Review and Progress in the Treatment of Renal Light Chain Amyloidosis / 罕见病研究

Renal light chain amyloidosis (AL amyloidosis) had poor prognosis before the 21st century. However, the treatment of AL amyloidosis has made great progress in the last decade. We reviewed traditional treatments of AL amyloidosis such as alkylating agents, proteasome inhibitors, and recent advances such as monoclonal antibodies. Bortezomib improved the hematological response and survival effectively of the patients, and the combination of Daratumumab brings faster and deeper hematological response, increasing the response rate of target organs such as the kidneys and heart. The renal response was significant higher in the patients with the therapy of Daratumumab, part of them could achieve very good partial response or better renal response. Autologous hematopoietic stem cell transplantation(auto-HSCT)improves hematological as well as organ response, and could be the first choice among eligible patients. Kidney transplantation is a feasible option for those with good hematological response.

Primary systemic amyloidosis in a 66-year old Filipina presenting with extracardiac uptake on Tc-99m pyrophosphate (Tc-99m PYP) scintigraphy

@#This is a case of a 66-year-old, Filipina, who presented with persistent proteinuria diagnosed with renal amyloidosis. In the presented case, the equivocal cardiac and incidental extracardiac findings in the Tc-99m pyrophosphate (Tc-99m PYP) scan aided in the diagnosis of primary systemic light chain amyloidosis (AL amyloidosis). Tc-99m PYP scan with single photon emission computed tomography (SPECT) is currently used as a non-invasive imaging modality to diagnose Transthyretin amyloidosis (ATTR amyloidosis) however its role in diagnosing AL amyloidosis is not well documented. The case highlights its role in detecting extracardiac amyloid burden and suggests possible biopsy sites. The researchers recommend an additional whole-body planar scan with possible SPECT/CT on the 3rd hour delay to survey other areas with possible amyloid protein deposit.

Review and Progress in the Treatment of Renal Light Chain Amyloidosis / 罕见病研究

Renal light chain amyloidosis (AL amyloidosis) had poor prognosis before the 21st century. However, the treatment of AL amyloidosis has made great progress in the last decade. We reviewed traditional treatments of AL amyloidosis such as alkylating agents, proteasome inhibitors, and recent advances such as monoclonal antibodies. Bortezomib improved the hematological response and survival effectively of the patients, and the combination of Daratumumab brings faster and deeper hematological response, increasing the response rate of target organs such as the kidneys and heart. The renal response was significant higher in the patients with the therapy of Daratumumab, part of them could achieve very good partial response or better renal response. Autologous hematopoietic stem cell transplantation(auto-HSCT)improves hematological as well as organ response, and could be the first choice among eligible patients. Kidney transplantation is a feasible option for those with good hematological response.

Effect of saikosaponin A regulating MLCK/MLC2 signaling pathway on intestinal injury in SAP rats / 国际检验医学杂志

Objective To investigate the effect of saikosaponin A regulating myosin light chain kinase(MLCK)/myosin light chain 2(MLC2)signaling pathway on intestinal injury in rats with severe acute pan-creatitis(SAP).Methods A total of 10 rats were randomly selected as sham operation group,and the other rats were injected with sodium taurine cholate solution to construct SAP rat model.SAP rat models were ran-domly divided into SAP group,saikosaponin A group(10.0 mg/kg intraperitoneal injection of saikosaponin A)and iE-DAP group(3.5 mg/kg intraperitoneal injection of MLCK/MLC2 pathway activator iE-DAP),sai-kosaponin A+iE-DAP group(intraperitoneally injected with 10.0 mg/kg saikosaponin A+3.5 mg/kg iE-DAP),10 rats in each group were injected once a day for 1 week,sham operation group and SAP group were injected with the same amount of normal saline.The serum levels of amylase(AMY),lipase(LIP),diamine oxidase(DAO),interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)were detected by enzyme-linked immunosorbent assay(ELISA).HE staining was used to detect the pathological morphology of ileum tis-sue in each group.The levels of oxidative stress indexes in ileum tissue were detected by ELISA.Intestinal barrier re-lated proteins and MLCK/MLC2 pathway related proteins were detected by western blot.Results Compared with SAP group,the levels of AMY,LIP,DAO,IL-1β,IL-6 and TNF-α in saikosonin A group were significantly de-creased,while the levels of AMY,LIP,DAO,IL-1β,IL-6 and TNF-α in iE-DAP group were significantly in-creased,with statistical significance(P＜0.05).Compared with SAP group,the structure of ileum tissue was improved and the pathological score of ileum tissue was significantly decreased in SA group(P＜0.05).Com-pared with SAP group,the levels of glutathione and superoxide dismutase in SA group were significantly in-creased,and the levels of malondialdehyde were significantly decreased in SA group,with statistical signifi-cance(P＜0.05).Compared with SAP group,the protein levels of MLCK,p-MLC2/MLC2 in SA group were significantly decreased,and the difference was statistically significant(P＜0.05).Conclusion Saikosaponin A may improve intestinal injury in SAP rats by down-regulating the MLCK/MLC2 signaling pathway.

Establishment and verification of a real-time quantitative PCR method for determination of exogenous antibody gene copy number in CHO cells / 中国生物制品学杂志

@#Objective To establish a real-time quantitative PCR method using SYBR GreenⅠto detect the copy numbers of light chain(LC)and heavy chain(HC)of exogenous antibody gene in CHO cells,and verify and preliminarily apply this method.Methods With the B2m(β2-microglobulin)expressed stably in CHO cells as the internal reference gene,suitable primers of LC,HC genes and internal reference gene were designed respectively,and the reaction system and program of the real-time quantitative PCR method were determined. The established method was verified for the specificity,linearity,precision and durability,and used to detect the copy numbers of LC and HC genes in the recombinant cell lines of working cell bank(WCB)and cells of different passages.Results The primers of exogenous genes and internal reference gene showed specific binding to the target fragments;The efficiency of primer amplification for the B2m gene,LC gene,and HC gene was 106. 7%,106. 3% and 99. 1%,respectively,and the correlation coefficients of the linear equations were all greater than 0. 99 with a good linear relationship;The relative standard deviations(RSDs)of precision verification were all less than 1%;Few cycles of freeze-thaw in a short period had little effect on the detection results. The copy numbers of LC and HC genes in different generations of recombinant cell lines detected by the established method showed no obvious changes.Conclusion A real-time quantitative PCR method for the determination of the copy number of exogenous genes in CHO cells was successfully established with good specificity,linearity,precision and durability,which provides a reference for detecting the copy number of exogenous genes expressed in other CHO cell lines

Effects of Shengxue Tongbian Granules on Colonic Myoelectric and Ca2+/CaM/MLCK Signaling Pathway in Rats with Slow Transit Constipation of Blood-deficiency and Intestinal Dryness Syndrome / 中国中医药信息杂志

Objective To observe the effects of Shengxue Tongbian Granules on colonic myoelectricity and Ca2+/CaM/MLCK signaling pathway in rats with slow transit constipation(STC)of blood deficiency and intestinal dryness syndrome;To explore its mechanism for the treatment of STC.Methods The STC model of blood deficiency and intestinal dryness syndrome was established by intragastric administration of loperamide combined with tail bloodletting.The rats were divided into control group,model group,mosapride citrate group and Shengxue Tongbian Granules group,with 8 rats in each group.The administration group was given corresponding drugs by gavage for 14 days.The general condition of rats before and after treatment was observed,the fecal water content was detected,the slow wave frequency,amplitude,and coefficient of variation of colonic electromyography were detected using a biological function experimental system,and intestinal propulsion rate was detected.HE staining was used to observe the pathological changes of colon tissue,the concentration of Ca2+ in colonic smooth muscle cells was detected by colorimetry,the expression of Cx43,calmodulin(CaM),myosin light chain kinase(MLCK)and p-MLC20 in smooth muscle tissue were detected by Western blot.Results Compared with the control group,the body mass,fecal water content and intestinal propulsion rate of rats in the model group decreased(P<0.01),the slow wave frequency of colonic electromyography slowed down,the coefficient of variation of frequency increased(P<0.01),and the amplitude and coefficient of variation of slow wave increased(P<0.01);colonic mucosal structure was damaged,with visible inflammatory changes and significant erosion,and the concentration of Ca2+ and the expressions of Cx43,CaM,MLCK,p-MLC20 proteins in colonic smooth muscle cells were significantly decreased(P<0.01).Compared with the model group,the body mass,fecal water content and intestinal propulsion rate of the rats in the mosapride citrate group and the Shengxue Tongbian Granules group significantly increased(P<0.05,P<0.01),the slow wave frequency of colonic electromyography increased and the coefficient of variation of frequency decreased(P<0.01),and the slow wave amplitude and amplitude variation coefficient decreased(P<0.05,P<0.01);the colonic mucosal structure was relatively intact,the erosion situation was improved,and the Ca2+ concentration,Cx43,CaM,MLCK and p-MLC20 protein expressions in colonic smooth muscle cells significantly increased(P<0.01).Conclusion Shengxue Tongbian Granules can improve defecation symptoms and promote colonic motility in STC rats with blood deficiency and intestinal dryness syndrome,which may be related to regulating colonic myoelectric rhythm and activating Ca2+/CaM/MLCK signaling pathway.

¿Qué hay de nuevo en el diagnóstico de amiloidosis? / What is new in diagnosis of amyloidosis?

La amiloidosis siempre ha representado un desafío diagnóstico. En el año 2020, el Grupo de Estudio de Amiloidosis (GEA), confeccionó la Guía de Práctica Clínica para el Diagnóstico de Amiloidosis. Nuevas líneas de investigación se han desarrollado posteriormente. Esta revisión narrativa tiene como intención explorar el estado del arte en el diagnóstico de la amiloidosis. En pacientes con amiloidosis se recomienda la tipificación de la proteína mediante espectrometría de masa, técnica de difícil ejecución por requerir de microdisectores láser para la preparación de la muestra. Algunas publicaciones recientes proponen otros métodos para obtener la muestra de amiloide que se va a analizar, permitiendo prescindir de la microdisección. Por otra parte, en pacientes con Amiloidosis ATTR confirmada, la recomendación de secuenciar el gen amiloidogénico se encontraba destinada a los casos sospechosos de ATTR hereditaria (ATTRv,), pero actualmente esta se ha extendido a todos los pacientes sin importar la edad. En lo que respecta a los estudios complementarios orientados al diagnóstico de compromiso cardíaco, se ha propuesto el uso de la inteligencia artificial para su interpretación, permitiendo la detección temprana de la enfermedad y el correcto diagnóstico diferencial. Para el diagnóstico de neuropatía, las últimas publicaciones proponen el uso de la cadena ligera de neurofilamento sérica, que también podría resultar un indicador útil para seguimiento. Finalmente, con referencia a la amiloidosis AL, la comunidad científica se encuentra interesada en definir qué características determinan el carácter amiloidogénico de las cadenas livianas. La N-glicosilación de dichas proteínas impresiona ser uno de los determinantes en cuestión. (AU)

Amyloidosis has always represented a diagnostic challenge. In 2020, the Amyloidosis Study Group (ASG) developed the "Clinical Practice Guideline for the Diagnosis of Amyloidosis". New lines of research have subsequently emerged. This narrative review aims to explore the state of the art in the diagnosis of amyloidosis diagnosis. In patients with amyloidosis, protein typing by mass spectrometry is recommended, a technique hard to perform because it requires laser microdissection for sample preparation. Recent publications propose other methods to obtain the amyloid sample to be analyzed, making it possible to dispense with microdissection. On the other hand, in patients with confirmed TTR amyloidosis (aTTR), the recommendation to sequence the amyloidogenic gene was intended for suspected cases of hereditary aTTR but has now been extended to all patients regardless of age. (AU)

Trasplante cardíaco y autólogo de células progenitoras hematopoyéticas en pacientes con amiloidosis AL / Cardiac and autologous stem cell transplantation hematopoietic in patients with AL amyloidosis

Resumen Objetivo: Describir la evolución de las cadenas livianas libres séricas (CLL) en el período comprendido entre el trasplante cardíaco ortotópico (TCO) y el trasplante de células progenitoras hematopoyéticas (TCPH), la respuesta hematológica al año tras el TCPH y el tratamiento quimioterápico e inmunosupresor en pacientes con amiloidosis AL. Método: Serie de casos de pacientes consecutivos con diagnóstico de amiloidosis AL que recibieron TCO seguido de TCPH del Registro Institucional de Amiloidosis del Hospital Italiano de Buenos Aires, entre enero de 2010 y noviembre de 2021. Se reportaron los valores de CLL entre trasplantes y al año del TCPH. Las variables cuantitativas se describieron como mediana e intervalo intercuartil, y las variables categóricas como frecuencias absolutas y relativas. Resultados: De 106 pacientes con amiloidosis AL, seis tuvieron TCO seguido de TCPH. La mediana de edad fue de 55 años. La mayoría eran hombres (n = 5). En el período entre trasplantes, la CLL involucrada disminuyó en dos pacientes y se mantuvo estable en tres. Todos lograron la remisión hematológica completa al año del TCPH. Un solo paciente presentó recaída en el órgano sólido trasplantado. Tacrolimus, micofenolato de mofetilo y corticoides fue el esquema inmunosupresor utilizado después del TCO. Conclusiones: El TCO representa una opción de tratamiento en pacientes con falla cardíaca grave por amiloidosis, permitiendo luego un tratamiento intensivo con quimioterapia de inducción y TCPH. Si bien faltan estudios, la terapia inmunosupresora después del TCO podría tener algún efecto sobre las células plasmáticas clonales.

Abstract Objective: To describe the evolution of serum free light chains (FLC) in the period between orthotopic heart transplantation (OHT) and autologous stem cell transplantation (ASCT), the hematological response one year after ASCT and chemotherapy and immunosuppressive treatment in patients with AL amyloidosis. Method: Case series of consecutive patients diagnosed with AL amyloidosis who received OHT followed by ASCT from the Institutional Registry of Amyloidosis of the Italian Hospital of Buenos Aires, between January 2010 and November 2021. FLC values between transplants and at year post ASCT. Quantitative variables were described with their median and interquartile range. Categorical variables as absolute and relative frequencies. Results: Of 106 patients with AL amyloidosis, 6 had an OHT followed by ASCT. The median age was 55 years. Most were men (n = 5). In the period between transplants, the involved CLL decreased in two patients and remained stable in three. All achieved complete hematologic remission 1 year after ASCT. A single patient presented relapse in the transplanted solid organ. Tacrolimus, mycophenolate mofetil, and corticosteroids were the immunosuppressive regimen used after OHT. Conclusions: OHT represents a treatment option in patients with severe heart failure due to amyloidosis, allowing later intensive treatment with induction chemotherapy and ASCT. Although studies are lacking, immunosuppressive therapy after OHT might have some effect on clonal plasma cells.

Clinical characterization and outcomes of a cohort of colombian patients with AL Amyloidosis

Background: Amyloid light chain (AL) amyloidosis is characterized by amyloid fibril deposition derived from monoclonal immunoglobulin light chains, resulting in multiorgan dysfunction. Limited data exist on the clinical features of AL amyloidosis. Objective: This study aims to describe the clinical characteristics, treatments, and outcomes in Colombian patients with AL amyloidosis. Methods: A retrospective descriptive study was conducted at three high-complexity centers in Medellín, Colombia. Adults with AL amyloidosis diagnosed between 2012 and 2022 were included. Clinical, laboratory, histological, treatment, and survival data were analyzed. Results: The study included 63 patients. Renal involvement was most prevalent (66%), followed by cardiac involvement (61%). Multiorgan involvement occurred in 61% of patients. Amyloid deposition was most commonly detected in renal biopsy (40%). Bortezomib-based therapy was used in 68%, and 23.8% received high-dose chemotherapy with autologous hematopoietic stem cell transplantation (HDCT-ASCT). Hematological response was observed in 95% of patients with available data. Cardiac and renal organ responses were 15% and 14%, respectively. Median overall survival was 45.1 months (95% CI: 22.2-63.8). In multivariate analysis, cardiac involvement was significantly associated with inferior overall survival (HR 3.27; 95% CI: 1.23-8.73; p=0.018), HDCT-ASCT had a non-significant trend towards improved overall survival (HR 0.25; 95% CI: 0.06-1.09; p=0.065). Conclusions: In this study of Colombian patients with AL amyloidosis, renal involvement was more frequent than cardiac involvement. Overall survival and multiorgan involvement were consistent with data from other regions of the world. Multivariate analysis identified cardiac involvement and HDCT-AHCT as possible prognostic factors.

Antecedentes: La amiloidosis por amiloide de cadenas ligeras (AL) se caracteriza por el depósito de fibrillas amiloides derivadas de cadenas ligeras de inmunoglobulinas monoclonales, lo que resulta en disfunción multiorgánica. Existen datos limitados sobre las características clínicas de la amiloidosis AL. Objetivo: Este estudio tiene como objetivo describir las características clínicas, tratamientos y desenlaces en pacientes colombianos con amiloidosis AL. Métodos: Se llevó a cabo un estudio descriptivo retrospectivo en tres centros de alta complejidad en Medellín, Colombia. Se incluyeron adultos con diagnóstico de amiloidosis AL entre 2012 y 2022. Se analizaron datos clínicos, de laboratorio, histológicos, de tratamiento y de supervivencia. Resultados: El estudio incluyó 63 pacientes. La afectación renal fue más prevalente (66%), seguida de la afectación cardíaca (61%). El 61% de los pacientes presentaron afectación multiorgánica. El depósito amiloide se detectó con mayor frecuencia en la biopsia renal (40%). El tratamiento basado en bortezomib se utilizó en el 68%, y el 23.8% recibió altas dosis de quimioterapia con trasplante autólogo de progenitores hematopoyéticos (ADQT-TAPH). Se observó respuesta hematológica en el 95% de los pacientes con datos disponibles. La respuesta de órgano cardíaca y renal fue del 15% y 14%, respectivamente. La mediana de la supervivencia global fue de 45.1 meses (IC del 95%: 22.2-63.8). En el análisis multivariado, la afectación cardíaca se asoció significativamente con una supervivencia global inferior (HR 3.27; IC del 95%: 1.23-8.73; p=0.018), ADQT-TAPH mostró una tendencia no significativa hacia una mejora en la supervivencia global (HR 0.25; IC 95%: 0.06-1.09; p=0.065). Conclusiones: En este estudio de pacientes colombianos con amiloidosis AL, la afectación renal fue más frecuente que la afectación cardíaca. La supervivencia global y la afectación multiorgánica fueron consistentes con datos de otras regiones del mundo. El análisis multivariado identificó la afectación cardíaca y ADQT-TAPH como posibles factores pronósticos.

A case report of multiple myeloma complicated with primary systemic light amyloidosis peripheral neuropa-thy / 中国神经精神疾病杂志

This paper reports a case of multiple myeloma complicated with Primary systemic light chain amyloidosis peripheral neuropathy.Patient was a male,60 years old with subacute onset of peripheral neuropathy.Patient had previous history of hypertension,coronary heart disease and stable angina pectoris.His urine routine examination 2 months ago showed urine protein(+++).The first clinical symptom with peripheral neuropathy characterized by progressive numbness and weakness of both lower limbs.The diagnosis was made after completing electromyography,lumbar puncture,renal puncture,bone puncture and other related examinations,and relevant treatment was given.As a rare disease of nervous system,the incidence rate is low.Early diagnosis is an important step to reduce mortality and improve prognosis.However,the disease is easily misdiagnosed as diabetic peripheral neuropathy and chronic inflammatory demyelinating polyradiculoneuropathy,thereby delaying diagnosis and treatment.

Generation of Mlk3 KO mice by CRISPR/Cas9 and its effect on blood pressure / 生物工程学报

To explore the effect of Mlk3 (mixed lineage kinase 3) deficiency on blood pressure, Mlk3 gene knockout (Mlk3KO) mice were generated. Activities of sgRNAs targeted Mlk3 gene were evaluated by T7 endonuclease I (T7E1) assay. CRISPR/Cas9 mRNA and sgRNA were obtained by in vitro transcription, microinjected into zygote, followed by transferring into a foster mother. Genotyping and DNA sequencing confirmed the deletion of Mlk3 gene. Real- time PCR (RT-PCR), Western blotting or immunofluorescence analysis showed that Mlk3KO mice had an undetectable expression of Mlk3 mRNA or Mlk3 protein. Mlk3KO mice exhibited an elevated systolic blood pressure compared with wild-type mice as measured by tail-cuff system. Immunohistochemistry and Western blotting analysis showed that the phosphorylation of MLC (myosin light chain) was significantly increased in aorta isolated from Mlk3KO mice. Together, Mlk3KO mice was successfully generated by CRISPR/Cas9 system. MLK3 functions in maintaining blood pressure homeostasis by regulating MLC phosphorylation. This study provides an animal model for exploring the mechanism by which Mlk3 protects against the development of hypertension and hypertensive cardiovascular remodeling.

Diagnosis and treatment progress of primary light-chain amyloidosis / 白血病·淋巴瘤

Primary light-chain (AL) amyloidosis is a rare and fatal plasma cell disease. In recent years, the treatment of AL amyloidosis has changed from the era of bortezomib to the era of daratumumab immunotherapy. However, for the treatment choice of advanced-staged patients, how to achieve organ responses at the early stage and how to monitor the disease are questions that need to be further explored. The 64th American Society of Hematology Annual Meeting in 2022 has reported advances in the diagnosis and treatment of AL amyloidosis, which are briefly reviewed in this article.

Critical Management Strategies for Cardiovascular Involvementin Light Chain Amyloidosis / 罕见病研究

Light chain amyloidosis (AL) is a kind of rare disease. The misfolding of the light chain of monoclonal immunoglobulin forms amyloid substances and deposit in different tissues and organs, resulting in organ dysfunction. The heart involvement is represented as light chain cardiac amyloidosis (AL-CA), leading to heart failure, arrhythmia, myocardial infarction, valvular and pericardial diseases. Cardiac involvement is crucial for the risk stratification and prognosis prediction of patients with AL. The median survival time of AL-CA patients is 24 months and only 6 months for the heart failure patients. In recent years, the prognosis of patients with AL-CA has been significantly improved, thanks to the advancement in diagnosis & treatment strategies of cardiac amyloidosis, the milestone in the breakthroughs in targeted therapy of AL, and the improvement of coordinated multidisciplinary treatment. This paper focuses on the cardiovascular involvement of AL and briefly illustrates the pathogenesis, clinical manifestations, and the diagnosis and treatment of AL-CA.

Construction and application of a fluorescent expression cell model of botulinum toxin type B light chain / 军事医学

Objective To construct a fluorescent expression cell model of botulinum toxin type B light chain(BoNT/B,BLC),and evaluate the effect of small molecule compounds with this model.Methods The BLC gene was inserted into the fluorescent expression vector pEGFP-N1 to construct a recombinant plasmid before being transfected into the neural cell line Neuro-2a cells for expression.The fluorescence expression level of BLC-EGFP protein in the cells was observed under a fluorescence microscope,and the enzyme digestion activity and stability of BLC-EGFP in the cells were detected by Western blotting.Furthermore,the model was used to evaluate the effect of SRC kinase inhibitor KX2-391 on the intracellular stability of BLC-EGFP protein.Results The recombinant expression plasmid pEGFP-N1-BLC was constructed.It was found that the expression level of BCL-EGFP protein in Neuro-2a cells gradually increased over time,and that the intracellular substrate vesicle-associated membrane protein-2(VAMP-2)was cleaved after plasmid transfection.CHX was added to terminate protein synthesis after the plasmid was transfected for 12 h,and the intracellular level of BLC-EGFP did not change significantly within 72 h.Twenty-four hours of treatment with KX2-391 could significantly promote the intracellular degradation of BLC-EGFP protein.Conclusions A cell model of fluorescent expression of botulinum toxin type B light chain has been established,which provides a technical reserve for the subsequent study onthe intracellular persistence mechanism and intracellular antidote screening of botulinum toxin type B light chain.

Predictive value of serum neurofilament light chain on neurologic function in out-of-hospital cardiac arrest patients / 中华急诊医学杂志

Objective:To determine the predictive value of serum neurofilament light chain (NfL) on neurologic function in out-of-hospital cardiac arrest (OHCA) patients.Methods:The clinical data of 96 OHCA patients admitted to Cangzhou Central Hospital from January 2018 to March 2022 were retrospectively analyzed. According to the Glasgow-Pittsburgh cerebral performance category (CPC) upon hospital discharge, the patients were divided into the favorable neurologic function (grade 1-2) and poor neurologic function (grade 3-5) groups. The difference of serum NfL was compared between the two groups, and the relationship between serum NfL and neurologic function was assessed using correlation analysis and logistic regression analysis. The area under the curve (AUC), sensitivity, and specificity of serum Nfl were calculated by receiver operating characteristic (ROC) curve. Hanley & McNeil method test was used to compare the difference of AUCs between serum NfL and neuron specific enolase (NSE).Results:Twenty-six percent (25/96) patients were discharged with favorable neurologic function. Serum NfL in the favorable neurological function group was significantly lower than that in the poor neurologic function group (47.6 pg/mL vs. 261.4 pg/mL, P<0.001). Correlation analysis showed that serum NfL was positively correlated with neurologic function ( r=0.69, P<0.001). Logistic regression analysis showed that serum NfL was independently associated with neurological function ( OR=0.92, 95% CI: 0.86-0.98; P=0.010). ROC curve indicated that the AUC of serum NfL in predicting poor neurologic function was 0.95 (95% CI: 0.92-0.99), with a sensitivity of 84.5% and a specificity of 100% at the cutoff value of 80.0 pg/mL. The AUC of serum NSE in predicting poor neurologic function was 0.79 (95% CI: 0.69-0.89), with a sensitivity of 67.6% and a specificity of 80.0% at the cutoff value of 45.1 ng/mL. A pairwise comparison using Hanley & McNeil method showed that the AUC of serum NfL in predicting poor neurologic function was higher than that of NSE ( Z=3.22, P=0.001). Conclusions:Serum NfL is helpful for clinician to predict neurologic function in OHCA patients.

Effect of Zhiwei Fuwei Pills on autophagy in gastric antrum tissue of rats with precancerous lesions of gastric cancer based on mTOR/Beclin1/LC3 signaling axis / 中国临床药理学与治疗学

To investigate the effect of Zhiwei Fuwei Pills (ZWFW) on the expression of mammalian target of rapamycin (mTOR)/autophagy key molecule yeast Atg6 homologue (Beclin1)/microtubuleassociated protein 1 light chain 3 (LC3) signaling axis key molecules in gastric antrum tissue of rats with precancerous gastric lesions (PLGC). METHODS: SPF SD rats were randomly divided into normal group, model group, folic acid group, ZWFW low-dose, medium-dose, high-dose group. In addition to the normal group, the model group, folic acid group, ZWFW low-dose, medium-dose and high-dose groups, were used to establish the PLGC rat model by five factors compound modeling methods: N-methyl-N ' - nitro-n-nitroguanidine (MNNG) combined with hunger and satiation, ethanol intragastric administration, free drinking of ammonia and ranitidine feed. The rats were treated with normal saline, folic acid tablet aqueous solution (0.002 g/kg), ZWFW low-dose, medium-dose, high-dose aqueous solution (0.42, 0.84, 1.67 g/kg) for 4 weeks, and the stomach was removed by laparotomy. Hematoxylineosin (HE) staining was used to observe the histopathological changes in the antrum of rats, and real-time polymerase chain reaction (real-time PCR), Western blot (WB) and immunohistochemistry (IHC) were used to detect the expression of mammalian target of rapamycin mTOR, yeast Atg6 homologue 1 (Beclin1), microtubule-associated protein 1 light chain 3β (LC3B) mRNA and protein in the antrum of rats. RESULTS: Compared with the normal group, the Gastric antrum tissue of the model group was distended, thinner gastric wall, palegastric mucosa, atrophic and flat folds, disordered course and nodules and vegetations were visible. HE staining showed that compared with the normal group, the gastric mucosal glands in the model group were crowded and disordered, and the cell morphology was different, including a large number of goblet cells, basophilic cytoplasm, large, hyper-chromatic and irregular nuclei, and mucosal muscle infiltration and destruction. Compared with the model group, treated by ZWFW can significantly improve the pathological manifestations of gastric mucosal gland structure disorder and cell atypia. Compared with the normal group, mTOR mRNA and protein expression were significantly increased (P< 0.05) and Beclin1 and LC3B mRNA and protein expression were significantly decreased (P<0.05) in the antral tissue of rats in the model group; compared with the model group, mTOR mRNA and protein expression were decreased (P<0.05) in the medium and high dose groups of ZWFW, Beclin1 and LC3B protein expression in the antral tissue of rats in the low dose group of ZWFW and Beclin1 and LC3B mRNA and protein expression were increased (P<0.05) in the medium and high dose groups. CONCLUSION: Zhiwei Fuwei Pills can significantly improve the abnormal histopathological findings of gastric mucosa in PLGC model rats, and the mechanism may be related to the down-regulation of mTOR expression, up-regulation of Beclin1 and LC3B expression and then promoting autophagy.

Performance and clinical evaluation of two serum free light chain detection platforms in the diagnosis of multiple myeloma / 中华检验医学杂志

Objective:To compare the detection performance of serum free light chain (sFLC) in two platforms and evaluate the comparability of serum free light chain results in patients with multiple myeloma (MM).Methods:To evaluate the detection performance (repeatability, accuracy, linear range, reference range, interfering substances, etc.) of sFLC kit based on polyclonal antibodies. Spearman correlation analysis and Bland-Altman were used to analyze 214 sFLC results obtained on two detection platforms at the same time to evaluate the correlation between the results of the two methods and analyze the causes of methodological bias. 119 cases with aMM and 23 cases of disease control group (AL, WM, POEMS syndrome, MGUS, diffuse large B-cell lymphoma) initially diagnosed in the hematology department of Zhongshan Hospital of Fudan University from March 2020 to March 2021 were all included. A retrospective analysis was conducted to calculate the area under the curve of receiver operating characteristic (AUC-ROC) and obtain the optimal sensitivity and specificity cut-off points for the diagnosis of MM patients on monoclonal antibody platform.Results:Repeatability, accuracy, linear range, reference interval and anti-interfering capacity of the detection platform based on polyclonal antibodies were verified to meet clinical needs. The overall consistency of FLC/κ, FLC/λ and κ/λ ratios in two methods was 89.3%, 84.1% and 77.1% respectively; but the correlation results were highly heterogeneous. The correlation coefficient of FLC/κ R 2 was 0.922( P<0.001), while the correlation coefficients R 2of the FLC/λ and κ/λ ratios were only 0.349 and 0.441( P<0.001). After segment analysis, it was found that the correlation of FLC/λ was improved within the linear range and R 2 could rise to 0.78( P<0.001). Compared with monoclonal antibody platform, the vast majority points of FLC/κ fell within the 95% limit by Bland Altman analysis. While the results of FLC/λ on polyclonal antibody platform showed significant positive bias. The AUC of MM diagnosis on monoclonal antibody platform was 0.751 ( P=0.001), and the optimal cutoff value was 24.67. Conclusion:The overall consistency between the two platforms was good, but there were significant differences between the results, so they were not comparable and could not be interchanged. For monitoring the prognosis of patients with multiple myeloma, the same platform should be selected for testing.

Role of plasma neurofilament protein light chain in diagnosing and differentiating PD and multisystem atrophy-Parkinson subtypes / 中华神经医学杂志

Objective:To investigate the role of plasma neurofilament light chain (NfL) in diagnosing and differentiating Parkinson's disease (PD) and multiple system atrophy-Parkinsonian subtype (MSA-P).Methods:Forty PD patients and 23 MSA-P patients admitted to Department of Neurology, Henan Provincial People's Hospital from June 2019 to December 2021 were recruited; 27 healthy subjects accepted physical examination during the same period were selected as controls. Ultrasensitive Simoa technology was used to measure the plasma NfL. Differences in clinical data and plasma NfL were compared among all subjects. Correlations of plasma NfL with clinical characteristics, such as disease course, Hoehn-Year (H-Y) staging, Unified Parkinson's Disease Rating Scale (UPDRS), Montreal Cognitive Assessment (MoCA), Scale for Outcomes in Parkinson's Disease for Autonomic Symptoms (SCOPA-AUT) and levodopa equivalent daily dosage (LEDD), were analyzed with Pearson correlations. Receiver operating characteristic (ROC) curve was used to analyze the value of plasma NfL in diagnosing and differentiating PD and MSA-P.Results:Compared with MSA-P group, PD group had significantly longer disease course and statistically lower scores of UPDRS-II and SCOPA-AUT ( P<0.05). The plasma NfL in MSA-P group, PD group and healthy control group was decreased successively ([37.69±10.47] pg/mL, [17.85±4.23] pg/mL, [12.86±3.14] pg/mL, respectively), with statistical differences ( P<0.05). In MSA-P patients, Pearson correlations showed positive correlation between plasma NfL and age ( r=0.442, P=0.035); and Partial correlations showed positive correlations between plasma NfL and scores of UPDRS-I and UPDRS-III ( P<0.05), and plasma NfL showed no significant correlation with H-Y staging, UPDRS-III, MoCA, LEDD or SCOPA-AUT scores ( P>0.05). In PD patients, Pearson correlations showed that plasma NfL was positively correlated with age ( r=0.342, P=0.031); partial correlations showed that plasma NfL was positively correlated with H-Y staging and UPDRS-III, and negatively correlated with MoCA scores ( P<0.05); plasma NfL showed no significant correlation with disease course, scores of UPDRS-I and UPDRS-II, LEDD, and SCOPA-AUT scores ( P>0.05). ROC curve showed that the area under the curve (AUC) of plasma NfL in diagnosing PD was 0.814 (95% CI: 0.712-0.920, P<0.001); AUC of plasma NfL in differentiating and diagnosing PD and MSA-P was 0.980 (95% CI: 0.954-1.000, P<0.001); AUC of plasma NfL in diagnosing MSA-P was 0.998 (95% CI: 0.993-1.000, P<0.001). Conclusions:Plasma NfL is correlated with severity of motor symptoms in MSA-P patients; plasma NfL is correlated with cognitive function and disease course in PD patients. Besides, plasma NfL has high sensitivity and specificity in differentiating PD and MSA-P, therefore, plasma NfL could serve as a biomarker to diagnosis and differentiate PD.

The Expression Level and Diagnostic Value of Serum Free Light Chain in B-Cell Non-Hodgkin Lymphoma / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression level and the diagnostic value of serum free light chain in B-cell non-Hodgkin's lymphoma (B-NHL).@*METHODS@#We retrospectively analyzed the results of serum free light chain (sFLC) of 394 newly treated B-NHL patients in our hospital from January 2014 to December 2021 and compared the secretion levels of sFLC among different subtypes of B-NHL. The value of sFLC secretion levels in the diagnosis of WM was evaluated using ROC.@*RESULTS@#Increased proportion of sFLC, abnormal ratio of sFLC (κ / λ) and the secretion levels of sFLC (κ+λ) were different in different B-NHL subtypes, Waldenstrom's macroglobulinemia (WM) had the highest proportion of elevated sFLC(82.68%) and abnormal sFLC(κ/ λ)(87.0%), the proportion of FL(18.0%) and DLBCL patients(12.8%) with elevated sFLC was lower (P<0.05). The expression levels of sFLC can helpful in the diagnosis of WM (AUC=0.874，P<0.001， 95% CI: 0.779-0.970). At the same time, higher sFLC levels and sFLC cloning patterns predicted the possibility of bone marrow infiltration of lymphoma.@*CONCLUSION@#The serum free light chains is common in patients with B-NHL. The elevated level and type of free light chain are associated with the type of lymphoma, and the patients with bone marrow infiltration have higher sFLC（κ+ λ） expression level.

Compromiso renal atípico en amiloidosis de cadena ligera

Introducción la amiloidosis de cadena ligera (AL) es una entidad desencadenada por la proliferación de un clon de células plasmáticas que genera la acumulación de cadenas ligeras, las cuales se depositan en forma de fibrillas amiloides generando una disfunción orgánica. El compromiso renal generalmente se manifiesta como síndrome nefrótico, con un deterioro lento y progresivo de la función renal que puede llevar a un requerimiento de terapia dialítica. Objetivo demostrar el compromiso renal agresivo y subagudo de la amiloidosis sistémica. Presentación del caso paciente masculino de 35 años que consulta por malestar general, visión borrosa, mareos y oliguria con elevación de azoados, y que progresó rápidamente hasta el requerimiento de hemodiálisis en aproximadamente tres meses. Asociado se documenta polineuropatía periférica, infiltración cardiaca y ligera elevación de transaminasas. Ante negatividad de estudios de extensión, se logra documentar la presencia histopatológica de depósitos amiloides con inmunofluorescencia positiva para AL. Actualmente, se encuentra recibiendo esquema de quimioterapia con adecuada estabilidad clínica y tolerancia. Discusión y conclusión la amiloidosis AL es una entidad infrecuente, con compromiso multiorgánico importante y altas tasas de morbilidad y mortalidad. Se recalca en este caso el compromiso subagudo con requerimiento temprano de terapia dialítica y además se enfatiza la importancia de una sospecha y un diagnóstico oportuno en pacientes con compromiso renal y otras manifestaciones sistémicas.

Introduction Light chain amyloidosis is an entity triggered by the proliferation of a clone of plasma cells that generates the accumulation of light chains, which are deposited in the form of amyloid fibrils generating organic dysfunction. Renal compromise generally manifests as nephrotic syndrome, with a slow and progressive decline of renal function that can lead to dialysis therapy. Purpose The objective of this case report is to demonstrate the aggressive and subacute renal involvement of systemic amyloidosis. Case presentation We present a case of a 35-year-old male patient who consulted for general malaise, blurred vision, dizziness and oliguria with elevated nitrogen levels that progressed fastly to the requirement of hemodialysis in approximately 3 months. It was also reveal peripheral polyneuropathy, cardiac infiltration, and slight elevation of transaminases. Given the negativity of extension studies, amyloid deposits were documented histopathologically with positive immunofluorescence for LA. He is currently receiving chemotherapy regimen with adequate clinical stability and tolerance. Conclusion and discussion To conclude, AL amyloidosis is a rare entity, with significant multi-organ involvement and high rates of morbidity and mortality. In this case, the subacute involvement with an early requirement for dialysis therapy is emphasized, and the importance of suspicion and timely diagnosis in patients with kidney involvement and other systemic manifestations is emphasized.