Objective:To explore differences in the efficacy and safety of ruxolitinib in patients with myelofibrosis by age and to identify prognostic factors by analyzing clinical features and characteristics of chromosomes and gene mutations.Methods:This study retrospectively analyzed 188 patients with myelofibrosis who received ruxolitinib in the Department of Hematology, Qilu Hospital, Shandong University from January 1, 2017, to July 1, 2024. According to age at diagnosis, the patients were divided into the middle-aged group (≤55 years), young elderly group (56-65 years), and elderly group (>65 years). Clinical features, the characteristics of chromosomes and gene mutations, and the efficacy and safety of ruxolitinib treatment were compared across the three age groups. Independent factors influencing overall survival were identified through Cox proportional risk regression analysis.Results:Before treatment, the elderly group had more underlying comorbidities, a heavier symptom burden, higher leukocyte count, higher proportion and frequency of JAK2 mutations, and lower proportion of CALR mutations. The incidence of nondriver gene mutations was significantly higher in the young elderly group. After ruxolitinib treatment, the degree of reduction in spleen size did not differ significantly among the three groups. The length of the palpable spleen below the left costal margin reduced by more than 50% from baseline in 50.9% (27/53) of the patients in the middle-aged group, 43.5% (27/62) in the young elderly group, and 45.5% (20/44) in the elderly group ( P=0.720). No significant difference was observed among the three groups in the degree of reduction in Myeloproliferative Neoplasm Symptom Assessment Form (10-item version) score ( P=0.153), with a reduction in total symptom score by more than 50% achieved by 54.0% (27/50), 60.3% (41/68), and 66.7% (34/51) of the patients from the three groups, respectively ( P=0.429). The most common hematological adverse events were anemia and thrombocytopenia, while the most common nonhematological adverse events were electrolyte disturbance, elevated transaminase activity, and pulmonary infection. Multivariate analysis indicated that in ruxolitinib-treated patients with myelofibrosis, poor overall survival was independently predicted by increased age, reduced hemoglobin, percentage of bone marrow blasts ≥ 1%, absence of JAK2 mutations, chromosomal abnormalities, ≥2 high-molecular-risk mutations, and TP53 mutations. Conclusions:Patients with myelofibrosis stratified by age exhibited heterogeneous clinical features and gene mutation profiles but similar efficacy of ruxolitinib treatment and occurrence of adverse events.

Background: Diabetic kidney disease (DKD) is recognized as a significant complication of diabetes mellitus and categorized into glomerular DKDs and tubular DKDs, each governed by distinct pathological mechanisms and biomarkers. Methods: Through the identification of common features observed in glomerular and tubular lesions in DKD, numerous differentially expressed gene were identified by the machine learning, single-cell transcriptome and mendelian randomization. Results: The diagnostic markers versican (VCAN) was identified, offering supplementary options for clinical diagnosis. VCAN significantly highly expressed in glomerular parietal epithelial cell and proximal convoluted tubular cell. It was mainly involved in the up-regulation of immune genes and infiltration of immune cells like mast cell. Mendelian randomization analysis confirmed that serum VCAN protein levels were a risky factor for DKD, while there was no reverse association. It exhibited the good diagnostic potential for estimated glomerular filtration rate and proteinuria in DKD. Conclusion VCAN showed the prospects into DKD pathology and clinical indicator.

Background: Diabetic kidney disease (DKD) is recognized as a significant complication of diabetes mellitus and categorized into glomerular DKDs and tubular DKDs, each governed by distinct pathological mechanisms and biomarkers. Methods: Through the identification of common features observed in glomerular and tubular lesions in DKD, numerous differentially expressed gene were identified by the machine learning, single-cell transcriptome and mendelian randomization. Results: The diagnostic markers versican (VCAN) was identified, offering supplementary options for clinical diagnosis. VCAN significantly highly expressed in glomerular parietal epithelial cell and proximal convoluted tubular cell. It was mainly involved in the up-regulation of immune genes and infiltration of immune cells like mast cell. Mendelian randomization analysis confirmed that serum VCAN protein levels were a risky factor for DKD, while there was no reverse association. It exhibited the good diagnostic potential for estimated glomerular filtration rate and proteinuria in DKD. Conclusion VCAN showed the prospects into DKD pathology and clinical indicator.

Background: Diabetic kidney disease (DKD) is recognized as a significant complication of diabetes mellitus and categorized into glomerular DKDs and tubular DKDs, each governed by distinct pathological mechanisms and biomarkers. Methods: Through the identification of common features observed in glomerular and tubular lesions in DKD, numerous differentially expressed gene were identified by the machine learning, single-cell transcriptome and mendelian randomization. Results: The diagnostic markers versican (VCAN) was identified, offering supplementary options for clinical diagnosis. VCAN significantly highly expressed in glomerular parietal epithelial cell and proximal convoluted tubular cell. It was mainly involved in the up-regulation of immune genes and infiltration of immune cells like mast cell. Mendelian randomization analysis confirmed that serum VCAN protein levels were a risky factor for DKD, while there was no reverse association. It exhibited the good diagnostic potential for estimated glomerular filtration rate and proteinuria in DKD. Conclusion VCAN showed the prospects into DKD pathology and clinical indicator.

Background: Diabetic kidney disease (DKD) is recognized as a significant complication of diabetes mellitus and categorized into glomerular DKDs and tubular DKDs, each governed by distinct pathological mechanisms and biomarkers. Methods: Through the identification of common features observed in glomerular and tubular lesions in DKD, numerous differentially expressed gene were identified by the machine learning, single-cell transcriptome and mendelian randomization. Results: The diagnostic markers versican (VCAN) was identified, offering supplementary options for clinical diagnosis. VCAN significantly highly expressed in glomerular parietal epithelial cell and proximal convoluted tubular cell. It was mainly involved in the up-regulation of immune genes and infiltration of immune cells like mast cell. Mendelian randomization analysis confirmed that serum VCAN protein levels were a risky factor for DKD, while there was no reverse association. It exhibited the good diagnostic potential for estimated glomerular filtration rate and proteinuria in DKD. Conclusion VCAN showed the prospects into DKD pathology and clinical indicator.

Objective To investigate the effects of saikosaponin D(SSD)on the growth of esophageal squamous cell carcinoma(ESCC)Eca109 cells and the hypoxia-inducible transcription factor-2α(HIF-2α)/nuclear factor-κB p65(NF-κB p65)pathway.Methods Different concentrations of SSD(0,3,6,9,12,15,18,21,24 μmol·L-1)were used to treat Het-1a normal esophageal squamous epithelial cells and Eca109 ESCC cells to screen the experimental concentration of SSD.Eca109 cells were separated into control(NC)group,HIF-2α inhibitor(PT2385)group,SSD low and high concentration(SSD-L,SSD-H)groups,SSD-H+empty lentivirus(LV-NC)group,and SSD-H+HIF-2α recombinant lentivirus(LV-HIF-2α)group.MTT assay was applied to detect cell viability.Flow cytometry was applied to detect cell apoptosis.ELISA was applied to detect the levels of TNF-α,IL-1β,and IL-8 in cell culture media.Western blot was applied to detect cell proliferation,apoptosis,and protein expression related to the HIF-2α/NF-κB p65 pathway.The Eca109 cell tumor bearing nude mouse model was established to verify the effect of SSD on the in vivo growth of Eca109 cells.Results 6 and 12 μmol·L-1 SSD were selected as experimental concentrations.Compared with the NC group,the cell viability,TNF-α,IL-1β,IL-8 levels in cell culture medium,PCNA,Bcl-2,HIF-2α,p-NF-κB p65/NF-κB p65 protein levels,subcutaneous transplant tumor volume and weight in nude mice,serum TNF-α,IL-1β,IL-8 levels,and positive expression of HIF-2α and NF-κB p65 proteins in the PT2385 group,SSD-L group,and SSD-H group were lower,the apoptosis rate,Bax,and Cleaved-caspase3 protein levels were higher(P<0.05 or P<0.01).Overexpression of HIF-2α prominently weakened the inhibitory effects of SSD on the proliferation of Eca109 cells and the growth of subcutaneous transplanted tumors in nude mice(P<0.05 or P<0.01).Conclusion SSD can inhibit the proliferation of Eca109 cells,induce cell apoptosis,and suppress the growth of subcutaneous transplanted tumors of Eca109 cells in nude mice.Its effect may be related to the inhibition of the activation of the HIF-2α/NF-κB p65 pathway.

Objective:To investigate the differences in clinical outcomes of septic children with varying serum zinc levels,and to analyze the relationship between reduced serum zinc levels and organ dysfunction as well as 28-day mortality in septic children.Methods:This study conducted a retrospective analysis of clinical data from pediatric patients diagnosed with sepsis or septic shock in the Department of critical care medicine of the children's Hospital of Soochow University between January 2017 and December 2022.Clinical characteristics,organ dysfunction,and prognosis were compared between two groups:children with low serum zinc levels and those with normal zinc levels.Results:The serum zinc level of septic children within 24 hours of admission was 9.60(5.52,13.80)μmol/L,with 50.54%(94/186)of the children exhibiting low serum zinc levels(<10.07 μmol/L).Compared to the normal serum zinc group,the low serum zinc group had a significantly lower Pediatric Critical Illness Score(PCIS)[(78.71±9.35)vs.(85.12±8.51),P=0.005]and higher 28-day mortality(46.80%vs.14.13%,P<0.001).The low serum zinc group also had a higher proportion of invasive mechanical ventilation(64.89%vs.47.82%,P=0.019),renal replacement therapy(15.59%vs.3.26%,P=0.003),and use of vasoactive drugs(56.38%vs.30.43%,P<0.001).The rate of underlying conditions in the low serum zinc group was significantly higher than that in the normal serum zinc group(57.44%vs.36.95%,P=0.005).Additionally,the low serum zinc group had a higher incidence of disseminated intravascular coagulation(DIC),respiratory failure,acute kidney injury,shock,and multiple organ dysfunction syndrome(MODS)compared to the normal serum zinc group(P<0.05).Serum zinc levels had predictive value for 28-day mortality in septic children(AUC=0.813;95%CI:0.725～0.902;P<0.001).A serum zinc level of less than 6.950 μmol/L predicted the death of septic children with a sensitivity of 0.618 and a specificity of 0.902.Conclusion:Sepsis in children is commonly associated with low serum zinc levels,especially in those with underlying conditions such as hematologic and oncologic disorders.Sepsis patients hypozincemia with a higher incidence of DIC,respiratory failure,acute kidney injury,shock,and MODS.A serum zinc level below 6.95 μmol/L serves as a significant predictor of 28-day mortality in children with severe sepsis.

Objective To investigate the effects of saikosaponin D(SSD)on the growth of esophageal squamous cell carcinoma(ESCC)Eca109 cells and the hypoxia-inducible transcription factor-2α(HIF-2α)/nuclear factor-κB p65(NF-κB p65)pathway.Methods Different concentrations of SSD(0,3,6,9,12,15,18,21,24 μmol·L-1)were used to treat Het-1a normal esophageal squamous epithelial cells and Eca109 ESCC cells to screen the experimental concentration of SSD.Eca109 cells were separated into control(NC)group,HIF-2α inhibitor(PT2385)group,SSD low and high concentration(SSD-L,SSD-H)groups,SSD-H+empty lentivirus(LV-NC)group,and SSD-H+HIF-2α recombinant lentivirus(LV-HIF-2α)group.MTT assay was applied to detect cell viability.Flow cytometry was applied to detect cell apoptosis.ELISA was applied to detect the levels of TNF-α,IL-1β,and IL-8 in cell culture media.Western blot was applied to detect cell proliferation,apoptosis,and protein expression related to the HIF-2α/NF-κB p65 pathway.The Eca109 cell tumor bearing nude mouse model was established to verify the effect of SSD on the in vivo growth of Eca109 cells.Results 6 and 12 μmol·L-1 SSD were selected as experimental concentrations.Compared with the NC group,the cell viability,TNF-α,IL-1β,IL-8 levels in cell culture medium,PCNA,Bcl-2,HIF-2α,p-NF-κB p65/NF-κB p65 protein levels,subcutaneous transplant tumor volume and weight in nude mice,serum TNF-α,IL-1β,IL-8 levels,and positive expression of HIF-2α and NF-κB p65 proteins in the PT2385 group,SSD-L group,and SSD-H group were lower,the apoptosis rate,Bax,and Cleaved-caspase3 protein levels were higher(P<0.05 or P<0.01).Overexpression of HIF-2α prominently weakened the inhibitory effects of SSD on the proliferation of Eca109 cells and the growth of subcutaneous transplanted tumors in nude mice(P<0.05 or P<0.01).Conclusion SSD can inhibit the proliferation of Eca109 cells,induce cell apoptosis,and suppress the growth of subcutaneous transplanted tumors of Eca109 cells in nude mice.Its effect may be related to the inhibition of the activation of the HIF-2α/NF-κB p65 pathway.

Objective:To explore differences in the efficacy and safety of ruxolitinib in patients with myelofibrosis by age and to identify prognostic factors by analyzing clinical features and characteristics of chromosomes and gene mutations.Methods:This study retrospectively analyzed 188 patients with myelofibrosis who received ruxolitinib in the Department of Hematology, Qilu Hospital, Shandong University from January 1, 2017, to July 1, 2024. According to age at diagnosis, the patients were divided into the middle-aged group (≤55 years), young elderly group (56-65 years), and elderly group (>65 years). Clinical features, the characteristics of chromosomes and gene mutations, and the efficacy and safety of ruxolitinib treatment were compared across the three age groups. Independent factors influencing overall survival were identified through Cox proportional risk regression analysis.Results:Before treatment, the elderly group had more underlying comorbidities, a heavier symptom burden, higher leukocyte count, higher proportion and frequency of JAK2 mutations, and lower proportion of CALR mutations. The incidence of nondriver gene mutations was significantly higher in the young elderly group. After ruxolitinib treatment, the degree of reduction in spleen size did not differ significantly among the three groups. The length of the palpable spleen below the left costal margin reduced by more than 50% from baseline in 50.9% (27/53) of the patients in the middle-aged group, 43.5% (27/62) in the young elderly group, and 45.5% (20/44) in the elderly group ( P=0.720). No significant difference was observed among the three groups in the degree of reduction in Myeloproliferative Neoplasm Symptom Assessment Form (10-item version) score ( P=0.153), with a reduction in total symptom score by more than 50% achieved by 54.0% (27/50), 60.3% (41/68), and 66.7% (34/51) of the patients from the three groups, respectively ( P=0.429). The most common hematological adverse events were anemia and thrombocytopenia, while the most common nonhematological adverse events were electrolyte disturbance, elevated transaminase activity, and pulmonary infection. Multivariate analysis indicated that in ruxolitinib-treated patients with myelofibrosis, poor overall survival was independently predicted by increased age, reduced hemoglobin, percentage of bone marrow blasts ≥ 1%, absence of JAK2 mutations, chromosomal abnormalities, ≥2 high-molecular-risk mutations, and TP53 mutations. Conclusions:Patients with myelofibrosis stratified by age exhibited heterogeneous clinical features and gene mutation profiles but similar efficacy of ruxolitinib treatment and occurrence of adverse events.

Objective To establish a inductively coupled plasma-mass spectrometry(ICP-MS)method for 7 elemental impurities,cadmium(Cd),lead(Pb),arsenic(As),mercury(Hg),cobalt(Co),vanadium(V)and nickel(Ni)in medicinal excipient potassium sorbate according to the latest requirement of The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use(ICH)Q3D.Methods After dissolving the sample with 2％nitric acid solution,the potassium sorbate sample was digested by closed ultra-high pressure microwave digestion technology,and the elements rhenium(Re),rhodium(Rh),germanium(Ge),scandium(Sc)and bismuth(Bi)were used as internal standards to correct the matrix interference and drift.Results The 7 elements showed good linearity in the specified concentration range(r＞0.998 0).The detection limits were between 0.002-0.042 ng/g,the recovery rates were between 85％-106％,and the RSDs of each element precision were＜5％.The content of elemental impurities in 13 batches of domestic and imported potassium sorbate samples from different manufacturers was determined legally,and none exceeded the limit,which was in line with the provisions of ICH.Conclusion The established method is simple,accurate,sensitive,specific and reproducible.In this test,the elemental impurity residues in potassium sorbate are screened,which can not only grasp the overall quality level of potassium sorbate,but also ensure the safety of potassium sorbate as medicinal excipients,and provides a methodological basis for the revision of the Chinese Pharmacopoeia.