEC1 cell lines, and ibrutinib-resistant cell lines (MR). Western blot was used to analyze changes in BIM protein expression during apoptosis in MR. shRNA knockdown was used to assess the effects of BIM on cell proliferation and apoptosis. RNA-seq and the autophagy inhibitor chloroquine treatment were used to study autophagy-related changes in MR.Results:BIM expression was significantly downregulated before and after drug resistance in CLL primary cells and MEC1 cell lines ( P<0.0001). Knockdown of BIM in CLL cells inhibited ibrutinib-induced apoptosis and promoted cell proliferation ( P<0.05 for both). In addition, protective autophagy was increased in MR and apoptosis was increased after administration of chloroquine and small interfering RNA. The increased expression of LC3-Ⅱ protein in BIM-knockdown cell lines ( P<0.01) suggested that reduction of BIM may mediate autophagy activation. Conclusion:Downregulation of BIM may be a key factor in promoting ibrutinib resistance in CLL by activating protective autophagy. These findings provided a potential target for improving CLL treatment.

Objective:This study aimed to assess the efficacy and safety of thioamide as a maintenance therapy for peripheral T-cell lymphoma (PTCL) .Methods:This study retrospectively analyzed the data from 58 patients with PTCL who were treated in the Department of Hematology at the First Affiliated Hospital of Nanjing Medical University from January 2015 to July 2022. Chidamide was orally administered as a maintenance therapy after first-line or salvage treatment. Progression-free survival (PFS), overall survival (OS), and safety were analyzed.Results:Among the 58 patients with PTCL, 43 were males and 15 were females, and the median age was 66 (range: 29-83) years. Thirty-nine patients received thioamide as first-line maintenance therapy, and 19 patients received thioamide as maintenance therapy after salvage treatment. The median maintenance therapy duration was 16 months (range: 1-72 months), with a median PFS time of 33 (2-74) months, and the median OS time had not been reached. Patients who received first-line maintenance therapy with thioamide demonstrated superior PFS and OS outcomes compared with patients who received thioamide maintenance therapy after salvage treatment (median PFS time: not reached vs 7 months, P<0.001; median OS time: not reached vs 67 months, P=0.009). The most prevalent adverse reaction was a hematologic adverse reaction (77.6%). Twelve (20.7%) patients underwent a dose reduction and three patients discontinued treatment. Conclusion:Patients receiving thioamide maintenance therapy demonstrated a promising PFS and OS with a manageable safety profile, especially as the first-line maintenance therapy.

Community-acquired respiratory infection is the commonest cause of sepsis presenting to emergency departments. Yet current experimental animal models simulate peritoneal sepsis with intraperitoneal (I.P.) injection of lipopolysaccharide (LPS) as the predominant route. We aimed to compare the progression of organ injury between I.P. LPS and intranasal (I.N.) LPS in order to establish a better endotoxemia murine model of respiratory sepsis. Eight weeks old male BALB/c mice received LPS-Escherichia coli doses at 0.15, 1, 10, 20, 40 and 100 mg per kg body weight (e.g. LPS-10 is a dose of 10 mg/kg body weight). Disease severity was monitored by a modified Mouse Clinical Assessment Score for Sepsis (M-CASS; range 0–21). A M-CASS score ≥ 10 or a weight reduction of ≥ 20%, was used as a criterion for euthanasia. The primary outcome was the survival rate (either no death or no need for euthanasia). The progression of disease was specified as M-CASS, body weight, blood glucose, histopathological changes to lung, liver, spleen, kidney, brain and heart tissues. Survival rate in I.P. LPS-20 mice was 0% (2/3 died; 1/3 euthanized with M-CASS > 10) at 24 h. Survival rate in all doses of I.N. LPS was 100% (20/20; 3–4 per group) at 96 h. 24 h mean M-CASS post-I.P. LPS-10 was 6.4/21 significantly higher than I.N. LPS-10 of 1.7/21 (Unpaired t test, P < 0.05). Organ injury was present at 96 h in the I.P. LPS-10 group: lung (3/3; 100%), spleen (3/3; 100%) and liver (1/3; 33%). At 24 h in the I.P. LPS-20 group, kidney injury was observed in the euthanized mouse. At 96 h in the post-I.N. LPS-20 group, only lung injury was observed in 2/3 (67%) mice (Kruskal-Wallis test with Dunn’s, P < 0.01). At 24 h in the post-I.N. LPS-100 group all (4/4) mice had evidence of lung injury. Variable doses of I.N. LPS in mice produced lung injury but did not produce sepsis. Higher doses of I.P. LPS induced multi-organ injury but not respiratory sepsis. Lethal models of respiratory virus, e.g., influenza A, might provide alternative avenues that can be explored in future research.

Objective:This study aimed to assess the efficacy and safety of thioamide as a maintenance therapy for peripheral T-cell lymphoma (PTCL) .Methods:This study retrospectively analyzed the data from 58 patients with PTCL who were treated in the Department of Hematology at the First Affiliated Hospital of Nanjing Medical University from January 2015 to July 2022. Chidamide was orally administered as a maintenance therapy after first-line or salvage treatment. Progression-free survival (PFS), overall survival (OS), and safety were analyzed.Results:Among the 58 patients with PTCL, 43 were males and 15 were females, and the median age was 66 (range: 29-83) years. Thirty-nine patients received thioamide as first-line maintenance therapy, and 19 patients received thioamide as maintenance therapy after salvage treatment. The median maintenance therapy duration was 16 months (range: 1-72 months), with a median PFS time of 33 (2-74) months, and the median OS time had not been reached. Patients who received first-line maintenance therapy with thioamide demonstrated superior PFS and OS outcomes compared with patients who received thioamide maintenance therapy after salvage treatment (median PFS time: not reached vs 7 months, P<0.001; median OS time: not reached vs 67 months, P=0.009). The most prevalent adverse reaction was a hematologic adverse reaction (77.6%). Twelve (20.7%) patients underwent a dose reduction and three patients discontinued treatment. Conclusion:Patients receiving thioamide maintenance therapy demonstrated a promising PFS and OS with a manageable safety profile, especially as the first-line maintenance therapy.

Objective:To compare the safety and efficacy of the " step-up approach" versus the " step-jump approach" in treatment of infected pancreatic necrosis (IPN).Method:The clinical data of IPN patients who underwent step-up strategy or step-jump strategy treatment at the Department of Pancreatic and Biliary Surgery of the First Affiliated Hospital of Harbin Medical University from December 2018 to November 2022 were analyzed retrospectively. Propensity score matching (PSM) was done based on the nearest neighbor matching method (1: 1 ratio). After matching the baseline data (the caliper value was 0.01), a total of 62 patients with IPN were included, including 41 males and 21 females, aged (41.1±13.1) years old. Patients who were treated with the step-up strategy were included in the step-up group, while patients who were treated with the step-jump strategy were included in the step-jump group. There were 31 patients in each group after PSM, and the treatment effect of the two groups were compared.Results:Of the 62 patients with IPN, 43 received surgical intervention, and 19 were managed successfully using symptomatic anti-inflammatory treatment or percutaneous catheter drainage. The total hospitalization cost of patients in the step-jump group was significantly higher than that in the step-up group [122 000 (73 000, 179 000) yuan vs. 88 000 (46 000, 144 000) yuan, P=0.034]. The overall cure rate of IPN patients in the step-jump group was 93.5%(29/31). The 2 patients who died had type Ⅲ IPN. In the IPN patients in the step-up group were all cured, and the overall cure rate was 100%(31/31), with no death. There were no statistical differences between the two groups in the rates of death, postoperative complications, residual infection, debridement ≥2 times, and positive bacterial culture in blood or drainage fluid (all P>0.05). A total of 19.4% (12/62) patients had postoperative complications, including 4 patients with abdominal bleeding, 3 patients with new organ dysfunction, 2 patients with gastrointestinal bleeding, 2 patients with gastrointestinal fistula, and 1 patient with venous thrombosis in both lower limbs. Conclusion:Both the step-up treatment strategy and the step-jump treatment strategy were safe and effective for treatment of IPN patients.

Objective:To analyze the clinical features and genetic basis for a child featuring elevated creatine kinase (CK).Methods:Next-generation sequencing (muscular dystrophy-related gene panel) was carried out for the proband. Candidate variants were verified by Sanger sequencing of the child and his parents.Results:The child was found to harbor compound heterozygous variants of the FKTN gene, including a missense c. 536G>C (p.R179T) variant from his father and a non-frameshift c. 1299_1301delGTG (p.W434del) variant from his mother. Both variants were predicted to be pathogenic. Conclusion:The compound heterozygous variants of the FKTN gene probably underlay the disease in this child. Above finding has expanded the mutation spectrum of congenital muscular dystrophy.

Objective To investigate the underlying drug enhancement mechanisms of the Chuanwu (Aconiti Radix) and Huangqi (Astragali Radix) combination and toxicity reduction of Chuanwu combined with Gancao (Glycyrrhizae Radix et Rhizoma) in Wutou Decoction (乌头汤, WTD), and to elucidate the compatibility principle. Methods The active compounds and potential effective targets of the selected combinations were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Traditional Chinese Medicines Integrated Database (TCMID). The toxicity of Chuanwu (Aconiti Radix) was investigated by selecting all five toxic compounds from the literature and the TCMSP database, and obtaining their targets through SwissTargetPrediction. Targets related to rheumatoid arthritis (RA) were searched using DisGeNET, GenCards, and Online Mendelian Inheritance in Man (OMIM). Mutual targets between the drug pairs and RA were selected as potential RA therapy targets. The medicinally active compound-target network was constructed using Cytoscape 3.9.0. Gene ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were performed using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) platform. Results We obtained 191 active compound targets for Gancao (Glycyrrhizae Radix et Rhizoma), 171 for Huangqi (Astragali Radix), and 103 for Chuanwu (Radix Aconiti) (hypoaconitine’s target was obtained through literature and SwissTargetPrediction). A total of 5872 genes were obtained for RA. A drug-active compound-target network involving 13 effect-enhancing and nine toxicity reduction targets was constructed. PGR was the main effect enhancement target, and KCNH2 was the main toxicity reduction target. The effect-enhancing targets were related to 23 GO terms (such as positive regulation of transcription from RNA polymerase II promoter, steroid hormone-mediated signaling pathway, plasma membrane, and protein binding) (P < 0.01), and 13 KEGG pathways related to synergism [such as estrogen signaling pathway, cholinergic synapse, and phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway]. The toxicity reduction targets were related to 28 GO terms (mainly involes G-protein coupled receptor signaling pathway, plasma membrane, and drug binding) (P < 0.01), and five KEGG pathways related to toxicity reduction (cholinergic synapse, calcium signaling pathway, regulation of actin cytoskeleton, neuroactive ligand-receptor interaction, and serotonergic synapse). Conclusion The combination of Chuanwu (Aconiti Radix) and Huangqi (Astragali Radix) plays an important effect-enhancing role in WTD and involves the estrogen and PI3K/Akt signaling pathways, with PGR as the core. The Chuanwu (Aconiti Radix) and Gancao (Glycyrrhizae Radix et Rhizoma) combination decreases toxicity in WTD and is associated with the cholinergic synapse and calcium signaling pathways, with KCNH2 as the core.

OBJECTIVE: To explore the genetic basis for two Chinese pedigrees affected with Huntington disease and provide prenatal diagnosis for them. METHODS: Peripheral venous blood samples were collected from the probands. PCR and capillary gel electrophoresis were used to determine the number of CAG repeats in their IT15 gene. Pre-symptomatic testing was offered to their children and relatives, and prenatal diagnosis was provided to three pregnant women from the two pedigrees. RESULTS: The two probands, in addition with three asymptomatic members, were found to have a (CAG)n repeat number greater than 40. Upon prenatal diagnosis, the numbers of CAG repeats in two fetuses from pedigree 1 were determined as (16, 19) and (18, 19), both were within the normal range. A fetus from pedigree 2 was found to have a CAG repeat number of (15, 41), which exceeded the normal range. CONCLUSION Genetic testing can facilitate the diagnosis of Huntington disease and avoid further birth of affected children.
Child ; Female ; Genetic Testing ; Humans ; Huntington Disease/genetics* ; Nerve Tissue Proteins/genetics* ; Pedigree ; Pregnancy ; Prenatal Diagnosis

OBJECTIVE: To explore the prevalence and characteristics of chromosomal abnormalities in abortuses during early pregnancy with single nucleotide polymorphism microarray (SNP-array). METHODS: For 520 abortuses, copy number variations (CNVs) in chorionic villi were analyzed with SNP-array. RESULTS: In 510 (98.1%) of the samples, the analysis was successful. Among these, 57.6% (294/510) of the samples were found to harbor clinically significant chromosomal abnormalities. 38.8% of the samples (198/510) had a normal result. 2.4% (12/510) of the samples harbored benign CNVs, and 1.2% (6/510) harbored variants of uncertain significance (VOUS). Aneuploidies, polyploidies, pathogenic CNVs and uniparental disomies (UPD) had accounted for 75.2% (221/294), 13.9% (41/294), 8.2% (24/294), and 2.7% (8/294) of the samples, respectively. 45,XO was the most common finding, which was followed by trisomy 16 and trisomy 22. 69,XXY was the most common polyploidy. CONCLUSION Chromosomal abnormalities are the main cause for early miscarriage, among which aneuploidies are most common. The prevalence of aneuploidies is significantly increased among women over 35. SNP-array analysis has the advantage of high success rate, high resolution and great accuracy, but the clinical significance of microdeletions/microduplications found by SNP-array can be difficult for interpretation.
Chorionic Villi ; Chromosome Aberrations ; Chromosome Disorders ; DNA Copy Number Variations ; Female ; Genetic Testing ; Humans ; Karyotyping ; Polymorphism, Single Nucleotide ; Pregnancy