Objective:To explore the clinical characteristics and genetic variants in three children with late-onset Multiple acyl-Coenzyme A dehydrogenase deficiency (MADD type Ⅲ).Methods:Clinical data of three children diagnosed with late-onset MADD at the Children′s Hospital Affiliated to Zhengzhou University between March 2020 and March 2022 were retrospectively analyzed. All children were subjected to whole exome sequencing (WES), and candidate variants were verified by Sanger sequencing. All children had received improved metabolic therapy and followed up for 1 ~ 3 years.Results:The children had included 2 males and 1 female, and aged from 2 months to 11 years and 7 months. Child 1 had intermittent vomiting, child 2 had weakness in lower limbs, while child 3 had no symptom except abnormal neonatal screening. Tandem mass spectrometry of the three children showed elevation of multiple acylcarnitines with short, medium and long chains. Children 1 and 2 showed increased glutaric acid and multiple dicarboxylic acids by urine Gas chromatography-mass spectrometry (GC-MS) analysis. All children were found to harbor compound heterozygous variants of the ETFDH gene, including a paternal c. 1211T>C (p.M404T) and a maternal c. 488-22T>G variant in child 1, a paternal c. 1717C>T (p.Q573X) and a maternal c. 250G>A (p.A84T) variant in child 2, and a paternal c. 1285+ 1G>A and maternal c. 629A>G (p.S210N) variant in child 3. As for the treatment, high-dose vitamin B2, levocarnitine and coenzyme Q 10 were given to improve the metabolism, in addition with a low fat, hypoproteinic and high carbohydrate diet. All children showed a stable condition with normal growth and development during the follow-up. Conclusion:The compound heterozygous variants of the ETFDH gene probably underlay the muscle weakness, remittent vomiting, elevated short, medium, and long chain acylcarnitine, as well as elevated glutaric acid and various dicarboxylic acids in the three children with type Ⅲ MADD.

Objective:To investigate the correlation between clinical phenotypes and genetic characteristics of children with ring chromosomes (RCs).Methods:Case series study.The clinical data of 11 434 children who received treatment and peripheral blood chromosome karyotype detection in Henan Children′s Hospital from October 2008 to October 2023 due to growth retardation, intellectual impairment or congenital malformation were analyzed retrospectively.A total of 25 children with RCs were selected.Their age at diagnosis, karyotype distribution, clinical manifestations, and genetic detection results were analyzed.Results:RCs were detected in 25 out of 11 434 children, with a detection rate of 0.21%.The genome-wide copy number variation (CNV) analysis was performed on 7 RCs cases, and it found that pathogenic variation existed in all of them.Among the 25 RC cases (11 males and 14 females of social gender), the age at diagnosis ranged from 2 months to 14 years; there were 20 autosomal rings and 5 sex chromosome rings; 13 cases had chimeric karyotypes, and 12 cases had non-chimeric karyotypes.Most of the 25 children showed clinical manifestations of mental or developmental retardation, and some also presented with specific clinical manifestations, such as short stature, congenital malformation, and epilepsy.Conclusions:The pathogenesis of RCs is complex.The clinical manifestations are determined by both RCs syndrome and specific phenotypes caused by the dose effect and exhibit high heterogeneity, so it is easy to miss or misdiagnose.The combined application of cellular and molecular genetic detection technology can facilitate early diagnosis and treatment of RCs, and the correlation analysis of phenotypes and genetic characteristics can provide guidance for genetic counseling.

Objective:To explore the predictive value of transrectal multimodal ultrasound and prostate specific antigen (PSA) in clinically organ-confined prostate cancer.Methods:It was a cross-sectional study. The clinical data of patients with suspected prostate nodules treated in the First Hospital of Shanxi Medical University from May 2014 to April 2020 were analyzed retrospectively. Of the patients, 48 cases of clinically organ-confined prostate cancer and 51 cases of benign prostatic hyperplasia confirmed by clinical data and pathology were selected as research objects. The characteristics of transrectal multimodal ultrasound in the two groups were compared. Combined with PSA, logistic regression analysis was applied to screen the statistically significant features, and then the diagnosis model was established, and odds ratio of the variables were compared. The receiver operating characteristic (ROC) curve was constructed to analyze the predicting ability of the diagnosis model.Results:Four features were obtained with logistic regression analysis finally, including enhancement type, enhancement degree, elastography mode and PSA. The odds ratio of enhancement degree was higher than those of the other independent variables. The area under ROC curve of the diagnosis model was 0.868 ( P<0.01), the cut-off value was 0.514. The sensitivity and specificity of the diagnosis model in predicting clinically organ-confined prostate cancer was 79.2% and 80.4%, respectively. Conclusions:This combined diagnosis model of transrectal multimodal ultrasound and PSA has a certain clinical value in predicting clinically organ-confined prostate cancer.

Objective:To establish a three-dimensional spheroid expansion model of human glioma stem cells with spontaneous sphere forming and multilineage differentiation potential in vitro and investigated the contents of the proteins and lipids and their secondary components, so as to lay a foundation for further study of sphere metabolism. Methods:Human glioma stem cells GSC23 and SU3 were cultured in serum-free stem cell culture medium, respectively, and the cell spheres were harvested for about 2-3 weeks. After fixation in paraformaldehyde solution, dehydration, paraffin embedding, and sectioning, glioma-associated marker proteins were detected by immunohistochemical staining, and the protein and lipid and their secondary components contents in the spheroid tissues were analyzed by Raman imaging. One-way ANOVA was used to compare the protein, lipid and phenylalanine contents in the large sphere, medium sphere and small sphere groups.Results:Both stem cells were able to form stem cell expansion spheres resembling solid tumors within the culture dish. Immunohistochemical staining showed that the regular marker proteins of glioblastoma multiforme, CD133, nestin, epidermal growth factor receptor (EGFR), S100, Olig2, p53, Ki-67, glial fibrillary acidic protein (GFAP), vimentin, CXC chemokine receptor 4 (CXCR4), and CD34, were all expressed. Raman imaging revealed that the constructed expanded spheres of human glioma stem cells contained protein (2 930, 1 685 and 1 586 cm -1), lipid (2 845 and 1 444 cm -1), phenylalanine (1 003 cm -1) amide Ⅲ (1 250 cm -1), while there were no significant differences of protein, lipid and phenylalanine contents among the large sphere, medium sphere and small sphere groups ( P>0.05). Conclusions:We have successfully established an expanded spheroid model of human glioma stem cells in vitro, which not only exhibits the topographical characteristics and unlimited expansion ability of three-dimensional solid tumors, but also has the ability to stably store metabolically obligatory energy sources of tumor cells such as proteins and lipids, and is expected to serve as a promising tool for human glioma research in vitro.

Objective:To establish a three-dimensional spheroid expansion model of human glioma stem cells with spontaneous sphere forming and multilineage differentiation potential in vitro and investigated the contents of the proteins and lipids and their secondary components, so as to lay a foundation for further study of sphere metabolism. Methods:Human glioma stem cells GSC23 and SU3 were cultured in serum-free stem cell culture medium, respectively, and the cell spheres were harvested for about 2-3 weeks. After fixation in paraformaldehyde solution, dehydration, paraffin embedding, and sectioning, glioma-associated marker proteins were detected by immunohistochemical staining, and the protein and lipid and their secondary components contents in the spheroid tissues were analyzed by Raman imaging. One-way ANOVA was used to compare the protein, lipid and phenylalanine contents in the large sphere, medium sphere and small sphere groups.Results:Both stem cells were able to form stem cell expansion spheres resembling solid tumors within the culture dish. Immunohistochemical staining showed that the regular marker proteins of glioblastoma multiforme, CD133, nestin, epidermal growth factor receptor (EGFR), S100, Olig2, p53, Ki-67, glial fibrillary acidic protein (GFAP), vimentin, CXC chemokine receptor 4 (CXCR4), and CD34, were all expressed. Raman imaging revealed that the constructed expanded spheres of human glioma stem cells contained protein (2 930, 1 685 and 1 586 cm -1), lipid (2 845 and 1 444 cm -1), phenylalanine (1 003 cm -1) amide Ⅲ (1 250 cm -1), while there were no significant differences of protein, lipid and phenylalanine contents among the large sphere, medium sphere and small sphere groups ( P>0.05). Conclusions:We have successfully established an expanded spheroid model of human glioma stem cells in vitro, which not only exhibits the topographical characteristics and unlimited expansion ability of three-dimensional solid tumors, but also has the ability to stably store metabolically obligatory energy sources of tumor cells such as proteins and lipids, and is expected to serve as a promising tool for human glioma research in vitro.

OBJECTIVE: To explore the genetic basis for a child with clinically suspected 3-methylcrotonyl-coenzyme A carboxylase deficiency (MCCD). METHODS: Genomic DNA was extracted from peripheral blood samples of the proband and her parents. Whole exome sequencing was used to screen pathogenic variant in the proband. Suspected variant was verified by Sanger sequencing. Impact of the variant on the structure and function of protein product was analyzed by using bioinformatic software. RESULTS: Sanger sequencing showed that the proband has carried homozygous missense c.1342G>A (p.Gly448Ala) variant of the MCCC2 gene, for which her mother was a heterozygous carrier. The same variant was not detected in her father. The variant was predicted to be pathogenic by PolyPhen-2 and Mutation Taster software, and the site was highly conserved among various species. Based on the American College of Medical Genetics and Genomics standards and guidelines, the c.1342G>A (p.Gly448Ala) variant of MCCC2 gene was predicted to be likely pathogenic(PM2+PP2-PP5). CONCLUSION The homozygous missense variant of the MCCC2 gene c.1342G>A (p.Gly448Ala) probably underlay the molecular pathogenesis of the proband. Genetic testing has confirmed the clinical diagnosis.
Carbon-Carbon Ligases/genetics* ; Child ; Female ; Humans ; Male ; Mutation, Missense/genetics* ; Pedigree ; Urea Cycle Disorders, Inborn/genetics*

OBJECTIVE: To carry out cyto- and molecular genetic testing for a child featuring facial dysmorphism and attention deficit and hyperactive disorder. METHODS: The child was subjected to routine peripheral blood lymphocyte chromosomal karyotyping, fluorescence in situ hybridization (FISH) and single nucleotide polymorphism array (SNP-array) analyses. RESULTS: The child's facial dysmorphism included low-set ears, curly ear auricle, protuberance of eyebrow arch, nostril notch, short and flat philtrum and thin upper lip. SNP-array revealed that he has carried a 4.883 Mb deletion at 2q37. His chromosomal karyotype was ultimately determined as 45, XY, der(2;21) (2pter→ 2q37.3::21p13→ 21p10::20p10→ 20pter), der(20) (21qter→ 21q10::20q10→ 20qter). CONCLUSION A rare case of 2q37 deletion syndrome involving three chromosomes was discovered. Combined use of various cyto- and molecular genetic techniques is crucial for the diagnosis of chromosomal abnormalities with complex structures.
Child ; Chromosome Deletion ; Chromosomes ; Chromosomes, Human, Pair 2 ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Translocation, Genetic

Under the background of technological assistance to prepare for the Beijing Winter Olympics in China, the biomechanical research highlights and the latest achievements related to competitive performance of alpine skiers in recent years were systematically analyzed in this paper, so as to determine biomechanical factors affecting competitive performance of alpine skiers, including aerodynamic drag, frictional forces, ground reaction force (GRF), energy dissipation, turn radius, trajectory of the skis and/or center of mass (COM). In addition, biomechanical differences in turn techniques, multiple turns connections and abilities of individuals were also considered as important factors affecting the alpine skiing performance. In the case of slalom and giant slalom events, the earlier initiation of turns, longer path length and trajectory, earlier and smoother application of GRF, and carbene technique carving to reduce the ski-snow friction and thereby dissipate energy should be used to improve sports performance. During speed skiing, minimizing the exposed frontal area and positioning the arms close to the body can reduce the energy loss caused by aerodynamic drag, thereby improving sports performance. Top-level alpine skiers will always perform well on different courses, terrains and snow conditions during the race. Excellent alpine ski performance from a biomechanical perspective includes the efficient use of potential energy, minimizing ski-snow friction and aerodynamic drag, choosing optimal trajectory and maintaining high-speed skiing. Individual tactics and techniques should be valued in training and competition. For better results, the same performance on multiple sections and on different terrains is more important than excellence in individual sections and specific conditions.

Objective:To study the characteristics of macrophage lineage model polarized to adapt to the tumor micro-environment (TME) for further research on the plasticity of macrophages in TME.Methods:Bone marrow cells from transgenic Foxn1 nu.B6-CAG-EGFP/SU mice were induced by colony-stimulating factor 1 (CSF-1), IFN-γ+ LPS and IL-4 to differentiate into M0, M1 and M2 macrophages, respectively. Green fluorescent protein (GFP) was observed under inverted fluorescence microscope. Immunocytochemical staining was used to detect the marker and polarization-related proteins of macrophages. Moreover, the macrophages were co-cultured with human glioma stem cell SU3 for further analysis. Results:The bone marrow-derived M0, M1 and M2 macrophages all showed strong green fluorescence under inverted fluorescence microscope. The inherent plasticity of the macrophages could be observed under ordinary microscope with Wright-Giemsa staining. Immunocytochemical staining showed that CD11C and CD206 markers were observed on M0, M1 and M2 macrophages, while CD68 was only expressed on M1 macrophages. Moreover, the staining was strongly positive for CSF-1 and CSF-1R on M0, M1 and M2 macrophages. Green fluorescent cell infiltration and phagocytic reaction were observed in the co-cultured stem cell spheres.Conclusions:The bone marrow-derived macrophage lineage including M0, M1 and M2 subtypes with the inherent plasticity was successfully prepared using transgenic nude mice expressing GFP. The three subtypes expressed the common marker and polarization-related proteins, and had the phagocytic activity, suggesting that they could be used to study the interaction between tumor cells and macrophages, especially in tracer studies.

Objective:To investigate the prognosis of intensive care unit(ICU)patients with carbapenem-resistant Klebsiella pneumoniae(CR-KP)infection.Methods:Clinical data of 86 elderly patients with KP infection admitted to our hospital from August 2017 to July 2019 were retrospectively analyzed.All patients were identified by the VITEK system and were tested by using the Kirby-Bauer paper diffusion method for drug susceptibility.Based on the existence of CR-KP, patients were divided into the non-resistance group(n=40)and the resistance group(n=46). The number of ventilator days, ICU days and clinical prognosis were compared between the two groups.Results:Specimens from sputum had the highest rate of KP isolation(46/86, 53.49%), followed by specimens from drainage fluid(16/86, 18.60%), blood(12/86, 13.95%), puncture wound fluid(3/86, 3.50%)and the tip of deep vein catheters(9/86, 10.46%). KP had high drug resistance to meropenem, and the drug-resistance rate was 53.49%(46/86). Compared with the non-resistance group, ventilator days and ICU days increased in the drug-resistance group[(60.75±72.86)d vs.(26.11±41.10)d, (73.41±63.32)d vs.(38.96±50.75)d, t=2.660 and 2.756, P=0.003 and 0.004]. The clinical effectiveness rate was higher in the non-resistance group than in the drug-resistance group(57.50% or 23/40 vs.23.91 % or 11/46, χ2=10.097, P=0.001). Conclusions:ICU patients with CR-KP have a higher number of ventilator days and longer ICU stays than those without CR-KP.In clinical practice, it is necessary to strictly follow the indications for antibiotic use, regularly perform bacterial identification and drug susceptibility tests, and diagnose KP infection as early as possible, in order to provide evidence for clinical diagnosis and treatment.