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

OBJECTIVE: To determine the types of genetic variants in six Chinese pedigrees affected with Marfan syndrome (MFS) and analyze their clinical characteristics and molecular pathogenesis. METHODS: Six MFS pedigrees presented at the Taizhou Enze Medical Center (Group) between 2017 and 2022 were selected as the study subjects. Clinical data of pedigrees were retrospectively analyzed. Peripheral blood samples were collected from the probands and their family members for the extraction of genomic DNA. Whole exome sequencing (WES) was carried out. Candidate variants of the FBN1 gene were verified by Sanger sequencing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), pathogenicity of the candidate variants was assessed. AlphaFold3 and PyMOL software were used for homology modeling of the FBN1 protein and analysis of its three-dimensional structure and amino acid sequence conservation. This study was approved by the Medical Ethics Committee of Taizhou Enze Medical Center (Group) (Ethics No. 20231002). RESULTS: Cardiovascular system abnormalities were noted in all pedigrees, ocular abnormalities were present in pedigrees 2 and 5, skeletal system abnormalities were presented in pedigrees 1, and 4 to 6. FBN1 gene mutations were identified in all pedigrees, including c.1957_1958dupGT (p.Asp654fs), c.5014T>A (p.Cys1672Ser), c.8135delC (p.Pro2712fs), c.2302G>T (p.Glu768*), c.3473A>G (p.Glu1158Gly) and c.6169C>T (p.Arg2057*), with each involving a different exon. Four variants were rated as pathogenic, one as likely pathogenic, and one as variant of uncertain significance. Among these, c.5014T>A (p.Cys1672Ser), c.1957_1958dupGT (p.Asp654fs), c.8135delC (p.Pro2712fs), and c.2302G>T (p.Glu768*) were unreported previously. Bioinformatic analysis with SIFT and PolyPhen-2 predicted that the c.5014T>A (p.Cys1672Ser) and c.3473A>G (p.Glu1158Gly) variants were deleterious. Protein homologous sequence alignment analysis revealed that the four novel mutation sites are highly conserved across various species. Homology modeling of the FBN1 protein three-dimensional structure indicated that the six variant sites in the amino acid sequence are all close to hydrogen bonds and may alter the secondary and tertiary structures to varying degrees, thereby confirmed the relationship between the variants and MFS. CONCLUSION Four novel variants of the FBN1 gene have been discovered in this study, which has enriched the mutational and phenotypic spectrum of MFS and provided a basis for disease diagnosis and genetic counseling.
Adolescent ; Adult ; Child ; Female ; Humans ; Male ; Middle Aged ; Young Adult ; China ; East Asian People/genetics* ; Exome Sequencing ; Fibrillin-1/genetics* ; Marfan Syndrome/genetics* ; Mutation ; Pedigree ; Retrospective Studies ; Adipokines

Objective:To evaluate the knowledge of autism among child health care professionals in primary health care institutions.Methods:The study was a cross-sectional survey. An online questionnaire survey was conducted from February to March 2023 in primary health care institutions in Guangzhou to investigate the knowledge on autism among medical staff engaged in children′s health services and the influencing factors.Results:A total of 341 questionnaires were returned and 312 questionnaires were valid with a recovery rate of 91.5%. The age of 312 respondents was (35.9±7.9) years, of which 303 (97.1%) were female. One hundred and fifty-two (48.7%) child health care professionals in primary health care institutions had received specialist training in assessing the psychological and behavioral development of children, and only 139 (44.6%) reported that they were aware of the"five no"principle for early identification of autism. The questionnaire scores were 88.1% pass (275/312) and 53.2% excellent (166/312). The three questions with low accuracy were: autism can be cured with drugs, autism has a genetic basis and rehabilitation training has no effect, and the accuracy for these questions was 42.6% (133/312), 52.2% (163/312) and 70.2% (219/312), respectively. The passing of autism-related knowledge was positively associated with receiving relevant training ( OR=2.585, 95% CI:1.200-5.569), and the excellence was positively associated with the highest education ( OR=1.939, 95% CI:1.220-3.083) and receiving relevant training ( OR=2.016, 95% CI:1.247-3.260). Conclusions:There is a need for more professional training in autism knowledge among child health care professionals in primary health care institutions.

Objective:To investigate the value of quantitative parameters of magnetic resonance imaging (MRI) in predicting the efficacy of chimeric antigen receptor T-cell (CAR-T) therapy for children and adolescents with mature aggressive B-cell non-Hodgkin lymphoma (NHL).Methods:It was a retrospective multicenter study.Clinical data of 44 children and adolescents diagnosed with mature aggressive B-cell NHL between January 2016 and January 2023 in Henan Cancer Hospital, Beijing Gaobo Boren Hospital, and the First Affiliated Hospital of Xinxiang Medical University were retrospectively analyzed.Patients were divided into complete response (CR) group and non-CR group based on the international criteria for the diagnosis of pediatric NHL.Quantitative parameters of MRI, including T2 signal intensity, the minimal apparent diffusion coefficient (ADCmin), maximal ADC (ADCmax), and the mean ADC (ADCmean) were measured before and within 2 weeks after CAR-T infusion.The correlation between the above parameters and the achievement of CR was analyzed.The intraclass correlation coefficient (ICC) was used to assess the inter-observer agreement among observers in measuring quantitative parameters of MRI.Differences between groups were analyzed using the independent sample t-test.Factors influencing CR were identified through the binary Logistic regression analysis, and a prediction model was established.Model performance was evaluated by plotting receiver operating characteristic (ROC) curves. Results:Significant differences were observed between the CR group and non-CR group in T2 signal intensity before CAR-T infusion (267±152 vs.364±160, P=0.048), and ADCmin (0.94±0.38 vs.0.53±0.28, P<0.05), ADCmax (1.73±0.69 vs.0.84±0.43, P<0.05), ADCmean (1.28±0.48 vs.0.67±0.33, P<0.05), and T2 signal intensity within 2 weeks after CAR-T infusion (198±139 vs.345±168, P=0.004). A univariate prediction model was created by introducing the above quantitative parameters.The area under the curve (AUC), specificity, sensitivity, and accuracy of T2 signal intensity before CAR-T infusion in predicting the efficacy on children and adolescents with mature aggressive B-cell NHL were 0.800, 84.0%, 57.9%, and 72.7%, respectively.The AUC, specificity, sensitivity, and accuracy of ADCmax within 2 weeks of CAR-T infusion were 0.958, 88.0%, 78.9%, and 84.1%, respectively.The AUC, specificity, sensitivity, and accuracy of T2 signal intensity within 2 weeks of CAR-T infusion were 0.869, 84.0%, 68.4%, and 77.3%, respectively. Conclusions:Quantitative parameters of MRI, including ADC values and T2 signal intensity, are of great significance in the early prediction of CAR-T therapy efficacy on children and adolescents with mature aggressive B-cell NHL.Among these parameters, ADCmax presents the strongest predictive performance and serves as a valuable indicator for predicting a complete response with CAR-T treatment.

Bioluminescence is a widespread phenomenon in nature, and luminescent organisms can be found both on land and in the ocean. Among them, luciferase based bioluminescence systems have been widely studied, inspiring the exploration of genetic and epigenetic aspects and the development of a series of related assays for in vivo and in vitro studies. This paper summarizes the recent developments of luciferase based bioluminescence assays in terms of bioluminescence systems, types of luciferases, and the development and application of luciferase bioluminescence assays.

Objective:To investigate the pathogenesis and prognosis of transformation of primary myelofibrosis (PMF) to B-cell acute lymphoblastic leukemia (B-ALL).Methods:The diagnosis and treatment process of a patient transferred from PMF to B-ALL in Affiliated Tumor Hospital of Zhengzhou University in November 2018 were retrospectively analyzed, and the relevant literature was reviewed.Results:The patient was a 64-year-old female, she was initially diagnosed with PMF, and then she developed B-ALL 17 months later after receiving treatment of prednisone, danazole, levamisole, aspirin, thalidomide and jaktinib. After induction therapy, the patient received 8 months of continuous remission, and then the reexamination showed relapse. There was no remission after reinduction therapy. The patient gave up treatment and was discharged 2 months later. JAK2 V617F gene mutation was positive before and after leukemia transformation.Conclusions:The patients with transformation of PMF to B-ALL have poor clinical prognosis and short survival time. The possible mechanism of its transformation may be related to additional genetic events or certain high-risk genes. However, the specific mechanism is still unclear, and further investigation of the etiology is needed to seek targeted treatment.

Objective:To investigate the prognostic value of proprotein convertase subtilisin/kexin type 9 (PCSK9) and blood lipid indexes in patients with sepsis.Methods:Patients with sepsis or septic shock who were ≥ 18 years old and met the Sepsis-3.0 diagnostic criteria admitted to the department of critical care medicine of Binzhou Medical University Hospital from January to October 2021 were enrolled. Healthy adults at the same period were selected as healthy control group. Baseline characteristics, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and sequential organ failure assessment (SOFA) score were recorded. Venous blood samples were collected within 24 hours after diagnosis, and serum PCSK9 was determined by enzyme-linked immunosorbent assay (ELISA) at 1, 3 days and 5 days. Meanwhile, high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), total cholesterol (TC), triglyceride (TG) and lipoprotein A were detected. The differences of each index between sepsis group (28-day death group and survival group) and healthy control group were compared. Meanwhile, the indexes of patients with different severity and 28-day prognosis in sepsis group were compared. Receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of PCSK9 and blood lipid for the prognosis of sepsis. Multivariate Logistic regression was used to analyze the influencing factors for the prognosis of sepsis, and the Kaplan-Meier survival curve at 28th day was drawn.Results:There were 50 patients in sepsis group (including 19 patients with sepsis, 31 patients with septic shock) and 27 patients in healthy control group. In the sepsis group, 19 patients died and 31 patients survived within 28 days. The serum PCSK9 in the sepsis group was significantly higher than that in the healthy control group [μg/L: 223.09 (198.47, 250.82) vs. 188.00 (165.27, 214.90), P < 0.01], and HDL-C, LDL-C, TC and lipoprotein A were significantly lower than those in the healthy control group [HDL-C (mmol/L): 0.82±0.35 vs. 1.45±0.24, LDL-C (mmol/L): 1.53 (1.14, 2.47) vs. 2.89 (2.55, 3.19), TC (mmol/L): 2.03 (1.39, 2.84) vs. 4.24 (3.90, 4.71), lipoprotein A (g/L): 8.80 (5.66, 17.56) vs. 27.03 (14.79, 27.03), all P < 0.01]. PCSK9 in the sepsis death group was higher than that in the survival group [μg/L: 249.58 (214.90, 315.77) vs. 207.01 (181.50, 244.95), P < 0.01], and the HDL-C, LDL-C and TC were lower than those in the survival group [HDL-C (mmol/L): 0.64±0.35 vs. 0.93±0.30, LDL-C (mmol/L): 1.32±0.64 vs. 2.08±0.94, TC (mmol/L): 1.39 (1.01, 2.23) vs. 2.69 (1.72, 3.81), all P < 0.01]. With the progression of the disease, the PCSK9 in the sepsis death group and the survival group was significantly lower than that within 1 day of diagnosis (all P < 0.05). ROC curve analysis showed that PCSK9 had higher predictive value of 28-day death than HDL-C, LDL-C, TC [area under ROC curve (AUC) and 95% confidence interval (95% CI): 0.748 (0.611-0.885) vs. 0.710 (0.552-0.868), 0.721 (0.575-0.867), 0.702 (0.550-0.854)]. Multivariate Logistic regression analysis showed that PCSK9 was an independent risk factor affecting the 28-day prognosis of sepsis (β value was 1.014, P = 0.020). Kaplan-Meier survival curve analysis showed that when PCSK9 ≥ 208.97 μg/L, with the increase of PCSK9, the 28-day survival rate of sepsis patients decreased significantly. Conclusions:PCSK9, HDL-C, LDL-C and TC can all predict the 28-day prognosis of patients with sepsis. The prognostic value of PCSK9 is the highest. PCSK9 is an independent risk factor affecting the prognosis of sepsis. In the early stage of the disease, PCSK9 may have a good predictive value for the prognosis of sepsis. When PCSK9 ≥ 208.97 μg/L, the 28-day survival rate decreased significantly.

Objective:To explore the relationship between the expression of neuropilin-1 (NRP-1) on Treg cells and its ligands semaphorins-3A (Sema3A) , transforming growth factor-β 1 (TGF-β 1) as well as the balance of type 1 helper T cells (Th 1) and type 2 helper T cells (Th 2) cells. Methods:This study enrolled 62 patients with immune thrombocytopenia (ITP; 33 and 29 newly diagnosed and chronic ITP, respectively) from March 2014 to May 2015. Consequently, 30 healthy people in the same period were selected as the normal control group. The expression of NRP-1 in Treg cells was detected via flow cytometry. The Sema3A, TGF-β 1, IFN-γ, and IL-4 levels in plasma were detected by enzyme-linked immunosorbent assay. The real-time polymerase chain reaction technique was used to detect the mRNA expression levels of NRP-1, Sema3A, and TGF-β 1. The one-way analysis of variance and independent sample t-test was used for comparison between three and two groups, respectively. Correlations among the mRNA expression levels of NRP-1, Sema3A, and TGF-β 1 were assessed via Spearman correlation coefficients. Results:Treg cells in the newly diagnosed ITP group significantly increased compared with those in the chronic ITP and normal control groups. The expression of NRP-1 decreased[ (0.15 ± 0.03) %, (0.33 ± 0.15) %, and (0.46 ± 0.06) %; P<0.01], the plasma Sema3A level increased[ (8.10 ± 1.32) μg/L, (7.41±1.30) μg/L, and (2.88±0.82) μg/L; P<0.01], and the plasma TGF-β 1 level decreased[ (16.50±3.36) μg/L, (35.17±10.26) μg/L, and (41.00±10.02) μg/L; P<0.01]. Moreover, the level of plasma IFN-γ increased[ (17.21+2.80) ng/L, (10.23+1.59) ng/L, and (8.18+3.27) ng/L; P<0.01], and the ratios of Th 1/Th 2 (IFN-γ/IL-4) increased (1.29±0.30, 0.72±0.16, and 0.61±0.27; P<0.01) . The mRNA expressions of NRP-1 and Sema3A in the newly diagnosed ITP and chronic ITP groups were lower than that in the normal control group ( P<0.01) . Consequently, the NRP-1 mRNA expression was positively correlated with Sema3A and TGF-β 1 mRNA expression in the newly diagnosed ITP group. Conclusion:NRP-1 played an essential role in the pathogenesis of ITP.

Objective:To study the effect of spleen and marrow strengthening method combined with CAG regimen on the quality of life(QOL) of elderly patients with acute myeloid leukemia with spleen-kidney Yang deficiency syndrome.Methods:From June 2017 to October 2018, 50 elderly patients with acute myeloid leukemia with spleen-kidney Yang deficiency were randomly divided into treatment group and control group by random number table method, with 25 patients in each group.The patients in the control group were treated with CAG regimen(Ara-C+ Acla+ G-CSF), while the patients in the treatment group were treated with CAG regimen and leukemia prescription I, which was the empirical prescription for spleen and kidney Yang deficiency syndrome in elderly patients with acute myeloid leukemia.The patients were treated for two courses.The traditional Chinese medicine (TCM) syndromes and QOL scores of patients in two groups were compared and observed.Results:Before treatment, there was no statistically significant difference in the score of TCM syndromes between the two groups ( P>0.05). After treatment, the improvement of TCM syndromes in the treatment group had statistically significant difference compared with before treatment[before treatment (9.29±4.22)points, after treatment (5.04±3.83)points, t=3.656, P=0.001], but that in the control group had no statistically significant difference ( P>0.05). The treatment group was better than the control group ( t=-2.081, P=0.044). The total effective rate of the treatment group was 58.33% (14/24), which was significantly higher than that of the control group[26.32% (5/19)], and the difference was statistically significant (χ 2=5.831, P<0.05). Before treatment, there was no statistically significant difference in the total score of QOL between the two groups ( P>0.05). After treatment, the scores of QOL in both two groups were improved, the differences were statistically significant[the control group: (40.37±2.93)points vs.(38.21±2.76)points, t=2.337, P=0.025; the treatment group: (41.46±2.57)points vs.(36.54±2.34)points, t=6.929, P=0.000], and the QOL score of the treatment group was better than that of the control group ( t=-2.145, P=0.038). Conclusion:The improvement of TCM syndromes and QOL of elderly patients with acute myeloid leukemia with spleen-kidney Yang deficiency syndrome treated by spleen and marrow strengthening method combined with CAG regimen is better than that treated by CAG chemotherapy alone.

Objective: To explore the effects of Yiqi Yangyin Liangxue method on platelets, interleukin-10(IL-10) and transforming growth factor beta (TGF-β) of immune thrombocytopenic purpura(ITP) model mice, and to analyze its curative effect and possible mechanism. Methods: A total of 100 ITP model mice were randomly divided into blank group, model group, single Chinese medicine group, single hormone group and Chinese medicine combined with hormone group.Drug intervention was started on the 8th day after the establishment of the model, and the drug was given for a total of 14 days.The blood of mice was collected and the levels of platelets, TGF-β and IL-10 in serum of mice in each group were detected. Results: There was no statistically significant difference in platelet count among all groups before modeling(F=0.556, P>0.05). Before administration, there was statistically significant difference between the model group and the blank group (t=28.207, P<0.01), there were no statistically significant differences between the model group and the traditional Chinese medicine group, hormone group, hormone+ traditional Chinese medicine group (t=-1.96, -0.464, -0.979, all P>0.05). After gastric administration, the platelet counts in the traditional Chinese medicine group[(710.45±124.52)×10⁹/L], hormone group[(768.10±127.42)×10⁹/L], hormone+ traditional Chinese medicine group[(908.05±89.66)×10⁹/L] were significantly higher than that in the model group[(413.55±38.84)×10⁹/L](t=-10.18, -11.90, -22.63, all P<0.01). After gastric administration, there was no statistically significant difference between the traditional Chinese medicine group and the hormone group(t=-1.447, P>0.05). After gastric administration, the platelet count of the hormone+ traditional Chinese medicine group was significantly higher than that of the hormone group and the traditional Chinese medicine group(t=-4.017, -5.759, all P<0.01). There was statistically significant difference in IL-10 level among groups after administration(F=32.20, P<0.01). IL-10 levels between the model group[(20.28±0.75)ng/L] and traditional Chinese medicine group[(21.41±1.40)ng/L], hormone group[(21.79±1.14)ng/L] and traditional Chinese medicine+ hormone group[(23.14±1.34)ng/L] had statistically significant differences (t=-3.18, -4.93, -8.33, all P<0.01). There was no statistically significant difference between the traditional Chinese medicine group and hormone group(t=-0.927, P>0.05). There were statistically significant differences between the traditional Chinese medicine+ hormone group and traditional Chinese medicine group, hormone group (t=-3.97, -3.43, all P<0.01). There was statistically significant difference in TGF-beta level among all groups after administration(F=31.16, P<0.01). The TGF-β levels between the model group[(82.32±3.42)ng/L] and Chinese medicine group[(88.10±8.51)ng/L], hormone group[(90.03±4.50)ng/L] and Chinese medicine combined with hormone group[(94.41±2.53)ng/L] had statistically significant differences (t=-2.82, -6.10, -12.70, all P<0.01). There was no significant difference in TGF-β level between the Chinese medicine group and hormone group(t=-0.90, P>0.05). There were statistically significant differences in in TGF-beta level between the traditional Chinese medicine+ hormone group and Chinese medicine group, hormone group (t=-3.18, -3.79, all P<0.01). Conclusion Yiqi Yangyin Liangxue method can reduce platelet destruction by regulating the levels of IL-10 and TGF-β and inhibiting immunity, and it has good therapeutic effect on ITP.