A retrospective analysis was conducted on the clinical characteristics, renal pathology, genetic testing, and treatment of five patients —two males and three females—diagnosed with primary atypical hemolytic uremic syndrome (aHUS) in the Department of Nephrology at the Affiliated Hospital of Qingdao University from February 2022 to June 2024. The patients′ ages at disease onset ranged from 14 to 29 years. Four patients experienced prodromal infection symptoms. At disease onset, serum creatinine levels ranged from 168.5 to 1 230.2 μmol/L. All patients presented with hematuria, proteinuria, hypertension, non-immune hemolytic anemia, thrombocytopenia, elevated lactate dehydrogenase (LDH), and fragmented red blood cells in peripheral blood (0.5%-6.0%). Serum haptoglobin levels were below the normal lower limit in all cases. Four patients demonstrated decreased serum complement C3, while one maintained normal serum complement C3 throughout the course of the disease. One patient exhibited serum factor H concentrations below the normal lower limit. Another patient tested positive for anti-factor H antibodies. Renal biopsies were performed on four patients. Electron microscopy revealed typical acute-phase pathological features of aHUS in three cases, including glomerular endothelial cell swelling and widened subendothelial spaces. One patient demonstrated ischemic and atrophic changes in the glomerular capillaries, while another had concurrent membranous nephropathy. Whole-exome high-throughput sequencing related to aHUS was performed in all five patients, revealing heterozygous gene mutations in each case. Complement-related gene mutations, typically occurring in a heterozygous state, are prevalent in aHUS patients. The eight heterozygous gene variations identified in this study were absent from existing databases of known aHUS-associated pathogenic mutations. Four patients received eculizumab treatment at varying time points following diagnosis, resulting in differing clinical outcomes. The patient positive for anti-factor H antibodies was treated with rituximab. The patient with membranous nephropathy initiated combination therapy with rituximab and eculizumab after six months of eculizumab monotherapy. Following treatment, all five patients achieved complete cessation of intravascular mechanical hemolysis, with normalization of LDH and platelet levels, as well as varying degrees of renal function recovery. From a pathophysiological perspective, the timely administration of the complement C5 inhibitor eculizumab can rapidly induce clinical remission, reduce the incidence of end-stage renal disease, and improve prognosis in patients with aHUS.

An efficient analytical method was developed for simultaneous detection of alkylamines and alkylamides in food packaging plastics using liquid chromatography-high resolution mass spectrometry(LC-HRMS).Based on the physicochemical properties of alkylamines and alkylamides,as well as the complexity of plastic samples,sample pretreatment and chromatographic-mass spectrometric parameters were optimized.The samples were extracted by vortex-ultrasonic extraction with a methanol-acetonitrile mixture for 15 min,followed by nitrogen evaporation to concentrate the extract,reconstitution,and analysis.The chromatographic mobile phase consisted of 0.1%formic acid aqueous solution and acetonitrile,and a gradient elution was used.The electrospray ionization(ESI)source was operated in positive ion mode,and mass spectrometry data were collected in full scan and data-dependent acquisition modes.Quantification was performed using an isotope-labeled internal standard method.The results showed that within the quantification range of 1-1000 ng/mL,the calibration curves exhibited good linearity(R2>0.99).Some compounds interfered with the validation experiments at higher concentrations,so only 10 kinds of target analytes were validated.Using a mixed food packaging plastic matrix,the recoveries at spiking levels of 40,400,and 4000 ng/g were mostly between 66.0%and 117.1%,with relative standard deviations ranging from 0.6%to 10.6%.The method was applied to detect 14 food packaging plastic samples,and the results showed that the concentrations of alkylamines and alkylamides ranged from not detected to 8924 ng/g.This method offered high sensitivity and accuracy,and was suitable for the screening and quantitative determination of alkylamines and alkylamides in plastics.

OBJECTIVE: This study aimed to investigate the prevalence of HIV pretreatment drug resistance (PDR) and the transmission clusters associated with PDR-related mutations in newly diagnosed, treatment-naive patients between 2020 and 2023 in Dehong prefecture, Yunnan province, China. METHODS: Demographic information and plasma samples were collected from study participants. PDR was assessed using the Stanford HIV Drug Resistance Database. The Tamura-Nei 93 model within HIV-TRACE was employed to compute pairwise matches with a genetic distance of 0.015 substitutions per site. RESULTS: Among 948 treatment-naive individuals with eligible sequences, 36 HIV subtypes were identified, with unique recombinant forms (URFs) being the most prevalent (18.8%, 178/948). The overall prevalence of PDR was 12.4% (118/948), and resistance to non-nucleotide reverse transcriptase inhibitors (NNRTIs), nucleotide reverse transcriptase inhibitors (NRTIs), and protease inhibitors (PIs) was 10.7%, 1.3%, and 1.6%, respectively. A total of 91 clusters were identified, among which eight showed evidence of PDR strain transmission. The largest PDR-associated cluster consisted of six CRF01_AE drug-resistant strains carrying K103N and V179T mutations; five of these individuals had initial CD4+ cell counts < 200 cells/μL. CONCLUSION The distribution of HIV subtypes in Dehong is diverse and complex. PDR was moderately prevalent (12.4%) between 2020 and 2023. Evidence of transmission of CRF01_AE strains carrying K103N and V179T mutations was found. Routine surveillance of PDR and the strengthening of control measures are essential to limit the spread of drug-resistance HIV strains.
Humans ; HIV Infections/virology* ; China/epidemiology* ; Drug Resistance, Viral ; Male ; Adult ; Female ; Middle Aged ; HIV-1/genetics* ; Anti-HIV Agents/therapeutic use* ; Myanmar/epidemiology* ; Young Adult ; Prevalence ; Adolescent ; Mutation

A retrospective analysis was conducted on the clinical characteristics, renal pathology, genetic testing, and treatment of five patients —two males and three females—diagnosed with primary atypical hemolytic uremic syndrome (aHUS) in the Department of Nephrology at the Affiliated Hospital of Qingdao University from February 2022 to June 2024. The patients′ ages at disease onset ranged from 14 to 29 years. Four patients experienced prodromal infection symptoms. At disease onset, serum creatinine levels ranged from 168.5 to 1 230.2 μmol/L. All patients presented with hematuria, proteinuria, hypertension, non-immune hemolytic anemia, thrombocytopenia, elevated lactate dehydrogenase (LDH), and fragmented red blood cells in peripheral blood (0.5%-6.0%). Serum haptoglobin levels were below the normal lower limit in all cases. Four patients demonstrated decreased serum complement C3, while one maintained normal serum complement C3 throughout the course of the disease. One patient exhibited serum factor H concentrations below the normal lower limit. Another patient tested positive for anti-factor H antibodies. Renal biopsies were performed on four patients. Electron microscopy revealed typical acute-phase pathological features of aHUS in three cases, including glomerular endothelial cell swelling and widened subendothelial spaces. One patient demonstrated ischemic and atrophic changes in the glomerular capillaries, while another had concurrent membranous nephropathy. Whole-exome high-throughput sequencing related to aHUS was performed in all five patients, revealing heterozygous gene mutations in each case. Complement-related gene mutations, typically occurring in a heterozygous state, are prevalent in aHUS patients. The eight heterozygous gene variations identified in this study were absent from existing databases of known aHUS-associated pathogenic mutations. Four patients received eculizumab treatment at varying time points following diagnosis, resulting in differing clinical outcomes. The patient positive for anti-factor H antibodies was treated with rituximab. The patient with membranous nephropathy initiated combination therapy with rituximab and eculizumab after six months of eculizumab monotherapy. Following treatment, all five patients achieved complete cessation of intravascular mechanical hemolysis, with normalization of LDH and platelet levels, as well as varying degrees of renal function recovery. From a pathophysiological perspective, the timely administration of the complement C5 inhibitor eculizumab can rapidly induce clinical remission, reduce the incidence of end-stage renal disease, and improve prognosis in patients with aHUS.

Objective:To explore the effects of hydroxychloroquine(HCQ)on immune mediators dysregulation in severe infection after chemotherapy in acute myeloid leukemia(AML)and its molecular mechanism.Methods:Bone marrow or peripheral blood samples of 36 AML patients with severe infection(AML-SI)and 29 AML patients without infection(AML-NI)after chemotherapy were collected from the First Affiliated Hospital of Gannan Medical University from August 2022 to June 2023.In addition,the peripheral blood of 21 healthy subjects from the same period in our hospital was selected as the control group.The mRNA expressions of CXCL12,CXCR4 and CXCR7 were detected by RT-qPCR technology,and the levels of IL-6,IL-8 and TNF-α were detected by ELISA.Leukemia-derived THP-1 cells were selected and constructed as AML disease model.At the same time,bone marrow mesenchymal stem cells(BM-MSCs)from AML-SI patients were co-cultured with THP-1 cells and divided into Mono group and Co-culture group.THP-1 cells were treated with different concentration gradients of HCQ.The cell proliferation activity was subsequently detected by CCK-8 method and apoptosis was detected by Annexin V/PI double staining flow cytometry.ELISA was used to detect the changes of IL-6,IL-8 and TNF-α levels in the supernatant of the cell co-culture system,RT-qPCR was used to detect the mRNA expression changes of the core members of the CXCL12-CXCR4/7 regulatory axis,and Western blot was used to detect the expressions of apoptosis regulatory molecules and related signaling pathway proteins.Results:CXCL12,CXCR4,CXCR7,as well as IL-6,IL-8,and TNF-α were all abnormally increased in AML patients,and the increases were more significant in AML-SI patients(P＜0.01).Furthermore,there were statistically significant differences between AML-NI patients and AML-SI patients(all P＜0.05).HCQ could inhibit the proliferation and induce the apoptosis of THP-1 cells,but the low concentration of HCQ had no significant effect on the killing of THP-1 cells.When THP-1 cells were co-cultured with BM-MSCs of AML patients,the levels of IL-6,IL-8 and TNF-α in the supernatance of Co-culture group were significantly higher than those of Mono group(all P＜0.01).After HCQ intervention,the levels of IL-6,IL-8 and TNF-α in cell culture supernatant of Mono group were significantly decreased compared with those before intervention(all P＜0.01).Similarly,those of Co-culture group were also significantly decreased(all P＜0.001).However,the expression of the core members of the CXCL12-CXCR4/7 regulatory axis was weakly affected by HCQ.HCQ could up-regulate the expression of pro-apoptotic protein Bax,down-regulate the expression of anti-apoptotic protein Bcl-2,as well as simultaneously promote the hydrolytic activation of Caspase-3 when inhibiting the activation level of TLR4/NF-κ B pathway,then induce the programmed death of THP-1 cells after intervention.Conclusion:The core members of CXCL12-CXCR4/7 axis and related cytokines may be important mediators of severe infectious immune disorders in AML patients.HCQ can inhibit cytokine levels to reverse immune mediators dysregulation and suppress malignant biological characteristics of leukemia cells.The mechanisms may be related to regulating the expression of Bcl-2 family proteins,hydrolytically activating Caspase-3 and inhibiting the activation of TLR4/NF-κ B signaling pathway.

Aim To establish a simple and practical method for visualizing neurovascular coupling in the mouse barrel cortex in vivo.Methods Male C57BL/6J mice received stereotaxic in-jections of pAAV-hSyn-jGCaMP7s-WPRE into the barrel cortex to monitor neuronal activity.Three weeks post-injection,a crani-al window was implanted,and TRITC-Dextran 155 ku was ad-ministered intravenously to visualize the vasculature and blood flow dynamics.A custom-built whisker stimulator was used to e-voke controlled neuronal excitation.Two-photon microscopy was employed to monitor neuronal and vascular responses to whisker stimulation in real-time.Results Neuronal calcium signals and plasma signals were clearly observed using two-photon microsco-py.Whisker stimulation led to a significant increase in neuronal calcium signals in the barrel cortex,indicating effective neuronal activation.This neuronal excitation was accompanied by a syn-chronous increase in blood vessel diameter,blood flow velocity and overall blood flow.Conclusions This study successfully establishes a three-dimensional visualization framework(spatial,temporal,and functional)for in vivo visualization of neurovascu-lar coupling in the mouse barrel cortex,which provides a useful tool for investigating the pathophysiological mechanisms of neuro-vascular dysfunction and evaluating the efficacy of potential ther-apies.

Objective·To analyze the molecular epidemiological characteristics of Klebsiella pneumoniae isolated from children,including the distribution of serotypes,resistance genes,and virulence genes,to provide a scientific basis for the prevention and treatment strategies of Klebsiella pneumoniae infections in children.Methods·A total of 133 non-duplicate strains of Klebsiella pneumoniae clinically isolated from Shanghai Children's Hospital,Shanghai Jiao Tong University School of Medicine,from April 2023 to April 2024,were collected.Antimicrobial susceptibility testing was performed by using the Vitek-2 Compact system.Capsular polysaccharide K antigen serotypes(K types)were determined by wzy-dependent initiator(wzi)gene sequencing.Resistance genes were detected by the colloidal gold method.Virulence genes and lipopolysaccharide O antigen serotypes(O types)were identified by PCR method.Results·Among the 133 strains of Klebsiella pneumoniae,a total of 50 strains of carbapenem-resistant Klebsiella pneumoniae(CRKP)were detected,mainly distributed in the neonatal ward and intensive care unit(ICU).CRKP exhibited high resistance to most antimicrobial agents,and the main type of carbapenemase gene was blaNDM(34/50,68.00%),followed by co-carriage of blaNDM+blaOXA-48(10/50,20.00%)and blaKPC(6/50,12.00%).Serotype analysis revealed that the 50 CRKP strains could be divided into 8 K types and 6 O types.The most common K type was KL17(30/50,60.00%),followed by KL105(10/50,20.00%)and KL47(5/50,10.00%).The most common O type was O4(30/50,60.00%),followed by O3b(7/50,14.00%,),O3/O3a(6/50,12.00%),and OL101(5/50,10.00%).Significant correlations were identified between KL47 and OL101 serotypes(KL47:OL101),between KL17 and O4 serotypes(KL17:O4),and between KL1/KL2 and O1 serotypes(KL1/KL2:O1).Conclusion·The CRKP strains infecting children primarily carry blaNDM-type carbapenemase genes,showing a significant difference compared to CRKP-infected adults.The detection rate of CRKP in the neonatal ward is significantly higher than those in other departments,suggesting that infection prevention and control measures in this ward need to be strengthened.

OBJECTIVES: To explore the efficacy and adverse reactions of nusinersen combined with risdiplam in the treatment of spinal muscular atrophy (SMA). METHODS: A retrospective analysis was conducted on the clinical data of 10 pediatric SMA patients treated with nusinersen combined with risdiplam at the Children's Medical Center of Xiangya Hospital, Central South University. RESULTS: Among the 10 SMA patients, there were 4 with type I, 4 with type II, and 2 with type III. Nine patients initially received nusinersen monotherapy, while 1 patient received nusinersen combined with risdiplam. The median duration of combination therapy with nusinersen and risdiplam for the 10 patients was 10.5 months (range: 0.5-20.0 months), with 6 patients undergoing combination therapy for more than 6 months, showing improvements in motor and/or respiratory function. The remaining 4 patients had combination treatment durations of 0.5, 1.0, 1.3, and 4.0 months, respectively, with no significant overall improvement. After combined treatment, 5 patients experienced skin hyperpigmentation, 2 had lumbar puncture site pain, 1 experienced vomiting, 1 had increased sputum production, and 1 had reduced total sleep time. All adverse reactions were mild and did not require medical intervention. CONCLUSIONS Nusinersen combined with risdiplam demonstrates efficacy in the treatment of SMA, and no significant adverse reactions have been observed.
Humans ; Oligonucleotides/adverse effects* ; Male ; Female ; Child, Preschool ; Retrospective Studies ; Infant ; Muscular Atrophy, Spinal/drug therapy* ; Drug Therapy, Combination ; Child ; Azo Compounds ; Pyrimidines

The patient was a girl, aged 10 years, who was admitted due to fever for 5 days and pancytopenia in peripheral blood for 2 days. Bone marrow examination showed the presence of phagocytic activity, and peripheral blood tests showed pancytopenia, an increase in ferritin, a reduction in fibrinogen, increases in triglyceride and sCD25, and a reduction in natural killer cell activity, which led to the diagnosis of hemophagocytic lymphohistiocytosis (HLH). On the day of admission, the child developed convulsions and rapidly progressed to refractory status epilepticus, which was consistent with the manifestations of febrile infection-related epilepsy syndrome. HLH was controlled after active immunotherapy, with the sequela of refractory epilepsy, and her cognitive function was essentially within normal limits. This article reports the condition of febrile infection-related epilepsy syndrome caused by HLH for the first time in China, in order to improve the awareness of this disease among clinicians.
Humans ; Lymphohistiocytosis, Hemophagocytic/complications* ; Female ; Child ; Epilepsy/etiology* ; Fever/etiology* ; Epileptic Syndromes/etiology*

Objective:To investigate the clinicopathological features of advanced-stage mycosis fungoides (MF).Methods:A retrospective case-series study was conducted. The clinical data of 5 cases diagnosed with advanced-stage MF in Chengdu Second People's Hospital between January 2015 and July 2023 were analyzed. The clinicopathological features of patients were summarized.Results:There were 2 males and 3 females in 5 MF patients, with the median age of 55 years (45-86 years) and the medical history of 2-16 years. The main symptoms were pruritus and erythema. The lesions were presented by erythema, scales, plaques, blisters, erosion, ulcers, pigmentation, nodules, and erythroderma. Histopathological examination showed different skin lesion patterns such as psoriasis-like, interfacial dermatitis, non-infectious granuloma, deep and shallow perivascular dermatitis, tumors. Among 5 patients, 1 case was mycosis fungoides bullosa, 2 cases were erythrodermic MF, 1 case was granulomatous MF, and 1 case was classical MF. Lymphocyte epidermis was found in 4 cases, cytoplasmic halos cells lined up along the basal layer of the epidermis and Pautrier microabscess were found in 3 cases, large-cell transformation was found in 1 case. Tumor cells were positive for CD3, CD4 and negative for CD8, CD56, ALK and CD20; EBER 1/2 hybridization in situ was negative. CD30 was positive in transformed large cells and T cell receptor gene rearrangement was positive. The tumor cells were detected in bone marrow and peripheral blood of 2 cases and in cerebrospinal fluid of 1 case. Head magnetic resonance imaging of 1 case indicated abnormal signal nodules in the right temporal region and the normal architecture of the lymph nodes in 2 cases was completed destroyed by malignant cells. TNMB stage: 2 cases were in stage Ⅱ B, 2 cases were in stage Ⅳ A2, and 1 case was in stage Ⅳ B. Interferon α-based systemic therapy was performed in 1 case, 2 cases received chemotherapy or combined with intrathecal injection and radiotherapy, and other 2 cases were not treated. All of them just achieved partial remission. Finally, 1 case died of sudden cardiac death, 2 cases died of lung infection, and 2 cases survived with tumors. Conclusions:Advanced-stage MF is presented with different skin lesion manifestations and histopathologic changes. Multidisciplinary combined management helps the diagnosis and treatment of MF.