OBJECTIVE: To explore the genetic variants and phenotypic characteristics of patients with Neurofibromatosis type I (NF1). METHODS: Twenty two NF1 patients who presented at Enze Medical (Center) Group in Taizhou between 2018 and 2024 were selected as the study subjects. Clinical phenotype and family history were collected for the patients. Whole exome sequencing (WES) was carried out for the 22 probands to screen the variants of NF1 gene. Candidate variants were verified by Sanger sequencing of their family members. This study was approved by the Medical Ethics Committee of the Hospital (Ethics No.: K20230902). RESULTS: The 22 probands were diagnosed between the age of 5 months to 47 years old, and have all shown cafe au lait spots on their skin. Seventeen patients exhibited the phenotype at birth, and 11 had various degrees of neurofibromatosis. Among them, probands 1 and 13 underwent surgical resection of the tumor but had recurred, while proband 12 had amputation due to the huge size and serious impact of the neurofibroma and had no recurrence. Five patients had various degrees of scoliosis. In total 22 germline mutations and one somatic mutation were identified among the 22 families, with 5 variants unreported previously, including 1 nonsense mutation c.1603C>T (Q535*), 3 frameshift mutations [c.7268_7269delCA (Thr2423fs), c.2293del (Arg765Alafs*26), and c.5433_5438delinsGC (Phe1812ArgfsTer50)], and 1 deletion involving exons 41-44 of the NF1 gene and adjacent introns. Proband 13 was found to harbor germline mutation c.6796C>T (Gln2266Ter) and somatic mutation c.1019_1020del (Ser340Cysfs Ter12) in the peripheral blood and tumor tissue, respectively. Among the 22 NF1 probands, 6 had received treatment due to severe illness. Proband 1 had tumor resection in the right upper limb, but was found to have malignant lung tumor and died during follow-up. Proband 12 had multiple recurrence of neurofibroma in the left ring finger. Proband 4 underwent spinal correction surgery due to severe scoliosis. Proband 11 had died due to a central nervous system disease. Among the 22 germline mutations, 6 had led to the occurrence of truncated proteins, which may have a more severe impact on the phenotype. CONCLUSION This study investigated the genetic variants and clinical phenotypes of 22 NF1 families and identified 5 novel variants of the NF1 gene, which has expanded the genotypic and phenotypic spectra of the NF1. Preliminary studies have identified an association between truncated mutations, young age, and severe phenotypes, which may provide important clues for prognosis evaluation. For the clinical diagnosis and treatment of NF1, it is necessary to consider the phenotypic characteristics and genetic testing in combination with genetic counseling and long-term follow-up.
Humans ; Neurofibromatosis 1/pathology* ; Male ; Female ; Pedigree ; Adult ; Child ; Child, Preschool ; Middle Aged ; Adolescent ; Infant ; Young Adult ; Neurofibromin 1/genetics* ; Phenotype ; Asian People/genetics* ; Mutation ; Exome Sequencing ; East Asian People

The liver regenerative capacity is crucial for patients with end-stage liver disease following partial hepatectomy (PHx). The specific bacteria and mechanisms regulating liver regeneration post-PHx remain unclear. This study demonstrated dynamic changes in the abundance of Parabacteroides distasonis (P. distasonis) post-PHx, correlating with hepatocyte proliferation. Treatment with live P. distasonis significantly promoted hepatocyte proliferation and liver regeneration after PHx. Targeted metabolomics revealed a significant positive correlation between P. distasonis and β-hydroxybutyric acid (BHB), as well as hyodeoxycholic acid and 3-hydroxyphenylacetic acid in the gut after PHx. Notably, treatment with BHB, but not hyodeoxycholic acid or 3-hydroxyphenylacetic acid, significantly promoted hepatocyte proliferation and liver regeneration in mice after PHx. Moreover, STAT3 inhibitor Stattic attenuated the promotive effects of BHB on cell proliferation and liver regeneration both in vitro and in vivo. Mechanistically, P. distasonis upregulated the expression of fatty acid oxidation-related proteins, and increased BHB levels in the liver, and then BHB activated the STAT3 signaling pathway to promote liver regeneration. This study, for the first time, identifies the involvement of P. distasonis and its associated metabolite BHB in promoting liver regeneration after PHx, providing new insights for considering P. distasonis and BHB as potential strategies for promoting hepatic regeneration.

[This corrects the article DOI: 10.1016/j.apsb.2024.03.030.].

Objective:To investigate the causes and management strategies of anesthetic complications during percutaneous spinal endoscopic surgery under local anesthesia.Methods:A total of 16 800 patients (8 625 males and 8 175 females) who underwent percutaneous spinal endoscopic surgery under local anesthesia (including intravenous basic anesthesia) in Tianjin Hospital, Shandong Public Health Clinical Center and Hebei General Hospital from February 2012 to February 2023 were retrospectively analyzed. The average age was 45.3±21.6 years (range, 12-84 years). There were 220 cases of posterior cervical keyhole endoscopic surgery, 50 cases of thoracic transforaminal endoscopic surgery, 70 cases of thoracic posterior interlaminar endoscopic surgery, 11 670 cases of lumbar transforaminal endoscopic surgery, and 4 790 cases of lumbar posterior interlaminar endoscopic surgery. The occurrence time, clinical manifestations, management of intraoperative anesthesia complications were recorded, as well as surgical segments, puncture sites, complication symptoms, signs, outcome and prognosis.Results:All patients received percutaneous water-mediated uniaxial spinal endoscopic surgery under local anesthesia. There were 9 patients experienced anesthesia complications, including 6 cases of epidural diffusion of anesthetics and 3 cases of anesthetics mistakenly entering the subarachnoid space. There were 4 males and 5 females, aged 48.4±18.2 years (range, 28-84 years). There were 1 case of T 12L 1 disc herniation, 1 case of C 5-6 disc herniation, 3 cases of L 4-5 disc herniation and 4 cases of L 5S 1 disc herniation. Surgical segments and procedures: 1 case of C 5-6 posterior Keyhole endoscopic surgery, 1 case of T 12L 1 transforaminal endoscopic surgery, 2 cases of L 4-5 transforaminal endoscopic surgery, 1 case of L 4-5 interlaminar endoscopic surgery, and 4 cases of L 5S 1 interlaminar endoscopic surgery. Anesthesia complications all appeared 5-10 min after injection of local anesthetics, with symptoms of decreased oxygen saturation, decreased blood pressure, altered consciousness, and sensory and motor dysfunction of limbs. 6 patients with epidural diffusion of anesthetics recovered completely after symptomatic treatment in 5 cases, and 1 case was left with foot drop. Three patients with anesthetics mistakenly entering the subarachnoid space were immediately converted to the supine position, of which one recovered by mask oxygenation; 1 patient improved after emergency tracheal intubation, rehydration, and application of vasoconstrictive medications; and 1 patient developed multiple complications such as multiorgan failure, rhabdomyolysis, and sepsis after tracheal intubation, and recovered at 3 months after surgery with symptomatic treatment. Conclusions:Epidural diffusion and entering into subarachnoid space of anesthetics are serious complications of local anesthesia in percutaneous spinal endoscopic surgery. In addition to sensory and motor dysfunction of the limbs, the functions of the respiratory and circulatory systems can also be affected. It is necessary to be alert to the occurrence of anesthesia-related complications during operation and early identification and treatment.

OBJECTIVE: To explore the clinical and genetic characteristics of a Chinese pedigree affected with Hypertrophic cardiomyopathy (HCM) due to a truncating variant of ALPK3 gene. METHODS: A 44-year-old male admitted to Taizhou Hospital of Zhejiang Province on December 29, 2018 was selected as the study subject. Whole-exome sequencing (WES) was carried out, and candidate variant was interpreted by following the guidelines from the American College of Medical Genetics and Genomics (ACMG). For ALPK3 was considered an autosomal recessive gene, the WES results was considered insufficient to explain his phenotype. In April 2023, the proband's WES data were re-analyzed using updated annotation pipelines, and peripheral blood samples were collected from his first-degree relatives (mother and brother) for Sanger sequencing validation. Conservation analysis and protein structural modeling were performed to assess the impact of the variant. Clinical evaluation and genetic counseling were provided to the proband's family members. Relevant literature on ALPK3tv-induced HCM patients were searched in Wanfang Data Knowledge Service Platform, CNKI, and PubMed database using "ALPK3" and "hypertrophic cardiomyopathy" as keywords. Clinical characteristics of HCM patients with heterozygous ALPK3tv variants were summarized and compared with the clinical characteristics of HCM patients with positive sarcomere-associated gene variants (SARC+). This study was approved by the Medical Ethics Committee of Taizhou Hospital of Zhejiang Province Affiliated to Wenzhou Medical University (Ethics No.: K20230314). RESULTS: The proband was a 44-year-old male who was transferred to our institution on December 29, 2018 due to "chest tightness and pain for 6 months, exacerbated for 2 days". Emergency coronary angiography was performed, which led to a preliminary diagnosis of "acute coronary syndrome", and the patient was admitted to the Cardiology Department for treatment. Based on electrocardiogram and echocardiogram findings, the diagnosis was revised as HCM. The patient's condition has stabilized post-coronary angiography, and he was discharged with improved condition. On January 2019, WES was conducted to determine the etiology of the proband's HCM. WES results identified a novel heterozygous c.2156dupC (p.Pro720ThrfsTer53) truncating variant in the ALPK3 gene. At that time, the inheritance pattern could not explain the phenotype. In 2022, a literature indicated that heterozygous ALPK3tv could lead to autosomal dominant HCM. Consequently, in April 2023, the proband's whole-exome data were re-annotated, revealing changes in the transcript and protein versions, with the updated site annotated as ALPK3 (NM_020778.5): c.1550dupC (p.Pro518ThrfsTer53). Sanger sequencing confirmed that the proband's mother and brother also carried this variant. The mother exhibited obstructive HCM, while the brother showed no related phenotype. Bioinformatics analysis demonstrated conservation of this site across multiple species, and the variant has resulted in the loss of a protein domain. Based on ACMG guidelines, the variant was classified as likely pathogenic. Literature review and Bayesian calculation further elevated the pathogenicity rating, indicating that this variant was the cause of HCM in the patient. Literature study revealed distinctions between HCM caused by this variant type and SARC+ HCM. The age of onset among heterozygous ALPK3tv patients was delayed by approximately 10 years compared to SARC+ patients. Both forms of HCM exhibited a male predominance, which was particularly marked in individuals with ALPK3tv. Electrocardiographic left ventricular hypertrophy was more prevalent in heterozygous ALPK3tv patients than in SARC+ patients. The incidence of apical or concentric hypertrophy patterns was higher in heterozygous ALPK3tv patients compared to asymmetric septal hypertrophy, which predominated in SARC+ patients. ALPK3tv patients exhibited lower penetrance and later onset compared to SARC+ patients. A positive correlation between left ventricular wall thickness and age was noted in female patients only. CONCLUSION In this pedigree, the proband has presented with HCM, characterized by echocardiographic evidence of apical left ventricular hypertrophy without significant outflow tract obstruction or extracardiac phenotypes. Although his mother and brother had carried the same heterozygous ALPK3 (NM_020778.5) c.1550dupC (p.Pro518ThrfsTer53), the mother exhibited severe obstructive HCM, while the brother was asymptomatic, suggesting incomplete or age-dependent penetrance within the family. This study has enriched the evidence for the pathogenicity of ALPK3tv among Chinese HCM pedigrees and underscored the importance of periodic literature reviews and genetic re-analysis for unresolved genetic testing results.
Humans ; Male ; Pedigree ; Adult ; Cardiomyopathy, Hypertrophic/genetics* ; Heterozygote ; Asian People/genetics* ; Exome Sequencing ; Mutation ; China ; Female ; East Asian People

Objective:To investigate the causes and management strategies of anesthetic complications during percutaneous spinal endoscopic surgery under local anesthesia.Methods:A total of 16 800 patients (8 625 males and 8 175 females) who underwent percutaneous spinal endoscopic surgery under local anesthesia (including intravenous basic anesthesia) in Tianjin Hospital, Shandong Public Health Clinical Center and Hebei General Hospital from February 2012 to February 2023 were retrospectively analyzed. The average age was 45.3±21.6 years (range, 12-84 years). There were 220 cases of posterior cervical keyhole endoscopic surgery, 50 cases of thoracic transforaminal endoscopic surgery, 70 cases of thoracic posterior interlaminar endoscopic surgery, 11 670 cases of lumbar transforaminal endoscopic surgery, and 4 790 cases of lumbar posterior interlaminar endoscopic surgery. The occurrence time, clinical manifestations, management of intraoperative anesthesia complications were recorded, as well as surgical segments, puncture sites, complication symptoms, signs, outcome and prognosis.Results:All patients received percutaneous water-mediated uniaxial spinal endoscopic surgery under local anesthesia. There were 9 patients experienced anesthesia complications, including 6 cases of epidural diffusion of anesthetics and 3 cases of anesthetics mistakenly entering the subarachnoid space. There were 4 males and 5 females, aged 48.4±18.2 years (range, 28-84 years). There were 1 case of T 12L 1 disc herniation, 1 case of C 5-6 disc herniation, 3 cases of L 4-5 disc herniation and 4 cases of L 5S 1 disc herniation. Surgical segments and procedures: 1 case of C 5-6 posterior Keyhole endoscopic surgery, 1 case of T 12L 1 transforaminal endoscopic surgery, 2 cases of L 4-5 transforaminal endoscopic surgery, 1 case of L 4-5 interlaminar endoscopic surgery, and 4 cases of L 5S 1 interlaminar endoscopic surgery. Anesthesia complications all appeared 5-10 min after injection of local anesthetics, with symptoms of decreased oxygen saturation, decreased blood pressure, altered consciousness, and sensory and motor dysfunction of limbs. 6 patients with epidural diffusion of anesthetics recovered completely after symptomatic treatment in 5 cases, and 1 case was left with foot drop. Three patients with anesthetics mistakenly entering the subarachnoid space were immediately converted to the supine position, of which one recovered by mask oxygenation; 1 patient improved after emergency tracheal intubation, rehydration, and application of vasoconstrictive medications; and 1 patient developed multiple complications such as multiorgan failure, rhabdomyolysis, and sepsis after tracheal intubation, and recovered at 3 months after surgery with symptomatic treatment. Conclusions:Epidural diffusion and entering into subarachnoid space of anesthetics are serious complications of local anesthesia in percutaneous spinal endoscopic surgery. In addition to sensory and motor dysfunction of the limbs, the functions of the respiratory and circulatory systems can also be affected. It is necessary to be alert to the occurrence of anesthesia-related complications during operation and early identification and treatment.

Objective:To explore the relapse-related genes and their clinicopathological connections of diffuse large B cell lymphoma (DLBCL).Methods:Targeted panel sequencing was conducted on 32 eligible DLBCL samples; the patients were diagnosed, treated, and went into complete remission at the First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2019, including 14 cases with recurrence (relapsed group) and 18 cases with long-term complete remission of over five years (remission group). Clinical and pathological data were further reviewed. Fisher′s exact test was employed to compare the differences in clinicopathological characteristics and mutation patterns between the two groups.Results:Among the 32 patients, there were 18 males and 14 females, with a male to female ratio of 1.3∶1.0 and a median age of 53 (45.5, 67.0) years. In the relapsed group, PIM1 (11/14), KMT2D (7/14), PRDM1 (6/14), MYD88 (6/14), DTX1 (6/14) emerged as the most frequently mutated genes. In the remission group, while recurrent PIM1, KMT2D and MYD88 mutations were also observed, the TP53 gene exhibited the highest mutation frequency (6/18). Compared to the remission group, relapsed group showed elevated mutation frequencies of PIM1 ( P=0.013) and FAT4 ( P=0.010), alongside a reduced incidence of TP53 mutations. In all 32 patients, DLBCL with CD79B, CCND3, DTX1, KMT2D and PRDM1 mutations demonstrated a propensity towards advanced clinicopathologic stage. Conclusions:Relapsed DLBCL has distinctive clinicopathological and genetic features. PIM1 and FAT4 may be served as potential biomarkers for screening relapsed DLBCL-NOS and as targets for novel therapeutic strategies.

Objective:To explore the relapse-related genes and their clinicopathological connections of diffuse large B cell lymphoma (DLBCL).Methods:Targeted panel sequencing was conducted on 32 eligible DLBCL samples; the patients were diagnosed, treated, and went into complete remission at the First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2019, including 14 cases with recurrence (relapsed group) and 18 cases with long-term complete remission of over five years (remission group). Clinical and pathological data were further reviewed. Fisher′s exact test was employed to compare the differences in clinicopathological characteristics and mutation patterns between the two groups.Results:Among the 32 patients, there were 18 males and 14 females, with a male to female ratio of 1.3∶1.0 and a median age of 53 (45.5, 67.0) years. In the relapsed group, PIM1 (11/14), KMT2D (7/14), PRDM1 (6/14), MYD88 (6/14), DTX1 (6/14) emerged as the most frequently mutated genes. In the remission group, while recurrent PIM1, KMT2D and MYD88 mutations were also observed, the TP53 gene exhibited the highest mutation frequency (6/18). Compared to the remission group, relapsed group showed elevated mutation frequencies of PIM1 ( P=0.013) and FAT4 ( P=0.010), alongside a reduced incidence of TP53 mutations. In all 32 patients, DLBCL with CD79B, CCND3, DTX1, KMT2D and PRDM1 mutations demonstrated a propensity towards advanced clinicopathologic stage. Conclusions:Relapsed DLBCL has distinctive clinicopathological and genetic features. PIM1 and FAT4 may be served as potential biomarkers for screening relapsed DLBCL-NOS and as targets for novel therapeutic strategies.

ObjectiveTo explore the mechanism and pathway of Gandou Fumu decoction （GDFMD） in the development of liver fibrosis in Wilson's disease （WD）. MethodFirst， 30 TX-j mice were randomly divided into the model group， high-dose， medium-dose， and low-dose GDFMD groups， and penicillamine group， with six mice in each group， and another six wild-type mice were used as the normal group. The high-dose， medium-dose， and low-dose GDFMD groups were intragastrically administered drugs of 13.92， 6.96， 3.48 g·kg^-1. In the penicillamine group， 0.1 g·kg^-1 of penicillamine was given by intragastric administration. The model group and the normal group were given equal volume of normal saline， once a day， for four consecutive weeks. Samples were collected four weeks after gavage， and enzyme-linked immunosorbent assay （ELISA） was used to detect type Ⅲ procollagen peptide （PCⅢ）， collagen type Ⅳ （Col Ⅳ）， hyaluronic acid （HA）， and laminin （LN）. Hematoxylin-eosin （HE）， Masson， and picric acid-Sirus red collagen （Sirus Red） staining were used to observe the histopathological changes of liver fibrosis. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， immunohistochemistry， and Western blot were used to observe the expressions of α-smooth muscle actin （α-SMA） and collagen type Ⅰ （Col Ⅰ）， which were related to the activation of hepatic stellate cells （HSCs）. The expression of miR-29b-3p was observed by Real-time PCR. The expression of Unc-51-like kinase 1 （ULK1） and its downstream-related factors were observed by Western blot. The downstream genes of miR-29b-3p were verified by the dual luciferase reporter gene detection method. ResultCompared with the normal group， the four items of liver fibrosis （PCⅢ， Col Ⅳ， HA， and LN） in the model group were significantly abnormal （P<0.01）， and the pathology was significantly abnormal. The expression of HSC activation-related indicators including α-SMA and Col Ⅰ， as well as α-SMA mRNA and Col Ⅰ mRNA was up-regulated （P<0.05， P<0.01）， and miR-29b-3p expression was down-regulated （P<0.01）. ULK1， p-ULK1， autophagy-related gene 13 （Atg13）， p-Atg13， Beclin-1， FAK family kinase-interacting protein of 200 kDa （FIP200）， activating molecule in BECN1-regulated autophagy protein 1 （AMBKA1）， and microtubule-associated protein 1 light chain 3Ⅱ/Ⅰ（LC3Ⅱ/Ⅰ） were up-regulated （P<0.05， P<0.01）. p62 protein expression was down-regulated （P<0.01）. Compared with the model group， the four items of liver fibrosis in the high-dose， medium-dose， and low-dose GDFMD groups and the penicillamine group were significantly improve （P<0.01）， and the pathological conditions were improved. The expression of HSC activation-related indicators including α-SMA and Col Ⅰ， as well as α-SMA mRNA and Col Ⅰ mRNA was down-regulated （P<0.05， P<0.01）， and the expression of miR-29b-3p was up-regulated （P<0.01）. ULK1， p-ULK1， Atg13， p-Atg13， Beclin-1， FIP200， AMBKA1， and LC3Ⅱ/Ⅰ were down-regulated （P<0.05， P<0.01）， and p62 protein expression was up-regulated （P<0.01）. The prediction software predicted that there was a binding site between miR-29b-3p and ULK1. The dual-luciferase reporter gene detection method indicated that the luciferase activity of the ULK1-WT plasmid-transfected cell group was reduced when miR-29b-3p mimics were co-cultured （P<0.01）. ConclusionGDFMD can regulate ULK1-mediated autophagy by up-regulating miR-29b-3p and further exert its anti-hepatic fibrosis effect in Wilson's disease.

As an active hydroxyanthraquinone ingredient, emodin is abundant in Chinese medicine herbs, such as Rheum palmatum, Polygonum cuspidatum and Polygonum multiflorum.Modern pharmacological studies have shown that emodin has a variety of pharmacological activities including anti-tumor, anti-inflammatory and immunoregulatory, antibacterial and anti-viral effects, myocardial protection, neuroprotection, renal protection, bone protection, antifibrosis and so on, which indicate its high medicinal value and broad application prospects.This article aims to summarize the progress in the pharmacological activity and mechanism of action of emodin published in domestic and international journals over the last 5 years and highlight the potential targets and molecular signaling pathways linked with emodin, so as to provide some clues and references for further development and clinical application of emodin.